CN107647090A - 高效抗仔猪腹泻微生态制剂 - Google Patents
高效抗仔猪腹泻微生态制剂 Download PDFInfo
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- CN107647090A CN107647090A CN201710598353.XA CN201710598353A CN107647090A CN 107647090 A CN107647090 A CN 107647090A CN 201710598353 A CN201710598353 A CN 201710598353A CN 107647090 A CN107647090 A CN 107647090A
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- probiotics
- piglet diarrhea
- wall material
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- preventing piglet
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Abstract
本发明提供一种高效抗仔猪腹泻微生态制剂,包括肠溶性壁材和包埋在所述肠溶性壁材中的益生菌,包埋技术能有效抵抗制粒过程中的严格考验,并能顺利通过胃部进入肠道中释放具有活力的菌株。所述益生菌总体有效含量为50*108CFU/g以上,其中嗜酸乳杆菌占30%‑40%,枯草芽孢杆菌占60%‑70%,耐热酵母菌占0.1%‑0.2%。特别添加益生元及植物活性物质,一方面保护微生态的生存条件,一方面协同作用于动物机体,综合调控动物的生长发育和生产需要,效果稳定可靠。解决养殖关键问题,如腹泻,高度耐药,免疫抑制,环境恶劣等。
Description
技术领域
本发明涉及领域食物添加剂领域,尤其涉及一种高效抗仔猪腹泻微生态制剂。
背景技术
仔猪阶段具有自身特殊的生理发育特点,例如胃酸分泌不足,肠道免疫屏障不健全,肠道消化酶系和微生物生态系统薄弱,是一个易感敏感期,也是生长发育的关键时期,需要经过哺乳、断奶这两个关键环节,容易受到各类疾病、环境、温度、应激、饲料等多种外因的影响,而且在中国,这两个阶段需要注射3-6种疫苗,导致其免疫应答不足,核苷酸及免疫物质产生不足等。它的食物来源主要为奶水和饲料,一旦发生变化或其它因素的攻击,就非常容易导致仔猪在不同阶段的腹泻,仔猪肠道会发生绒毛萎缩、脱落、隐窝增生、肠黏膜通透性增强等系列变化,导致仔猪死亡的直接结果一方面是营养不足和脱水,而另一方面则是各种毒素和自由基进入血液后导致仔猪脏器衰竭而亡。
近年来,仔猪腹泻成为养猪行业中前三名的重大流行的疾病,各类腹泻因素导致死亡的仔猪每年估计达到了6000万头,是当前死亡率最高损失最大的疾病,直接导致行业损失近150亿元。导致腹泻的因素一类是来自病源性的影响,一是来自非病源性的影响。病源性导致仔猪腹泻的主要包括寄生虫、病毒性和细菌性疾病大致20余种,目前具有特异性治疗或预防病源性腹泻的疫苗大致有 7-8种,且效果不理想,因腹泻疾病之间无交叉保护力,且经常发生变异,使用疫苗和药物用量越多,它们的变异就会越频繁,这是有害微生物自我保护的生存法则,而疫苗的研制总是赶不上微生物的变异,这就是多年来我国病源性腹泻爆发的核心原因。而非病源性腹泻的问题则十分突出,主要因素有温度、饲料、奶水不足、断奶应激等,常常因温度而导致消化不良、仔猪膘情损失、断奶应激而产生体内酶类不足、酸度偏低、血液中能量不足,另外饲料中常常因霉菌毒素也损坏肠道及器官而紊乱消化机能。目前针对腹泻特点,除了疫苗以外,还有抗生素、中草药和益生素三大类产品,其中抗生素预防和治疗效果不好,形成耐药性和中毒症状,且对病毒无效,而中草药总体来讲存在资源性短缺的现状加上制药工艺的问题,往往理论上可行,而行之有效的产品不多,最具潜力的仍然是微生态制剂类的产品。
