CN111808786A - 植物性酵母乳酸菌的制备方法及应用 - Google Patents
植物性酵母乳酸菌的制备方法及应用 Download PDFInfo
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- CN111808786A CN111808786A CN202010805373.1A CN202010805373A CN111808786A CN 111808786 A CN111808786 A CN 111808786A CN 202010805373 A CN202010805373 A CN 202010805373A CN 111808786 A CN111808786 A CN 111808786A
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Abstract
本发明涉及植物性酵母乳酸菌的制备方法,包括以下步骤:(1)制备植物性酵母乳酸菌培养基;(2)接种植物性乳酸菌及酵母菌菌种;(3)制备植物性酵母乳酸菌发酵培养基;(4)制备植物性酵母乳酸原液A;(5)制备植物性酵母乳酸菌二次培养原液B,将植物性酵母乳酸菌原液A稀释2‑5倍,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10‑20%,在35‑50oC温度中培养1‑5天。所述植物性酵母乳酸菌二次培养原液B能够在医药保健、农业生产、环境保护领域中广泛应用,功用涵盖平衡人体胃肠道菌群、提高免疫力、生产益生菌饲料、生产有机肥料、土壤改良、动植物保护、绿色生态农业种植和养殖、农业废弃物无害化处理、食用菌生产等多个领域。
Description
技术领域
本发明涉及微生物发酵领域,具体是涉及植物性酵母乳酸菌的制备方法及应用。
技术背景
乳酸菌分为“动物乳酸菌”和“植物乳酸菌”。生活在诸如酸奶和奶酪等动物性物质中的乳酸菌被称为“动物乳酸菌”,并与诸如哺乳动物牛奶之类的动物性物质分离。生活在谷物、蔬菜和水果等植物中的乳酸菌被称为“植物乳酸菌”,并与谷物,蔬菜,水果和种子等植物分离。与富含营养并在诸如牛奶等细菌的稳定环境中生长的“动物乳酸菌”相比,“植物乳酸菌”具有各种特性,能够在缺乏营养源的恶劣环境中生存。大多数“动物乳酸菌”主要利用乳糖进行发酵,并且倾向于与其他糖类一起生长。另一方面,“植物乳酸菌”是具有耐酸性和耐盐性等特性的物质,可以使用除乳糖以外的各种糖,并且具有比其他细菌更强壮地生长的机制。“植物乳酸菌”可以适应各种严酷环境,因此具有更广泛的应用前途。近年来,“植物乳酸菌”受到世界各国关注,并且逐渐被广泛应用于各个领域。
以往植物性乳酸菌和酵母菌等的使用只作为微生物技术应用于土壤和环境改良。但是即使是在最优化的状态下,它们还是很难充分完全地发挥作用。
发明内容
为了弥补上述现有技术的不足,本发明提出一种植物性酵母乳酸菌的制备方法及应用,建立完整植物性酵母乳酸菌发酵工艺体系,充分发挥植物性乳酸菌和酵母菌的作用,活性强,能够在医药保健、农业生产、环境保护领域中广泛应用,具有杀菌、除臭、水质净化等作用。
本发明的目的是通过以下技术方案实现的,一种植物性酵母乳酸菌的制备方法,其技术要点是:包括以下步骤:
一种植物性酵母乳酸菌的制备方法,包括以下步骤:
(1)制备植物性酵母乳酸菌培养基
所述植物性酵母乳酸菌培养基原料组分按重量比包括如下:所述大豆提取物蛋白质(包括雪花菜)70-95,糯米5-30,米糠0-30;
将上述培养基加纯净水调和成含水分40%-60%粘稠状,用搅拌机充分搅拌均匀,得到植物性酵母乳酸菌培养基;
(2)接种植物性乳酸菌及酵母菌菌种
所述植物性乳酸菌菌种为Lactobacillus fermentum(发酵乳酸杆菌),Lactobacillus paracasei(副干酪乳杆菌), Lactobacillus acidophilus(嗜酸乳杆菌),Lactobacillus casei(干酪乳杆菌),Lactobacillus delbrueckii(保加利亚乳杆菌),Lactobacillus arabinosus(阿拉伯乳杆菌),Lactobacillus caucasicus(高加索乳杆菌), Lactobacillus lactis(乳酸乳球菌), Lactobacillus Leichmanni(莱氏乳杆菌),Lactobacillus musicus(穆氏乳杆菌), Lactobacillus thermophilus(嗜热乳杆菌),Lactobacillus plantarum(植物乳杆菌),Streptococcus thermophilus(嗜热链球菌),Streptococcus cremoris(乳脂链球菌), Sterptococcusfaecalis(粪肠链球菌),Streptococcus lactis(乳酸链球菌),Leuconostocmesenteroides(肠膜明串珠菌)中的一至六种;
所述酵母菌菌种为Pichiakluyveri(克鲁弗毕赤酵母),Saccharomyces intermedius(间型酵母), Saccharomyces validus(酿酒酵母),Saccharomyces ellipsoideus(椭圆酿酒酵母), Saccharomyces malirisler(马里利斯勒酵母), Saccharomyces mandshuricus(东北酵母), Saccharomyces Vordermannii(沃尔酵母),Saccharomyces Peka(佩卡酵母), Saccharomyces shasshing(沙欣酵母), Saccharomyces piriformis(梨形酵母),Saccharomyces anamensis(越南酵母),Saccharomyces cartilaginosus(软骨状酵母),Saccharomyces Awamori(泡盛酒酵母), Saccharomyces Batatae(甘薯酒酵母),Saccharomyces Coreanus(朝鲜酵母), Saccharomyces robustus(粗壮酵母),Saccharomyces Carlsbergensis(卡尔酵母),Saccharomyces Monacensis(摩拿酵母),Saccharomyces Marxianus(马克斯酵母),Zygo Saccharomyces major(大接合酵母),Saccharomyces lactis(乳酸酵母), Saccharomyces Roouxii(鲁氏酵母),Hansenulaanomala(汉氏酵母),Zygosacckromyceskoiuxii (Boutroux) yaitow(鲁氏结合酵母),Pichiuahomala( E. C. Hahshe) Kurzmiah(异构毕赤酵母)的一种至五种;
