CN111685238A - 无菌大小鼠的无菌饲料及其制备方法 - Google Patents
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Abstract
本发明属于实验动物技术领域,具体涉及无菌大小鼠的无菌饲料的制备方法,精确称取基础饲料、3.5%质量比的矿物质预混料和0.1%质量比复合维生素进行混合,加入6.99%质量比的多不饱和脂肪酸进行搅拌,加入水分搅拌混合,通过制粒机进行制粒,再进行低温烘干水分、辐照灭菌和高温高压灭菌,获得无菌大小鼠的无菌饲料。本发明在无菌大小鼠的无菌饲料的制备过程中,在基础饲料上额外添加矿物质和维生素,保证经辐射灭菌、高压蒸汽灭菌后,对损失的营养物质的补充,从而保证无菌大小鼠生长情况的稳定。本发明还提供了利用上述方法制备的无菌大小鼠的无菌饲料经过辐照灭菌和高温高压灭菌的双重灭菌,能够完全满足无菌实验大小鼠的无菌饲料的需求。
Description
技术领域
本发明属于实验动物技术领域,具体涉及无菌大小鼠的无菌饲料及其制备方法。
背景技术
无菌动物是不携带包括细菌、病毒、真菌和寄生虫等任何外源生命形式的动物。普通的环境中存在大量的微生物,无菌动物必须饲养在一个完全不含有微生物的空间内。这样的空间在自然界内是不存在的,必须人为建造。这种人为创造的并能够保证无菌的装置命名为隔离器。随着无菌动物研究的开展,无菌动物的饲料和饲养装置也不断地改进。无菌动物体内外不携带任何生物体,饲育于和外界隔离的装置,这样排除生物对于实验结果的干扰,明显提高了实验的敏感性、重复性。有利于对生命活动基本规律的研究和分析,已在生物、医学等方面有广泛的应用。无菌饲料是实验室培育合格实验动物的重要条件之一,只有达到标准的饲料才能保证实验动物健康生长,达到试验预期效果,否则将直接影响实验动物的生长和试验结果的评定。常见的饲料灭菌方式包括:高温高压灭菌和辐照灭菌。目前我国应用较多的实验室灭菌方法为高压蒸汽灭菌法,该方法能够达到灭菌的目的,但对饲料组织的结构和营养成分破坏较大,不能满足实验室对无菌、营养饲料的要求。饲料辐照加工是采用60Co-γ射线照射饲料,以达到灭菌的目的,不会产生辐照残留,如同食品的加热除菌和水的加氯消毒一样,是一个物理加工过程,不会对动物体造成损害,但是辐照灭菌存在灭菌不彻底和剂量难以控制的情况。
发明内容
针对上述现有技术的不足,本发明提供了无菌大小鼠的无菌饲料的制备方法,目的是为了解决现有技术中高压蒸汽灭菌法对饲料组织的结构和营养成分破坏较大,不能满足实验室对无菌、营养饲料的要求;辐照灭菌存在灭菌不彻底和剂量难以控制的情况的技术问题。本发明还提供了上述方法制备的无菌大小鼠的无菌饲料。
本发明提供的无菌大小鼠的无菌饲料的制备方法,具体技术方案如下:
无菌大小鼠的无菌饲料的制备方法,包括如下步骤:
S1,精确称取基础饲料、3.5%质量比的矿物质预混料和0.1%质量比复合维生素进行混合,获得混合粉料;
S2,步骤S1中的混合粉料加入6.99%质量比的多不饱和脂肪酸进行搅拌,获得油料混合物;
S3,步骤S2中的油料混合物加入水分搅拌混合,通过制粒机进行制粒,获得软颗粒饲料;
S4,将步骤S3中的软颗粒饲料进行低温烘干水分、辐照灭菌和高温高压灭菌,获得无菌大小鼠的无菌饲料。
