CN111802515A - 一种生物补钙素及其制备和应用方法 - Google Patents
一种生物补钙素及其制备和应用方法 Download PDFInfo
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- CN111802515A CN111802515A CN202010706334.6A CN202010706334A CN111802515A CN 111802515 A CN111802515 A CN 111802515A CN 202010706334 A CN202010706334 A CN 202010706334A CN 111802515 A CN111802515 A CN 111802515A
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Classifications
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- A—HUMAN NECESSITIES
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- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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Abstract
本发明公开了一种生物补钙素,属于饲料添加剂技术领域。生物补钙素的原料质量份数组成为:益生菌发酵菌粉20~30份、牡蛎粉30~50份、环糊精5~10份、维生素2~5份、葡萄糖酸钙1~2份、乳钙蛋白粉5~10份、增稠剂3~6份。本发明利用现代生物技术和水产养殖营养调控技术制备高效生物补钙素,兼有补钙和益生菌发酵饲料的双重功效,具有显著改良水质、钙质吸收快等优点,并通过益生菌发酵作用显著提升补钙素的补钙效果,提升水产养殖动物的机体免疫力,从而显著提升养殖经济效益,降低养殖风险,促进水产养殖业可持续发展。
Description
技术领域
本发明涉及一种生物补钙素,尤其是一种利用现代生物技术和水产养殖营养调控技术制备的高效生物补钙素,属于饲料添加剂技术领域。
背景技术
为了满足不断增长的水产品的需求,解决因水产养殖规模不断扩大和生长周期缩减而导致的生理机能紊乱、疾病频发成为水产养殖的重点问题,其中补钙饲料添加剂的研发及应用成为水产养殖的关键环节。钙是养殖水体中附有植物生长所需营养元素之一,钙是植物细胞壁的中药组成成份,在缺乏钙元素的情况下,会限制藻类繁殖速度,会出现肥水困难,水色容易变清等问题。同时,钙是水产养殖品种骨骼、鳞片、甲壳等组成重要成份,水产养殖品种在缺钙情况下会出现蜕壳时间延迟,硬壳速度减慢,蜕壳死亡现象。钙的作用不仅体现在水产养殖品种生理需要上,同时也体现在养殖环境修复与调节上。水产养殖池塘,如果长时间养殖一个品种的水产品,那么就容易出现钙磷等营养元素缺乏,进而对水产品的质量产生不良影响。
水产养殖品种,特别是虾蟹类养殖品种,其生长形态的变化,器官的发育和断肢再生等,都要通过蜕皮与蜕壳未完成。钙质是关键,钙是虾蟹甲壳的必须组成部分。当虾蟹体内钙量得到满足的情况下,其蜕壳速度、蜕壳成功率及蜕壳后个体的大小都会有所增加,提高虾蟹的抗病能力,最终保证养殖的经济效益。向水体中投放钙,可以增加水体的含钙量,同时有效地促进藻类的繁殖生长,使水质达到肥活爽嫩的要求,为养殖水体及养殖品种提供天然的产氧源以及天然的抗应激保护。
目前,补钙饲料添加剂在水产养殖中的应用愈加广泛,现有技术中,专利201610869220.7公开了一种水产养殖用的补钙剂,包括以下重量份的原料组分:珍珠层粉12-18份、乳钙蛋白粉15-25份、蛋壳粉 5-15份、骨粉20-30份、骨胶原蛋白4-6份、维生素A6-8份、维生素E 20-30份;该发明的补钙剂添加可以用于人工水产养殖虾蟹鱼快速补钙、防治钙缺乏症,保护养殖动物肝肾功能、提高免疫力、及时纠正酸中毒、维持体内电解质平衡,促进虾蟹鱼新存代谢顺利脱壳固壳,促进生长;专利201811097108.1公开了一种用于水产养殖的补钙溶液及其制备和使用方法,由以下重量份组分组成:葡萄糖酸钙20-40份;乳酸钙10-30份;磷酸氢钙20-40份;维生素A 5-10份;维生素D3 5-10份;维生素B2 1-10份;酸化剂10-30份;增稠稳定剂5-15份;该补钙溶液能够增强水产品的活力,促进机体钙质吸收,同时能够有效降低死亡率。但目前补钙饲料添加剂存在补钙效果差、持续药效时间短、生产成本高等,制约了补钙饲料添加剂的应用。
发明内容
为了克服上述现有技术的不足,本发明提供一种生物补钙素,利用现代生物技术和水产养殖营养调控技术制备高效生物补钙素,具有显著改良水质、钙质吸收快等优点,并通过益生菌发酵作用显著提升补钙素的补钙效果,提升水产养殖动物的机体免疫力,从而显著提升养殖经济效益,降低养殖风险,促进水产养殖业可持续发展。
本发明通过下述技术方案实现上述技术效果:
一种生物补钙素,其原料质量份数组成为:益生菌发酵菌粉20~30份、牡蛎粉30~50份、环糊精5~10份、维生素2~5份、葡萄糖酸钙1~2份、乳钙蛋白粉5~10份、增稠剂3~6份。
优选的,一种生物补钙素,其原料质量份数组成为:益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份。
进一步的,上述益生菌包括芽孢杆菌和乳杆菌的混合菌剂,其中所述芽孢杆菌为凝结芽孢杆菌,所述乳杆菌为嗜酸乳杆菌。
优选的,所述凝结芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009中的一种或两种的混合菌剂,其中凝结芽孢杆菌GDMCC1.645购于广东省微生物菌种保藏中心,凝结芽孢杆菌CGMCC1.2009购自中国微生物菌种保藏管理委员会普通微生物中心。
进一步的,所述嗜酸乳杆菌购于广东省微生物菌种保藏中心,编号分别为GDMCC1.208。