发明内容
本发明提供一种高效抗仔猪腹泻微生态制剂,包括肠溶性壁材和包埋在所述肠溶性壁材中的益生菌,所述益生菌总体有效含量为50*108CFU/g以上,其中嗜酸乳杆菌占30%-40%,枯草芽孢杆菌占60%-70%,耐热酵母菌占0.1%-0.2%;
另外,所述益生菌中配制甘露寡糖100-120g/kg,乳酸70-80g/kg,维他命 B1:40-50mg/kg,维他命B2:60-100mg/kg,维他命B6:20-30mg/kg,维他命 B12:15-20mg/kg,叶酸8-15mg/kg,耐热短乳杆菌素50-70g/kg、氧化锌 100-120g/kg,载体为葡萄糖和乳酸钙组成,成分比例为葡萄糖30%-50%,乳酸钙70%-50%。
可选的,包埋材质为蛋白包埋层及磷脂类物质,辅以氧化锌。
可选的,包埋材料为阿拉伯胶或麦芽糊精溶。
可选的,益生菌包埋在所述肠溶性壁材中的步骤包括:
将制备好的菌悬液加入等体积无菌的1%海藻酸钠溶液,将二者充分混合均均后,滴入含0.2%吐温的80的植物油中,200r/min搅拌10min,滴加4%的氯化钙溶液,固化30min后,制备出海藻酸钠-菌剂包埋后的微颗粒液体;
以脱脂乳作为壁材的成分之一,利用响应面分析法进行优化得到微胶囊最佳制备工艺参数。
可选的,所述微胶囊最佳制备工艺参数为:壁材浓度35%、壁材配比1:9、脱脂奶粉添加量1.5%、喷雾干燥的进口温度160℃、出口温度87℃、进料速度 45ml/min、固形物含量27%。
可选的,分别对益生菌中的芽孢菌、酵母菌和乳酸菌进行菌种纯度控制和优化。
可选的,乳酸菌实施双层包埋技术予以保护,并在配方中进行电解质和PH 值的调节,以与仔猪饲料配制特点相适宜。
可选的,采取涂布平板法或平板划线法进行菌种的纯度控制。
可选的,利用液态半连续发酵技术进行菌株筛选和培育。
可选的,所述包埋为双层微胶囊包埋。
本发明的高效抗仔猪腹泻微生态制剂包括肠溶性壁材和包埋在所述肠溶性壁材中的益生菌,包埋技术能有效抵抗制粒过程中的严格考验,并能顺利通过胃部进入肠道中释放具有活力的菌株。所述益生菌总体有效含量为50*108CFU/g 以上,其中嗜酸乳杆菌占30%-40%,枯草芽孢杆菌占60%-70%,耐热酵母菌占 0.1%-0.2%。特别添加益生元及植物活性物质,一方面保护微生态的生存条件,一方面协同作用于动物机体,综合调控动物的生长发育和生产需要,效果稳定可靠。解决养殖关键问题,如拉稀,高度耐药,免疫抑制,环境恶劣等。
具体实施方式
针对现有技术存在的问题,本发明针对养殖重大问题通过仔猪生理学和微生态定向发酵工艺的研究,筛选设计出了最佳的益生菌组方,应用双层包埋技术和电解质平衡调节的方法,确保微生态制剂能够在保存、加工和过胃酸等环节保持一定比例的存活率,并在小肠段进行分批精准释放,配以微生物最佳营养需要而迅速发挥功效或定植生长,达到了调节肠道微生态环境而促进益生菌生长的目标,是一个多功能高效的复合型微生态添加剂。