CN114946998B - 一种混合益生菌发酵米糠饲料及其制备方法 - Google Patents
一种混合益生菌发酵米糠饲料及其制备方法 Download PDFInfo
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Abstract
本发明提出了一种混合益生菌发酵米糠饲料的制备方法,按0.4‰~0.8‰接种混合益生菌发酵种子液至米糠固态发酵培养基中于33‑37℃发酵24‑48h后得混合益生菌发酵米糠饲料;所述米糠固态发酵培养基为将将米糠与无菌水按1.0:0.5‑0.7比例混匀制得;本发明制得的米糠发酵后粗纤维和植酸含量降低,营养价值高且具有酸香味,pH值降低,可以提高饲料的适口性;此外,米糠发酵过程中益生菌代谢产生大量酶和其他有益因子,且富含益生菌,有助于改善动物肠道微生态平衡,抑制动物肠道内有害菌的生长,降低发病率,提高饲料利用率。
Description
技术领域
本发明涉及生物发酵技术领域,尤其涉及一种混合益生菌发酵米糠饲料的及其制备方法。
背景技术
米糠是糙米碾制成精米过程中产生,主要由果皮、外胚乳和糊粉层组成,约占糙米重量的5%-12%,是一种量大而面广的优质可再生资源。米糠营养丰富,含有糙米中64%以上的营养物质,富含碳水化合物、蛋白质、脂肪(80%由不饱和脂肪酸组成)和维生素。但米糠含有大量脂肪酶,易酸败,不利于储存,限制了其使用,此外,米糠中也含有植酸、胰蛋白酶抑制因子、非淀粉多糖等抗营养因子,钙少磷多,且大部分是不能利用的植酸磷,直接饲用消化利用率低,不利于动物健康,并造成资源浪费和环境污染。
微生物发酵法是利用乳酸菌、芽孢杆菌、酵母等一些益生菌,通过它们之间的协同作用进行饲料发酵,在生产中被广泛应用。发酵后的米糠具有酸香味,可提高饲粮的适口性,促进胃肠道蠕动,提高动物对蛋白质和各种矿物元素的吸收利用。大量研究表明发酵米糠有潜在的抗氧化、抑菌抗炎、降血脂以及维持肠道健康的功能。我国作为拥有丰富米糠资源的大国,更应该合理地生产加工米糠和利用米糠资源,优化我国的米糠应用前景,缓解资源短缺等问题。
发明内容
本发明要解决的技术问题是克服现有技术存在的缺陷,提出了一种混合益生菌发酵米糠饲料及其制备方法,提供一种营养价值高,可提高饲粮适口性,改善动物肠道微生态平衡,提高机体抗氧化水平的发酵米糠及其制备方法。
为解决上述技术问题,本发明采用的技术方案是:一种混合益生菌发酵米糠饲料的制备方法,按0.4‰~0.8‰接种混合益生菌发酵种子液至米糠固态发酵培养基中,于33-37℃发酵 24-48h后得混合益生菌发酵米糠饲料;所述米糠固态发酵培养基为将将米糠与无菌水按 1.0:0.5-0.7比例混匀制得;
所述混合益生菌发酵种子液为枯草芽孢杆菌发酵种子液以及植物乳杆菌发酵种子液的混合物,所述枯草芽孢杆菌发酵种子液和植物乳杆菌发酵种子液比例为2-4:8-6;
所述枯草芽孢杆菌发酵种子液自枯草芽孢杆菌培养基发酵所得,于所述枯草芽孢杆菌培养基中包括牛肉浸粉3.00g,氯化钠5.00g,胰蛋白胨10.00g,葡萄糖1.00g,pH为7.2±0.2,以去离子水定容至1000mL;
所述植物乳杆菌发酵种子液自植物乳杆菌培养基发酵所得,于所述植物乳杆菌培养基包括牛肉浸粉10.00g,胰蛋白胨10.00g,葡萄糖20.00g,酵母浸粉5.00g,无水乙酸钠5.00g,磷酸氢二钾2.00g,硫酸镁0.20g,柠檬酸二铵2.00g,硫酸锰0.04g,pH为6.5±0.2,以去离子水定容至1000mL;
所述枯草芽孢杆菌ZJUAF-3为自有菌株,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO.20513;所述植物乳杆菌购买于中国微生物菌种保藏管理委员会农业微生物中心,菌株编号:ACCC 11016。
