CN113249255B - 一种防控禽肠炎的枯草芽孢杆菌及其应用 - Google Patents
一种防控禽肠炎的枯草芽孢杆菌及其应用 Download PDFInfo
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Abstract
本发明涉及功能微生物筛选与应用技术领域,提供了一种防控禽肠炎的枯草芽孢杆菌及其应用。所述枯草芽孢杆菌已于2019年6月6日保藏于中国武汉 武汉大学的中国典型培养物保藏中心,保藏编号为CCTCC NO:M2019437。所述枯草芽孢杆菌对产气荚膜梭菌和球虫导致的肉鸡肠道性炎症有良好的预防效果,可广泛应用于家禽养殖领域。
Description
技术领域
本发明涉及功能微生物筛选与应用技术领域,具体涉及一种防控禽肠炎的枯草芽孢杆菌及其应用。
背景技术
肠道健康是家禽高效生产的重要保障,但在细菌耐药性日益严重和无抗养殖的双重压力下,家禽肠道问题更加突出。尤其是在我国家禽养殖业转型升级时期,饲养管理与疫病防控水平相对滞后于规模化和集约化的进程,家禽养殖场肠道疫病发生率居高不下。危害家禽肠道健康的病原菌主要包括大肠杆菌、沙门菌、葡萄球菌、空肠弯曲菌、产气荚膜梭菌等,这些病原引发的疫病造成的急性死亡或隐性感染严重危害家禽健康,导致生产能力降低,甚至导致死亡。
益生菌是一类具有生物活性和对宿主健康有益的微生物,并因其无污染、无毒及无不良反应、抗菌保健和促生长等特点,成为继抗生素之后的新型添加剂。益生菌可通过调节肠道微生物组成发挥抑制病原菌作用。益生菌通常能与肠黏膜上皮细胞结合,占据致病菌肠黏膜上皮细胞结合位点,竞争性抑制宿主肠道病原菌的黏附,从而能够保护肠粘膜,降低肠道致病菌感染的风险。益生菌定植于肠道,刺激肠粘膜上皮细胞,促进局部粘膜组织生长发育,有助于维持肠黏膜屏障功能。
胡勇等从健康鸡粪便中共分离出41株益生菌株,并从中筛选出对肠道沙门氏菌有较强颉颃作用的益生菌株4株,其中1菌株同时具有较强的耐酸、耐胆盐特性和较好的疏水性能,有望作为潜在的颉颃性饲用益生菌用于控制鸡的肠炎沙门氏菌感染。王佰魁等进行了益生菌对肉鸡产气荚膜梭菌进行攻毒的保护效果研究,发现植物乳杆菌和多粘芽孢杆菌可通过抑制产气荚膜梭菌的生长,提高鸡巨噬细胞的吞噬和免疫功能,改善肉鸡肠道菌群结构、微生物功能及黏膜结构等,降低产气荚膜梭菌感染引起的负面效应。黄晨轩等研究表明饲粮中添加1‰复合微生态制剂可提高蛋鸡后期的养分消化率和免疫能力,改善肠道菌群结构,改善机体健康。张雷等研究发现通过饮水给与复合微生态制剂能够提高雏鸡的平均体重,显著增加鸡的胸腺指数和法氏囊指数,并促进十二指肠和盲肠内乳酸杆菌和双歧杆菌增殖。
研究显示微生态制剂的主要作用是维持肠道内菌群平衡,而肠道也是体内淋巴细胞主要聚集的器官,在机体免疫中起到了至关重要的作用,特别是对肠道感染的细菌具有显著的抑制作用。微生态制剂在家禽生产中已经广泛使用,但目前仅作为改善家禽肠道健康,提升饲料报酬和防治疾病的辅助手段。因为微生态制剂只能部分替代抗生素作用,和敏感抗菌药相比,其抗感染作用还有一定差距。并且微生态制剂在使用过程中起效慢及需长时间连续使用等缺点。因此,益生菌优良菌株的筛选、多种活性成份的配合、益生菌在机体消化代谢、免疫调节、疫病防治等方面的作用机制等仍需深入研究。
发明内容
本发明的目的是提供一种用于防控禽肠炎的枯草芽孢杆菌及其应用。所述枯草芽孢杆菌对产气荚膜梭菌和球虫导致的肉鸡肠道性炎症有良好的预防效果,可广泛应用于家禽养殖领域。
