CN111154677B - 嗜酸乳杆菌及其应用 - Google Patents
嗜酸乳杆菌及其应用 Download PDFInfo
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- CN111154677B CN111154677B CN202010033981.5A CN202010033981A CN111154677B CN 111154677 B CN111154677 B CN 111154677B CN 202010033981 A CN202010033981 A CN 202010033981A CN 111154677 B CN111154677 B CN 111154677B
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- lactobacillus acidophilus
- salmonella
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- bacteria
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Abstract
本发明提供一种嗜酸乳杆菌及其应用。本发明的嗜酸乳杆菌Ma.SSRGJ1为革兰氏阳性菌,可在pH值3.0以上的酸性环境生长,耐胆盐能力强,其对革兰氏阴性菌有抑菌能力,将嗜酸乳杆菌Ma.SSRGJ1制成菌剂后用于饲喂动物效果安全可靠,在促进营养物质消化吸收,提高饲料转化效率,促进生长方面具有积极作用。
Description
技术领域
本发明属于微生物学和饲用益生菌领域,具体地说,涉及一种嗜酸乳杆菌及其应用。
背景技术
在畜牧养殖业,微生态制剂作为绿色环保型饲料添加剂正逐步替代饲用抗生素,它可以改善肠道微生态环境,促进畜禽肠道有益菌的增殖,阻止或抑制有害菌繁殖,调整肠道菌群平衡,明显减轻动物粪便的氨臭,减少蚊蝇虫害,改善和优化畜禽饲养生态环境,减少环境污染,可以提高动物的抗应激能力和免疫力,能够提高动物饲料转化率,降低生产成本增加经济效益。
微生态制剂理想的可直接饲用的菌种应该是:①不会使人和动物致病,不与病原微生物产生杂交种;②在体内外易于繁殖,体外繁殖速度快;③在低pH和胆汁中可以存活,并能植入肠黏膜;④在发酵过程中能产生乳酸和过氧化氢等物质;⑤能合成对大肠杆菌、沙门氏菌、葡萄球菌、梭状芽孢杆菌等肠道致病菌的抑制物而不影响自己的活性;⑥经加工后活菌存活率高,混入饲料后高温下稳定性好;⑦最好来自动物自身肠道中;⑧有利于促进宿主的生长发育及提高抗病能力。
微生态制剂由于具有重要的益生作用已经在畜牧养殖业中广泛应用,但饲用嗜酸乳杆菌微生态制剂的研究少见。饲用嗜酸乳杆菌微生态制剂既有益生菌的功能,又具有活性高和稳定性优良的特点。随着制备工艺的提高及研究的深入,微生态制剂必将会得到越来越广泛的应用。
嗜酸乳杆菌属于美国FDA(1989)和我国农业部(1999)公布的对动物无致病性可直接用于动物饲料的细菌。国内外大量的试验也证实嗜酸乳杆菌有利于动物体内正常菌群的建立,有助于抵抗有害微生物侵袭,能提高动物的生产性能和防病治病。嗜酸乳杆菌不仅可以调整肠道菌群平衡,而且能分泌嗜酸乳菌、嗜酸杆菌素、乳酸菌素等抗生素类物质,从而对肠道致病菌产生拮抗作用,抑制肠道不良微生物的增殖。目前,关于嗜酸乳杆菌的应用主要集中在乳品行业、医药行业及其他工业上的应用。嗜酸乳杆菌在乳品上的应用主要是发酵乳制品,在医药行业的应用主要是由于其特殊的保健作用。
发明内容
本发明的目的是提供一种兼具益生性和抑菌功能的饲用嗜酸乳杆菌及其应用,本发明提供的嗜酸乳杆菌耐酸、耐胆盐、抑菌效果佳。