在某些实施方式中,步骤S1中,所述基础饲料按质量百分比计包括19.99%酪氨酸、0.3%胱氨酸、39.68%玉米淀粉、13.19%麦芽糊精、11%蔗糖、5%膳食纤维、0.0014%抗氧化剂和0.25%酒石酸氢胆碱。
在某些实施方式中,步骤S2中,所述多不饱和脂肪酸包括1.08%SFA、1.62MUFA和4.29%的PUFA。
在某些实施方式中,步骤S3中,所述软颗粒饲料的粒径为10-15mm。
在某些实施方式中,步骤S4中,所述烘干的温度为26摄氏度,时间为48h;辐照灭菌的剂量50kgy;高温高压灭菌的温度为121℃,时间为30min。
本发明还提供了根据上述的方法制备的无菌大小鼠的无菌饲料。
本发明具有以下有益效果:本发明在无菌大小鼠的无菌饲料的制备过程中,在基础饲料上额外添加矿物质和维生素,保证对50kgy辐照剂量下饲料粗蛋白、蛋氨酸均明显降低,维生素A、E、B1、B6均有不同程度的损失,以及饲料再次经高压蒸汽灭菌后,其粗蛋白、精氨酸、赖氨酸、蛋氨酸明显减少和维生素A、B1、B2、B6损失的补充。此外,本发明制备的无菌大小鼠的无菌饲料经过辐照灭菌和高温高压灭菌的双重灭菌,能够完全满足无菌实验大小鼠的无菌饲料的需求,由于额外兼顾对营养的补充,从而保证无菌大小鼠生长情况的稳定;同时,本发明提供的无菌饲料使大小鼠自由采食,不需要灌胃,如此不仅符合动物伦理,也节约实施灌胃的人力成本。
附图说明
图1是本发明提供的无菌大小鼠的无菌饲料的制备方法的流程图;
图2是实施例中实施例组和对照组的生长曲线图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明白,以下结合具体实施例,并参照附图1-2,对本发明进一步详细说明。
实施例
本实施例提供的无菌大小鼠的无菌饲料的制备方法,具体技术方案如下:
无菌大小鼠的无菌饲料的制备方法,包括如下步骤:
S1,精确称取如表1所示的原料,按质量百分比计,将19.99%酪氨酸、0.3%胱氨酸、39.68%玉米淀粉、13.19%麦芽糊精、11%蔗糖、5%膳食纤维、0.0014%抗氧化剂、0.25%酒石酸氢胆碱、3.5%矿物质预混料和0.1%复合维生素进行混合,获得混合粉料;
S2,步骤S1中的混合粉料加入1.08%SFA、1.62MUFA和4.29%的PUFA进行搅拌,获得油料混合物;
S3,步骤S2中的油料混合物加入水分搅拌混合,通过制粒机进行制粒,获得粒径为10-15mm的软颗粒饲料,本实施例中软颗粒饲料的粒径为12mm左右;
S4,将步骤S3中的软颗粒饲料进行低温烘干水分(温度为26摄氏度,时间为48h)、辐照灭菌(辐照灭菌的剂量50kgy)和高温高压灭菌(温度为121℃,时间为30min),获得无菌大小鼠的无菌饲料。
表1本实施例提供的无菌大小鼠的无菌饲料的称量原料
成分 | 重量(g) |
酪蛋白 | 200 |
L-胱氨酸 | 3 |
玉米淀粉 | 397 |
麦芽糊精 | 132 |
蔗糖 | 110 |
膳食纤维 | 50 |
抗氧化剂 | 0.014 |
矿物质预混料 | 35 |
酒石酸氢胆碱 | 2.5 |
复合维生素 | 1 |
SFA | 10.8227 |
MUFA | 16.17 |
PUFA | 42.9436 |
合计 | 1000.4503 |
