CN114591863B - 用于提高断奶仔猪生长性能和免疫功能的藏猪源菌剂组合 - Google Patents
用于提高断奶仔猪生长性能和免疫功能的藏猪源菌剂组合 Download PDFInfo
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- CN114591863B CN114591863B CN202210294554.1A CN202210294554A CN114591863B CN 114591863 B CN114591863 B CN 114591863B CN 202210294554 A CN202210294554 A CN 202210294554A CN 114591863 B CN114591863 B CN 114591863B
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Abstract
本发明公开了用于提高断奶仔猪生长性能和免疫功能的藏猪源菌剂组合,从藏猪体内分离出乳杆菌(保藏号为CGMCC No.23773)和嗜热嗜酸双歧杆菌(CGMCC No.23772),这两株菌具有强产乙酸能力、强耐酸和耐胆盐能力以及具有广泛的碳源利用谱等特点,可显著提高断奶仔猪饲料表观消化率、生长性能、降低腹泻率和提高免疫功能,将两株菌组合使用可降低菌株代谢负担,菌株之间协同发挥作用具有更好的效果。
Description
技术领域
本发明属于微生物技术领域,具体涉及可防治断奶仔猪腹泻、提高断奶仔猪生长性能和免疫功能的藏猪源乳杆菌、嗜热嗜酸双歧杆菌及其组合物。
背景技术
中国是世界第一养猪大国和猪肉消费国,集约化饲养单一外来猪品种(比如瘦肉型猪种长白猪、大白猪和杜洛克猪)是应对生猪产品巨大需求的主要策略。但是,集约化单一品种的饲养加剧了动物疫病防治的压力。目前动物疫病防控方面主要是依赖抗生素的使用。据调查,2013年我国抗生素总使用量约为16.2万吨,其中人用抗生素占到总量16%,其余为兽用抗生素。正由于抗生素过度使用,在中国,细菌整体的耐药率,要远远高于欧美国家,大约在45%左右。有学者进行了探索性的研究,发现长期喂食抗生素,生猪肠道微生物的空间稳态就会被打破,不仅影响了肠道功能菌群的特异性定植,也影响了调控肠菌稳态的噬菌体病毒的定植,其后果必然会影响动物自身的健康,甚至导致疾病的频繁发生。相应地,有研究人员发现断奶仔猪常常伴随着严重的肠道感染和抗生素的过度使用,加剧了肠道微生物的失调,导致仔猪腹泻的频繁发生。
针对特定生理代谢功能,研发新型功能菌群制剂和无抗饲料,辅助提升生猪生长和免疫功能等重要关键性状,不仅符合国家和地方的双重需求,也有助于在我国养殖行业的无抗养殖时代,实现绿色、高效的畜牧业养殖模式。肠道微生物应用于人类、动物健康方面,当前经典的方式是几类益生菌单用或联用,比如乳酸菌、双歧杆菌、嗜热链球菌、粪肠球菌、屎肠球菌、芽孢杆菌等。中国地方品种猪资源丰富,为新型功能菌群的研究提供了天然的肠道微生物资源。藏猪(Tibetan pig)素有“高原之珍”之美誉,它是世界上少有的半驯化状态的高原型猪种,也是中国国家级重点保护品种中唯一的高原性猪种。2004年,藏猪正式列入《中国畜牧品种志》,被正式确定为地方原始猪种;其中云南省已经把藏猪养殖作为3大新特色农业之一。藏猪能适应恶劣的高寒气候、终年放牧和低劣的饲养管理条件。这种极强的适应能力和抗逆性,是其他猪种所不具备的独特种质特性。研究发现从中国抗性地方品种猪发现了乳酸菌代谢产生的细菌素可以减轻或阻止早期断奶胁迫诱导产生的腹泻问题,从实践方面证实了挖掘具有优良性状地方品种猪肠道微生物资源,并将其用于生猪生产以改善其生产性能和免疫功能是可行的。目前,单株益生菌作为饲料添加剂应用于断奶仔猪已经非常广泛,然而其效果有限。为了进一步挖掘益生菌在断奶仔猪上的应用潜力,微生物菌株的组合应用成为潜在的新方法。与单菌株相比,微生物菌株的组合应用可降低菌株代谢负担,菌株之间协同发挥作用具有更好的效果。
发明内容
本发明从藏猪体内分离出两株菌种,这两株菌具有强产乙酸能力、强耐酸和耐胆盐能力以及具有广泛的碳源利用谱等特点,可显著提高断奶仔猪饲料表观消化率、生长性能、降低腹泻率和提高免疫功能,将两株菌组合使用可降低菌株代谢负担,菌株之间协同发挥作用具有更好的效果。