为了实现本发明目的,第一方面,本发明提供一种嗜酸乳杆菌(Lactobacillusacidophilus)Ma.SSRGJ1,该菌是从北京市某养猪实验基地的猪肠道中分离并经过紫外诱变后筛选得到的。经16S rRNA基因序列分析,菌株Ma.SSRGJ1为嗜酸乳杆菌(Lactobacillusacidophilus)。该菌株已于2019年11月12日保藏在中国微生物菌种保藏管理委员会普通微生物中心(简称CGMCC,地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,邮编100101),分类命名为嗜酸乳杆菌Lactobacillus acidophilus,保藏编号为CGMCCNO.18939。
嗜酸乳杆菌Ma.SSRGJ1的微生物学特性为:革兰氏阳性菌,细胞形态为杆状,杆末端呈圆形,无芽孢;单菌落大小≤1mm,颜色呈乳白色,不透明,菌落表面光滑湿润,边缘整齐,有光泽。菌体可在pH值3.0以上的酸性环境生长,耐胆盐能力强。且具有一定的抑菌能力,特别是抑制大肠杆菌和沙门氏菌效果明显。
第二方面,本发明提供含有所述嗜酸乳杆菌Ma.SSRGJ1的菌剂。
第三方面,本发明提供含有所述嗜酸乳杆菌或其菌剂的饲料添加剂或动物饲料。
所述饲料添加剂中嗜酸乳杆菌Ma.SSRGJ1的活菌数为1×106CFU/g~1×1010CFU/g;优选地,含有的嗜酸乳杆菌Ma.SSRGJ1饲料添加剂的活菌数为1×108CFU/g~1×109CFU/g。
所述动物饲料中嗜酸乳杆菌Ma.SSRGJ1的活菌数为1×105CFU/kg~1×108CFU/kg,优选1×107CFU/kg~1×108CFU/kg。
本发明通过体外法鉴定了嗜酸乳杆菌Ma.SSRGJ1的益生效果,结果表明,嗜酸乳杆菌Ma.SSRGJ1能够耐酸、酸胆盐,能抵抗胃肠道的内环境,具备益生菌的潜力。
基于嗜酸乳杆菌Ma.SSRGJ1具有突出的抑菌特性,含有嗜酸乳杆菌Ma.SSRGJ1或其菌剂的药物或组合物,特别是抑菌类药物属于本发明的保护范围。
第四方面,本发明提供含有所述嗜酸乳杆菌或其菌剂的防腐剂。
第五方面,本发明提供所述嗜酸乳杆菌或其菌剂在抑菌方面的应用,包括非治疗目的的应用。
前述的应用,所述菌为革兰氏阴性菌。
优选地,所述菌包括但不限于埃希氏菌属(Escherichia)细菌或沙门氏菌属(Salmonella)细菌,更优选大肠杆菌、沙门氏菌。如大肠杆菌O157、大肠杆菌K99、鸡白痢沙门氏菌(Salmonella pullorum)CVCC1791。
第六方面,本发明提供所述嗜酸乳杆菌或其菌剂的以下任一应用:
1)用于制备饲料添加剂;
2)用于提高饲料转化率;
3)用于促进动物生长、提高动物体重及抗病能力;
4)用于食品、药品或保健品领域;
5)用于防腐剂领域。
第七方面,本发明提供嗜酸乳杆菌菌剂的制备方法,所述方法包括:发酵培养嗜酸乳杆菌Ma.SSRGJ1,发酵液离心取沉淀,得到菌泥,与冻干保护剂混合后,冷冻干燥。
优选地,所用发酵培养基为:乳清粉30g/L、豆粕30g/L、葡萄糖5g/L、氯化钠4g/L、硫酸锌0.1g/L、硫酸锰0.6g/L、硫酸镁1g/L和消泡剂0.05%v/v。
发酵条件为:35℃,转速200rpm搅拌条件下,通入氮气和氢气体积比为9:1的混合气体,发酵培养16h。
所述冻干保护剂可以是:10%脱脂奶粉+6%乳糖。菌泥与冻干保护剂的用量比例按g:mL计为3:1。
本发明提供了嗜酸乳杆菌Ma.SSRGJ1或其菌剂在抑制革兰氏阴性菌中的应用。其中,所述革兰氏阴性菌包括但不限于大肠杆菌、沙门氏菌。在本发明的一个具体实施方式中,显示了嗜酸乳杆菌Ma.SSRGJ1对大肠杆菌O157和沙门氏菌CVCC1791具有很强的抑制能力,抑菌圈直径分别达到2.80cm和2.17cm。