本实施例还提供了根据上述的方法制备的无菌大小鼠的无菌饲料。经过检测,显示本实施例提供的无菌大小鼠的无菌饲料菌落为阴性,无菌检测方法:取饲料溶解液,以无菌操作将该供试品分别接种于需气菌、厌气菌培养基6管,其中1管接种金黄色葡萄球菌对照用菌液1ml,作阳性对照,另接种于真菌培养基5管。轻轻摇动,使供试品与培养基混合。需气菌、厌气菌培养基管置30~35℃、真菌培养基管置20~25℃培养7日。在培养期间应逐日观察并记录是否有菌生长。阳性对照管在24小时内应有菌生长,如在加入供试品后,培养基出现浑浊,培养7天后,不能从外观上判断有无微生物生长,可取该培养液适量转种至同种新鲜培养基中或斜面培养基上继续培养,细菌培养2日,真菌培养3日,观察是否再出现浑浊或斜面有无菌生长,或用接种环取培养液涂片,染色,用显微镜观察是否有菌。
取健康、清洁级wistar大鼠共60只,均为雄性,周龄6周龄,体重(150±20)g,动物室为清洁级,室内温度控制在(25±2)℃之间,相对湿度为50%-70%,自然光照,避免噪音及其它干扰,自由进食和饮水。将wistar大鼠随机分为2组,即对照组和实施例组,对照组20只,喂食普通饲料。实施例组40只,喂食本实施例提供的无菌大小鼠的无菌饲料。实验期间大鼠自由饮水,电子秤称重,并记录每只大鼠体重,制作生长曲线图。如图2所示,无菌饲料喂养组(实施例组)和普通饲料喂养组(对照组)裸鼠分别饲养在无菌环境和普通清洁级环境中,通过长期的喂养两组的生长曲线几乎一致,如此说明本实施例提供的无菌饲料适合无菌大小鼠生长繁殖所需。
上述仅本发明较佳可行实施例,并非是对本发明的限制,本发明也并不限于上述举例,本技术领域的技术人员,在本发明的实质范围内,所作出的变化、改型、添加或替换,也应属于本发明的保护范围。
Claims (6)
1.无菌大小鼠的无菌饲料的制备方法,其特征在于,包括如下步骤:
S1,精确称取基础饲料、3.5%质量比的矿物质预混料和0.1%质量比复合维生素进行混合,获得混合粉料;
S2,步骤S1中的混合粉料加入6.99%质量比的多不饱和脂肪酸进行搅拌,获得油料混合物;
S3,步骤S2中的油料混合物加入水分搅拌混合,通过制粒机进行制粒,获得软颗粒饲料;
S4,将步骤S3中的软颗粒饲料进行低温烘干水分、辐照灭菌和高温高压灭菌,获得无菌大小鼠的无菌饲料。
2.根据权利要求1所述的无菌大小鼠的无菌饲料的制备方法,其特征在于,步骤S1中,所述基础饲料按质量百分比计包括19.99%酪氨酸、0.3%胱氨酸、39.68%玉米淀粉、13.19%麦芽糊精、11%蔗糖、5%膳食纤维、0.0014%抗氧化剂和0.25%酒石酸氢胆碱。
3.根据权利要求1所述的无菌大小鼠的无菌饲料的制备方法,其特征在于,步骤S2中,所述多不饱和脂肪酸包括1.08%SFA、1.62MUFA和4.29%的PUFA。
4.根据权利要求1所述的无菌大小鼠的无菌饲料的制备方法,其特征在于,步骤S3中,所述软颗粒饲料的10-15mm。
5.根据权利要求1所述的无菌大小鼠的无菌饲料的制备方法,其特征在于,步骤S4中,所述烘干的温度为26摄氏度,时间为48小时;辐照灭菌的剂量50kgy;高温高压灭菌的温度为121℃,时间为30min。
6.无菌大小鼠的无菌饲料,其特征在于,根据权利要求1-5任一项所述的方法制备的无菌大小鼠的无菌饲料。
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