为了实现上述目的,本发明采用以下技术方案:
本发明提供一株乳杆菌(Lactobacillus sp.)23-2,分离自藏猪新鲜粪便,保藏号CGMCC No.23773,保藏日期为2021年11月10日,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心。
本发明提供一株嗜热嗜酸双歧杆菌(Bifidobacterium thermacidophilum)11-1-1,分离自藏猪新鲜粪便,保藏号CGMCC No.23772,保藏日期为2021年11月10日,保藏单位为中国微生物菌种保藏管理委员会普通微生物中心。
所述乳杆菌23-2和嗜热嗜酸双歧杆菌11-1-1均具有一定的耐酸和耐胆盐能力,两株菌在pH4.0条件下处理120分钟以及0.05%胆盐处理90分钟的过程中始终能维持90%以上的存活率。
所述乳杆菌23-2和嗜热嗜酸双歧杆菌11-1-1均具有产乙酸和丙酸能力,以添加了低聚木糖的GAM培养基中作为发酵培养基,乳杆菌23-2和嗜热嗜酸双歧杆菌11-1-1产乙酸量分别为134.03μg/mL和339.61μg/mL,产丙酸量分别为1.82μg/mL和5.63μg/mL。
所述乳杆菌23-2和嗜热嗜酸双歧杆菌11-1-1均具有广泛的碳源利用谱,以缺乏碳源的寡营养MM培养基为基础培养基,添加不同碳源,发现乳杆菌23-2和嗜热嗜酸双歧杆菌11-1-1对34种植物、动物来源的单糖、多糖都具有利用能力。
乳杆菌23-2或嗜热嗜酸双歧杆菌11-1-1在防治断奶仔猪腹泻、促进断奶仔猪生长及提高免疫功能中的应用:添加乳杆菌23-2或嗜热嗜酸双歧杆菌11-1-1可以提高断奶仔猪生长性能,降低断奶仔猪料肉比和腹泻率,提高断奶仔猪的免疫功能。因此,乳杆菌23-2或嗜热嗜酸双歧杆菌11-1-1可用于制备防治断奶仔猪腹泻的产品(比如药物、饲料添加剂等),也可以用于促进断奶仔猪生长及提高免疫功能。
一种菌剂组合,包括乳杆菌23-2和嗜热嗜酸双歧杆菌11-1-1,与单株菌相比,这两株菌组合可以显著提高断奶仔猪的日增重,降低料肉比和腹泻率,提高免疫球蛋白含量,降低炎症因子含量,提高饲料营养物质的表观消化率等,具有完全替代抗生素的作用。因此,该菌剂组合可用于制备防治断奶仔猪腹泻的产品(比如药物、饲料添加剂等),也可以用于促进断奶仔猪生长及提高免疫功能。
与现有技术相比,本发明具有以下优点及有益效果:
本发明所述的乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1均分离自藏猪,在猪上应用不会产生排斥作用以及副作用,安全性高。所述两株菌生产成本低,生产应用简单,对环境无污染,适合推广应用。所述两株菌均具有强耐酸和耐胆盐的能力,可以过胃酸到达猪肠道发挥作用,提高断奶仔猪的生长性能并降低腹泻率;两株菌具有强产乙酸能力,可以有效提高断奶仔猪的免疫功能;两株菌均具有广泛的碳源利用谱,可有效提高断奶仔猪的饲料表观消化率,降低养殖成本。微生物菌株的组合应用菌株之间可以协同发挥作用具有更好的效果。
附图说明
图1为乳杆菌23-2的显微照片。
图2为嗜酸嗜热双歧杆菌11-1的显微照片。
图3乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1的酸、胆盐耐受能力检测结果。A为嗜酸嗜热双歧杆菌11-1-1的酸耐受能力检测,B为嗜酸嗜热双歧杆菌11-1-1的胆盐耐受能力检测,C为乳杆菌23-2的酸耐受能力检测,D为乳杆菌23-2的胆盐耐受能力检测。
具体实施方式
为了更好的理解本发明,下面结合实施例进一步阐明本发明的内容,但本发明的内容不仅仅局限于下面的实施例。
实施例1乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1的分离与鉴定