进一步的,所述益生菌发酵菌粉的制备工艺具体包括如下步骤:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉。
上述芽孢杆菌种子液的制备工艺为:将活化后的芽孢杆菌接种至种子培养基,所述种子培养基的组分为:葡萄糖2%、蛋白胨2%、酵母膏1%、硫酸镁0.01%、硫酸锰0.003%,pH6.5~7.0,在温度28~37℃、转速150~3000rpm条件下,培养16~24h,要求种子液的有效活菌数≥109cfu/mL。
上述芽孢杆菌活化的工艺为:采用本领域常用培养基LB固体培养基,在温度28~37℃条件下,培养24h。
最优的,所述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为2:3。
进一步的,所述增稠剂为黄原胶、海藻胶和纤维素纳中的一种或多种,增稠稳定剂,能够是制备的体系长期稳定存在。
本发明还提供一种生物补钙素的制备工艺,具体包括如下步骤:
(1)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(2)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(3)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(4)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素。
本发明提供一种生物补钙素,具有补钙、保健、治疗和调节免疫等多种功效,是水产养殖补钙的理想、绿色、环保、安全、新型饲料添加剂。该发明饲料添加剂以牡蛎粉、葡萄糖酸钙和乳钙蛋白粉为主要钙剂,辅以牡蛎粉,通过微生物的发酵作用将难以吸收利用的钙粉转化为易于吸收利用的有机钙,促进水产品对钙的吸收,显著提升补钙效果。同时,通过益生菌的发酵作用,可以高效调理南美白对虾等水产动物肠道健康,标本兼治,固本培元,保持肠道微生态平衡和最佳免疫状态,提高营养物质的消化吸收率,提高动物机体免疫力。
本发明提供一种生物补钙素,所述补钙素产品在水产饲料全价料中使用的质量比例为0.1~0.5%:或所述的饲料添加剂在预混料、浓缩料以全价料中所需的比例添加使用。
本发明提供一种生物补钙素,与现有技术相比,具有以下显著优势:
(1)本发明兼有补钙和益生菌发酵饲料的双重功效,利用好氧菌(凝结芽孢杆菌)和厌氧菌(嗜酸乳杆菌)共同发酵制备生物补钙素,通过微生物间的协同作用显著提升补钙素的补钙效果,增强免疫力功效,进入动物肠道后,对促进双歧杆菌等肠道益生菌的生产,抑制病害菌生长,恢复肠道菌群平衡,增强肠道免疫功能和生理功能具有重要作用,可以直接替代益生菌发酵饲料添加剂;
(2)本申请将牡蛎壳粉应用于饲料添加剂,牡蛎壳含大量2-10μm微孔结构,含有多种生物活性的氨基多糖及特性蛋白,具有较强的吸附能力,能容纳一定大小的分子,牡蛎壳粉与β-环糊精交联,能够很好的吸附和包裹益生菌、维生素、葡萄糖酸钙和乳钙蛋白粉,构建了稳定的补钙反应体系,延长了饲料添加剂的作用时间;同时牡蛎粉也是较好的补钙原料,实现了资源化高效利用;
(3)本发明实验结果表明益生菌发酵菌粉对产品的补钙效果影响显著,改变益生菌发酵菌粉的菌种对产品的补钙效果具有显著的影响作用,较直接添加补钙原料补钙效果差异显著;
(4)本申请生物补钙素应用工艺简单,便于规模化推广应用。
具体实施方式
在以下实施例中,芽孢杆菌种子液的制备工艺为:将活化后的芽孢杆菌接种至种子培养基,所述种子培养基的组分为:葡萄糖2%、蛋白胨2%、酵母膏1%、硫酸镁0.01%、硫酸锰0.003%,pH6.5~7.0,在温度28~37℃、转速150~3000rpm条件下,培养16~24h,要求种子液的有效活菌数≥109cfu/mL;其中芽孢杆菌活化的工艺为:上述芽孢杆菌活化的工艺为:采用本领域常用培养基LB固体培养基,在温度28~37℃条件下,培养24h。
所述凝结芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009中的一种或两种的混合菌剂,其中凝结芽孢杆菌GDMCC1.645购于广东省微生物菌种保藏中心,凝结芽孢杆菌CGMCC1.2009购自中国微生物菌种保藏管理委员会普通微生物中心;所述嗜酸乳杆菌购于广东省微生物菌种保藏中心,编号分别为GDMCC1.208。
实施例1
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(6)配料:按益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份比例准备配料;
(7)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(8)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(9)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(10)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为2:3;上述增稠剂为海藻胶。
实施例2
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(6)配料:按益生菌发酵菌粉30份、牡蛎粉30份、环糊精10份、维生素2份、葡萄糖酸钙2份、乳钙蛋白粉5份、增稠剂6份比例准备配料;
(7)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(8)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(9)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(10)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为2:3;上述增稠剂为海藻胶。