所述植物性乳酸菌菌种和酵母菌菌种调制成混合菌种,所述混合菌种中乳酸菌、酵母菌孢子浓度比例为10-150:1-10;将混合菌种接种至植物性酵母乳酸菌培养基中,接种后乳酸菌、酵母菌的孢子总浓度为1x106-4x109cfu/kg;
(3)制备植物性酵母乳酸菌发酵培养基
上述接种植物性乳酸菌及酵母菌菌种的植物性酵母乳酸菌培养基置于4-10oC环境中培养45-180天,获得植物性酵母乳酸菌发酵培养基,所述植物性酵母乳酸菌发酵培养基中的植物性乳酸菌数为≧1×106-108cfu/g,酵母菌数为≧2.0×106cfu/g;
(4)制备植物性酵母乳酸菌原液A
将上述植物性酵母乳酸菌发酵培养基和纯净水按重量比例 1:5-20进行混合搅拌,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10-20%, 35-50oC培养1-5天,获得植物性酵母乳酸菌原液A;所述植物性酵母乳酸原液A中乳酸菌、酵母菌孢子总浓度为1×106-2×108 cfu/ml;
(5)制备植物性酵母乳酸菌二次培养原液B
二次培养原液B制备方法:将植物性酵母乳酸菌原液A稀释 2-5倍,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10-20%,在35-50oC温度中培养1-5天,获得植物性酵母乳酸菌二次培养原液B,所述植物性酵母乳酸菌二次培养原液B乳酸菌、酵母菌孢子总浓度为1×106-2×108cfu/ml。
进一步的,所述植物性酵母乳酸菌原液A、植物性酵母乳酸菌二次培养原液B可采用植物性酵母乳酸菌培养装置进行制备:
所述植物性酵母乳酸菌培养装置,包括搅拌发酵罐,固液分离装置,过滤装置,所述搅拌发酵罐设有投料口和出料口,所述出料口是通过连接管Ⅰ与固液分离装置上口连接,所述连接管Ⅰ设有阀门Ⅰ;所述固液分离装置下口通过连接管Ⅱ连接过滤装置的滤液投入口,所述连接管Ⅱ上设有阀门Ⅱ;所述过滤装置设有出料口,所述出料口连接有出料管,所述出料管上设有阀门Ⅲ;
所述搅拌发酵罐包括罐体,搅拌电机;所述罐体主体为柱形,底部为倒圆锥形,所述罐体侧部设有水位计;所述投料口设置于罐体的顶部,所述出料口设置于罐体的底部中心;所述搅拌电机下端固定连接有转轴,所述转轴从上至下固定连接有多个搅拌折叶组,所述搅拌折叶组设置于罐体主体和圆锥形底部,所述搅拌折叶组包括周向多个均匀分布的折叶,置于罐体底部倒圆锥内折叶组中的折叶长度短于置于罐体主体内折叶组中折叶的长度;所述罐体内设有温度感应器、酸碱度感应器、电导率感应器、糖度计传感器,所述温度感应器、酸碱度感应器、电导率感应器、糖度计传感器均连接于自动检测控制计算机,所述自动检测控制计算机连接搅拌电机,控制搅拌电机电源、转速;
所述固液分离装置内设有过滤板,所述过滤板上设有滤膜Ⅰ,所述滤膜Ⅰ孔径为30-40微米;
所述过滤装置包括过滤罐体,所述过滤罐体为柱形体,过滤罐体内部设有过滤筒,所述过滤筒将过滤罐体分隔为内、外两个腔体,所述滤液投入口置于过滤罐体内部腔体上方,所述出料口置于过滤罐体外部腔体下方;所述过滤筒上设有滤膜Ⅱ,所述滤膜Ⅱ孔径为3-5微米;
所述植物性酵母乳酸菌培养装置用于植物性酵母乳酸菌培养,主要步骤为:
1)投料
原料为植物性酵母乳酸菌发酵培养基按比例添加低聚糖或蔗糖,向搅拌发酵罐投料时80-100 rpm/min高速均匀搅拌1-2分钟后10-30 rpm/min低速均匀搅拌;
2)固液分离
所述固液分离装置为压力式分离器,固液分离过滤板的滤膜Ⅰ孔径是30-40微米;
3)过滤
所述过滤装置5中滤膜Ⅱ的孔径是3-5微米;过滤得到2微米酵母菌和小于2微米的乳酸菌,获得植物性酵母乳酸菌原液A;
将植物性酵母乳酸菌原液A稀释 2-5倍,重复上述步骤,按比例添加低聚糖或蔗糖,扩增培养获得二次植物性酵母乳酸菌培养原液B。
本发明还提出了植物性酵母乳酸菌的应用,其技术要点是:所述植物性酵母乳酸菌二次培养原液B用于家禽、家畜或水产养殖饲料的益生菌添加剂或不含酒精的安全性杀菌剂。
进一步的,所述用于家禽、家畜或水产养殖饲料的益生菌添加功效的植物性酵母乳酸菌二次培养原液B制备中:植物乳酸菌菌种为:Lactobacillus fermentum(发酵乳酸杆菌), Lactobacillus paracasei(副干酪乳杆菌),Lactobacillus plantarum(植物乳杆菌),Lactobacillus lactis(乳酸乳球菌)Streptococcus lactis(乳酸链球菌);酵母菌菌种为:Zygosacckromyceskoiuxii (Boutroux) yaitow(鲁氏结合酵母菌),Pichiuahomala( E. C. Hahshe) Kurzmiah(异构毕赤酵母),Pichiakluyveri(克鲁弗毕赤酵母),Saccharomyces lactis(乳酸酵母),Saccharomyces validus(酿酒酵母)。
所述用于家禽、家畜或水产养殖饲料的益生菌添加功效的植物性酵母乳酸菌二次培养原液B的使用方法包括:
(1)投喂:饲养家禽、家畜(例如牛、猪、羊、鸡、鸭、鹅等)及其宠物(猫、狗等)投喂植物性酵母乳酸菌二次培养原液B和饲料的混合物,所述混合物中植物性酵母乳酸菌二次培养原液B和饲料重量比为:1-10:100;