进一步地,所述枯草芽孢杆菌培养基于121℃灭菌15min后,于室温下接种枯草芽孢杆菌菌粉至枯草芽孢杆菌培养基中,接种量为0.4%,于35℃环境下,150rpm震荡培养15-18h 得到枯草芽孢杆菌发酵种子液。
进一步地,所述植物乳杆菌培养基于121℃灭菌15min,于室温下接种植物乳杆菌菌粉至植物乳杆菌培养基中,接种量为0.4%,于35℃环境下,静置培养15-18h的植物乳杆菌发酵种子液。
进一步地,所述米糠于发酵之前经稳定化处理,所述稳定化处理过程具体如下:称取一定量的米糠置于托盘中,铺平至2±0.3cm厚,置于鼓风干燥箱中120℃烘干30min,晾干回潮后装在密封袋中,置于阴凉避光处保存。
进一步地,一种混合益生菌发酵米糠饲料,混合益生菌发酵米糠饲料的制备方法制得,于所述混合益生菌发酵米糠饲料中,粗蛋白含量≥14.50%,酸溶蛋白含量≥32.10%,粗纤维含量≤9.46%,总膳食纤维含量≥33.97%,植酸含量≤31.39mg/g,总酚含量≥5.92ug/g,pH ≤4.52,乳酸含量≥80.64mmol/L,枯草芽孢杆菌活菌数≥1.01×108cfu/g,植物乳杆菌活菌数≥5.35×108cfu/g。
与现有技术相比,本发明的有益效果包括:制得的米糠发酵后粗纤维和植酸含量降低,营养价值高且具有酸香味,pH值降低,可以提高饲料的适口性;此外,米糠发酵过程中益生菌代谢产生大量酶和其他有益因子,且富含益生菌,有助于改善动物肠道微生态平衡,抑制动物肠道内有害菌的生长,降低发病率,有助于提高饲料利用率。
具体实施方式
容易理解,根据本发明的技术方案,在不变更本发明实质精神下,本领域的一般技术人员可以提出可相互替换的多种结构方式以及实现方式。因此,以下具体实施方式仅是对本发明的技术方案的示例性说明,而不应当视为本发明的全部或者视为对本发明技术方案的限定或限制。
实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件,或者按照产品说明书进行。所用仪器等未注明生产厂商者,均为可通过正规渠道商购买得到的常规产品。所述方法如无特别说明均为常规方法,所述原材料如无特别说明者均能从公开商业途径获得。实施例中未注明具体技术条件者,按照本领域内的文献所描述的技术或条件,或者按照产品说明书进行。
实施例1,干热处理对米糠的脂肪酸值和其中的过氧化物酶残余活力的影响。
对照组即米糠样品不做任何处理,热处理米糠的制备方法为:称取一定量的米糠置于托盘中,铺平约2cm厚,置于鼓风干燥箱中120℃烘30分钟,中间翻搅1次,晾干回潮后装在密封袋中备用。
表1:干热处理对米糠脂肪酸值和其中的过氧化物酶残余活力的影响
从上述预试验结果可以看出,干热处理可以显著降低米糠物料的过氧化物酶残余活力,但不影响其脂肪酸值。
实施例2,枯草芽孢杆菌培养基的配制与菌种活化。
枯草芽孢杆菌(Bacillus subtilis)ZJUAF-3为自有菌株,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏日期为2020年8月10日,保藏编号为:CGMCCNo.20513。
活化和发酵培养基(营养肉汤):牛肉浸粉3.0,氯化钠5.00,胰蛋白胨10.00,葡萄糖 1.00,调整pH为7.2±0.2,加入去离子水定容到1000mL,121℃高压灭菌15min。
菌种活化与种子液制备:从-80℃冰箱中取一支枯草芽孢杆菌冻干粉(约2-3g),在超净工作台内接种至500ml营养肉汤培养基中,置于摇床中35℃、150rpm震荡培养15h得到枯草芽孢杆菌发酵种子液,取5mL种子液至营养肉汤,摇床中35℃、150rpm培养15-18h,至活菌数不低于2×1011cfu/mL。