实施例3
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(6)配料:按益生菌发酵菌粉20份、牡蛎粉50份、环糊精5份、维生素5份、葡萄糖酸钙1份、乳钙蛋白粉10份、增稠剂3份比例准备配料;
(7)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(8)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(9)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(10)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为2:3;上述增稠剂为海藻胶。
实施例4
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(6)配料:按益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份比例准备配料;
(7)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(8)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(9)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(10)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为3:2;上述增稠剂为海藻胶。
实施例5
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(6)配料:按益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份比例准备配料;
(7)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(8)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(9)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(10)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌GDMCC1.645;上述增稠剂为海藻胶。
实施例6
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(6)配料:按益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份比例准备配料;
(7)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(8)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(9)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(10)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌CGMCC1.2009;上述增稠剂为海藻胶。
对比实施例1
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)厌氧发酵:接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(3)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(4)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(5)配料:按益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份比例准备配料;
(6)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(7)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(8)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(9)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述增稠剂为海藻胶。
对比实施例2
一种生物补钙素,具体制备工艺步骤包括:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)低温浓缩:发酵结束后对发酵液进行低温浓缩;
(4)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉;
(5)配料:按益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份比例准备配料;
(6)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(7)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(8)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(9)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素;
上述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为2:3;上述增稠剂为海藻胶。
实施例7
实验动物:南美白对虾;