2018年3月在云南昆明解剖藏猪后,将0.5g藏猪肠道内容物浸没于5mL PBS(除氧),涡旋5min,低强度超声波处理5min;使用细胞筛过滤浊液,除去固形物;利用梯度稀释法分离藏猪肠道细菌,对滤液进行10倍梯度稀释,取10-4至10-8梯度的稀释液,涡旋混匀后,吸取100μL涂布于平板,每一梯度三个重复;37℃倒置厌氧恒温培养(混合气体配比为N285%、H2 10%、CO2 5%);将分离到的单菌落用划线分离法进行纯化。
菌株标号为23-2的菌落形态呈不规则状,透明,表面干燥,边缘不规则,中央无凸起,直径1-2mm。在光学显微镜下,23-2菌体呈典型长杆状菌,成串排列,无分支(如图1所示);菌株标号为11-1的菌落形态呈圆形,不透明白色,表面干燥,边缘前期光滑,后期呈不规则状,中央无凸起,直径1-2mm。在光学显微镜下,11-1有明显的多形态性呈双叉状的细杆菌、一端或两端膨大呈棒状、空泡状、球状以及无双叉的杆菌(如图2所示)。
23-2和11-1-1的16S rRNA基因进行了PCR扩增并测序,使用T载体试剂盒将得到的DNA片段与T载相连,筛选阳性克隆子,送北京六合华大基因科技有限公司进行序列测序,序列如SEQ ID NO:1和SEQ ID NO:2所示。根据引物截选出目的片段,在EzbioCloud中进行序列相似性比对得到结果,菌株23-2与德氏乳酸杆菌(Lactobacillus delbrueckii)有95.28%的相似性,菌株11-1-1与嗜酸嗜热双歧杆菌(Bifidobacteriumthermacidophilum)有100%的相似性。
结合形态特点和16S rRNA基因测序和序列比对分析结果,将分离菌株23-2鉴定为乳杆菌(Lactobacillus sp.),分离菌株11-1-1鉴定为嗜酸嗜热双歧杆菌(Bifidobacterium thermacidophilum)。申请人已于2021年11月10日交由中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)进行保藏,乳杆菌(Lactobacillus sp.)23-2的保藏号为CGMCC No.23773,嗜酸嗜热双歧杆菌(Bifidobacterium thermacidophilum)11-1-1的保藏号为CGMCC No.23772。
实施例2乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1耐酸和耐胆盐实验
1、挑取新鲜活化的待测菌株单菌落接种于5mL液体培养基,37℃静置培养48-72h至稳定期,液体培养基为对应菌株的最适培养基;所述乳杆菌23-2最适培养基组成如下:蛋白胨10.0g,牛肉浸膏10.0g,酵母膏5.0g,葡萄糖20.0g,Tween 80 1.0g;MgSO4·7H2O0.05g,K2HPO4 2.0g,MnSO4·7H2O 0.2g,醋酸钠5.0g,柠檬酸三铵2.0g,加入1L蒸馏水,121℃灭菌15min。所述双歧杆菌11-1-1最适培养基组成如下:蛋白胨15.0g,酪蛋白胨10.0g,牛肉浸膏2.0g,酵母膏5.0g,葡萄糖3.0g,NaCl 3.0g,可溶性淀粉0.3g,血清粉13.5g,K2HPO42.5g,巯基乙酸钠2.0g,调节pH至7.0,加入1L蒸馏水,121℃灭菌15min。按体积比1:100转接5mL液体培养基,37℃静置或震荡培养12-24h;取菌液,经涡旋仪混匀后,分别吸取800μL菌液于1.5mL无菌离心管,PBS洗菌两次后,用800μL PBS重悬菌体。
2、酸耐受:分别使用800μL PBS、pH为2.0、3.0、4.0的培养基重悬菌体,37℃孵育并开始计时,在30min、60min、90min、120min分别对培养基pH为2.0、3.0、4.0的菌体进行10×梯度稀释涂板,取10-5梯度的稀释液,经涡旋仪混匀后,吸取100μL涂布于平板,重复三次;胆盐耐受:分别使用800μL PBS、胆汁盐(bile salt)浓度为0.05%、0.1%、0.3%的培养基重悬菌体,37℃孵育并开始计时,在30min、60min、90min对胆汁盐(bile salt)浓度为0.05%、0.1%、0.3%处理的菌体进行梯度稀释涂板,取10-5梯度的稀释液,经涡旋仪混匀后,吸取100μL涂布于平板,重复三次;在0min对PBS重悬的菌体进行10倍梯度稀释,取10-5梯度的稀释液,经涡旋仪混匀后,吸取100μL涂布于平板,每一梯度三个重复。37℃厌氧倒置培养24h,对平板上菌落计数。
