CN116731939B - 一种植物乳杆菌及其应用 - Google Patents
一种植物乳杆菌及其应用 Download PDFInfo
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- CN116731939B CN116731939B CN202311022692.5A CN202311022692A CN116731939B CN 116731939 B CN116731939 B CN 116731939B CN 202311022692 A CN202311022692 A CN 202311022692A CN 116731939 B CN116731939 B CN 116731939B
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Abstract
本发明提供了一种植物乳杆菌及其应用,属于功能微生物发酵与应用技术领域,本发明提供的植物乳杆菌(Lactobacillus plantarum)命名为LBR‑001,保藏编号为CGMCC No.26774,该菌对金黄色葡萄球菌、大肠杆菌、沙门氏菌等均具有较强的抑制作用,具有较强的耐酸能力、产酸能力,对中药具有极强的发酵转化能力。应用效果确切、安全可靠,适用于中药发酵、饲料添加剂、发酵饲料开发。
Description
技术领域
本发明属于功能微生物发酵与应用技术领域,涉及一种植物乳杆菌及其应用。
背景技术
随着中国家禽养殖规模的持续扩大和家禽养殖密度的不断上升,家禽呼吸道疾病发病率也随之升高,给养殖业带来了严重的经济损失。上呼吸道感染作为一种常见的急性呼吸道感染性疾病,若不及时治疗,炎症反应会蔓延至其他器官并引发相应症状。对于上呼吸道感染的治疗,临床上通常采取退热、补液、化痰止咳、抗感染等针对性治疗措施,虽然能够缓解家禽的临床症状,但治疗周期较长,家禽恢复较慢。细菌是引发上呼吸道感染的主要病原体,其中金黄色葡萄球菌、大肠杆菌和沙门氏菌是常见的细菌类型。每年冬季是高发季节,防治不及时易导致大规模死亡,造成巨大的经济损失。
目前,预防和治疗家禽呼吸道感染的主要途径是使用抗生素。然而,随着抗生素的长期使用,畜禽产品质量安全和环境污染问题已成为制约我国畜牧业可持续发展的关键因素。此外,抗生素的滥用也严重威胁到食品安全和人类健康。为了解决这些问题,寻找合适的抗生素替代产品已成为行业研究的重点。
植物乳杆菌(Lactobacillus plantarum)广泛存在于蔬菜、各类发酵产品以及肠道中,具有抗炎、抗氧化、调节肠道菌群平衡、提高机体免疫力和促进营养物质吸收等多种益生作用,对动物胃肠道环境具有良好的耐受能力,有利于动物对其吸收利用。对金黄色葡萄球菌、大肠杆菌、沙门菌属具有很好的抑制作用。但市场上乳酸菌制剂种类和来源繁杂,有效成分难以证实,并且植物乳杆菌菌株较多,不同菌株在不同的动物种类中的作用效果及添加剂量尚不明确。因此,针对不同的菌株,研究植物乳杆菌菌株在畜禽生产中应用,形成在特定畜禽中使用的具体应用方案及方法,可以将植物乳杆菌的应用效果最大化。
中药作为药物或饲料添加剂具有高安全性、无耐药性、不易残留等优点,可作为抗生素的替代品用于家禽呼吸道疾病的预防和治疗,对家禽的健康生长具有积极作用。然而,中药起效较慢,且家禽直接利用中药的效果有待加强,因此对于防治发病快、感染快、死亡率高的家禽呼吸道感染存在较大的行业挑战。
发明内容
本发明的目的是提供一种植物乳杆菌,该植物乳杆菌对金黄色葡萄球菌、大肠杆菌、沙门氏菌等具有显著的抑制作用,具有较强的耐酸能力和产酸能力,对发酵中药具有极强的发酵、转化能力,可用于制备发酵中药、饲料添加剂、发酵饲料。