本发明一方面提供了一株枯草芽孢杆菌VB236(Bacillus subtilis VB236),已于2019年6月6日保藏于中国武汉 武汉大学的中国典型培养物保藏中心,保藏编号为CCTCC M2019437。
本发明一方面提供了一种用于预防或治疗家禽肠炎的组合物。
所述组合物包含上述枯草芽孢杆菌VB236。
所述组合物还包含地衣芽孢杆菌、解淀粉芽孢杆菌,凝结芽孢杆菌、短小芽孢杆菌、丁酸梭菌、屎肠球菌、乳酸菌、粪肠球菌、植物乳杆菌、干酪乳杆菌、鼠李糖乳杆菌中的任意一种或多种的组合。
所述组合物中枯草芽孢杆菌VB236的活菌量不低于109CFU/g。
本发明还提供了一种饲料添加剂,包含上述组合物。
本发明还提供了所述饲料添加剂在家禽养殖中的应用。
本发明所述枯草芽孢杆菌VB236对大肠杆菌、产气荚膜梭菌、金黄色葡萄球菌、沙门氏菌和鸡源产气荚膜梭菌等多种病原菌均具有显著的抑制效果,尤其是对鸡源产气荚膜梭菌的抑制能力最强,抑菌圈直径达到23.5mm。该菌株对球虫的抑制效果非常突出,能够显著降低球虫引起的鸡胚死亡,抑制球虫在鸡胚内的增殖,每胚卵囊量比对照组减少了94.3%,取得了意料不到的效果。
本发明提供的枯草芽孢杆菌VB236抗逆性强,在人工胃液和肠液中1h和2h后的存活率均为100%。该菌株可有效防控鸡源产气荚膜梭菌、大肠杆菌等常致病菌引起的家禽肠道疾病以及鸡球虫病,显著降低肠道损伤,同时能够改善家禽的粪便状态、提高生产性能,可广泛应用于家禽养殖领域。
说明书附图
图1为VB236菌株的菌落形态图;
图2为VB236菌株蛋白质谱峰图;
图3 为VB236菌株基因指纹图谱。
具体实施方式
下面结合具体实施例进一步阐述本发明。对于实施例中所用到的具体方法或材料,本领域技术人员可以在本发明技术思路的基础上,根据已有的技术进行常规的替换选择,而不仅限于本发明实施例的具体记载。
本发明所选用的设备和试剂可以选自市售任意一种。
实施例1 菌株分离与筛选
1、样品来源:山东省潍坊市高密养殖场肉鸡肠道粪便。
2、菌株分离与筛选:
称取粪便样品1g,置于9mL无菌生理盐水中,在30℃,200rpm,处理10min,得到菌悬液;将菌悬液置于80℃水浴锅中,处理10min,取出,迅速冷却;将处理后的菌悬液以10倍梯度稀释至合适浓度,取100uL于营养琼脂平板上,涂抹均匀,37℃培养16-24h;挑取营养琼脂平板上已培养好的不同单菌落,进一步划线2次,纯化获得单菌株。申请人从中选取了5株单菌,分别命名为A、B、C、D、E。
将柔嫩艾美耳球虫子孢子液分别与5株菌的菌液上清等体积混合,37℃孵育1h后接种9日龄SPF鸡胚,尿囊腔注射,每枚鸡胚0.2mL,每个样品接种10枚,置于37℃培养箱中进行孵育。每天照胚检查鸡胚存活情况。存活鸡胚啄壳后,收集胚体的绒毛尿囊膜于100 mL2.5%重酪酸钾溶液中,用高通量组织研磨器破碎后,经100目纱网过滤,取10 mL滤液,3000r/min离心10 min ,弃去上清,加入4 mL饱和盐水,用麦氏虫卵计数板计卵囊数。具体结果见表1。
表1 不同菌株抑菌圈及球虫卵囊计数结果
菌株 | 存活/总胚数 | 卵囊总量 | 每胚卵囊量 |
A | 6/10 | 1.5×10<sup>5</sup> | 25×10<sup>3</sup> |
B | 6/10 | 1.38×10<sup>5</sup> | 23×10<sup>3</sup> |
C | 5/10 | 1.15×10<sup>5</sup> | 23×10<sup>3</sup> |