(2)制备有机肥:将植物性酵母乳酸菌二次培养原液B直接稀释100倍,制备植物性酵母乳酸菌原液B稀释液,所述植物性酵母乳酸菌原液B稀释液与家畜的屎尿混合,二者重量比为10-20:100;发酵方法为夏季发酵一周,春秋季节发酵两周,冬季发酵两至三周。
所述用于不含酒精的安全杀菌的植物性酵母乳酸菌二次培养原液B制备中:
植物性乳酸菌菌种为:Lactobacillus fermentum(发酵乳酸杆菌), Lactobacillusparacasei(副干酪乳杆菌),Lactobacillus plantarum(植物乳杆菌),Lactobacillusacidophilus(嗜酸乳杆菌) ,Lactobacillus thermophilus(嗜热乳杆菌),Streptococcus lactis(乳酸链球菌);所述杀菌功效中主要选择的酵母菌菌种为:Zygosacckromyceskoiuxii(Boutroux) yaitow(鲁氏结合酵母菌),Pichiuahomala( E. C.Hahshe) Kurzmiah(异构毕赤酵母),Saccharomyces lactis(乳酸酵母), SaccharomycesRoouxii(鲁氏酵母),Pichiakluyveri(克鲁弗毕赤酵母)。
所述用于不含酒精的安全杀菌的植物性酵母乳酸菌二次培养原液B的使用方法包括:
(1)水质净化:将植物性酵母乳酸菌二次培养原液B放入装有活性炭或二氧化硅等吸附材料的细网吊袋中放入流水、养殖鱼水箱;使用用量为3吨水中加入5 kg装有活性炭或二氧化硅等吸附剂,所述吸附剂均匀喷淋1升稀释100倍的植物性酵母乳酸菌原液B,用于去除不流动水中霉菌和异味;
(2)杀菌、除味:将植物性酵母乳酸菌二次培养原液B直接稀释100倍,制备植物性酵母乳酸菌原液B稀释液,所述植物性酵母乳酸菌原液B稀释液喷淋到厨房灶台台面、畜舍中用于杀菌、除味、消臭。
本发明的有益效果:
所述植物性酵母乳酸菌的制备方法能够在医药保健、农业生产、环境保护领域中广泛应用,功用涵盖平衡人体胃肠道菌群、提高免疫力、生产益生菌饲料、生产有机肥料、土壤改良、动植物保护、绿色生态农业种植和养殖、农业废弃物无害化处理、食用菌生产等多个领域。
附图说明
图1为植物性酵母乳酸菌培养装置的结构示意图;
图2为搅拌发酵罐结构示意图;
图3为搅拌发酵罐俯视图;
图4为搅拌折叶俯视图;
图5为过滤装置结构示意图;
图6为过滤装置俯视图;
图7为搅拌折叶平面与转轴平面连接示意图;
图8为搅拌折叶边缘内折平面示意图;
图9为搅拌发酵罐中各感应结构与计算机连接结构示意图;
图10为过滤筒或过滤板截面示意图;
图11 LABBIO植物性酵母乳酸菌LBF原液的工艺流程图;
图12 LABBIO植物性酵母乳酸菌LBF原液的杀菌试验结果。
图1-图10中各结构的具体名称为:搅拌发酵罐1,搅拌电机1-1,水位计1-2,自动检测控制计算机1-3,投料口1-4,出料口1-5,罐体1-6,转轴1-7,搅拌折叶组1-8,折叶1-9,温度感应器1-10、酸碱度感应器1-11、电导率感应器1-12、糖度计传感器1-13,阀门Ⅰ2,固液分离装置3,过滤板3-1,下口3-2,滤膜Ⅰ3-3,过滤孔Ⅰ3-4,上口3-5,阀门Ⅱ4,过滤装置5,滤液投入口5-1,过滤筒5-2,过滤罐体5-3,过滤孔Ⅱ5-4,出料口5-5,滤膜Ⅱ5-6,阀门Ⅲ6,连接管Ⅰ7,连接管Ⅱ8,出料管9。
具体实施方式
实施例1
本实施例的菌种均可在市场购买得到;
一种植物性酵母乳酸菌的制备方法,所述植物性酵母乳酸菌命名为:LABBIO植物性酵母乳酸菌,制备包括以下步骤:
(1)制备植物性酵母乳酸菌培养基
所述植物性酵母乳酸菌培养基原料组分按重量比包括如下:所述大豆蛋白质提取物(包括雪花菜)70-95,糯米5-30,米糠0-30;
将上述培养基加纯净水调和成含水分40%-60%粘稠状,用搅拌机充分搅拌均匀,得到植物性酵母乳酸菌培养基;
(2)接种植物性乳酸菌及酵母菌菌种
所述植物性乳酸菌菌种为Lactobacillus fermentum(发酵乳酸杆菌),Lactobacillus paracasei(副干酪乳杆菌), Lactobacillus acidophilus(嗜酸乳杆菌),Lactobacillus casei(干酪乳杆菌),Lactobacillus delbrueckii(保加利亚乳杆菌),Lactobacillus arabinosus(阿拉伯乳杆菌),Lactobacillus caucasicus(高加索乳杆菌), Lactobacillus lactis(乳酸乳球菌), Lactobacillus Leichmanni(莱氏乳杆菌),Lactobacillus musicus(穆氏乳杆菌), Lactobacillus thermophilus(嗜热乳杆菌),Lactobacillus plantarum(植物乳杆菌),Streptococcus thermophilus(嗜热链球菌),Streptococcus cremoris(乳脂链球菌), Sterptococcusfaecalis(粪肠链球菌),Streptococcus lactis(乳酸链球菌),Leuconostocmesenteroides(肠膜明串珠菌)中的一至六种;