为实现本发明的目的,本发明的植物乳杆菌(Lactobacillus plantarum)LBR-001是从四川酸白菜中分离的78株乳酸菌菌株经过层层筛选获得的,该菌株LBR-001为植物乳杆菌(Lactobacillus plantarum)。该菌已于2023年3月9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,分类命名为Lactobacillusplantarum,保藏编号为CGMCCNo.26774。
本发明提供的植物乳杆菌(Lactobacillus plantarum)LBR-001的微生物学特性为:革兰氏阳性菌,细胞形态为短杆菌,不规则形状。菌落呈圆形,色白,偶尔呈浅黄色,直径1-2μm,表面光滑湿润,中间略凸起,不透明,边缘整齐呈锯齿状,无晕环。
本发明通过16S rDNA序列分析方法将菌株LBR-001鉴定为植物乳杆菌(Lactobacillus plantarum)。
本发明提供一种植物乳杆菌(Lactobacillus plantarum)LBR-001,其特征在于,适宜生长温度为28-40℃;pH值为5.5-7.5;发酵时间24—72h。
进一步地,本发明提供一种植物乳杆菌(Lactobacillus plantarum)LBR-001,其特征在于,适宜生长温度为37℃;pH值为6.5;发酵时间60h。
本发明提供含有该植物乳杆菌(Lactobacillus plantarum)LBR-001的复合菌剂。该复合菌剂由植物乳杆菌(Lactobacillus plantarum)LBR-001与丁酸梭菌、屎肠球菌、粪肠球菌、嗜热乳杆菌、嗜酸乳杆菌、酿酒酵母以质量比为3:2:1:1:1:1:1复配而成。
本发明通过体外模拟鉴定,发现植物乳杆菌(Lactobacillus plantarum)LBR-001具有耐酸性和抵抗胃肠道内环境的特性,具备开发为益生菌的潜力。
本发明还发现植物乳杆菌(Lactobacillus plantarum)LBR-001在pH值为3.0时,活菌存活率仍高达80%,同时,对发酵中药具有极强的发酵、转化能力;并且对金黄色葡萄球菌、大肠杆菌、沙门氏菌均具有较强的抑制作用,因此本发明提供了含有植物乳杆菌LBR-001抑制金黄色葡萄球菌、大肠杆菌、沙门氏菌的发酵中药制剂,以及提供了植物乳杆菌LBR-001在降低饲料中金黄色葡萄球菌、大肠杆菌、沙门氏菌污染中的应用。
本发明提供一种含有植物乳杆菌(Lactobacillus plantarum)LBR-001或其发酵产物的发酵中药制剂。
本发明提供一种发酵中药制剂,由甘草、蒲公英、鱼腥草、癞蛤蟆草、板蓝根、黄芩、淫羊藿、山豆根、金银花以质量比为2:2:2:1:1:1:1:1:1构成的中药制剂经植物乳杆菌(Lactobacillus plantarum)LBR-001发酵制成。
本发明提供了一种含有该植物乳杆菌(Lactobacillus plantarum)LBR-001的饲料添加剂。
本发明提供了一种含有该植物乳杆菌(Lactobacillus plantarum)LBR-001的发酵饲料。
本发明的有益效果主要体现在:
1.植物乳杆菌LBR-001具有出色的耐酸能力,在胃肠道中表现出良好的耐受性,能耐受胃肠道中的低酸环境,并迅速在肠道内定植,有助于调节肠道菌群。此外,它还能在胃肠道中发挥显著的抑菌作用,具有开发为防治腹泻和肠炎的益生菌制剂的潜力。
2.植物乳杆菌LBR-001展现出显著的产酸能力,产出大量有机酸,对于发酵中药的转化拥有卓越的潜力。这种菌株可增强发酵中药活性成分的含量,将大分子物质转化为小分子。这种能力同样能提升饲料的品质,并改善饲料的营养价值。此外,植物乳杆菌LBR-001还具备天然防腐功效,有助于防止发酵中药和饲料的腐败变质。因此,这种菌株在中药发酵以及饲料发酵领域具有广泛的应用潜力。
3.植物乳杆菌LBR-001对金黄色葡萄球菌、大肠杆菌、沙门氏菌等均具有显著的抑制作用,具备开发为微生态中药制剂的潜力,可用于抑制致病菌和降低饲料中上述细菌的污染。
4.本发明的植物乳杆菌LBR-001的发酵产物作为益生菌和饲料添加剂,不存在抗生素类饲料添加剂的耐药性和残留问题,应用安全。