在菌株筛选和培育上条件上进行了大量摸索,利用液态半连续发酵技术,在产生高效的菌群的同时,定向生产符合肠道健康的中间代谢产物,例如酶、细菌素、乳酸等,每一类菌单独进行设计和发酵,后经喷雾干燥保持活性,对产物进行测量后进行复配。公司研发的这一产品与市场上同类产品的最大差异就是菌株的筛选与组方,浓度精确,能够有效补充仔猪产酸及产酶的不足,缓解应激带来的影响,对革兰式阳性菌和阴性菌同时具备杀灭和抑制作用,考虑了饲料制粒的加工环境导致的微生态损失,采用组方之一的乳酸菌实施双层包埋技术予以保护,并在配方中进行了电解质和PH值的调节,与仔猪饲料配制特点相适宜,可以充分保障益生菌能进入小肠指定部位后迅速扩增繁殖,在仔猪食糜停留期间充分发挥作用,还有特定的成分不但可以满足和补充微生物的繁殖,而且对肠道具有一定的保护和修复作用,对病源性腹泻具有一定的抑制或杀灭作用,并对可能存在的霉菌毒素具有一定能力的吸附作用,减少对肠道的损伤和毒害。使用公司研制的产品加上必要的生物安全措施和管理方法,能从根本上降低带毒率,切断传播途径,有效控制多种非病源性导致的腹泻,在预防和治疗上都有其重要是价值。
以下结合具体实施例对本发明作进一步详细说明。根据下面说明和权利要求书,本发明的优点和特征将更清楚。
本发明包括肠溶性壁材和包埋在所述肠溶性壁材中的益生菌,所述益生菌总体有效含量为50*108CFU/g以上,其中嗜酸乳杆菌(Lactobacillus)占 30%-40%,枯草芽孢杆菌(Bacillus subtilis)占60%-70%,耐热酵母菌占 0.1%-0.2%;
另外,配制甘露寡糖100-120g/kg,乳酸70-80g/kg,维他命B1:40-50mg/kg,维他命B2:60-100mg/kg,维他命B6:20-30mg/kg,维他命B12:15-20mg/kg,叶酸8-15mg/kg,耐热短乳杆菌素50-70g/kg、氧化锌100-120g/kg,载体为葡萄糖和乳酸钙组成,成分比例为葡萄糖30%-50%,乳酸钙70%-50%。
1.益生菌的设计及其作用
本发明主要设计了两个产酸和产酶菌,辅以酵母菌。
其中,枯草芽孢杆菌可耐高温,在120℃温度(饲料制粒温度最高为85度, 15秒)下能存活20分钟以上,是耐加工和储存的一类菌群。进入仔猪小肠后可产生枯草菌素、多粘菌素、制霉菌素、短杆菌肽等活性物质,这些活性物质对致病菌或内源性感染的条件致病菌有明显的抑制作用,特别是大多数药物无法抑制的大肠杆菌和革兰式阴性菌等有害菌。她在使用初期可迅速消耗肠道中的游离氧,造成厌氧环境促进外源性及内源性乳酸菌生长,并产生乳酸等有机酸类,降低肠道PH值,间接抑制其它致病菌生长。通过补充部分乳酸可以尽早模拟其生态系统环境,而快速启动益生菌的增殖。它可以直接刺激小肠的绒毛生长发育,激活T、B淋巴细胞,提高免疫球蛋白和抗体水平,增强细胞免疫和体液免疫功能,提高群体免疫力。在体内合成α-淀粉酶、蛋白酶、脂肪酶、纤维素酶等酶类,补充仔猪消化酶不足的现象而发挥作用。利用肠道营养和配方提供的营养合成维生素B1、B2、B6、烟酸等多种B族维生素,提高体内干扰素和巨噬细胞的活性。
嗜酸乳杆菌是一种容易受温度和胃酸影响的菌群,经双包埋技术处理后,加上酸度调节技术的应用,可以使外层的胶质在小肠中、后段完全溶解,内层的蛋白质可帮助益生菌定植在肠壁上并定点增殖,有效调节肠道微生态。嗜酸乳杆菌能分泌抗生物素类物质(嗜酸乳菌素(acidolin)、嗜酸杆菌素(acidophilin)、乳酸菌素(1aetocidon)),对肠道致病菌产生的拮抗作用。它不仅可以覆盖小肠段,还可以覆盖至大肠前段,维持肠道生态系统,它可产生少量的乳酸和醋酸,提高钙、磷、铁的利用率,促进铁和维生素D3的吸收,产生维生素K及少量的维生素B族,以提升消化能力和细胞免疫能力,对肠道固有层细胞的发育起到重要作用,分泌的乳糖酶可以弥补奶水中酶的不足而达到仔猪哺乳期间奶水能量消化率的提升,而增加断奶重。利用冻干干酵母和破壁酵母,起到仔猪诱食的作用,补充免疫多糖,促进益生菌生存和增殖,同时补充了核酸、谷胱甘肽、生物素、泛酸、硫胺素、吡哆醇和肌醇等,起到营业性调节剂的作用。
