CN104814278A - 一种提高奶牛产奶量的微生态制剂 - Google Patents

一种提高奶牛产奶量的微生态制剂 Download PDF

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CN104814278A
CN104814278A CN201510178564.9A CN201510178564A CN104814278A CN 104814278 A CN104814278 A CN 104814278A CN 201510178564 A CN201510178564 A CN 201510178564A CN 104814278 A CN104814278 A CN 104814278A
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季烽
王秀颖
温洪宇
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Jiangsu Normal University
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Abstract

一种提高奶牛产奶量的微生态制剂,属于提高奶牛产奶量的制剂。该微生态制剂采用免培养技术结合统计学的方法筛选高产奶量奶牛瘤胃的优势菌,利用纯培养技术获得优势菌的纯种菌株,经发酵培养配伍组合成微生态制剂;具体步骤为:(1)测序、(2)筛选、(3)分离纯化菌种、(4)培养、(5)发酵、(6)组合(7)包装。本微生态制剂包含帮助分解多糖和纤维素的拟杆菌和瘤胃球菌,可以提高奶牛瘤胃拟杆菌和瘤胃球菌含量,保证其优势地位,提高对纤维素和多糖的利用率,进而提高奶牛对饲料中营养成分的吸收利用,提高产奶量,带来经济效益。拟杆菌帮助宿主吸收多糖、提高营养利用率和宿主的免疫力,维持胃肠道微生态平衡,针对性、专一性强。