所述酵母菌菌种为Pichiakluyveri(克鲁弗毕赤酵母),Saccharomyces intermedius(间型酵母), Saccharomyces validus(酿酒酵母),Saccharomyces ellipsoideus(椭圆酿酒酵母), Saccharomyces malirisler(马里利斯勒酵母), Saccharomyces mandshuricus(东北酵母), Saccharomyces Vordermannii(沃尔酵母),Saccharomyces Peka(佩卡酵母), Saccharomyces shasshing(沙欣酵母), Saccharomyces piriformis(梨形酵母),Saccharomyces anamensis(越南酵母),Saccharomyces cartilaginosus(软骨状酵母),Saccharomyces Awamori(泡盛酒酵母), Saccharomyces Batatae(甘薯酒酵母),Saccharomyces Coreanus(朝鲜酵母), Saccharomyces robustus(粗壮酵母),Saccharomyces Carlsbergensis(卡尔酵母),Saccharomyces Monacensis(摩拿酵母),Saccharomyces Marxianus(马克斯酵母),Zygo Saccharomyces major(大接合酵母),Saccharomyces lactis(乳酸酵母), Saccharomyces Roouxii(鲁氏酵母),Hansenulaanomala(汉氏酵母),Zygosacckromyceskoiuxii(Boutroux) yaitow(鲁氏结合酵母),Pichiuahomala( E. C. Hahshe) Kurzmiah(异构毕赤酵母)的一种至五种;
所述植物性乳酸菌菌种和酵母菌菌种调制成混合菌种,所述混合菌种中乳酸菌、酵母菌孢子浓度比例为10-150:1-10;将混合菌种接种至植物性酵母乳酸菌培养基中,接种后乳酸菌、酵母菌的孢子总浓度为1x106-4x109cfu/kg;
(3)制备植物性酵母乳酸菌发酵培养基
上述接种植物性乳酸菌及酵母菌菌种的植物性酵母乳酸菌培养基置于4-10oC环境中培养45-180天,获得植物性酵母乳酸菌发酵培养基,所述植物性酵母乳酸菌发酵培养基中的植物性乳酸菌数为≧1×106-108cfu/g,酵母菌数为≧2.0×106cfu/g;
(4)制备植物性酵母乳酸菌原液A(图11中一次培养)
将上述植物性酵母乳酸菌发酵培养基和纯净水按重量比例 1:5-20进行混合搅拌,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10-20%,35-50oC培养1-5天,获得植物性酵母乳酸菌原液A;所述植物性酵母乳酸菌原液A中乳酸菌、酵母菌孢子总浓度为1×106-2×108cfu/ml;
(5)制备植物性酵母乳酸菌二次培养原液B(图11中二次培养)
二次培养原液B制备方法:将植物性酵母乳酸菌原液A稀释 2-5倍,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10-20%,在35-50oC温度中培养1-5天,获得植物性酵母乳酸菌二次培养原液B,所述植物性酵母乳酸菌二次培养原液B乳酸菌、酵母菌孢子总浓度为1×106-2×108cfu/ml。
此外:所述植物性酵母乳酸菌原液A、植物性酵母乳酸菌二次培养原液B可采用植物性酵母乳酸菌培养装置进行制备:
所述培养装置命名为:LABBIO植物性酵母乳酸菌培养装置,包括搅拌发酵罐1,固液分离装置3,过滤装置5,所述搅拌发酵罐1设有投料口1-4和出料口1-5,所述出料口1-5是通过连接管Ⅰ7与固液分离装置3上口3-5连接,所述连接管Ⅰ7设有阀门Ⅰ2;所述固液分离装置下口3-2通过连接管Ⅱ8连接过滤装置5的滤液投入口5-1,所述连接管Ⅱ8上设有阀门Ⅱ4;所述过滤装置5设有出料口5-5,所述出料口5-5连接有出料管9,所述出料管9上设有阀门Ⅲ6;
所述搅拌发酵罐1包括罐体1-6,搅拌电机1-1;所述罐体1-6主体为柱形,底部为倒圆锥形,所述罐体1-6侧部设有水位计1-2;所述投料口1-4设置于罐体1-6的顶部,所述出料口1-5设置于罐体1-6的底部中心;所述搅拌电机1-1下端固定连接有转轴1-7,所述转轴1-7从上至下固定连接有多个搅拌折叶组1-8,所述搅拌折叶组1-8设置于罐体1-6主体和圆锥形底部,所述搅拌折叶组1-8包括周向多个均匀分布的折叶1-9,置于罐体底部倒圆锥内折叶组中的折叶长度短于置于罐体1-6主体内折叶组中折叶的长度;所述罐体1-6内设有温度感应器1-10、酸碱度感应器1-11、电导率感应器1-12、糖度计传感器1-13,所述温度感应器、酸碱度感应器、电导率感应器、糖度计传感器均连接于自动检测控制计算机1-3,所述自动检测控制计算机连接搅拌电机1-1,控制搅拌电机1-1电源、转速;所述搅拌折叶基部平面与转轴垂直中心面呈45度夹角,所述折叶外部边缘斜向内折,所述内折的折叶边缘与折叶主体呈135度夹角;
所述固液分离装置3内设有过滤板3-1,所述过滤板上设有滤膜Ⅰ3-3,所述滤膜Ⅰ3-3孔径为30-40微米;所述过滤板3-1可为双层滤板,所述双层滤板上均设有同心过滤孔Ⅰ3-4,双层滤板中间夹装滤膜Ⅰ3-3;
所述过滤装置5包括过滤罐体5-3,所述过滤罐体5-3为柱形体,过滤罐体5-3内部设有过滤筒5-2,所述过滤筒5-2将过滤罐体5-3分隔为内、外两个腔体,所述滤液投入口5-1置于过滤罐体5-3内部腔体上方,所述出料口置于过滤罐体5-3外部腔体下方;所述过滤筒5-2上设有滤膜Ⅱ5-6,所述滤膜Ⅱ5-6孔径为3-5微米;过滤筒5-2可为双层过滤筒,所述双层过滤筒上均设有同心过滤孔Ⅱ5-4,所述双层过滤筒中间夹装滤膜Ⅱ5-6,所述滤膜Ⅱ5-6为纳米过滤膜,孔径为3-5微米。
所述LABBIO植物性酵母乳酸菌培养装置用于植物性酵母乳酸菌培养,主要步骤为:
1)投料
原料为植物性酵母乳酸菌发酵培养基,按比例添加低聚糖或蔗糖,向搅拌发酵罐投料时80-100 rpm/min高速均匀搅拌1-2分钟后10-30 rpm/min低速均匀搅拌;
2)固液分离
搅拌之后的固液混合物通过出料口1-5、连接管Ⅰ7,进入固液分离装置3的上口3-5,所述固液分离装置3为压力式分离器,过滤板的滤膜Ⅰ3-3孔径是30-40微米;分离所得植物性酵母乳酸菌培养液通过过滤板3-1后从下口3-2流出,进入连接管Ⅱ8;
3)过滤
所述过滤装置5中滤膜Ⅱ5-6的孔径是3-5微米;过滤得到2微米酵母菌和小于2微米的乳酸菌,获得植物性酵母乳酸菌原液A;
将一次培养植物性酵母乳酸菌原液A稀释 2-5倍,按比例添加低聚糖或蔗糖,重复上述步骤,扩增培养获得二次植物性酵母乳酸菌培养原液B。