试验结果如下:如图3所示,乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1在pH4.0条件下处理120分钟以及0.05%胆盐处理90分钟的过程中始终能维持90%以上的存活率。说明乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1具有一定的耐酸、胆盐能力,具备在仔猪肠道中定植的基础,可在实际生产中推广应用。
实施例3乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1碳源利用能力测定
1、挑取新鲜活化的待测菌株单菌落接种于5mL GAM液体培养基,37℃静置培养48-72h至稳定期;按1:100转接5mL GAM液体培养基,37℃静置或震荡培养24h;在超净台中进行96孔板碳源布板,每孔分装100μL 2×碳源溶液,碳源溶液以寡营养MM培养基为基础培养基,碳源浓度为0.5g/100mL,每种碳源两个重复且相邻碳源之间需间隔一孔。最后添加两100μL蒸馏水的孔作为阴性对照。布板完成后转移到厌氧箱中静置12h(置换板中气体)。
2、在厌氧箱中,吸取1ml菌液于1.5ml离心管,使用1×MM培养基洗菌两次;使用1mL2×MM培养基重悬菌体,按1:100转接到50ml 2×MM培养基,充分混匀后,在每个添加了碳源溶液的孔中补加100μL菌悬液至终体积为200μL;在96孔板正面覆盖一层透光聚氨酯薄膜并使用放置在厌氧箱中的酶标仪在48h内每隔2h对96孔板OD600进行一次测量。
表1碳源利用能力检测结果
如表1所示,△OD值表示的是菌株对碳源的利用能力,△OD值越大,表明该菌利用该碳源的能力越强;乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1具有广泛的碳源利用谱,有利于其在肠道中生存。对支链淀粉、果糖、半乳糖和葡萄糖的利用能力较强,便于工业生产。
实施例4乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1短链脂肪酸产生能力测定
1、挑取新鲜活化的待测菌株单菌落接种于5mL GAM液体培养基,37℃静置培养48-72h至稳定期;按1:100转接5mL含有低聚木糖0.5g/100mL的GAM液体培养基,37℃静置培养48h;取2mL菌液,10000rpm离心5min,取上清;在10mL厌氧管中依次加入1200μL菌液上清,400μL 50%硫酸(必须使用玻璃移液管量取)、2mL色谱级乙醚(国产乙醚需经过0.45μm有机滤膜过滤)后迅速密封;将厌氧管置于28℃摇床200rpm萃取45min;室温静置分层,使用玻璃吸管将上清快速转移到气相色谱样品瓶中并密封以免样品挥发。
2、对样品进行GC-MS检测。短链脂肪酸测定采用外标法。仪器为岛津公司GCMS QP2010Ultra色谱柱:(30m×0.25mm i.d×0.25μm);色谱柱升温程序:初始温度35℃,以5℃/min升至130℃,以30℃/min升至230℃后保持12min;进样口温度:230℃;进样体积:1μl;分流比5:1;载气:高纯氮(99.999%以上);流速:1mL/min;采用70eV电子束进行电子轰击电离,每个样本检测时间均为34.3min。
表2短链脂肪酸(SCFA)产生能力检测结果
研究表明,可溶性可发酵碳水化合物(FCS)的摄入增加了模式动物肠道中的SCFA的含量。据此我们将细菌发酵碳水化合物产生SCFAs能力作为益生菌筛选的指标。并将添加了低聚木糖的GAM培养基中作为发酵培养基,进行SCFAs的检测,从表2中数据可以看出嗜酸嗜热双歧杆菌11-1-1和乳杆菌23-2可以发酵木聚糖产生较高水平的乙酸,以添加了低聚木糖的GAM培养基中作为发酵培养基,乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1产乙酸量分别为134.03μg/mL和339.61μg/mL,也具有产生少量丙酸的能力,产丙酸量分别为1.82μg/mL和5.63μg/mL。这证实这两株细菌对低聚木糖具有较强的代谢能力。