本发明提供的嗜酸乳杆菌Ma.SSRGJ1具有提高饲料利用率,促进饲料中营养物质的消化吸收;增强动物免疫功能,提高日增重,降低料肉比;无污染,无残留,生物环保等特点,在促进动物生长和提高动物体重方面具有显著的效果。
附图说明
图1为本发明实施例1中嗜酸乳杆菌Ma.SSRGJ1的菌落形态图。
图2为本发明实施例1中嗜酸乳杆菌Ma.SSRGJ1的革兰氏染色图。
图3为本发明实施例2中嗜酸乳杆菌Ma.SSRGJ1的耐酸性检测结果。
图4为本发明实施例2中嗜酸乳杆菌Ma.SSRGJ1的耐胆盐检测结果。
图5为本发明实施例3中嗜酸乳杆菌Ma.SSRGJ1的生长曲线。
图6为本发明实施例2中嗜酸乳杆菌Ma.SSRGJ1的抑菌试验结果。;
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。若未特别指明,实施例中所用的技术手段为本领域技术人员所熟知的常规手段,所用原料均为市售商品。
以下实施例中使用的MRS培养基配制如下:称取蛋白胨10g,牛肉膏粉5g,酵母膏粉4g,葡萄糖20g,吐温-80 1.08g,磷酸氢二钾2g,乙酸钠5g,柠檬酸三铵2g,硫酸镁(MgSO4·7H2O)0.2g,硫酸锰(MnSO4·4H2O)0.05g,琼脂15g,用蒸馏水定容至1L,调节pH至6.2。放入灭菌锅中121℃,灭菌20分钟。
实施例1嗜酸乳杆菌(Lactobacillus acidophilus)Ma.SSRGJ1的分离与鉴定
一、菌株Ma.SSRGJ1的分离
1、菌株的分离培养
取1g来自北京市海淀区中国农业大学养猪实验基地的猪的肠道食糜样品装入有9ml生理盐水的试管中,漩涡器震荡混匀,即为1:10稀释液,再取稀释液进行10倍递增稀释,然后选择3个适宜梯度的稀释液各1ml涂布于MRS培养基上。37℃培养48-72小时,观察并记录菌落形态,挑取长势良好的单菌落,进行划线分离纯化。用生理盐水配制菌悬液。
2、菌株的紫外诱变与筛选
将灭菌后的MRS培养基倒入培养皿中,待凝固后,取步骤1所得的菌悬液涂布于平板上,每个培养皿控制菌落在50-80个左右,培养12小时后,距离紫外灯20cm,诱变30s。
挑取诱变后的菌株,接种于MRS液体培养基中,培养24小时,测定OD值,选取生长速度较快的菌株,取适量的菌悬液涂布于MRS平板上,在37℃恒温培养箱中培养至24小时,进行下一步革兰氏染色。
3、菌株的革兰氏染色
在载玻片上滴一滴经灭菌的蒸馏水,挑取一个诱变后生长速度较快的单菌落(菌落形态图见图1)在水中溶解,经刮片后,在酒精灯上烘干固定。滴加结晶紫染色液,染2min,水洗,自然晾干;滴加碘液媒染2min,水洗,自然晾干;滴加碱性品红乙醇溶液50S,水洗,自然晾干;在普通光学显微镜上观察,若菌体呈紫色为阳性,菌体呈红色为阴性,结果见图2。选取革兰氏阳性杆菌进行下一步芽孢染色实验。
4、菌株的芽孢染色
取经步骤2诱变后生长速度较快的菌株,在载玻片上滴一滴经灭菌的蒸馏水,挑取一个单菌落在水中溶解,经刮片后,在酒精灯上烘干固定。滴加5%孔雀石绿溶液3-5滴,在酒精灯上加热3-5min,注意不能使液体沸腾或者干枯,水洗,自然晾干;滴加蕃红溶液染色2min,水洗,自然晾干;在普通光学显微镜下观察,芽孢呈绿色,菌体呈红色。通过步骤1-4的分离筛选,最终获得一株革兰氏染色阳性,无芽孢的菌株。将该菌株编号为Ma.SSRGJ1。
二、菌株Ma.SSRGJ1的鉴定
1、形态学鉴定
处于对数生长期且菌落大小稳定的菌株Ma.SSRGJ1的单菌落描述如下:单菌落大小为≤1mm,圆形,颜色呈乳白色,不透明,菌落表面湿润光滑,边缘规则。