本发明还提出了植物性酵母乳酸菌的应用,其技术要点是:所述植物性酵母乳酸菌二次培养原液B用于家禽、家畜或水产养殖饲料的益生菌添加剂或不含酒精的安全性杀菌剂。
进一步的,所述用于家禽、家畜或水产养殖饲料的益生菌添加功效的植物性酵母乳酸菌二次培养原液B制备中:植物乳酸菌菌种为:Lactobacillus fermentum(发酵乳酸杆菌), Lactobacillus paracasei(副干酪乳杆菌), Lactobacillus plantarum(植物乳杆菌),Lactobacillus lactis(乳酸乳球菌)Streptococcus lactis(乳酸链球菌);酵母菌菌种为:Zygosacckromyceskoiuxii(Boutroux) yaitow(鲁氏结合酵母菌),Pichiuahomala(E. C. Hahshe) Kurzmiah(异构毕赤酵母),Pichiakluyveri(克鲁弗毕赤酵母),Saccharomyces lactis(乳酸酵母),Saccharomyces validus(酿酒酵母);
所述用于家禽、家畜或水产养殖饲料的益生菌添加功效的植物性酵母乳酸菌二次培养原液B的使用方法包括:
(1)投喂:饲养家禽、家畜(例如牛、猪、羊、鸡、鸭、鹅等)及其宠物(猫、狗等)投喂植物性酵母乳酸菌二次培养原液B和饲料的混合物,所述混合物中植物性酵母乳酸菌二次培养原液B和饲料重量比为:1-10:100;
(2)制备有机肥:将植物性酵母乳酸菌二次培养原液B直接稀释100倍,制备植物性酵母乳酸菌原液B稀释液,所述植物性酵母乳酸菌原液B稀释液与家畜的屎尿混合,二者重量比为10-20:100;发酵方法为夏季发酵一周,春秋季节发酵两周,冬季发酵两至三周;
所述用于用于不含酒精的安全杀菌的植物性酵母乳酸菌二次培养原液B制备中:
植物性乳酸菌菌种为:Lactobacillus fermentum(发酵乳酸杆菌),Lactobacillusparacasei(副干酪乳杆菌),Lactobacillus plantarum(植物乳杆菌),Lactobacillusacidophilus(嗜酸乳杆菌) ,Lactobacillus thermophilus(嗜热乳杆菌),Streptococcus lactis(乳酸链球菌);所述杀菌功效中主要选择的酵母菌菌种为:Zygosacckromyceskoiuxii(Boutroux) yaitow(鲁氏结合酵母菌),Pichiuahomala( E. C.Hahshe) Kurzmiah(异构毕赤酵母),Saccharomyces lactis(乳酸酵母), SaccharomycesRoouxii(鲁氏酵母),Pichiakluyveri(克鲁弗毕赤酵母);
所述用于不含酒精的安全杀菌的植物性酵母乳酸菌二次培养原液B的使用方法包括:
(1)水质净化:将植物性酵母乳酸菌二次培养原液B放入装有活性炭或二氧化硅等吸附材料的细网吊袋中放入流水、养殖鱼水箱;使用用量为3吨水中加入5 kg装有活性炭或二氧化硅等吸附剂,所述吸附剂均匀喷淋1升稀释100倍的植物性酵母乳酸菌原液B,用于去除不流动水中霉菌和异味;
(2)杀菌、除味:将植物性酵母乳酸菌二次培养原液B直接稀释100倍,制备植物性酵母乳酸菌原液B稀释液,所述植物性酵母乳酸菌原液B稀释液喷淋到厨房灶台台面、畜舍中用于杀菌、除味、消臭。
实施例2
将实施例1获得的植物性酵母乳酸菌二次培养原液B记录为LABBIO植物性酵母乳酸菌LBF,所述LABBIO植物性酵母乳酸菌LBF的菌株安定,具有良好的活性,而且具有耐热,耐酸、耐盐等强性菌株的特性。
(1)LABBIO植物性酵母乳酸菌LBF物理性质
a. 耐酸:LABBIO植物性酵母乳酸菌LBF在pH2.0~4.2强酸环境下可以存活;
b. 耐盐:LABBIO植物性酵母乳酸菌LBF在海水中可以存活;
c. 耐热:LABBIO植物性酵母乳酸菌LBF经过短时间高温处理后仍有个体存活。
(2)LABBIO植物性酵母乳酸菌LBF的杀菌及抑菌作用(见图12)
a. 试验菌:Escherichia coli (大肠杆菌),Escherichia coli O157:H7(肠出血性大肠杆菌),Staphylococcus aureus (金黄色葡萄球菌)。
b.试验方法:将上述大肠杆菌以及金黄色葡萄球菌以105~106cfu/ml浓度分别接种于LABBIO植物性酵母乳酸菌LBF原液,10倍,100倍,300倍,500倍,1000倍稀释的LBF菌液。同时,将同等量的细菌接种于灭菌生理食盐水中,作为对照组。15分钟后,分别取200ul细菌与乳酸菌液的混合液铺于LB寒天培养基,并置于37度培养箱。12小时后取出平板计数细菌菌落。
c. 试验结果:接种细菌的灭菌蒸馏水出现了大量的大肠杆菌以及金黄色葡萄球菌的菌落,LABBIO植物性酵母乳酸菌LBF原液,10倍,100倍,300倍,500倍稀释的LBF菌液中均未检测到菌落。1000被稀释菌液虽然检测到菌落,但是较蒸馏水菌落明显减少。
d. 试验结论:LABBIO植物性酵母乳酸菌LBF原液具有较强的杀菌作用。
(3)LABBIO植物性酵母乳酸菌LBF生产代谢物质
目前常见的乳酸菌产品有Yupiter(尤比特)菌、EM菌、生源菌、Arpis(阿比斯)菌、(Virutus)缅甸菌、发酵菌、酵母菌等。一般多为动物性乳酸菌,例如酸奶制品,仅供食用。但是对人体和动植物最有效果的乳酸菌被认为是植物性乳酸菌。植物性乳酸菌与酵母经过适当的培养基发酵、熟成,可生成大量具有亲和性,易于生物及人体吸收的机能性成分。LABBIO植物性酵母乳酸菌LBF由以下多种乳酸菌生产代谢物质构成,是具有极其广泛应用价值的机能性物质。
a. 短链脂肪酸,乳酸-醋酸,丁酸,丙酸,乳酸等;
b. 菌体成分:胞壁酰二肽,肽聚糖,酵母聚糖等;
c. 丙烯酸类异黄酮黄豆苷元,染料木黄酮等;
d. 氨基酸—缬氨酸、亮氨酸、异亮氨酸、甘氨酸、丙氨酸、胱氨酸、丝氨酸、甲硫氨酸、苏氨酸、谷氨酰胺,天冬酰胺、苯丙氨酸、酪氨酸、色氨酸、组氨酸、赖氨酸、精氨酸、脯氨酸,天冬胺酸、谷氨酸、GABA(γ-氨基酸)等;