2.肠道功能性修复和治疗作用
利用蛋白包埋层及磷脂类物质可帮助益生菌定植在肠道粘膜表面,修复并重建被破坏的肠道生物屏障,阻止致病菌及毒素的入侵,诱导肠粘膜产生各种抗体,特别是IgA的分泌,营造好的肠道发育环境,减少饲料霉菌毒素及细菌外毒素的影响,促进隐窝细胞的发育,代替已经受损的绒毛膜细胞,促进上皮细胞的移动和修复,减少腹泻损伤。同时辅以具有吸附作用的氧化锌,在针对一般脱水性腹泻方面有很好的抑制作用,维持肠道营养浓度和体液渗透压平衡,维持益生菌及其代谢产物的作用,同时对上皮细胞通透性有抑制收敛的作用,保障谷氨酰胺(Gln)参与上皮细胞的能量代谢,促进淋巴细胞和巨噬细胞的发育,并组织有害菌产生的含氮类毒素进入血液而造成机体脏器损害,并同时促进细胞分泌黏蛋白到外围而保护肠道,从而达到一定的治疗性作用。通过酵母外源补充谷胱甘肽(GSH)成分,可以增强肠道细胞的抗氧化能力,清除肠道内产生的氧自由基(OFR)和黄嘌呤氧化物(XOD),也能起到在发病状态下减少自由基对肠道及机体的损害。
制剂的包埋:
背景:益生菌在机体内发挥益生作用必须满足两个条件:
一、益生菌在被摄入时必须是活的。
二、必须要有足够数量的益生菌定植于肠道中。目前大多数活性益生菌制剂存在的问题就是:益生菌被摄入后,对消化道中低值的胃酸、胆汁酸、消化酶等的耐受性较差,致使进入肠道中定植的活菌数量低于理论上能够发挥生理作用的最小值。
利用包埋技术,将益生菌包埋在肠溶性壁材中,一方面能增强菌体对消化道逆环境的抵抗力,另一方面在肠道适当条件的作用下,包埋后的益生菌又可以被释放出来,显著提高了菌体在到达肠道后的存活率,从而使尽可能多的活菌到达肠道,真正发挥益生菌的健康作用。
使用的药品与试剂:
工艺设计:
包埋材料的选择:选择合适的包埋材料可以提高微胶囊性能,降低生产成本,提高生产效果。。选择的壁材要具有以下特性:较好的溶解性:良好的乳化性:干燥性能好:其溶液粘度低:成膜性好:无毒:生物相容性好:阻氧性能较好:阻湿性能良好:阻热性能较好。具有以上特性的壁材可以很好的保护植物乳杆菌的活性,提高菌种存活率。制备出的微胶囊表面光洁,速溶性好,安全性好,对菌种的储藏也很有帮助。阿拉伯胶安全性高,具有非常好的亲水性,而且是一种天然乳化稳定剂,乳化稳定性较好。麦芽糊精溶不但溶解解性好、安全性高,而且粘度低,具有很好的乳化效果,不易吸潮,稳定性好,难以变质。
经过对该菌剂的包埋的方法进行研究,确定了最佳工艺条件,得出以下结论:
第一步:将制备好的菌悬液加入等体积无菌的1%海藻酸钠溶液,将二者充分混合均均后,滴入含0.2%吐温的80的植物油中,200r/min搅拌10min,滴加 4%的氯化钙溶液,固化30min。制备出海藻酸钠-菌剂包埋后的微颗粒液体。
第二步:利用乳清蛋白中的乳球蛋白在低pH值及胃蛋白酶存在的情况下依然能够保持结构完整的特性,以脱脂乳作为壁材的成分之一,利用响应面分析法进行优化得到的微胶囊最佳制备工艺参数,即壁材浓度35%、壁材配比(阿拉伯胶:麦芽糊精)1:9、脱脂奶粉添加量1.5%、喷雾干燥的进口温度160℃、出口温度87℃、进料速度45ml/min、固形物含量27%在此工艺参数下菌种存活率为79.21%。