接着,对处于对数生长期的菌株Ma.SSRGJ1进行染色,采用光学显微镜观察菌体形态。分离并筛选到的菌株Ma.SSRGJ1,革兰氏染色呈阳性,细胞形态为杆状,无芽孢。
2、16S rRNA基因序列同源性分析
细菌总DNA的提取采用天根生化科技有限公司的细菌基因组DNA提取试剂盒提取。提取后的样品送到上海美吉生物医药科技有限公司进行测序。将测定结果在GenBank数据库中进行BLAST同源性比对,确定菌种类别为嗜酸乳杆菌(Lactobacillus acidophilus)。测序结果见SEQ ID NO:1。
经16S rRNA基因测序,根据测序结果以及上述微生物学特征及理化特性,将该菌鉴定为嗜酸乳杆菌Lactobacillus acidophilus。
实施例2嗜酸乳杆菌Ma.SSRGJ1的抗逆性检测
1、耐热性检测
将嗜酸乳杆菌Ma.SSRGJ1菌液至于水浴锅内20分钟,分别用60℃、80℃、100℃处理,每个处理3个重复,处理结束后采用倾注法测定其活菌数。
8lg(cfu/ml)嗜酸乳杆菌Ma.SSRGJ1在60℃处理20分钟后,活菌数为5.86lg(cfu/ml),在80℃处理20min后,活菌数接近于0,说明嗜酸乳杆菌不耐受高温,在工业化生产中需要对其进行包被处理。
2、耐酸性检测
将108CFU/ml嗜酸乳杆菌Ma.SSRGJ1分别接种到pH值为2.0、3.0、4.0、5.0的MRS培养基中,分别在1h、2h、3h、4h采用平板倾注法测定其活菌数。
当处于酸性环境即pH为5.0、4.0、3.0时,嗜酸乳杆菌可以正常生长,当pH为2.0时,对嗜酸乳杆菌的生长略有抑制作用,但活菌数仍然能维持在7.48lg(cfu/ml)。说明该菌种对酸的耐受能力较强。结果显示嗜酸乳杆菌Ma.SSRGJ1可以耐受胃酸的影响(图3)。
3、耐胆盐检测
将活化好的嗜酸乳杆菌Ma.SSRGJ1用无菌生理盐水做倍比稀释,选取合适的稀释梯度并吸取200微升稀释液放于无菌培养皿里,做6个重复,然后用含不同浓度牛胆酸钠(0.1%、0.2%、0.3%、0.4%)的MRS培养基倾注平板,37℃培养4小时,每隔1小时菌落计数,同时用不含牛胆酸钠的MRS培养基倾注平板,37℃培养48小时,菌落计数,作为对照组。图4结果显示,不同胆盐浓度下的活菌数随着时间的延长而下降的趋势不明显。0.1%、0.2%、0.3%的胆盐作用对嗜酸乳杆菌Ma.SSRGJ1的影响比较微弱,几乎不影响其正常生长。在0.4%的胆盐作用4小时后,活菌数略有下降,但不具有显著差异。说明嗜酸乳杆菌Ma.SSRGJ1的耐胆盐能力较强。
4、抗生素敏感性检测
将适宜浓度的嗜酸乳杆菌Ma.SSRGJ1涂布于MRS培养基上,每个培养皿中均匀的贴附1个药敏纸片,培养36小时,观察抑菌圈大小(表1)。
表1嗜酸乳杆菌Ma.SSRGJ1对不同抗生素敏感性结果
名称 | 抑菌圈直径(mm) | 敏感度 |
氨苄西林 | 12 | 中敏 |
多西环素 | 15 | 高敏 |
青霉素 | 23 | 极敏 |
卡那霉素 | 8 | 低敏 |
庆大霉素 | 9 | 低敏 |
红霉素 | 22 | 极敏 |
头孢氨苄 | 22 | 极敏 |
环丙沙星 | 8 | 低敏 |
氯霉素 | 29 | 极敏 |
以上实验结果表明,嗜酸乳杆菌Ma.SSRGJ1 CGMCC No.18939并不具有良好的耐药性,因此作为饲用益生菌使用是安全可靠的。
5、嗜酸乳杆菌Ma.SSRGJ1对革兰氏阴性菌的抑菌实验
在培养皿中倒入配置好的MRS培养基,待培养基凝固后,放置牛津杯,上层倒入混有1%致病菌(大肠杆菌O157、大肠杆菌K99、沙门氏菌CVCC1791)的LB培养基,凝固后即可使用。在牛津杯中分别加入200微升的菌体,菌液和上清,小心放入37℃恒温培养箱正置培养24小时,查看抑菌圈大小。结果显示嗜酸乳杆菌MA.SSRGJ1对大肠杆菌O157和沙门氏菌CVCC1791的抑菌效果十分明显,抑菌圈大小分别为2.8cm和2.17cm,对大肠杆菌K99也具有一定的抑菌效果,抑菌圈达到2cm(图6)。