D | 7/10 | 3.23×10<sup>4</sup> | 4.7×10<sup>3</sup> |
E | 9/10 | 1.26×10<sup>4</sup> | 1.4×10<sup>3</sup> |
从表1的结果可以看出,本发明筛选到的五株菌中E菌株接种的鸡胚成活率最高,达90%,而且每胚中球虫的卵囊数最少。从而说明该菌株对球虫有很强的抑制作用,能够显著降低球虫引起的鸡胚死亡,抑制球虫在鸡胚内的增殖。
申请人将E菌株命名为VB236,并对其进一步评价。
实施例2 VB236菌株的鉴
2.1 菌落形态鉴定
如图1所示,VB236菌株在营养琼脂培养基上的菌落呈微黄色,表面粗糙不透明,无粘液。
2.2 16S rRNA分子鉴定
采用试剂盒提取VB236菌株的基因组。然后以该基因组为模板,利用特异性引物对其16S rRNA进行扩增。将扩增得到的PCR产物进行1%的琼脂糖凝胶电泳检测,并送测序公司进行测序。
测序结果显示,PCR扩增产物的序列为SEQ ID NO:1。通过将该序列在NCBI数据库中进行BLAST比对,发现其与枯草芽孢杆菌(Bacillus subtilis)的相似性最高。因此,初步确定VB236菌株为枯草芽孢杆菌(Bacillus subtilis)。
2.3 MALDI-TOF-MS蛋白质谱鉴定
取少量VB236单菌落以薄膜的形式涂布于靶板上; 加1μL质谱样本预处理试剂盒中的裂解液,室温下自然晾干;加1μL质谱样本预处理试剂盒中的基质溶液覆盖样品,室温下自然晾干;将样品靶放入质谱仪进行鉴定。结果如图2所示,经与蛋白质图谱库中标准菌株比对鉴定结果为枯草芽孢杆菌(Bacillus subtilis)。
2.4 RiboPrinter全自动微生物基因指纹鉴定
根据全自动微生物基因指纹鉴定系统操作说明对菌株VB236进行了上机鉴定,得到其rRNA基因指纹图,如图3所示。通过与已知标准菌株库指纹图进行比对,发现VB236菌株与枯草芽孢杆菌相似度高达90%以上。因此,鉴定该菌株为枯草芽孢杆菌(Bacillus subtilis)。
申请人利用16S rRNA测序、MALDI-TOF-MS蛋白质谱、RiboPrinter全自动微生物基因指纹鉴定系统三种分子生物学手段对VB236菌株进行鉴定,鉴定结果一致。再结合VB236菌株的菌落形态特征,申请人确定该菌株为枯草芽孢杆菌(Bacillus subtilis),命名为枯草芽孢杆菌VB236(Bacillus subtilis VB236)。
申请人已于2019年6月6日将上述枯草芽孢杆菌VB236(Bacillus subtilisVB236)保藏于中国武汉 武汉大学的中国典型培养物保藏中心,保藏编号为CCTCC M2019437。
实施例2枯草芽孢杆菌VB236的抑菌能力
1、菌液制备
将枯草芽孢杆菌VB 236先在平板上培养,将培养好的枯草芽孢杆菌VB 236取1接种环接种到营养肉汤培养基中,37 ℃,220r/min培养14h,制备成供试菌液,菌量为108~109CFU/ml备用。
2、病原菌的准备
将收集到的大肠杆菌、金黄色葡萄球菌、沙门氏菌等病原菌甘油管或斜面接种于营养肉汤培养基中,37℃,220r/min 培养 16~18h后备用;产气荚膜梭菌接种于营养肉汤培养基中,37℃厌氧培养 24h后备用。