附图说明
图1为不同起始pH值对植物乳杆菌LBR-001生长的影响。
图2为不同发酵温度对植物乳杆菌LBR-001生长的影响。
图3为不同发酵时间对植物乳杆菌LBR-001生长的影响。
图4为植物乳杆菌LBR-001革兰氏染色镜检形态图。
图5为菌株在不同pH值培养基中的生长状况和培养后培养基pH值的变化。
图6为LBP-012菌株在不同培养时间时培养基pH值的变化情况。
图7为植物乳杆菌LBR-001对发酵中药部分有效成分的影响。
图8为植物乳杆菌LBR-001对发酵饲料品质的影响。
图9为植物乳杆菌LBR-001及其发酵中药抑菌情况。
图10为植物乳杆菌LBR-001发酵中药在五图高家庄某养殖场使用效果反馈。
图11为植物乳杆菌LBR-001发酵中药在潍坊市安丘市兴安街道某养殖场使用效果反馈图。
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。下述实施例中所使用的材料、试剂等如无特殊说明,均可从商业途径得到。本发明实施例采用了牛津杯法检测植物乳杆菌LBR-001及其发酵中药的抑菌活性。
发酵中药培养基:将组合中药甘草、蒲公英、鱼腥草、癞蛤蟆草、板蓝根、黄芩、淫羊藿、山豆根、金银花,其质量比例为:2:2:2:1:1:1:1:1:1,粉碎并过40目筛,加水淹没中药,浸泡1个小时,熬至两次浓缩,中药提取液体终浓度0.5g/ml,按照体积按比例加入补充培养基成分,121℃、20min高压蒸汽灭菌备用。
补充培养基成分(g/L):
蛋白胨10.0,牛肉浸粉5.0,酵母浸粉4.0,葡萄糖20.0,磷酸氢二钾2.0,柠檬酸三铵2.0,醋酸钠5.0,硫酸镁0.2,硫酸锰0.05。
实施例1植物乳杆菌LBR-001的筛选、分离与鉴定:
1、植物乳杆菌LBR-001的筛选、分离与纯化
从四川酸白菜中刮取1g样品,加入至9ml无菌蒸馏水中,旋涡震荡2min,制成悬液,再将其稀释后涂布于乳酸菌选择性培养基MRS培养基上,37℃厌氧培养36h,挑取平板上单菌落,在MRS培养基平板上再次划线,37℃厌氧培养,将得到的菌落多次划线纯化培养,得到纯的菌落。将所述单菌落依次命名为LBP-001、LBT-002、LBT-003、……、LBT-078。将分离纯化获得的上述单菌落进行活化并培养至24h,以体积的1%为接种量分别接种至pH值为6.5、5.0、3.5和2.0的MRS液体培养基中,37℃厌氧培养24h,同时以相应pH值的无菌培养基为空白对照组,分别测定各组培养基600nm吸光度值及pH值。
实验结果表明,申请人筛选到的78株菌中只有LBP-012、LBP-054和LBP-068三株菌可在pH值为3.5—4.0的MRS液体培养基中正常生长,具有较强的耐酸性,其中LBP-012菌株的耐酸性最强,pH值为3.0时,活菌存活率仍高达80%,同时产酸能力也最强。具体结果如图5所示。
2、植物乳杆菌LBP-012的鉴定:
(1)形态学特征和革兰氏染色
将LBP-012菌株多次划线培养,观察菌落形态为圆形,色白,偶尔呈浅黄色,直径1-2μm,表面光滑湿润,中间略凸起,不透明,边缘整齐呈锯齿状,无晕环。进行革兰氏染色,镜检显示为圆端直杆状。菌株LBP-012镜检见图4。
(2)生理、生化特征鉴定:
通过葡萄糖产气试验、过氧化氢酶活性测定、硝酸盐还原试验、硫化氢产生试验、明胶液化试验、吲哚试验及乳酸菌生化鉴定条(HBIG11)对菌株LBP-012进行生理、生化特征测定。
鉴定结果显示:菌株LBP-012发酵葡萄糖产酸不产气,且过氧化氢酶阴性属于同型发酵,硝酸盐、硫化氢、明胶和吲哚均为阴性,七叶苷、纤维二糖、麦芽糖、甘露醇、水杨苷、山梨醇、蔗糖、棉子糖、菊糖和乳糖均为阳性,初步判定菌株LBP-012为植物乳杆菌(Lactobacillus plantarum)。
(3)分子生物学鉴定:
以所述的菌株LBP-012的DNA为模板,使用16S rRNA通用引物进行扩增,扩增片段进行序列测定,将得到的菌株LBR-001的16S rDNA测序结果与GenBank中的序列进行比对分析,结果显示,菌株LBP-012与Lactobacillus plantarum同源性最高,因此确定该菌株LBR-001为植物乳杆菌。
申请人将筛选到LBP-012菌株命名为植物乳杆菌(Lactobacillus plantarum)LBR-001进行菌种保藏,保藏单位:中国微生物菌种保藏管理委员会普通微生物中心(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;保藏日期:2023年03月09日;植物乳杆菌(Lactobacillusplantarum)LBR-001的保藏编号为:CGMCCNo.26774。
实施例2植物乳杆菌LBR-001生长性能测定:
将植物乳杆菌LBR-001以1%接种量接种于MRS液体培养基中以不同起始pH值,以OD600为测定指标绘制生长曲线见图1;将植物乳杆菌LBR-001以3%接种量接种于MRS液体培养基中,以不同温度、发酵时间,以OD600为测定指标分别绘制生长曲线见图2、图3。由图1可知,植物乳杆菌LBR-001在pH值3.5-7.5均可生长,生长最适宜初始pH值为6.5;由图2可知,植物乳杆菌LBR-001在28℃—40℃均可生长,最适宜生长温度为37℃;由图3可知,植物乳杆菌LBR-001生长迟滞期为培养初始~6h左右,随后进入对数生长期,此期间菌体呈指数倍增加,当培养60h时OD600达到最高值,24~60h为植物乳杆菌LBR-001生长的稳定期,菌体生长缓慢,60h后逐渐进入衰亡期。
实施例3植物乳杆菌LBR-001产酸能力评价:
为了进一步验证植物乳杆菌LBR-001的产酸能力,申请人将植物乳杆菌LBR-001和植物乳杆菌ATCC 14917菌株(对照菌株)分别等量接种于pH值6.5的MRS液体培养基液中,37℃条件下培养,每2h测定培养液的pH值,具体结果如图6所示。
由图6的数据可以看出,培养12h后,接种对照菌株植物乳杆菌ATCC 14917的培养基pH值由6.50降低到4.67,而接种植物乳杆菌LBR-001的培养基pH值由6.50降低到3.68;培养24h后,接种对照菌株植物乳杆菌ATCC 14917的培养基pH降低到4.28,而接种植物乳杆菌LBR-001的培养基pH值降低到3.16;从而说明,本发明提供的植物乳杆菌LBR-001比对照菌株,具有更强的产酸能力、更快的产酸速度。
实施例4植物乳杆菌LBR-001对中药发酵转化能力评价:
(1)发酵中药培养基制备:
将组合中药:甘草、蒲公英、鱼腥草、癞蛤蟆草、板蓝根、黄芩、淫羊藿、山豆根、金银花,其使用比例为:2:2:2:1:1:1:1:1:1,粉碎并过40目筛,加水淹没中药,浸泡1个小时,熬至两次浓缩,中药提取液体终浓度0.5g/ml,按照体积按比例加入补充培养基成分,121℃、20min高压蒸汽灭菌备用。
补充培养基成分(g/L):
蛋白胨10.0,牛肉浸粉5.0,酵母浸粉4.0,葡萄糖20.0,磷酸氢二钾2.0,柠檬酸三铵2.0,醋酸钠5.0,硫酸镁0.2,硫酸锰0.05。
(2)将植物乳杆菌LBR-001按体积的5%接种于发酵中药培养基中,用30%氢氧化钠溶液调节pH值为6.5,发酵温度为37℃,100r/min振荡发酵60h。
发酵中药培养基中蒲公英、鱼腥草、山豆根、金银花中含有黄酮类活性成分,淫羊藿、黄芩含有苷类活性成分,以不接种任何菌株的发酵中药培养基为对照组,添加植物乳杆菌LBR-001的发酵中药培养基为处理组,对照组的pH值为5.19,处理组pH值为3.76,对发酵中药培养基中有效成分进行含量测定,具体结果见图7。
从图7中我们可以看出,与对照组相比,经植物乳杆菌LBR-001发酵后的中药有效成分比未发酵的中药提高1-2倍,苷类活性成分提高幅度最大,从而说明,本发明提供的植物乳杆菌LBR-001对中药具有极强的发酵转化能力,将中药中难以溶解的有效成分释放出来。