实施例3嗜酸乳杆菌Ma.SSRGJ1的生长曲线测定
生长曲线代表了细菌在新的适宜的环境中生长繁殖直至衰老死亡全过程的动态变化。将嗜酸乳杆菌Ma.SSRGJ1按10%(v/v)的接种量,接种到MRS培养基中,37℃培养20小时,以不加菌液的MRS培养基作为空白对照,每隔2小时测定OD600值,从而计算活菌数。实验设三次重复,结果取其平均值,记录数据并绘制生长曲线。如图5所示,在2-16小时,嗜酸乳杆菌Ma.SSRGJ1处于对数生长阶段,繁殖速率较高。在16-20小时嗜酸乳杆菌Ma.SSRGJ1数目趋于稳定,处于平台期。
实施例4嗜酸乳杆菌制剂的制备
1、发酵培养基配方:乳清粉30g/L、豆粕30g/L、葡萄糖5g/L、氯化钠4g/L、硫酸锌0.1g/L、硫酸锰0.6g/L、硫酸镁1g/L、消泡剂0.05%(v/v),加水充分溶解制成发酵培养基。
2、115℃高温蒸汽灭菌30min。
3、待发酵培养基降温至35℃时,接种菌龄20小时的菌液5%(v/v)。
4、在35℃条件下,保持转速200rpm搅拌,通入氮气和氢气的混合气体,二者混合体积比为9:1,发酵培养16小时,放罐,得到嗜酸乳杆菌的活菌数大于1×109cfu/ml的发酵液。发酵液4℃离心取沉淀,得到菌泥。
5、向300g菌泥中加入100mL冻干保护剂,用振荡器混匀制成菌悬液。-80℃预冷1.5小时,迅速将冻结样品转移至冷冻干燥机中冻干24小时,使菌粉含水量达3%左右。冻干保护剂配方为:10%脱脂奶粉+6%乳糖。
实施例5嗜酸乳杆菌Ma.SSRGJ1制剂的安全性评价
本实施例以小鼠作为实验动物,采用灌胃试验的方法,评价嗜酸乳杆菌的安全性,具体方法如下:
1、实施例4方法制得的嗜酸乳杆菌菌剂的冻干粉,经平板计数测定,其嗜酸乳杆菌Ma.SSRGJ1菌数为1×109cfu/g。
2、选取8周龄左右的小鼠72只,随机分为4组(A组为对照组灌服无菌生理盐水,B组为高剂量组按照1×109cfu/只的量灌服菌液,C组为中剂量组按照1×108cfu/只的量灌服菌液,D组为低剂量组按照1×107cfu/只的量灌服菌液),每组3个重复,每个重复6只鼠。
3、每天上午九点灌胃一次,连续灌服21天。
实验鼠房控制温度湿度恒定,自然光照,小鼠自由采食、饮水,每7天清扫鼠笼一次。实验过程中,每天观察并记录小鼠的状态,存活情况,有无临床异常症状等。
检测指标:
(1)于实验结束当天采用心脏取血的方式,取得实验小鼠血液样品,经静止离心后获得血清,用于检测血清中白蛋白、总蛋白、高密度脂蛋白、低密度脂蛋白、甘油三酯、胆固醇、尿素、肿瘤细胞坏死因子等血液生化指标。
(2)取完整的心、肝、脾、肾(双侧)称湿重,分别计算心脏指数=(心脏湿重/体重)×100%、肝脏指数=(肝脏湿重/体重)×100%、脾脏指数=(脾脏湿重/体重)×100%、肾脏指数=(肾脏湿重/体重)×100%。
表2不同处理组小鼠存活情况
从表2可以看出,使用嗜酸乳杆菌Ma.SSRGJ1 CGMCC No.18939给小鼠灌胃21天后,实验各处理组小鼠均存活,说明上述嗜酸乳杆菌对动物安全。
表3不同处理组小鼠的脏器系数
A组 | B组 | C组 | D组 | |
心脏 | 0.59 | 0.68 | 0.65 | 0.63 |
肝脏 | 5.64 | 5.58 | 5.53 | 5.61 |
脾脏 | 0.44 | 0.43 | 0.46 | 0.44 |
肾脏 | 1.31 | 1.37 | 1.32 | 1.35 |