取无菌平皿,倾注无菌营养琼脂培养基 18~20mL,使其在平皿内均匀摊布,放置水平台面上使凝固,作为底层,将平板倒置平均划分区域并标记待添加物。另取适量无菌半固体营养琼脂培养基(琼脂含量为 1%),放冷至 48~50 ℃,每 50~100mL 培养基加入病原菌菌悬液 1~2mL,在每个平皿中分别加入5mL半固体营养琼脂培养基,使其在底层上均匀摊布,作为菌层。放置在水平台上冷却后,在每 1 平皿中以等距离均匀安置牛津杯 4~6个。每个双层平皿中的牛津杯中分别滴装扩培好的枯草芽孢杆菌VB 236菌液0.2mL,37℃培养24h后,测量各抑菌圈直径,具体结果见表2。
表2 枯草芽孢杆菌VB 236对不同病原菌的抑菌效果
病原菌 | 抑菌圈直径 mm |
大肠杆菌O1 | 21±1.0 |
大肠杆菌O2 | 20±1.0 |
大肠杆菌O78 | 21±1.0 |
金黄色葡萄球菌 | 21±1.0 |
沙门氏菌 | 19±1.0 |
鸡源产气荚膜梭菌 | 23.5±1.0 |
从表2的结果可以看出,本发明筛选到的枯草芽孢杆菌VB236对大肠杆菌、产气荚膜梭菌、金黄色葡萄球菌、沙门氏菌和鸡源产气荚膜梭菌等多种病原菌均具有显著的抑制效果,尤其是对鸡源产气荚膜梭菌的抑制能力最强,抑菌圈直径达到23.5mm,取得了意料不到的技术效果。
实施例3 枯草芽孢杆菌VB236抗球虫能力评价
1、柔嫩艾美耳球虫子孢子的制备
将纯化后的柔嫩艾美耳球虫孢子化卵囊悬液移入灭菌的匀浆器中进行研磨,使其释放孢子囊,将研磨完悬液移入锥形瓶中,加入0.25%胰蛋白酶、5%新鲜鸡胆汁,加入无菌PBS至20mL,置于41℃水浴锅中震荡孵育30min。镜检95%子孢子溢出时,离心去除上清,用PBS反复洗涤,去除胆汁和胰蛋白酶。用含血清、双抗DMEM培养基稀释至0.5×104备用。
2、供试菌株枯草芽孢杆菌 VB 236代谢产物的准备
将枯草芽孢杆菌 VB 236接种到营养琼脂液体培养基,37℃震荡培养56h,镜检观察芽孢数量,1000 r/min离心5min,取上清过滤器。
3、鸡胚接种
将柔嫩艾美耳球虫子孢子液与236菌液上清,等体积混合于37℃孵育1h后接种鸡胚,同时设立柔嫩艾美耳球虫与无菌水等体积混合作为对照组。取发育良好的9日龄SPF鸡胚,照胚划出气室,用碘液消毒后,75%酒精进行脱碘,尿囊腔注射,每枚鸡胚0.2mL,封蜡,置于37℃培养箱中进行孵育。每天照胚检查鸡胚存活情况。
4、卵囊收集及计数
存活鸡胚啄壳后,收集胚体的绒毛尿囊膜于100 mL 2.5%重酪酸钾溶液中,用高通量组织研磨器破碎后,经100目纱网过滤,取10 mL滤液,3000 r/min离心10 min ,弃去上清,加入4 mL饱和盐水,用麦氏虫卵计数板计卵囊数。具体结果见表3。
表3 枯草芽孢杆菌VB 236抗球虫效果实验
组别 | 接种 | 鸡胚死亡率 | 卵囊总量 | 每胚卵囊量 |
实验组 | 菌液上清+球虫子孢子 | 5% | 2.28×10<sup>4</sup> | 1.2×10<sup>3</sup> |
对照组 | 无菌水+球虫子孢子 | 20% | 3.36×10<sup>5</sup> | 2.1×10<sup>4</sup> |
从表3可以看出,本发明筛选到的枯草芽孢杆菌VB236对球虫的抑制效果非常突出,能够显著降低球虫引起的鸡胚死亡,抑制球虫在鸡胚内的增殖,每胚卵囊量比对照组减少了94.3%,取得了意料不到的技术效果。
实施例4 枯草芽孢杆菌VB236产酶能力评价
1、供试菌液的制备
将枯草芽孢杆菌VB 236先在平板上培养,将培养好的枯草芽孢杆菌 VB 236取1接种环接种到LB培养基中,37 ℃,220r/min培养14h,制备成供试菌液备用。