耐受性实验结果:
本试验对包埋后的菌剂应用性能进行了研究,通过对成品进行人工胃液和人工肠液处理及贮存稳定性能的研究,得出以下结论:经过包埋后处理,有利于提高菌体对胃酸环境的抵抗能力,在人工胃液中处理后,可达到和,菌体存活率可以达到9.15×109CFU/g左右,而对于没有经过微胶囊化处理的菌体而言,经过胃液处理后的存活率仅有23%。经过人工胃液(pH1.3)处理后,包埋后的菌剂能够达到9.15×109CFU/g,存活率达到80%。
按本工艺制备得到具备较好的肠溶性。在-18°条件下贮存60d后,菌体存活率接近68%,活菌数依然可以达到6.22×109CFU/g。在2°条件下贮存60d 后,仍然有接近65%的存活率,活菌数可达5.49×109CFU/。说明釆用本工艺获得包埋方法可以有效提高菌剂在低温下的贮存稳定性。在人工胃液中具有较好的耐受性,在人工肠液中具有良好的肠溶性,并且具有贮存稳定性。可以认为这种包埋方法可以提高微生态菌剂在应用过程中的菌体存活率,具有积极的生产意义。
高效抗仔猪腹泻微生态制剂中涉及的各菌种的形成方式如下
1,芽孢菌:
菌种生物学特征与菌种纯化
将芽抱杆菌接种于培养基37℃培养,观察菌落的形态、大小、边缘、表面、凹凸度、透明度。革兰氏染色观察观察菌体形态。菌株在培养基上生长良好,形成直径左右大小的圆形菌落,白色而有光泽,表面湿润、平坦,边缘整齐。油镜观察, 菌株菌体形态为长杆状,单个、成对或链状排列,芽抱椭圆形近中生,抱囊不膨大。
2,酵母菌:
生物学特性与优化
产朊假丝酵母(Candidautilis)是一种形态结构简单,以芽殖为主的真菌,其有细胞较大,代谢旺盛,生长繁殖快,能形成较好的絮体,易适用于多种不同的生物反应器,对环境适应性强等特点。产朊假丝酵母是目前人类使用量较大的一种微生物。因产朊假丝酵母体内含有丰富的蛋白质、脂质、糖和各种维生素等,以及发酵过程中会产生酶、辅酶、核糖核酸、甾醇等和一些新陈代谢的中间产物而被广泛应用于食品、医药、实验研究和畜牧业中。
优化后的培养基:
培养基:葡萄糖5.0%、酵母粉2.0%、磷酸二氢钾0.2%、硫酸镁0.1%、硫酸铵0.1%、脂1.5%,p H5.5;
优化后的发酵参数:
对产朊假丝酵母(Candida utilis),通过用YPD培养基进行生长曲线的测定,得出12小时是此种酵母菌的生长最优时间。
对产朊假丝酵母进行单因素条件筛选,包括培养温度、摇床转速、碳氮比,实验得出温度在30℃、转速180r/min、碳氮比(葡萄糖和蛋白胨的质量比)为3:1 时菌体密度最高。
通过采用L9(34)正交试验设计确定了各因素对酵母菌生长的影响。
优化条件为培养时间13h、温度30℃、转速160r/min、碳氮比为3:1时菌体密度最高,各因素对酵母菌生长无显著影响(P>0.05)。
通过对产朊假丝酵母菌的紫外诱变,筛选出蛋白质含量较高的菌株,实验得出在15W紫外线(30cm)处,照射120S,致死率为89%,得到了蛋白质含量为31.4%的诱变菌株,比诱变前的22.9%提高了37.12%。
3、乳酸菌
生物学特征与优化:
乳酸菌(lactobacillus),直或弯的杆菌,形态各异,从长的和细长杆状到短的球杆状,一般形成链,特别是在对数生长期的后期。通常不运动,如运动则借助于周生鞭毛。不形成芽抱。一般是革兰氏阳性菌,随菌龄和培养基酸度的增加会由革兰氏阳性变为阴性。有些菌株当用革兰氏染色或次甲基蓝染色时呈现出两极体和内部颗粒,或呈现出横线条。在新陈代谢方面,是异养厌氧型,有些菌株在分离时是专性厌氧的。乳酸菌具有强抗酸能力,如食物制作中,有很多菌类生长,但因乳酸菌不断地产生乳酸使得生存环境改变而杀死多种不耐酸的细菌。葡萄糖发酵时值 pH值会降低。在碳的最终产物中至少有一半是乳酸盐。最适合pH值通常是 5.5-6.8。在pH值为中性或碱性时,可能延长生长周期,或者总的生长量减少。