Description

一种提高奶牛产奶量的微生态制剂
技术领域
本发明涉及一种提高奶牛产奶量的制剂,特别是一种提高奶牛产奶量的微生态制剂。
背景技术
微生态制剂是一种绿色、安全的饲料添加剂,无毒副作用、无耐药性,能够调整或维持动物胃肠道内微生态平衡,增强机体免疫,促进营养物质的消化吸收。
目前,国内外反刍动物普遍使用的饲用微生态制剂有乳酸菌类(乳酸杆菌、双歧杆菌等)、芽孢杆菌类(枯草芽孢杆菌、蜡样芽孢杆菌等)等细菌类,曲霉类(黑曲霉、米曲霉)和酵母类(啤酒酵母、和石油酵母及其培养物)等单一菌制剂及复合菌制剂。这些市场上销售的微生态制剂针对性、专一性不强,影响了饲用微生态制剂在动物上的应用效果。
发明内容
本发明的目的是要提供一种提高奶牛产奶量的微生态制剂,解决现有微生态制剂针对性、专一性不强的问题。
本发明是按如下技术方案实现的:采用免培养技术结合统计学的方法筛选高产奶量奶牛瘤胃的优势菌,利用纯培养技术获得优势菌的纯种菌株,经发酵培养配伍组合成微生态制剂;按如下操作步骤制备:
(1)测序:抽提荷斯坦奶牛瘤胃液DNA进行PCR扩增,经基因组测序仪高通量测序和分类学分析获得奶牛瘤胃微生物群落组分;
(2)筛选:经统计学分析筛选出高产奶牛瘤胃优势菌,即拟杆菌和瘤胃球菌;
(3)分离纯化菌种:提取奶牛瘤胃液,利用亨盖特滚管法培养瘤胃微生物,选取拟杆菌和瘤胃球菌,进一步提纯并获得大量菌种;
(4)培养:将拟杆菌和瘤胃球菌菌种分别接种于亨盖特滚管法的滚管中37℃恒温培养2-3天;
(5)发酵:将滚管中拟杆菌和瘤胃球菌分别接种于相对应的厌氧发酵罐37℃恒温培养3-5天后,使发酵液降温至20℃;
(6)组合:收集拟杆菌和瘤胃球菌的发酵菌悬液混合均匀;
(7)包装:根据需要加入其他营养成分后密封包装或直接密封包装。
所述的拟杆菌培养基配方:葡萄糖1g/L、蛋白胨10g/L、牛肉膏3g/L、NaCl 5g/L、维生素B 10ug/L、抗坏血酸1g/L,并调节PH至7.0-7.5;瘤胃球菌培养基的配方:尿素5g/L、KH2PO41g/L、K2HPO41g/L、FeSO4·7H2O 0.05g/L、MnSO4·H2O 0.02g/L、MgSO4·7H2O 0.5g/L、酵母浸出液0.2g/L、稻草秆20g/L,并调节PH至6.0-7.0;
拟杆菌和瘤胃球菌量达1.0×108cfu/ml–2.5×108cfu/ml,并且比例在0.5-1.5之间组合。
有益效果,由于采用了上述方案,本微生态制剂包含帮助分解多糖和纤维素的拟杆菌和瘤胃球菌,可以提高奶牛瘤胃拟杆菌和瘤胃球菌含量,保证其优势地位,提高对纤维素和多糖的利用率,进而提高奶牛对饲料中营养成分的吸收利用,提高产奶量,带来经济效益。同时,解决了微生态制剂针对性、专一性不强的问题,实现了本发明的目的。
优点:本发明所含的拟杆菌和瘤胃球菌能够帮助宿主分解纤维素和多糖。拟杆菌帮助宿主吸收多糖、提高营养利用率和宿主的免疫力,维持肠道微生态平衡。拟杆菌在对多糖的利用方面的作用尤其突出,其在代谢多糖过程中产生的中间产物,可以为宿主提供营养,减少对营养源的竞争。瘤胃球菌对纤维素有很强的附着力,能够附着于饲料颗粒上,通过粘附、穿透等一系列作用,并分泌一系列纤维素酶,实现对纤维物质的降解。与现有微生态制剂相比,仅针对奶牛瘤胃微生态,针对性、专一性强。
附图说明:
图1是本发明的工艺流程图。
具体实施方式
下面结合实例对本发明进一步描述
实施例1:采用免培养技术结合统计学的方法筛选高产奶量奶牛瘤胃的优势菌,利用纯培养技术获得优势菌的纯种菌株,经发酵培养配伍组合成微生态制剂;按如下操作步骤制备:
(1)测序:抽提荷斯坦奶牛瘤胃液DNA进行PCR扩增,经基因组测序仪高通量测序和分类学分析获得奶牛瘤胃微生物群落组分;所述的基因组测序仪英文表达Miseq;
(2)筛选:经统计学分析筛选出高产奶牛瘤胃优势菌,即拟杆菌和瘤胃球菌;
(3)分离纯化菌种:提取奶牛瘤胃液,利用亨盖特滚管法培养瘤胃微生物,选取拟杆菌和瘤胃球菌,进一步提纯并获得大量菌种;所述的亨盖特滚管法英文表达为Hungate氏滚管法;
(4)培养:将拟杆菌和瘤胃球菌菌种分别接种于亨盖特滚管法的滚管中37℃恒温培养2-3天;
(5)发酵:将滚管中拟杆菌和瘤胃球菌分别接种于相对应的厌氧发酵罐37℃恒温培养3-5天后,使发酵液降温至20℃;
(6)组合:收集拟杆菌和瘤胃球菌的发酵菌悬液混合均匀;
(7)包装:根据需要加入其他营养成分后密封包装或直接密封包装。
所述的拟杆菌培养基配方:葡萄糖1g/L、蛋白胨10g/L、牛肉膏3g/L、NaCl 5g/L、维生素B 10ug/L、抗坏血酸1g/L,并调节PH至7.0-7.5;瘤胃球菌培养基的配方:尿素5g/L、KH2PO41g/L、K2HPO41g/L、FeSO4·7H2O 0.05g/L、MnSO4·H2O 0.02g/L、MgSO4·7H2O 0.5g/L、酵母浸出液0.2g/L、稻草秆20g/L,并调节PH至6.0-7.0;
拟杆菌和瘤胃球菌量达1.0×108cfu/ml–2.5×108cfu/ml,并且比例在0.5-1.5之间组合。
制备方法如图1所示
(1)分离纯化菌种:提取奶牛瘤胃液,利用Hungate氏滚管法培养瘤胃微生物,选取拟杆菌和瘤胃球菌,进一步提纯并获得大量菌种;
(2)培养:将拟杆菌和瘤胃球菌菌种分别接种于Hungate氏滚管中37℃恒温培养2-3天;(3)发酵:将滚管中拟杆菌和瘤胃球菌分别接种于相对应的厌氧发酵罐37℃恒温培养3-5天后,使发酵液降温至20℃;
(4)组合:收集拟杆菌和瘤胃球菌的发酵菌悬液按1:1的比例混合均匀;
(5)包装:根据需要加入其他营养成分后密封包装或直接密封包装。
含拟杆菌和瘤胃球菌各1.0×108cfu/ml–2.5×108cfu/ml。
应用效果试验
为了更好地了解本复合微生态制剂的作用功效,选取的60头体况、泌乳年龄,胎次等相似荷斯坦奶牛进行喂养试验,试验周期为30天,试验相关情况如下。
1材料与方法
1.1材料
供试动物:荷斯坦奶牛
供试微生态制剂:本复合微生态制剂
1.2试验设计
60头荷斯坦奶牛平均分为2组,分别为实验组和对照组,再将对照组和实验组各分为3小组,记录实验数据。奶牛基础日粮由羊草、青贮饲料、混合精料组成,每头牛每天喂30kg青贮饲料、20kg羊草和10kg混合精料。对照组的饲料中不添加微生态制剂,实验组的饲料中按精料的0.5%添加复合微生态制剂,添加时加少量水与精料混合均匀后添加到供试牛日粮中,确保奶牛完全采食。每日于9:00和15:00饲喂2次,喂料结束后散放到运动场自由运动,饮水槽保证足够清洁的饮水。每日于7:00和17:00人工挤奶2次,测定产奶量。
2试验结果与分析
2.1本复合微生态制剂对荷斯坦奶牛产奶量的影响,结果见表1。
表1复合微生态制剂对荷斯坦奶牛产奶量的影响
2.2分析
由表1可知:本复合微生态制剂作用功效显著,可将荷斯坦奶牛产奶量提高10%左右,效果显著,从而能够增加经济效益。