地址:青岛贝宝海洋科技有限公司琅邪养殖基地;
实验组:分别采用实施例1-6、对比例1-2及对照组1-2所述的饲料添加剂按0.5%的比例添加到饲料中,混合均匀,进行投喂,其中对照1组生物补钙素不含益生菌发酵菌粉,其余同实施例1;对照2组为空白对照组,饲料中不含补钙素;
每组实验设3个平行,池子大小为4.2×5.4×1.2m,每池投放1000尾软壳虾虾苗,喂养时间3个月,观察虾的生长情况,并测定虾的增重率(%);经检测,各实验组的有关实验结果如表1所示:
表1 养殖实验结果
组别 | 增重率(%) | 活力 | 软壳虾恢复情况 |
实施例1 | 32.17±0.79 | 强 | 壳硬光滑 |
实施例2 | 30.21±1.73 | 强 | 壳硬光滑 |
实施例3 | 31.17±2.02 | 强 | 壳硬光滑 |
实施例4 | 29.68±1.98 | 较强 | 壳硬光滑 |
实施例5 | 26.85±1.62 | 较强 | 甲壳粗厚 |
实施例6 | 27.38±1.55 | 较强 | 甲壳粗厚 |
对比实施例1 | 26.19±0.94 | 较弱 | 甲壳粗厚 |
对比实施例2 | 28.53±1.15 | 较弱 | 甲壳粗厚 |
对照1组 | 24.76±1.08 | 弱 | 甲壳粗厚 |
对照2组 | 21.86±1.34 | 弱 | 甲壳粗厚 |
上述实验结果表明,实施例1-4具有较好的补钙效果,软壳虾活力强,且软壳虾的壳硬光滑,这说明了实施例1-4生物补钙素取得了较好的补钙效果;同时,实验结果表明生物补钙素显著促进了软壳虾的生长,与空白对照组相比差异显著,其中实施例1为本发明最佳实施例。实施例5-6的实验结果表明,好氧发酵过程中菌剂对生物补钙和促生长具有显著的影响;对比实施例1-2实验结果表明本发明中好氧发酵和厌氧发酵对促进软壳虾的生长取得了“1+1>2”的技术效果。综合比较,实施例1为本发明最佳实施例。
以上实施例仅用于说明本发明的技术方案,而非对其进行限制;尽管参照前述实施例对被发明进行了详细的说明,但对于本领域的普通技术人员来说,依然可以对前述实施例所记载的技术方案进行修改,或者对其中部分技术特征进行等同替换;而对这些修改或者替换,并不使相应技术方案的本质脱离本发明所要求保护的技术方案的精神和范围。
Claims (9)
1.一种生物补钙素,其特征在于所述补钙素原料质量份数组成为:益生菌发酵菌粉20~30份、牡蛎粉30~50份、环糊精5~10份、维生素2~5份、葡萄糖酸钙1~2份、乳钙蛋白粉5~10份、增稠剂3~6份。
2.根据权利要求1所述的一种生物补钙素,其特征在于所述生物补钙素原料质量份数组成为:益生菌发酵菌粉28份、牡蛎粉45份、环糊精6份、维生素3份、葡萄糖酸钙1.8份、乳钙蛋白粉8份、增稠剂4份。
3.根据权利要求1所述的一种生物补钙素,其特征在于所述益生菌发酵菌粉由芽孢杆菌和乳杆菌发酵制备,其中所述芽孢杆菌为凝结芽孢杆菌,所述乳杆菌为嗜酸乳杆菌;所述所述凝结芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009中的一种或两种的混合菌剂,其中凝结芽孢杆菌GDMCC1.645购于广东省微生物菌种保藏中心,凝结芽孢杆菌CGMCC1.2009购自中国微生物菌种保藏管理委员会普通微生物中心;所述嗜酸乳杆菌购于广东省微生物菌种保藏中心,编号分别为GDMCC1.208。
4.根据权利要求1所述的一种生物补钙素,其特征在于所述益生菌发酵菌粉的制备工艺具体包括如下步骤:
(1)发酵液制备:按照质量百分比麸皮2%、豆粕2%、蛋白胨1%、玉米浆1%、磷酸二氢钾0.02%、磷酸氢二钾0.01%、硫酸镁0.02%、氯化钙0.01%,余量比为水的比例配制液体发酵液,调节pH至6.5~7.0;
(2)好氧发酵:接种凝结芽孢杆菌种子液,接种量分别为2%,控制发酵温度28~37℃,转速150~300rpm;在此发酵条件下发酵24h;
(3)厌氧发酵:好氧发酵结束后,接种嗜酸乳杆菌菌粉,接种量为2%,发酵温度28~37℃,控温厌氧发酵12h;
(4)低温浓缩:厌氧发酵结束后对发酵液进行低温浓缩;
(5)吸附干燥:浓缩后用0.5倍质量比的麸皮吸附,在40~60℃鼓风干燥机中干燥,水分控制在8~10%,即得益生菌发酵菌粉。
5.根据权利要求4所述的一种生物补钙素,其特征在于所述芽孢杆菌种子液的制备工艺为:将活化后的芽孢杆菌接种至种子培养基,所述种子培养基的组分为:葡萄糖2%、蛋白胨2%、酵母膏1%、硫酸镁0.01%、硫酸锰0.003%,pH6.5~7.0,在温度28~37℃、转速150~3000rpm条件下,培养16~24h,要求种子液的有效活菌数≥109cfu/mL;所述芽孢杆菌活化的工艺为:采用本领域常用培养基LB固体培养基,在温度28~37℃条件下,培养24h。
6.根据权利要求4所述的一种生物补钙素,其特征在于所述芽孢杆菌为凝结芽孢杆菌GDMCC1.645和凝结芽孢杆菌CGMCC1.2009的混合菌剂,其接种比例为2:3。
7.根据权利要求1所述的一种生物补钙素,其特征在于所述所述增稠剂为黄原胶、海藻胶和纤维素纳中的一种或多种,增稠稳定剂,能够是制备的体系长期稳定存在。
8.根据权利要求1所述的一种生物补钙素,其特征在于所述生物补钙素的制备工艺具体包括如下步骤:
(1)制备牡蛎粉颗粒,先将牡蛎壳粉进行超声震荡,分别形成40~80nm的颗粒;
(2)将牡蛎粉加入质量比10倍的去离子水,然后加入环糊精,搅拌分散均匀,得到牡蛎粉与环糊精的混合溶液;
(3)将益生菌发酵菌粉、维生素、葡萄糖酸钙、乳钙蛋白粉、增稠剂按比例加入到牡蛎粉与环糊精的混合溶液;
(4)混合后在60~65℃下加热、超声振荡1~2h后,冷却、过滤、低温真空烘干,得到生物补钙素。
9.根据权利要求1所述的一种生物补钙素,其特征在于所述补钙素产品在水产饲料全价料中使用的质量比例为0.1~0.5%:或所述的饲料添加剂在预混料、浓缩料以全价料中所需的比例添加使用。
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