菌种的形态特征是鉴别菌种的重要参考,乳酸菌多为杆菌和球菌,不同培养基或不同培养条件下菌种形态变化很大,革兰氏染色多为阳性。所筛选的产酸较高的株菌℃下平板培养基上培养的菌落和镜检形
在平板培养基的菌落形态
菌落形态:圆形,较光滑,突起,乳白,不透明
从中筛选出来的株菌株菌落的形态特征,可以看出,不同菌落形态有一定的差异,主要表现在菌落表面光滑度,在不同的培养基或培养条件下的形态可能有一定的变化。
该菌株发酵与性能参数:
具有很强的耐酸性,可以耐受6%的食盐,耐热性很差,温度较高时就会马上死亡。生长曲线:0-6h是延滞期,6-18h是对数生长期,18-45h是稳定生长期,45-48是衰亡期。
产酸曲线:0-9h,pH值由6.03降到4.28之后,pH值逐渐降低,最终降至3.44。
最佳离心条件:6000r/min,离心10min。
培养基优化:
培养基组方:1.32%葡萄糖、0.68%蔗糖、1.5%酵母浸粉、0.5%麸皮,0.005%硫酸锰,0.02%硫酸镁,0.05%无水乙酸钠、0.2%磷酸氢二钾。在优化培养基的基础上,分析了培养时间、温度、接种量、装液量对产抗菌物质的影响。结果显示,在接种量4%、装液量40%于33℃静置养30h后,共培养上清液的抑菌活性最强,其抑菌圈直径可达24.87mm。
在上述菌种的纯度控制上,采取涂布平板法或平板划线法进行控制。
1、涂布平板法
首先把微生物悬液通过适当的稀释,取一定量的稀释液放在无菌的已经凝固的营养琼脂平板上,然后用无菌的玻璃刮刀把稀释液均匀地涂布在培养基表面上,经恒温培养便可以得到单个菌落。
2、平板划线法
最简单的分离微生物的方法是平板划线法。用无菌的接种环取培养物少许在平板上进行划线。划线的方法很多,常见的比较容易出现单个菌落的划线方法有斜线法、曲线法、方格法、放射法、四格法等。当接种环在培养基表面上往后移动时,接种环上的菌液逐渐稀释,最后在所划的线上分散着单个细胞,经培养,每一个细胞长成一个菌落。
本发明的高效抗仔猪腹泻微生态制剂采用了专用系列化方案,针对不同动物和生长阶段的特点,围绕养殖关键问题采用全新思路进行配伍设计。达到以下目标:
1、解决养殖关键问题,如拉稀,高度耐药,免疫抑制,环境恶劣等;
2、提高综合效益,如饲料转化率,减少淘汰,增产,逐步降低药物的用量等;
3、多种功效,使用简便,如耐储存和加工,直接添加,兼具保健、营养、改善饲料品质和肉质的多重作用。
本系列产品的微生态菌种在畜禽动物方面主选枯草芽孢、双岐杆菌、乳酸菌、粪链球菌、酵母菌等组合而成,特别添加益生元及植物活性物质,一方面保护微生态的生存条件,一方面协同作用于动物机体,综合调控动物的生长发育和生产需要,效果稳定可靠。
温度敏感菌株采用高效双层微胶囊包埋技术,能有效抵抗制粒过程中的严格考验(高温和压力),并能顺利通过胃部进入肠道中释放具有活力的菌株。经测试在90度温度10分钟,保留率超过80%。由于产品经过包埋,在预混料中能完全耐受高浓度的抗生素和微量元素。常温干燥贮存12个月,活菌存活率达到标示量70%。为保证其混入饲料后的有效性,我们建议已添加本产品的饲料,6 个月内用完为佳。