Claims (3)

1.一种提高奶牛产奶量的微生态制剂,其特征是:采用免培养技术结合统计学的方法筛选高产奶量奶牛瘤胃的优势菌,利用纯培养技术获得优势菌的纯种菌株,经发酵培养配伍组合成微生态制剂;按如下操作步骤制备:
(1)测序:抽提荷斯坦奶牛瘤胃液DNA进行PCR扩增,经基因组测序仪高通量测序和分类学分析获得奶牛瘤胃微生物群落组分;
(2)筛选:经统计学分析筛选出高产奶牛瘤胃优势菌,即拟杆菌和瘤胃球菌;
(3)分离纯化菌种:提取奶牛瘤胃液,利用亨盖特滚管法培养瘤胃微生物,选取拟杆菌和瘤胃球菌,进一步提纯并获得大量菌种;
(4)培养:将拟杆菌和瘤胃球菌菌种分别接种于亨盖特滚管法的滚管中37℃恒温培养2-3天;
(5)发酵:将滚管中拟杆菌和瘤胃球菌分别接种于相对应的厌氧发酵罐37℃恒温培养3-5天后,使发酵液降温至20℃;
(6)组合:收集拟杆菌和瘤胃球菌的发酵菌悬液混合均匀;
(7)包装:根据需要加入其他营养成分后密封包装或直接密封包装。
2.根据权利要求1所述的一种提高奶牛产奶量的微生态制剂,其特征是:所述的拟杆菌的培养基配方:葡萄糖1 g/L、蛋白胨10 g/L、牛肉膏3 g/L、NaCl 5 g/L、维生素B 10 ug/L、抗坏血酸1 g/L,并调节PH至7.0-7.5;瘤胃球菌的培养基配方:尿素 5g/L 、KH2PO4  1g/L 、K2HPO4 1g/L、FeSO4·7H2O 0.05g/L 、MnSO4·H2O 0.02g/L 、MgSO4·7H2O 0.5 g/L 、酵母浸出液 0.2 g/L 、稻草秆 20 g/L,并调节PH至6.0-7.0。
3.根据权利要求1所述的一种提高奶牛产奶量的微生态制剂,其特征是:拟杆菌和瘤胃球菌量达1.0×108cfu/ml–2.5×108cfu/ml,并且比例在0.5-1.5之间组合。
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