2、枯草芽孢杆菌 VB 236产淀粉酶能力评价
将培养好的枯草芽孢杆菌 VB 236的菌液接种到含有淀粉的琼脂培养基上,37℃培养48h,可以看到菌株VB 236周围有透明圈产生,透明圈直径为22mm,枯草芽孢杆菌 VB236可以产生一定的淀粉酶。
3、枯草芽孢杆菌 VB 236产蛋白酶能力评价
将培养好的枯草芽孢杆菌 VB 236的菌液接种到含有脱脂奶粉的琼脂培养基上,37℃培养48h,可以看到菌株VB236周围有透明圈产生,透明圈直径为23 mm,枯草芽孢杆菌VB 236可以产生一定的蛋白酶。
4、枯草芽孢杆菌VB236产纤维素酶能力评价
将培养好的枯草芽孢杆菌VB236的菌液接种到含有羧甲基纤维素的琼脂培养基上,37℃培养48h,可以看到菌株VB236周围有透明圈产生,透明圈直径为21mm。
实施例5 枯草芽孢杆菌VB236对人工胃液、肠液的耐受性测试
1、供试菌液的制备
将枯草芽孢杆菌VB 236先在平板上培养,将培养好的枯草芽孢杆菌 VB236取1接种环接种到LB培养基中,37 ℃,220r/min培养14h,分装于试管中,做为供试菌液。
2、胃液、肠液耐受性测定
取一个含供试菌试管用盐酸溶液调整pH为2.5,加入胃蛋白酶使其终浓度为1%,模拟人工胃液;取一个含供试菌试管调整pH为7.2,加入胰蛋白酶使其终浓度为1%,模拟人工肠液。37℃震荡培养,分别于1h和2h后取样,10倍倍比稀释后进行菌落计数,同时设立正常对照组,计算存活率(%)。
结果显示,枯草芽孢杆菌VB236在人工胃液中1h和2h后的存活率均为100%,在人工肠液中1h和2h后的存活率也均为100%。从而说明,本发明提供的枯草芽孢杆菌VB236对人工胃肠液有较强的耐受性。
实施例6 枯草芽孢杆菌VB236对肉鸡感染产气荚膜梭菌和球虫的预防效果
1、枯草芽孢杆菌VB 236菌粉制备
将枯草芽孢杆菌VB 236在5吨的发酵罐中进行液体发酵,镜检芽孢率达到90%以上时停止发酵,然后将发酵液进行离心后再喷雾干燥,制得菌量为10亿/g的菌粉。
2、攻毒株的准备
球虫疫苗购自佛山正典生物科技有限公司,4℃保存备用;将A型产气荚膜梭菌接种于营养肉汤培养基进行厌氧培养,37 ℃静置培养24h,制备成菌液备用。
3、肉鸡感染产气荚膜梭菌预防效果实验
(1)实验地点:青岛农业大学动物房;
(2)实验设计:针对产气荚膜梭菌导致的肉鸡肠道性炎症,将1日龄健康AA肉鸡180只,按照体重随机分为3组,每个组3个重复,每个重复20只鸡。
空白对照组:饲喂无抗日粮,不使用枯草芽孢杆菌VB 236菌粉,不感染球虫及产气荚膜梭菌,14日龄及21日龄口腔灌服生理盐水;
阳性对照组:饲喂无抗日粮,不使用枯草芽孢杆菌VB 236菌粉,14日龄时口腔灌服浓度为10倍剂量的球虫疫苗进行感染,在肉鸡的21日龄时口腔灌服产气荚膜梭菌菌液进行感染;
实验处理组:饲喂按600g/吨添加枯草芽孢杆菌VB 236菌粉(1010CFU/g)的无抗日粮,14日龄时口腔灌服浓度为10倍剂量的球虫疫苗进行感染,在肉鸡的21日龄时口腔灌服产气荚膜梭菌菌液进行感染。
分别在24日龄、35日龄时,每个重复随机抽取10只实验肉鸡,称重扑杀后,解剖取出十二指肠、空肠、回肠肠段,对小肠各肠段进行病理学分析,根据各肠段损害的严重程度及观察结果进行评分(评分标准见表4),记分为0~4分。
表4肠道病变评分标准
肠道病变描述 | 评分 |
肠道正常,无肉眼病变 | 0 |
肠道中段有散在的针点状出血点和白色斑点,肠腔中有少量桔黄色黏液,黏膜损伤不明显 | 1 |