实施例5乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1联合添加与单株菌在断奶仔猪上的应用比较实验
试验方案:选择21日龄断奶的杜长大健康仔猪40头,随机分成4组,每组10头仔猪,试验周期14天。对照组仔猪饲喂基础日粮;试验组1仔猪添加乳杆菌23-2,最终含量为0.5×106cfu/g(乳杆菌组);试验组2仔猪添加嗜酸嗜热双歧杆菌11-1-1,最终含量为0.5×106cfu/g(双歧杆菌组);试验组3仔猪添加乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1组合,两种菌含量分别为0.5×106cfu/g(两菌联合组)。基础日粮及营养水平见表3。试验第0和14天记录仔猪体重,同时记录仔猪全程采食量和腹泻情况。试验结束时计算仔猪日增重,日采食量、料肉比以及腹泻指数(每组仔猪腹泻评分总和/(总头数×总天数))等。腹泻评分规则标准如下:干粪,1分;无腹泻,正常干粪,2分;轻度腹泻,粪表面稍微有点湿,半干粪,3分;中度腹泻,软粪,稍微有点稀,4分;严重腹泻,水样粪,5分。
表3基础日粮组成及营养水平
1.单独添加益生菌以及添加两株菌组合物对断奶仔猪生长性能的影响
如表4所示,乳杆菌组和双歧杆菌组仔猪第14天体重和平均日增重显著高于对照组仔猪,两菌联合组仔猪第14天体重和平均日增重显著高于其他三个试验组;乳酸杆菌组和双歧杆菌组仔猪料肉比显著低于对照组仔猪,两菌联合组仔猪料肉比显著低于其他三个试验组。
表4生长性能
注:BW,体重;ADG,平均日增重;ADFI,平均日采食量;F/G,料肉比
2.单独添加益生菌以及添加两株菌组合物对断奶仔猪腹泻指数的影响
如表5所示,乳杆菌组和双歧杆菌组仔猪腹泻指数显著低于对照组仔猪,两菌联合组仔猪腹泻指数显著低于其他三个试验组。
表5腹泻指数
3.单独添加益生菌以及添加两株菌组合物对断奶仔猪血清抗体和炎症因子水平的影响
如表6所示,乳杆菌组和双歧杆菌组仔猪血清中IgG含量显著高于对照组仔猪,两菌联合组仔猪血清中IgG含量显著高于其他三个试验组;乳杆菌组和双歧杆菌组仔猪TNF-α和IL-1β含量显著低于对照组仔猪,两菌联合组仔猪TNF-α和IL-1β含量显著低于其他三个试验组。
表6血清抗体和炎症因子水平
4.小结
添加单株益生菌可以一定程度上提高断奶仔猪生长性能,降低断奶仔猪料肉比和腹泻率,提高断奶仔猪的免疫功能;益生菌组合物的上述作用优于单株益生菌。
实施例6乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1组合物在断奶仔猪上替抗效果实验
试验方案:选择21日龄断奶的杜长大健康仔猪30头,饲喂玉米豆粕型基础日粮一周后,随机分成3组,每组10头仔猪,试验周期28天;对照组仔猪饲喂基础日粮;抗生素组仔猪基础日粮上添加金霉素75g/T;益生菌组仔猪基础日粮上添加乳杆菌23-2和嗜酸嗜热双歧杆菌11-1-1组合,两种菌含量分别为0.5×106cfu/g。基础日粮及营养水平同表3。试验第0、14、28天记录仔猪体重,同时记录仔猪全程采食量和腹泻情况。试验结束时计算仔猪日增重,日采食量、料肉比以及腹泻指数(每组仔猪腹泻评分总和/(总头数×总天数))等。腹泻评分规则同实施例5。
试验结果如下:
1.乳杆菌和双歧杆菌组合物与抗生素对断奶仔猪生长性能的影响比较
如表7所示,体重方面,与对照组相比,抗生素组和益生菌组仔猪末重(28日龄)显著升高;增重方面,与对照组相比,抗生素组和益生菌组仔猪0-14天以及0-28天的平均日增重显著升高,而15-28天日增重无显著差异;采食量方面,益生菌组仔猪全程日采食量显著高于抗生素组和对照组;抗生素组和益生菌组仔猪0-14天以及0-28天的料肉比显著低于对照组。
表7生长性能
注:BW,体重;ADG,平均日增重;ADFI,平均日采食量;F/G,料肉比.
2.乳杆菌和双歧杆菌组合物与抗生素对断奶仔猪腹泻指数的影响
如表8所示,抗生素组和益生菌组仔猪腹泻指数显著低于对照组,抗生素组和益生菌组仔猪腹泻指数无显著差异。