实施例3,植物乳杆菌培养基的配制与菌种活化。
植物乳杆菌(Lactobacillus plantarum)ACCC11016,购买于中国微生物菌种保藏管理委员会农业微生物中心。
活化和发酵培养基(MRS肉汤)(g/L):牛肉浸粉10.0,胰蛋白胨10.0,葡萄糖20.0,酵母浸粉5.0,无水乙酸钠5.0g,磷酸氢二钾2.0,硫酸镁0.2,柠檬酸二铵2.0,硫酸锰0.04,调整pH为pH6.5±0.2,加入去离子水定容到1000mL,121℃灭菌15min。
菌种活化:从-80℃冰箱中取出一支植物乳杆菌冻干粉(2mL),在超净工作台内接种至 500mlMRS肉汤培养基中,然后35℃,150rpm震荡培养15-18h得到植物乳杆菌发酵种子液。
发酵菌液制备:在无菌接种室内接种5mL种子液至培养基后置于摇床中35℃、150rpm 培养15-18h,至菌含量≥109cfu/mL。
实施例4
一种混合益生菌发酵米糠,制备方法为:
(1)将未处理米糠和实施例1得到的的热处理米糠分别与无菌水按1.0:0.5~0.7比例混匀制成米糠固态发酵培养基;
(2)将实施例2得到的枯草芽孢杆菌和实施例3得到的植物乳杆菌发酵菌液按2~4:8~6 比例混合均匀得到混合益生菌发酵菌液;
(3)将步骤(2)得到的混合益生菌发酵种子液按接种量为0.4‰~0.8‰接种至米糠固态发酵培养基中,搅拌混匀后装进发酵袋里,置于恒温恒湿培养箱中35℃左右发酵24~48h,得到发酵未处理米糠和发酵热处理米糠。
表2:米糠发酵前后相关指标的变化(%)
由表2可知,米糠经热处理后粗纤维有降低的趋势,植酸含量显著降低了20.32%、不溶性膳食纤维降低了25.84%,肠杆菌数量减少,真菌毒素含量降低,其中呕吐毒素降低了 18.93%。与未发酵米糠相比,米糠发酵后粗蛋白含量没有显著变化,但酸溶蛋白含量显著提高了44.97%;饱和脂肪酸和不饱和脂肪酸比例发生了改变,发酵后饱和脂肪酸比例下降,不饱和脂肪酸含量增加,整体脂肪酸含量没有变化;米糠发酵后粗纤维降低了7.12%,植酸降低了30.88%;总糖和还原糖含量分别提高了1.62倍和1.70倍,膳食纤维提高了22.7%;γ- 谷维素提高了2.06倍,总酚含量提高了21.61%;蛋白酶、纤维素酶、木聚糖酶活力分别提高了3.06、3.97和2.15倍;米糠发酵后平均pH降低到4.58,平均乳酸含量达到87.02mmol/L;枯草芽孢杆菌和植物乳杆菌的活菌数分别达到1.47×108cfu/g和5.35×108cfu/g,肠杆菌的数量降低了80.15%;黄曲霉毒素B1、玉米赤霉烯酮和呕吐毒素分别降低了6.69%、19.49%和 53.87%。综上所述,枯草芽孢杆菌和植物乳杆菌协同发酵米糠可有效改善米糠的营养水平和生理活性物质的含量,发酵后的米糠具有酸香味,富含益生菌及其代谢产物,毒素含量显著降低。
实施例5,发酵米糠在蛋鸡上的应用
参照《鸡饲养标准(NY/T33-2004)产蛋鸡营养需要》配制蛋鸡粉状基础日粮,其中包括55~60%比例玉米,25%~30%豆粕,5.0%~10%石粉,2.5%~5.05热处理米糠,1%大豆油和 5%左右的蛋鸡复合预混料。根据等量原则,将实施例4制备的发酵米糠(干基重量)以2.5%和5.0%的比例等水平替代蛋鸡饲料中的热处理米糠,在混料机混合均匀后装袋备蛋鸡饲用。
将上述饲料饲喂给蛋鸡,预试期12天,正式试验进行56天,饲养期间记录蛋鸡产蛋量、采食量等,计算1~8周的平均生产数据,试验结果如表4:
表4:实施例4制备的不同水平发酵米糠对蛋鸡生产性能的影响