部分菌种采用了二次固体发酵技术,以产生大量代谢产物,主要有消化酶类、溶菌酶、B族维生素、杆菌肽、细菌素、有机酸、多种小肽、核酸和未知促生长因子等。杆菌肽主要包括短杆菌肽、环多肽及其他抑菌物质,这些小肽可以清除埋藏在肠道粘膜深处的病原菌,抑制其他有害菌的生长,其特有的细菌素和溶菌酶能广谱杀灭有害菌群和病毒。还含有内源性蛋白酶菌种和纤维分解的酶类,以及小分子有机酸等,对调节肠道菌群平衡和促进消化吸收有重要的生理作用。
显然,本领域的技术人员可以对发明进行各种改动和变型而不脱离本发明的精神和范围。这样,倘若本发明的这些修改和变型属于本发明权利要求及其等同技术的范围之内,则本发明也意图包括这些改动和变型在内。
Claims (10)
1.一种高效抗仔猪腹泻微生态制剂,其特征在于,包括肠溶性壁材和包埋在所述肠溶性壁材中的益生菌,所述益生菌总体有效含量为50*108CFU/g以上,其中嗜酸乳杆菌占30%-40%,枯草芽孢杆菌占60%-70%,耐热酵母菌占0.1%-0.2%;
另外,所述益生菌中配制甘露寡糖100-120g/kg,乳酸70-80g/kg,维他命B1:40-50mg/kg,维他命B2:60-100mg/kg,维他命B6:20-30mg/kg,维他命B12:15-20mg/kg,叶酸8-15mg/kg,耐热短乳杆菌素50-70g/kg、氧化锌100-120g/kg,载体为葡萄糖和乳酸钙组成,成分比例为葡萄糖30%-50%,乳酸钙70%-50%。
2.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,包埋材质为蛋白包埋层及磷脂类物质,辅以氧化锌。
3.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,包埋材料为阿拉伯胶或麦芽糊精溶。
4.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,益生菌包埋在所述肠溶性壁材中的步骤包括:
将制备好的菌悬液加入等体积无菌的1%海藻酸钠溶液,将二者充分混合均均后,滴入含0.2%吐温的80的植物油中,200r/min搅拌10min,滴加4%的氯化钙溶液,固化30min后,制备出海藻酸钠-菌剂包埋后的微颗粒液体;
以脱脂乳作为壁材的成分之一,利用响应面分析法进行优化得到微胶囊最佳制备工艺参数。
5.如权利要求4所述的高效抗仔猪腹泻微生态制剂,其特征在于,所述微胶囊最佳制备工艺参数为:壁材浓度35%、壁材配比1:9、脱脂奶粉添加量1.5%、喷雾干燥的进口温度160℃、出口温度87℃、进料速度45ml/min、固形物含量27%。
6.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,分别对益生菌中的芽孢菌、酵母菌和乳酸菌进行菌种纯度控制和优化。
7.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,乳酸菌实施双层包埋技术予以保护,并在配方中进行电解质和PH值的调节,以与仔猪饲料配制特点相适宜。
8.如权利要求6所述的高效抗仔猪腹泻微生态制剂,其特征在于,采取涂布平板法或平板划线法进行菌种的纯度控制。
9.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,利用液态半连续发酵技术进行菌株筛选和培育。
10.如权利要求1所述的高效抗仔猪腹泻微生态制剂,其特征在于,所述包埋为双层微胶囊包埋。
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