肠道中段有大量出血点,肠腔中可见大量桔黄色黏液,也可见肠管稍肿胀 | 2 |
肠道出血,浆膜面出血点或白斑,肠管明显肿胀,内含黏液及血液 | 3 |
肠道严重出血,肠管明显肿胀,肠壁增厚,内充满红褐色黏液 | 4 |
4、肉鸡感染产气荚膜梭菌预防效果实验结果及分析
肉鸡感染产气荚膜梭菌预防效果24日龄时和35日龄时肠道病变评分见表5和表6。
表5 24日龄时肠道病变评分表
组别 | 十二指肠 | 空肠 | 回肠 | 盲肠 |
实验处理组 | 0.39±0.11 | 0.44±0.056 | 0.33±0.083 | 0.17±0.083 |
阳性对照组 | 0.83±0.083 | 0.94±0.056 | 0.78±0.088 | 0.28±0.088 |
空白对照组 | 0.056±0.056 | 0.28±0.087 | 0.17±0.083 | 0 |
表6 35日龄时肠道病变评分表
组别 | 十二指肠 | 空肠 | 回肠 | 盲肠 |
实验处理组 | 0.44±0.056 | 0.28±0.12 | 0.33±0.083 | 0.056±0.056 |
阳性对照组 | 0.78±0.088 | 0.72±0.088 | 0.50±0.083 | 0.44±0.13 |
空白对照组 | 0.28±0.088 | 0.17±0.083 | 0 | 0 |
从表5、表6的实验数据可以看出,与阳性对照组相比,日粮中添加枯草芽孢杆菌VB236菌粉的实验处理组肉鸡的肠道损伤得到显著降低。从而说明,本发明提供的枯草芽孢杆菌VB236对产气荚膜梭菌和球虫导致的肉鸡肠道性炎症有良好的预防效果。
本发明所述枯草芽孢杆菌VB236可有效防控鸡源产气荚膜梭菌、大肠杆菌等常致病菌引起的家禽肠道疾病以及鸡球虫病,同时能够改善家禽的粪便状态、提高生产性能,可广泛应用于家禽养殖领域。
序列表
<110> 山东康地恩生物科技有限公司
山东蔚蓝生物科技有限公司
<120> 一种防控禽肠炎的枯草芽孢杆菌及其应用
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1397
<212> DNA
<213> 枯草芽孢杆菌(Bacillus subtilis)
<400> 1
tacctcaccg acttcgggtg ttacaaactc tcgtggtgtg acgggcggtg tgtacaaggc 60
ccgggaacgt attcaccgcg gcatgctgat ccgcgattac tagcgattcc agcttcacgc 120
agtcgagttg cagactgcga tccgaactga gaacagattt gtgggattgg cttaacctcg 180
cggtttcgct gccctttgtt ctgtccattg tagcacgtgt gtagcccagg tcataagggg 240
catgatgatt tgacgtcatc cccaccttcc tccggtttgt caccggcagt caccttagag 300
tgcccaactg aatgctggca actaagatca agggttgcgc tcgttgcggg acttaaccca 360
acatctcacg acacgagctg acgacaacca tgcaccacct gtcactctgc ccccgaaggg 420
gacgtcctat ctctaggatt gtcagaggat gtcaagacct ggtaaggttc ttcgcgttgc 480