从表3可以看出,处理组小鼠的脏器指数与对照组相比没有明显变化,说明上述嗜酸乳杆菌没有造成小鼠各脏器异常。
利用生化分析仪检测小鼠血清中白蛋白、总蛋白、高密度脂蛋白、低密度脂蛋白、甘油三酯、胆固醇、尿素、肿瘤细胞坏死因子等,结果均显示正常,说明本发明实施例4提供的嗜酸乳杆菌制剂对小鼠的各项生理指标不构成影响。
实施例6嗜酸乳杆菌Ma.SSRGJ1制剂的应用
本实验选取28日龄杜长大三元杂交断奶仔猪72头,实验期45天,按照随机区组设计分为2个组,每组6个重复,每个重复6头猪。A组为对照组(基础日粮组),B组为处理组(基础日粮中添加260g/t的实施例4制得的嗜酸乳杆菌制剂,有效活菌数为1×109cfu/g)。
试验期间仔猪饲养在全封闭式保育猪舍内,温度控制在25-27℃,自由采食、饮水。基础日粮中不含任何抗生素,仔猪免疫按照常规免疫程序进行。
测定指标:各处理组断奶仔猪的生产性能,具体包括以下指标:
1、每天记录仔猪的采食量,试验结束后计算平均日采食量。
2、在试验开始和结束的当天记录仔猪体重,计算平均日增重。
3、通过A、B的试验结果计算料肉比,其计算方式为平均日采食量/平均日增重。
4、试验期间每天早上10:00观察并记录仔猪的粪便状况,计算断奶仔猪腹泻率,腹泻率(%)=(腹泻头数×腹泻天数)/(猪只数×试验天数)×100%。
表4基础日粮中添加嗜酸乳杆菌制剂对断奶仔猪生产性能的影响
从表4可以看出,处理组仔猪平均日采食量和平均日增重都显著高于对照组(P<0.05),料肉比低于对照组,说明添加嗜酸乳杆菌制剂的饲料效益更好。处理组与对照组相比腹泻率显著降低,说明本发明的嗜酸乳杆菌具有改善仔猪腹泻的作用。
虽然,上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述,但在本发明基础上,可以对之做一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
序列表
<110> 中国农业大学
<120> 嗜酸乳杆菌及其应用
<130> KHP201110099.3
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1161
<212> DNA
<213> 嗜酸乳杆菌(Lactobacillus acidophilus)
<400> 1
actgcgggcg gcgtgctata catgcaagtc gaacgcttct ttttccaccg gagcttgctc 60
caccggaaaa agaggagtgg cgaacgggtg agtaacacgt gggtaacctg cccatcagaa 120
ggggataaca cttggaaaca ggtgctaata ccgtataaca atcaaaaccg catggttttg 180
atttgaaagg cgctttcggg tgtcgctgat ggatggaccc gcggtgcatt agctagttgg 240
tgaggtaacg gctcaccaag gccacgatgc atagccgacc tgagagggtg atcggccaca 300
ttgggactga gacacggccc aaactcctac gggaggcagc agtagggaat cttcggcaat 360
ggacgaaagt ctgaccgagc aacgccgcgt gagtgaagaa ggttttcgga tcgtaaaact 420
ctgttgttag agaagaacaa ggatgagagt aactgttcat cccttgacgg tatctaacca 480