项目 | 2.5%未发酵米糠 | 2.5%发酵米糠 | 5.0%未发酵米糠 | 5.0%发酵米糠 |
产蛋率,% | 67.63 | 68.25 | 68.66 | 70.03 |
平均蛋重,g | 53.53 | 53.64 | 54.51 | 55.66 |
日采食量,g/hen·d | 97.64 | 98.72 | 98.15 | 98.60 |
料蛋比,g/g | 1.79 | 1.69 | 1.74 | 1.67 |
由表4可知,和其他组别相比,5.0%发酵热处理米糠组的产蛋率、平均蛋重和日增重均高于未发酵米糠和2.5%发酵米糠组,破损率、料蛋比及料重比显著降低,说明给蛋鸡饲喂5.0%发酵米糠组的饲料,能有效提高产蛋前期蛋鸡的生产性能。
本发明的技术范围不仅仅局限于上述说明中的内容,本领域技术人员可以在不脱离本发明技术思想的前提下,对上述实施例进行多种变形和修改,而这些变形和修改均应当属于本发明的保护范围内。
Claims (2)
1.一种混合益生菌发酵米糠饲料的制备方法,其特征在于,按0.4‰~0.8‰接种混合益生菌发酵种子液至米糠固态发酵培养基中,于33-37℃发酵24-48h后得混合益生菌发酵米糠饲料;所述米糠固态发酵培养基为将米糠与无菌水按1.0:0.5-0.7比例混匀制得;
所述混合益生菌发酵种子液为枯草芽孢杆菌发酵种子液以及植物乳杆菌发酵种子液的混合物,所述枯草芽孢杆菌发酵种子液和植物乳杆菌发酵种子液比例为2-4:8-6;
所述枯草芽孢杆菌发酵种子液自枯草芽孢杆菌培养基发酵所得,所述枯草芽孢杆菌培养基中包括牛肉浸粉3.00g,氯化钠5.00g,胰蛋白胨10.00g,葡萄糖1.00g,pH为7.2±0.2,以去离子水定容至1000mL;
所述植物乳杆菌发酵种子液自植物乳杆菌培养基发酵所得,所述植物乳杆菌培养基包括牛肉浸粉10.00g,胰蛋白胨10.00g,葡萄糖20.00g,酵母浸粉5.00g,无水乙酸钠5.00g,磷酸氢二钾2.00g,硫酸镁0.20g,柠檬酸二铵2.00g,硫酸锰0.04g,pH为6.5±0.2,以去离子水定容至1000mL;
所述枯草芽孢杆菌ZJUAF-3为自有菌株,保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC NO.20513;所述植物乳杆菌购买于中国微生物菌种保藏管理委员会农业微生物中心,菌株编号:ACCC 11016;
其中,所述米糠于发酵之前经稳定化处理,所述稳定化处理过程具体如下:称取一定量的米糠置于托盘中,铺平至2.0±0.3cm厚,置于鼓风干燥箱中120℃烘干30min,晾干回潮后装在密封袋中,置于阴凉避光处保存;
所述枯草芽孢杆菌培养基于121℃灭菌15min后,于室温下接种枯草芽孢杆菌菌粉至枯草芽孢杆菌培养基中,接种量为0.4%,于35℃环境下,150rpm震荡培养15-18h得到枯草芽孢杆菌发酵种子液;
所述植物乳杆菌培养基于121℃灭菌15min,于室温下接种植物乳杆菌菌粉至植物乳杆菌培养基中,接种量为0.4%,于35℃环境下,静置培养15-18h得到植物乳杆菌发酵种子液。
2.一种混合益生菌发酵米糠饲料,其特征在于,基于权利要求1所述的混合益生菌发酵米糠饲料的制备方法制得,于所述混合益生菌发酵米糠饲料中,粗蛋白含量≥14.50%,酸溶蛋白含量≥32.10%,粗纤维含量≤9.46%,总膳食纤维含量≥33.97%,植酸含量≤31.39mg/g,总酚含量≥5.92ug/g,pH≤4.52,乳酸含量≥80.64mmol/L,枯草芽孢杆菌活菌数≥1.01×108cfu/g,植物乳杆菌活菌数≥5.35×108cfu/g。
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