同时对两组进行加速试验:温度40℃±2℃、相对湿度75%±5%的同等条件下放置6个月。对照组初始为深褐色均匀液体,具有中药蒸煮的涩味,第1月出现絮状物,出现大量沉积物,第2个月颜色明显改变,出现分层现象,并出现腐败气味,第3月腐败、轻微出现青色霉菌状物质,第4个月完全腐败,出现大量青色菌癍。处理组初始为浅褐色均匀液体,具有乳酸味,带有中药的香味,第1个月出现轻微一摇即散的沉淀,颜色、气味无变化,第2个月沉淀物、颜色、气味无明显变化。第3个月,第6个月均无明显变化。从加速试验结果我们可以看出,植物乳杆菌LBR-001对发酵中药具有防腐保质作用,其可以作为一种饲料添加剂使用。
根据对照试验与加速试验结果来看,植物乳杆菌(Lactobacillus plantarum)LBR-001具有较强的产酸能力,产生大量的有机酸,对发酵中药中具有极强的发酵转化能力,可以有效提高发酵中药活性成分的含量,将大分子转化为小分子。同时具有天然防腐剂作用,有效防止发酵中药腐败变质,可广泛应用于中药发酵领域。
实施例5植物乳杆菌LBR-001对发酵饲料品质的影响:
将植物乳杆菌LBR-001悬液划线于MRS固体培养基中,分别于37℃静置培养36h,分离得到单一菌落。挑取单一菌落接种于MRS液体培养基中,37℃、100r·min-1培养2d,获得发酵用种子液;将植物乳杆菌LBR-001以1%的接种量接种到MRS液体培养基中,在37℃条件下静置培养24h;按照质量分数计,将上述菌液以5%的接种量接种在由麦麸30%、米糠15%、豆粕10%组成的粗饲料中,加无菌水调整至最终水分40%—45%。将混合物放于密闭发酵袋中,置于37℃条件下培养。以不接种任何菌株的上述粗饲料为对照组,发酵60h后取样,分别测定发酵饲料的pH值、活菌数、总酸、粗蛋白和酸溶蛋白含量,具体结果见图8。
从图8的数据可以看出,与对照组相比,经植物乳杆菌LBR-001发酵后的发酵组饲料中粗蛋白、酸溶蛋白含量分别提高了76.77%、64.44%,有机酸含量显著提高,pH显著降低。从而说明,本发明提供的植物乳杆菌LBR-001可降解部分多蛋白质、脂肪等大分子物质,生成有机酸、可溶性多肽等小分子物质,形成营养丰富、适口性好、活菌含量高的生物饲料,显著提高发酵饲料的适口性及营养价值。
实施例6植物乳杆菌LBR-001及其发酵中药抑菌性能评价:
1、材料和试验仪器:
(1)材料:
LB液体培养基,琼脂粉,采购自青岛海博生物技术有限公司。
LB固体培养基:LB液体培养基中加15.0g琼脂粉,121℃高压灭菌20min即得。
MRS液体培养基:蛋白胨10.0g,牛肉浸取物10.0g,酵母提取液5.0g,葡萄糖5.0g,乙酸钠5.0g,柠檬酸二胺2.0g,吐温-80 1.0g,磷酸氢二钾2.0g,七水硫酸镁0.2g,七水硫酸锰0.05g,碳酸钙20.0g,琼脂20.0g,蒸馏水定容至1000ml,pH6.8,121℃高压灭菌20min即得。
无菌吸管:1ml(具0.01ml刻度)、10ml(具0.1ml刻度)、微量移液器及吸头。
无菌锥形瓶:容量250ml、500ml。
无菌培养皿:直径90mm。
无菌试管:18cm。
试剂:无菌生理盐水。
仪器:高压灭菌器、离心机、混匀器、恒温培养箱、水浴锅、超净操作台、酸度计等。
2、试验方法:
(1)病原菌菌悬液的制备:
将金黄色葡萄球菌ATCC 29213、大肠杆菌ATCC 259922和沙门氏菌ATCC 12002这3株病原菌作为测试菌株,用LB培养基37℃培养。
(2)植物乳杆菌LBR-001菌悬液的制备:
在无菌条件将纯种的植物乳杆菌LBR-001接种于100mlMRS液体培养基中,37℃静置培养24h,菌液浓度达到108CFU/ml。
(3)植物乳杆菌LBR-001发酵中药培养基制备:
发酵温度为37℃,接种量为5%,初始pH6.5,发酵时间为60h,最终含植物乳杆菌LBR-001活菌为5.32×109CFU/ml。将植物乳杆菌LBR-001发酵中药培养基无菌操作稀释50倍,未发酵中药培养基灭菌后稀释50倍,备用。