gaaagccacg gctaactacg tgccagcagc cgcggtaata cgtaggtggc aagcgttgtc 540
cggatttatt gggcgtaaag cgagcgcagg cggtttctta agtctgatgt gaaagccccc 600
ggctcaaccg gggagggtca ttggaaactg ggagacttga gtgcagaaga ggagagtgga 660
attccatgtg tagcggtgaa atgcgtagat atatggagga acaccagtgg cgaaggcggc 720
tctctggtct gtaactgacg ctgaggctcg aaagcgtggg gagcaaacag gattagatac 780
cctggtagtc cacgccgtaa acgatgagtg ctaagtgttg gagggtttcc gcccttcagt 840
gctgcagcta acgcattaag cactccgcct ggggagtacg accgcaaggt tgaaactcaa 900
aggaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga agcaacgcga 960
agaaccttac caggtcttga catcctttga ccctctagag atagagcttc cccttcgggg 1020
gcaaagtgac aggtggtgca tggttgtcgt cagctcgggc gtgaaatgtt gggttaagtc 1080
ccgcaacgag cgcaccctta ttgttattgc ctcattcagt tgggcctcta gcagaatgcc 1140
ggtgaaaacc ggaggaaggg g 1161
Claims (11)
1.嗜酸乳杆菌(Lactobacillus acidophilus)Ma.SSRGJ1,保藏编号为CGMCCNO.18939。
2.含有权利要求1所述嗜酸乳杆菌的菌剂。
3.含有权利要求1所述嗜酸乳杆菌或权利要求2所述菌剂的饲料添加剂或动物饲料。
4.含有权利要求1所述嗜酸乳杆菌或权利要求2所述菌剂的防腐剂。
5.含有权利要求1所述嗜酸乳杆菌或权利要求2所述菌剂的药物。
6.权利要求1所述嗜酸乳杆菌或权利要求2所述菌剂在非治疗目的抑菌中的应用。
7.根据权利要求6所述的应用,其特征在于,所述菌为革兰氏阴性菌。
8.根据权利要求7所述的应用,其特征在于,所述革兰氏阴性菌为埃希氏菌属(Escherichia)细菌或沙门氏菌属(Salmonella)细菌。
9.根据权利要求8所述的应用,其特征在于,所述埃希氏菌属(Escherichia)细菌为大肠杆菌,所述沙门氏菌属(Salmonella)细菌为沙门氏菌。
10.权利要求1所述嗜酸乳杆菌或权利要求2所述菌剂的以下任一应用:
1)用于制备饲料添加剂;
2)用于提高饲料转化率;
3)用于制备食品、药品或保健品;
4)用于制备防腐剂。
11.嗜酸乳杆菌菌剂的制备方法,其特征在于,所述方法包括:发酵培养权利要求1所述嗜酸乳杆菌,发酵液离心取沉淀,得到菌泥,与冻干保护剂混合后,冷冻干燥;
所述发酵培养所用的发酵培养基为:乳清粉30-35g/L、豆粕30-35g/L、葡萄糖4-6g/L、氯化钠3-5g/L、硫酸锌0.1-0.2g/L、硫酸锰0.6-0.7g/L、硫酸镁0.8-1.2g/L和消泡剂0.05-0.08%v/v;
发酵条件为:35℃,转速200rpm搅拌条件下,通入氮气和氢气体积比为9:1的混合气体,发酵培养16h。
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