e. 天然维他命B 1、B 2、B 6、E、K1、H等;
f. 天然矿物质-磷、钙、镁、钠、钾等;
g. 大豆多肽 BB 1、露那辛(Lunasin多肽)等;
h. 卵磷脂(大豆磷脂)-磷脂酰胆碱,磷脂酰丝氨酸等;
i. 皂苷-大豆皂苷A,B,E,DDMP等;
j. 核酸-脱氧核糖核酸,核糖核酸等。
(4)LABBIO植物性酵母乳酸菌LBF的应用
a. 在家禽家畜等养殖业中的应用:
饲养家禽、家畜(例如牛、猪、羊、鸡、鸭、鹅等)及其宠物(猫、狗等)投喂LABBIO植物性酵母乳酸菌二次培养原液B之后,不仅会显著改善家禽、家畜健康,而且减轻环境粪尿异味,杀菌消臭,而且家畜的粪便经LABBIO植物性酵母乳酸菌LBF二次培养原液B喷淋发酵之后会得到良好的益生菌有机堆肥。喂食植物性酵母乳酸菌后的饲养家禽、家畜(例如牛、猪、羊、鸡、鸭、鹅等)及其宠物(猫、狗等),不仅杀灭病原菌,改善胃肠道功能,更有利于健康,提高成活率,而且改变了饲养家禽、家畜(例如牛、猪、羊、鸡、鸭、鹅等)肉质的呈味成分和氨基酸成分,使肉质更加鲜嫩。
在鸡舍中使用稀释100倍的LABBIO植物性酵母乳酸菌二次培养原液B进行喷淋,并且按照常规方法在鸡粪中添加稀释100倍的LABBIO植物性酵母乳酸菌二次培养原液B进行堆肥,如表1所示,监测了其中含有的氨、甲基硫醇等恶臭成分的浓度(单位:mg/m3),测定方法采用“三点比较式臭袋法”进行测定(国家检测标准编号为GB/T 14675-93 );结果显示可以大幅度地减少恶臭对环境的污染程度。通过这种添加LABBIO植物性酵母乳酸菌的方式,我们能够生产出臭味极小,易于土壤改良和植物吸收的有机肥料。同时,使鸡舍充分杀菌减少了家禽的发病率,并且能够有效提高家禽的成活率。
表1所述培养原液B对氨、甲基硫醇等恶臭成分的浓度的影响
对于饲养家禽、家畜(例如牛、猪、羊、鹅、鸭等)及宠物(猫、狗等)同样适用。例如,猫和狗等宠物,作为营养添加剂将LABBIO植物性酵母乳酸菌二次培养原液B的给予量应为每千克体重约1~2克。另外,将LABBIO植物性酵母乳酸菌二次培养原液B稀释到约1%后,可以作为家禽、家畜及宠物的饮用水使用。结果显示,不但家禽、家畜及其宠物的健康状况变得更好,而且非常明显地减少了家禽、家畜及其宠物的体臭和粪便的异臭味。
b. 在水质净化及改善生态平衡的作用
在河流水路(宽约2m)中用铁网划分约20m长作为试验水路的区域,将约5kg LABBIO植物性酵母乳酸菌LBF放入有细网的吊袋中,在试验水路上游的铁网中悬挂并浸入水中。另外,在试验水路内放养了鲤鱼,鲫鱼,鲑鱼和鳟鱼。实验开始后约20天,在试验水路内侧产生藻类。这种藻类为仅可在清澈水流中能观察到的藻类。此外,鲤鱼和鲫鱼生长状态非常健康。特别是当这几种鱼被煮熟并食用时,内脏完全无异味,可以食用。另外,众所周知鲑鱼和鳟鱼只能在水质良好的水域中生长。在实验经过约半年后,可以观察到被放生到试验水路中的所有鲑鱼和鳟鱼的生长状况都非常良好。实验开始约6个月后,在实验水路下游的水中检测到大约1x102cfu/ml植物性酵母乳酸菌。根据这些实验表明,在河流中通过投放LABBIO植物性酵母乳酸菌LBF可以净化河川水质并改善生态平衡。
c. 养殖观赏鱼的杀菌效果试验
在养殖观赏鱼如热带鱼或金鱼的水箱中设置净化装置,每45至60cm的间隔放置一个LABBIO植物性酵母乳酸菌二次培养原液B(用多孔质海绵物吸附)进行杀菌,同时分解由于观赏鱼的排泄物而产生的毒素氨类。在水箱中长期养殖的观赏鱼无病害,健康状况良好,可以减少管理的时间与精力。此外,可减少在水箱中由于水质恶化而易发生的蓝藻及水质污染,保持良好水质状态。
d. 公共浴池及家庭浴盆中去除霉菌和异味
公共浴池中通常需要使用强力清洗剂定期进行清洁,但是难以彻底冲洗附着在热水管道中的水垢及污泥。由于水垢容易积存在过滤槽中,因此需要经常清洗。对于公共浴池及家庭浴盆,尽管每天都进行清洁,但污垢往往会积聚在瓷砖的缝隙中,因此难以彻底消除浴池中令人不愉快的异味。污泥附着在使用后排出的热水管道中,容易发出难闻的恶臭,也容易产生蓝藻或蚊虫及细菌。虽然将除臭剂放在浴池中,可以被视为有香味的热水,但每天都需要补充三到四次,否则就无法维持香味等问题。为此,可将LABBIO植物性酵母乳酸菌二次培养原液B混合在公共浴池中。即,将1吨热水中混入5kg LABBIO植物性酵母乳酸菌二次培养原液B,或者将适量的原菌包装在网袋中,将袋子悬浮在公共浴池中。能有效地将通过热水管道中的污泥清除,并且之后很少堆积污泥。由于水垢变得难以堆积,清洗过滤罐的次数可以减少,在浴池中不会闻到不愉快的异味,并且浴缸内的水垢会减少、下水道的异味变得非常少,并且可以抑制发生蓝藻与蚊虫及细菌,当放入除臭剂时,可以达到香味突出、持续保持的效果。
f.放射性污水处理
将被放射性污染的土壤或污泥收集到处理槽中,加入处理液搅拌后脱水,脱水机的处理速率约每小时10立方米,处理后的脱水污泥含水率约为25%。然后将含有放射性的污水进行前处理,和一般的污水处理方法相同,经过絮凝及沉淀等前处理后,再将含有放射性污水导入到污水处理罐中。在污水处理罐中首先填充处理后的吸附材料,主要的吸附材料是植物性二氧化硅、沸石、活性炭及LABBIO植物性酵母乳酸菌二次培养原液B。其吸附材料的处理方法是将植物性二氧化硅用微孔网袋包扎好之后,浸泡在稀释100倍的LABBIO植物性酵母乳酸菌二次培养原液B中吸附饱和,同时将活性炭和沸石按照1:10的比例混合均匀。最后按照各种材质的比例将植物性二氧化硅:沸石:活性炭=1:5:0.5的比例搅拌均匀,填充到污水处理罐的处理层中,并且必须保证使污水的处理滞留时间保持在20分钟以上,这样处理后的污水即可达到无色、无味、杀菌、去除放射性污染率可达94%以上,能够使被污染的放射性污水处理后达到国家排放水标准。
表2及LABBIO植物性酵母乳酸菌二次培养原液B对放射性污染土壤降解的影响
检测项目 | 处理前污泥 | 处理后排放水 | 单位 | 检出限 | 半衰期 |
放射性碘(I-131) | 未检出 | 未检出 | Bq/kg | 40 | 8日 |