ttcgaattaa accacatgct ccaccgcttg tgcgggcccc cgtcaattcc tttgagtttc 540
agtcttgcga ccgtactccc caggcggagt gcttaatgcg ttagctgcag cactaagggg 600
cggaaacccc ctaacactta gcactcatcg tttacggcgt ggactaccag ggtatctaat 660
cctgttcgct ccccacgctt tcgctcctca gcgtcagtta cagaccagag agtcgccttc 720
gccactggtg ttcctccaca tctctacgca tttcaccgct acacgtggaa ttccactctc 780
ctcttctgca ctcaagttcc ccagtttcca atgaccctcc ccggttgagc cgggggcttt 840
cacatcagac ttaagaaacc gcctgcgagc cctttacgcc caataattcc ggacaacgct 900
tgccacctac gtattaccgc ggctgctggc acgtagttag ccgtggcttt ctggttaggt 960
accgtcaagg tgccgcccta tttgaacggc acttgttctt ccctaacaac agagctttac 1020
gatccgaaaa ccttcatcac tcacgcggcg ttgctccgtc agactttcgt ccattgcgga 1080
agattcccta ctgctgcctc ccgtaggagt ctgggccgtg tctcagtccc agtgtggccg 1140
atcaccctct caggtcggct acgcatcgtc gccttggtga gccgttacct caccaactag 1200
ctaatgcgcc gcgggtccat ctgtaagtgg tagccgaagc caccttttat gtctgaacca 1260
tgcggttcag acaaccatcc ggtattagcc ccggtttccc ggagttatcc cagtcttaca 1320
ggcaggttac ccacgtgtta ctcacccgtc cgccgctaac atcagggagc aagctcccat 1380
ctgtccgctc gactgca 1397
Claims (5)
1.一株枯草芽孢杆菌(Bacillus subtilis),其特征在于,所述枯草芽孢杆菌的保藏编号为CCTCC NO:M2019437。
2.一种用于预防或治疗家禽肠炎的组合物,其特征在于,所述的组合物包含权利要求1所述的枯草芽孢杆菌。
3.如权利要求2所述的组合物,其特征在于,所述的组合物还包含地衣芽孢杆菌、解淀粉芽孢杆菌,凝结芽孢杆菌、短小芽孢杆菌、丁酸梭菌、屎肠球菌、乳酸菌、粪肠球菌、植物乳杆菌、干酪乳杆菌、鼠李糖乳杆菌中的任意一种或多种的组合。
4.如权利要求2或3所述的组合物,其特征在于,所述的组合物中枯草芽孢杆菌的活菌量不低于109CFU/g。
5.一种饲料添加剂,其特征在于,所述的饲料添加剂包含权利要求2-4任一所述的组合物。
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