表8腹泻指数
3.乳杆菌和双歧杆菌组合物与抗生素对断奶仔猪血清抗体和炎症因子水平的影响比较
如表9所示,抗生素组和益生菌组仔猪血清中IgG含量显著高于对照组,而抗生素组和益生菌组仔猪血清中炎症因子TNF-α和IL-1β含量显著低于对照组。抗生素组和益生菌组仔猪血清中IgG、TNF-α和IL-1β含量无显著差异。
表9血清抗体和炎症因子水平
4.乳杆菌和双歧杆菌组合物与抗生素对断奶仔猪饲料表观消化率的影响比较
如表10所示,抗生素组和益生菌组仔猪干物质、粗脂肪、粗蛋白、酸性洗涤纤维和中性洗涤纤维的表观消化率显著高于对照组仔猪。抗生素组和益生菌组仔猪干物质、粗脂肪、粗蛋白、酸性洗涤纤维和中性洗涤纤维的表观消化率无显著差异。
表10饲料表观消化率
5.乳杆菌和双歧杆菌组合物与抗生素对断奶仔猪肠道乙酸含量的影响比较
如表11所示,益生菌组仔猪盲肠和结肠中乙酸含量显著高于对照组和抗生素组仔猪。
表11肠道乙酸含量
6.小结
乳杆菌和双歧杆菌组合物可以显著提高断奶仔猪的日增重,降低料肉比和腹泻率,提高免疫球蛋白含量,降低炎症因子含量,提高饲料营养物质的表观消化率等,且这些方面的作用效果与抗生物相似;此外,乳杆菌和双歧杆菌组合物显著提高了断奶仔猪盲肠和结肠中乙酸的含量,这是本实验益生菌组合物提高机体免疫功能的主要原因。因为大量研究报道,肠道微生物产生的乙酸具有显著的抗炎作用和促进中性粒细胞的趋化作用。这些结果表明乳杆菌和双歧杆菌组合物可提高断奶仔猪生长性能和免疫功能,提高养殖效率,具有完全替代抗生素的作用。
序列表
<110> 云南大学
中国科学院微生物研究所
中国科学院亚热带农业生态研究所
<120> 用于提高断奶仔猪生长性能和免疫功能的藏猪源菌剂组合
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1439
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
atgcaagtcg agcgagctga attcaaagat cccttcgggg tgatttgttg gacgcgagcg 60
gcggatgggt gagtaacacg tgggcaacct gccctaaaga ctgggatacc acttggaaac 120
aggtgctaat accggataac aacaaaagct gcatggcttt tgtttgaaag gcggctttaa 180
gctgtcactt taggatgggc ccgcggtgca ttagctagtt ggtggggtaa cggcccacca 240
aggcaatgat gcatagccga gttgagagac tgatcggcca cattgggact gagacacggc 300
ccaaactcct acgggaggca gcagtaggga atcttccaca atggacgcaa gtctgatgga 360
gcaacgccgc gtgagtgaag aaggttttcg gatcgtaaag ctctgttgtt ggtgaagaag 420
gatagtggca gcaactggcc attatttgac ggtaatcaac cagaaagtca cggctaacta 480
cgtgccagca gccgcggtaa tacgtaggtg gcaagcgttg tccggattta ttgggcgtaa 540
agcgagcgca ggcggaaaga taagtctgaa gtgaaagccc ccggcttaac cggggaactg 600
cttcggaaac tgtccttctt gagtgcagaa gaggagagtg gaactccatg tgtagcggtg 660
gaatgcgtag atatatggaa gaacaccagt ggcgaaggcg gctctctggt ctgcaactga 720
cgctgaggct cgaaagcatg ggtagcgaac aggattagat accctggtag tccatgccgt 780