放射性铯(Cs-134) | 1900 | 未检出 | Bq/kg | 20 | 2.1年 |
放射性铯(Cs-137) | 2600 | 未检出 | Bq/kg | 20 | 30.2年 |
g.家庭用天然杀菌剂
家庭清洁对家人的健康至关重要,但是目前市场上销售的家庭用消毒剂和消臭剂等都含有酒精等化学成分,对人体都有或多或少的伤害及副作用,尤其是不利于婴幼儿使用。LABBIO植物性酵母乳酸菌LBF菌液经发酵后,产生大量乳酸,能够有效的抑制细菌等病原微生物的生长,而且菌液来源于纯天然植物,对人体没有任何毒副作用。本发明将LABBIO植物性酵母乳酸菌LBF菌液喷洒于厨房灶台台面,对喷洒前,喷洒后,以及喷洒24小时后台面细菌进行检测,发现经LBF菌液喷洒后,灶台台面几乎无细菌残留。因此LABBIO植物性酵母乳酸菌LBF发酵产生的乳酸可用于制作家庭绿色纯天然消毒剂和消臭剂等。
综上所述,本发明中的LABBIO植物性酵母乳酸菌LBF在医药保健、食品工业、机能性化妆品、农业生产、环境保护等领域中应用广泛,功效涵盖平衡人体胃肠道菌群、清洁皮肤、杀菌,提高免疫力、生产益生菌饲料、生产有机肥料、土壤改良、动植物保护、绿色生态农业种植和养殖、农业废弃物无害化处理、食用菌生产等多个领域。不仅有益人体健康,而且可以应用于家禽、家畜的动物和水产养殖类等水生生物。更有利于土壤改良,污染水质改善,减少化肥和农药使用,改善农作物种植等。在上述所有领域中,应用都得到了非常好的验证结果。
Claims (7)
1.植物性酵母乳酸菌的制备方法,其特征是:包括以下步骤:
(1)制备植物性酵母乳酸菌培养基
所述植物性酵母乳酸菌培养基原料组分按重量比包括如下:大豆提取物蛋白质70-95,糯米5-30,米糠0-30;将上述原料加纯净水调和成含水分40%-60%粘稠状,用搅拌机充分搅拌均匀,得到植物性酵母乳酸菌培养基;
(2)接种植物性乳酸菌及酵母菌菌种
所述植物性乳酸菌菌种为Lactobacillus fermentum(发酵乳酸杆菌),Lactobacillus paracasei(副干酪乳杆菌), Lactobacillus acidophilus(嗜酸乳杆菌),Lactobacillus casei(干酪乳杆菌),Lactobacillus delbrueckii(保加利亚乳杆菌),Lactobacillus arabinosus(阿拉伯乳杆菌),Lactobacillus caucasicus(高加索乳杆菌), Lactobacillus lactis(乳酸乳球菌), Lactobacillus Leichmanni(莱氏乳杆菌),Lactobacillus musicus(穆氏乳杆菌), Lactobacillus thermophilus(嗜热乳杆菌),Lactobacillus plantarum(植物乳杆菌),Streptococcus thermophilus(嗜热链球菌),Streptococcus cremoris(乳脂链球菌), Sterptococcusfaecalis(粪肠链球菌),Streptococcus lactis(乳酸链球菌),Leuconostocmesenteroides(肠膜明串珠菌)中的一至六种;
所述酵母菌菌种为Pichiakluyveri(克鲁弗毕赤酵母),Saccharomyces intermedius(间型酵母), Saccharomyces validus(酿酒酵母),Saccharomyces ellipsoideus(椭圆酿酒酵母), Saccharomyces malirisler(马里利斯勒酵母), Saccharomyces mandshuricus(东北酵母), Saccharomyces Vordermannii(沃尔酵母),Saccharomyces Peka(佩卡酵母), Saccharomyces shasshing(沙欣酵母), Saccharomyces piriformis(梨形酵母),Saccharomyces anamensis(越南酵母),Saccharomyces cartilaginosus(软骨状酵母),Saccharomyces Awamori(泡盛酒酵母), Saccharomyces Batatae(甘薯酒酵母),Saccharomyces Coreanus(朝鲜酵母), Saccharomyces robustus(粗壮酵母),Saccharomyces Carlsbergensis(卡尔酵母),Saccharomyces Monacensis(摩拿酵母),Saccharomyces Marxianus(马克斯酵母),Zygo Saccharomyces major(大接合酵母),Saccharomyces lactis(乳酸酵母), Saccharomyces Roouxii(鲁氏酵母),Hansenulaanomala(汉氏酵母),Zygosacckromyceskoiuxii (Boutroux) yaitow(鲁氏结合酵母),Pichiuahomala( E. C. Hahshe) Kurzmiah(异构毕赤酵母)的一种至五种;
所述植物性乳酸菌菌种和酵母菌菌种调制成混合菌种,所述混合菌种中乳酸菌、酵母菌孢子浓度比例为10-150:1-10;将混合菌种接种至植物性酵母乳酸菌培养基中,接种后乳酸菌、酵母菌的孢子总浓度为1x106-4x109cfu/kg;
(3)制备植物性酵母乳酸菌发酵培养基
上述接种植物性乳酸菌及酵母菌菌种的植物性酵母乳酸菌培养基置于4-10oC环境中培养45-180天,获得植物性酵母乳酸菌发酵培养基,所述植物性酵母乳酸菌发酵培养基中的植物性乳酸菌数为≧1×106-108cfu/g,酵母菌数为≧2.0×106cfu/g;
(4)制备植物性酵母乳酸原液A
将上述植物性酵母乳酸菌发酵培养基和纯净水按重量比例 1:5-20进行混合搅拌,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10-20%, 35-50oC培养1-5天,获得植物性酵母乳酸原液A;所述植物性酵母乳酸原液A中乳酸菌、酵母菌孢子总浓度为1×106-2×108 cfu/ml;
(5)制备植物性酵母乳酸菌二次培养原液B
将植物性酵母乳酸菌原液A稀释 2-5倍,添加低聚糖或蔗糖,所述低聚糖或蔗糖的质量浓度为10-20%,在35-50oC温度中培养1-5天,获得植物性酵母乳酸菌二次培养原液B,所述植物性酵母乳酸菌二次培养原液B乳酸菌、酵母菌孢子总浓度为1×106-2×108cfu/ml。
2.根据权利要求1所述的植物性酵母乳酸菌的制备方法,其特征是:所述植物性酵母乳酸菌二次培养原液B使用植物性酵母乳酸菌培养装置进行制备:所述培养装置包括搅拌发酵罐,固液分离装置,过滤装置,所述搅拌发酵罐通过连接管Ⅰ与固液分离装置连接;所述固液分离装置通过连接管Ⅱ连接过滤装置;所述过滤装置连接有出料管,所述连接管Ⅰ、连接管Ⅱ、出料管上均设有阀门;
所述搅拌发酵罐包括罐体,搅拌电机;所述罐体主体为柱形,底部为倒圆锥形,所述罐体侧部设有水位计;所述投料口设置于罐体的顶部,所述出料口设置于罐体的底部中心;所述搅拌电机下端固定连接有转轴,所述转轴从上至下固定连接有多个搅拌折叶组,所述搅拌折叶组设置于罐体主体和圆锥形底部,所述搅拌折叶组包括周向多个均匀分布的折叶;所述固液分离装置内设有过滤板,所述过滤板上设有滤膜Ⅰ,所述滤膜Ⅰ孔径为30-40微米;所述过滤装置包括过滤罐体,所述过滤罐体为柱形体,过滤罐体内部设有过滤筒,所述过滤筒将过滤罐体分隔为内、外两个腔体,所述滤液投入口置于过滤罐体内部腔体上方,所述出料口置于过滤罐体外部腔体下方;所述过滤筒上设有滤膜Ⅱ,所述滤膜Ⅱ孔径为3-5微米;
所述植物性酵母乳酸菌培养装置用于植物性酵母乳酸菌培养的主要步骤为:
1)投料
原料为植物性酵母乳酸菌发酵培养基、纯净水,按比例添加低聚糖或蔗糖,向搅拌发酵罐投料时80-100 rpm/min高速均匀搅拌1-2分钟后10-30 rpm/min低速均匀搅拌;
2)固液分离
所述固液分离装置为压力式分离器,固液分离过滤板的滤膜Ⅰ孔径是30-40微米;
3)过滤
所述过滤装置中滤膜Ⅱ的孔径是3-5微米;过滤得到2微米酵母菌和小于2微米的乳酸菌,获得植物性酵母乳酸菌原液A;
将植物性酵母乳酸菌原液A稀释 2-5倍,重复上述步骤,按比例添加低聚糖或蔗糖,扩增培养获得二次植物性酵母乳酸菌培养原液B。
3.根据权利要求1所述的植物性酵母乳酸菌的应用,其特征是:所述植物性酵母乳酸菌二次培养原液B用于家禽、家畜或水产养殖饲料的益生菌添加剂或不含酒精的安全性杀菌剂。
4.根据权利要求3所述的植物性酵母乳酸菌的应用,其特征是:所述用于家禽、家畜或水产养殖饲料的益生菌添加功效的植物性酵母乳酸菌二次培养原液B制备中:植物乳酸菌菌种为:Lactobacillus fermentum(发酵乳酸杆菌), Lactobacillus paracasei(副干酪乳杆菌), Lactobacillus plantarum(植物乳杆菌),Lactobacillus lactis(乳酸乳球菌)Streptococcus lactis(乳酸链球菌);酵母菌菌种为:Zygosacckromyceskoiuxii(Boutroux) yaitow(鲁氏结合酵母菌),Pichiuahomala( E. C. Hahshe) Kurzmiah(异构毕赤酵母),Pichiakluyveri(克鲁弗毕赤酵母),Saccharomyces lactis(乳酸酵母),Saccharomyces validus(酿酒酵母)。
5.根据权利要求3所述的植物性酵母乳酸菌的应用,其特征是:所述用于家禽、家畜或水产养殖饲料的益生菌添加功效的植物性酵母乳酸菌二次培养原液B的使用方法包括:
(1)投喂:饲养家禽、家畜(例如牛、猪、羊、鸡、鸭、鹅等)及其宠物(猫、狗等)投喂植物性酵母乳酸菌二次培养原液B和饲料的混合物,所述混合物中植物性酵母乳酸菌二次培养原液B和饲料重量比为:1-10:100;
(2)制备有机肥料:将植物性酵母乳酸菌二次培养原液B直接稀释100倍,制备植物性酵母乳酸菌原液B稀释液,所述植物性酵母乳酸菌原液B稀释液与家畜的屎尿混合,二者重量比为10-20:100;发酵方法为夏季发酵一周,春秋季节发酵两周,冬季发酵两至三周。
6.根据权利要求3所述的植物性酵母乳酸菌的应用,其特征是:所述用于不含酒精的安全杀菌的植物性酵母乳酸菌二次培养原液B制备中:
植物性乳酸菌菌种为:Lactobacillus fermentum(发酵乳酸杆菌), Lactobacillusparacasei(副干酪乳杆菌),Lactobacillus plantarum(植物乳杆菌),Lactobacillusacidophilus(嗜酸乳杆菌) ,Lactobacillus thermophilus(嗜热乳杆菌),Streptococcus lactis(乳酸链球菌);所述杀菌功效中主要选择的酵母菌菌种为:Zygosacckromyceskoiuxii(Boutroux) yaitow(鲁氏结合酵母菌),Pichiuahomala( E. C.Hahshe) Kurzmiah(异构毕赤酵母),Saccharomyces lactis(乳酸酵母), SaccharomycesRoouxii(鲁氏酵母),Pichiakluyveri(克鲁弗毕赤酵母)。
7.根据权利要求3所述的植物性酵母乳酸菌的应用,其特征是:所述用于不含酒精的安全杀菌的植物性酵母乳酸菌二次培养原液B的使用方法包括:
(1)水质净化:将植物性酵母乳酸菌二次培养原液B放入装有活性炭或二氧化硅等吸附材料的细网吊袋中放入流水、养殖鱼水箱;使用用量为3吨水中加入5 kg装有活性炭或二氧化硅等吸附剂,所述吸附剂均匀喷淋1升稀释100倍的植物性酵母乳酸菌原液B,用于去除不流动水中霉菌和异味;
(2)杀菌、除味:将植物性酵母乳酸菌二次培养原液B直接稀释100倍,制备植物性酵母乳酸菌原液B稀释液,所述植物性酵母乳酸菌原液B稀释液喷淋到厨房灶台台面、畜舍中用于杀菌、除味、消臭。
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