aaacgatgag tgctaggtgt tggggacttt ccggttctca gtgccgcagc taacgcaata 840
agcactccgc ctggggagta cgaccgcaag gttgaaactc aaaggaattg acgggggccc 900
gcacaagcgg tggagcatgt ggtttaattc gaagcaacgc gaagaacctt accaggtctt 960
gacatcctgc gccaagccta gagataggcc gttcccttcg gggacgcaga gacaggtggt 1020
gcatggctgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 1080
ccttgtcact agttgccagc attaagttgg gcactctagt gagactgccg gtgacaaacc 1140
ggaggaaggt ggggatgacg tcaagtcatc atgcccctta tgacctgggc tacacacgtg 1200
ctacaatggg cagtacaacg agaagcgaac ccgcgagggc aagcggatct cttaaagctg 1260
ttctcagttc ggactgcagg ctgcaactcg cctgcacgaa gctggaatcg ctagtaatcg 1320
tggatcagca cgccacggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca 1380
tggaagtctg caatgcccga agtcggtggg ataaccttaa tggaatcagc cgcactaag 1439
<210> 2
<211> 1537
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
gtggagggtt cgattctggc tcaggatgaa cgctggcggc gtgcttaaca catgcaagtc 60
gaacgggatc ccgcgggctt tgcttgcggg tgagagtggc gaacgggtga gtaatgcgtg 120
accaacctgc cccatgctcc ggaatagctc ctggaaacgg gtggtaatgc cggatgtgcc 180
gggctcctgc atgggggtcc gggaaagctc tggcggcgtg ggatggggtc gcgtcctatc 240
agcttgttgg cggggtgagg gcccaccaag gcttcgacgg gtagccggcc tgagagggcg 300
accggccaca ttgggactga gatacggccc agactcctac gggaggcagc agtggggaat 360
attgcacaat gggcgcaagc ctgatgcagc gacgccgcgt gcgggatgga ggccttcggg 420
ttgtaaaccg cttttgtttg ggagcaagcc cttcggggtg agtgtacctt tcgaataagc 480
accggctaac tacgtgccag cagccgcggt aatacgtagg gtgcgagcgt tatccggatt 540
tattgggcgt aaagggctcg taggcggttc gtcgcgtccg gtgtgaaagt ccatcgccta 600
acggtggatc tgcgccgggt acgggcgggc tggagtgcgg taggggagac tggaattccc 660
ggtgtaacgg tggaatgtgt agatatcggg aagaacacca atggcgaagg caggtctctg 720
ggccgttact gacgctgagg agcgaaagcg tggggagcga acaggattag ataccctggt 780
agtccacgcc gtaaacggtg gatgctggat gtggggccct tccacgggtc ccgtgtcgga 840
gccaacgcgt taagcatccc gcctggggag tacggccgca aggctaaaac tcaaagaaat 900
tgacgggggc ccgcacaagc ggcggagcat gcggattaat tcgatgcaac gcgaagaacc 960
ttacctgggc ttgacatgtt cccgacgacc gtggagacac ggtttccctt cggggcgggt 1020
tcacaggtgg tgcatggtcg tcgtcagctc gtgtcgtgag atgttgggtc aagtcccgca 1080
acgagcgcaa ccctcgcccc gtgttgccag cgcgtcatgg cgggaactca cgggggaccg 1140
ccggggttaa cccggaggaa ggtggggatg acgtcagatc atcatgcccc ttacgtccag 1200
ggcttcacgc atgctacaat ggccggtaca acgggatgcg acatggtgac atggagcgga 1260
tccctgaaaa ccggtctcag ttcggatcgg agcctgcaac ccggctccgt gaaggcggag 1320
tcgctagtaa tcgcggatca gcaacgccgc ggtgaatgcg ttcccgggcc ttgtacacac 1380
cgcccgtcaa gtcatgaaag tgggcagcac ccgaagccgg tggcctgacc agttctgctg 1440
gggggagccg tctaaggtga ggctcgtgat tgggactaag tcgtaacaag gtagccgtac 1500
cggaaggtgc ggctggatca cctcctttct acggaga 1537
Claims (8)
1.乳杆菌(Lactobacillus sp.),其特征在于,保藏号为CGMCC No.23773。
2.嗜热嗜酸双歧杆菌(Bifidobacterium thermacidophilum),其特征在于,保藏号为CGMCC No.23772。
3.一种菌剂组合,其特征在于,包括保藏号为CGMCC No.23773的乳杆菌和保藏号为CGMCC No.23772的嗜热嗜酸双歧杆菌。
4.权利要求1所述的乳杆菌或权利要求2 所述的嗜热嗜酸双歧杆菌在制备防治断奶仔猪腹泻产品中的应用。
5.权利要求1所述乳杆菌或权利要求2 所述的嗜热嗜酸双歧杆菌在制备促进断奶仔猪生长及提高免疫功能的产品中的应用。
6.权利要求3所述的菌剂组合在制备防治断奶仔猪腹泻产品中的应用。
7.权利要求3所述的菌剂组合在制备促进断奶仔猪生长及提高免疫功能的产品中的应用。
8.权利要求3所述的菌剂组合在制备断奶仔猪饲料添加剂中的应用。
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CN113288919A (zh) * | 2021-06-17 | 2021-08-24 | 浙江大学 | 解淀粉芽孢杆菌在预防断奶仔猪腹泻中的应用 |
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RU2062038C1 (ru) * | 1993-06-29 | 1996-06-20 | Тараканов Борис Васильевич | Способ выращивания поросят |
CN107858310A (zh) * | 2017-12-04 | 2018-03-30 | 云南大学 | 一株产香放线菌及其对诺邓火腿增香的应用 |
CN112266880A (zh) * | 2020-10-20 | 2021-01-26 | 青岛普罗百世生物科技有限公司 | 一种嗜酸乳杆菌及其在断奶仔猪饲料中的应用 |
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