(4)植物乳杆菌LBR-001及其发酵中药抑菌性能测定:
采用牛津杯法,取培养好的病原菌,稀释菌液浓度约为105CFU/ml,涂布于固体LB培养基并依次放置牛津杯;将活化好的植物乳杆菌LBR-001及其发酵中药培养基,未发酵中药培养基于4℃、10000×g离心10min,取上清液用0.22μm微孔滤膜过滤除菌,随后取上清液100μL加入牛津杯中,4℃静置3h,后转入恒温培养箱37℃培养,24h后观察结果,测量其抑菌圈直径。对照组加入100μL无菌LB液体培养基,每组3个重复,试验重复3次,结果如图9所示。
由图9可知,植物乳杆菌LBR-001对金黄色葡萄球菌ATCC 29213、大肠杆菌ATCC259922和沙门氏菌ATCC 12002均有明显的抑制作用,植物乳杆菌LBR-001发酵中药培养基明显高于未发酵中药培养基,植物乳杆菌LBR-001及其发酵中药具有协同增效的作用,从临床微生物检验常用药敏抑菌圈直径解释标准来看,与抗生素相比,可替代部分抗生素,可应用于开发预防细菌性致病菌引起疾病的微生态中药制剂。
实施例7植物乳杆菌LBR-001发酵中药在畜禽养殖中的应用。
1、潍坊市昌乐县五图高家庄某养殖场,20日龄600只麻鸡,2021年12月23日,出现精神沉郁、委顿,有呼吸困难,啰音,咳嗽等呼吸道症状,经兽医诊断为大肠杆菌感染引起的急性呼吸道气囊炎,当日死亡50只,后连用植物乳杆菌LBR-001发酵中药5天,每L兑水1000L,全天集中3—4小时饮完,第1天死亡48只;第2天死亡35只,呼吸道症状明显减轻;第3天死亡20只;第4天死亡5只;第5天死亡0只,呼吸道症状基本没有,只有零星地喘咳声。由以上及图10可知,使用植物乳杆菌LBR-001发酵中药能快速、有效缓解家禽的呼吸道症状。
2、潍坊市安丘市兴安街道某养殖场,上鸭9000只,2021年10月3日,33日龄出现感冒、咳嗽等呼吸道症状,食欲减退,一天吃料45包,用植物乳杆菌LBR-001发酵中药每L兑水1000L,全天集中3—4小时饮完,见图11,用两天后呼吸道症状减轻,增料明显,一天吃料52包,用完三天后,咳嗽症状明显减轻,无继发感染,无伤亡。由以上及图11可知,使用植物乳杆菌LBR-001发酵中药不但能快速、有效缓解家禽的呼吸道感染,同时提高肉鸭的采食量,促进肉鸭生长。
3、安丘市凌河某养殖场,上鸭3万只,3个棚舍,2021年10月3日,23日龄,两个棚约18000只出现精神沉郁、感冒、咳嗽、食欲减退症状,白色稀粪。将两个棚分为西药组和植物乳杆菌LBR-001发酵中药实验组,两组饲料、管理条件一样,西药组使用抗生素疗程药,实验组使用本发明每L兑水1000L,全天集中3—4小时饮完。第2天,实验组相较于西药组大群采食恢复正常,精神状态好,粪便明显改善,西药组与实验组大群咳嗽症状无发展,病鸭伤亡开始减少。第3天西药组和实验组两组大群咳嗽症状数量减少,西药组出现减料情况,实验组增料。第4天,两组大群咳嗽症状几乎很少,无伤亡,西药组出现减料,实验组仍继续增料。由此可知,使用植物乳杆菌LBR-001发酵中药可替代抗生素防治家禽呼吸道感染,同时提神增料,无副作用。
本发明公开了一种植物乳杆菌及其应用,以上实施例仅用以说明本发明的技术方案,而非对其进行限制;尽管参照前述实施例对本发明进行了详细的说明,对于本领域的普通技术人员来说,依然可以对前述实施例所记载的技术方案进行修改,或者对其中部分技术特征进行等同替换;而这些修改或替换,并不使相应技术方案的本质脱离本发明所要求保护的技术方案的精神和范围。
Claims (6)
1.一种植物乳杆菌(Lactobacillus plantarum)LBR-001发酵中药制剂,其特征在于采用植物乳杆菌(Lactobacillus plantarum)LBR-001发酵得到的发酵中药制剂,所述发酵中药制剂由甘草、蒲公英、鱼腥草、癞蛤蟆草、板蓝根、黄芩、淫羊藿、山豆根、金银花以质量比为2:2:2:1:1:1:1:1:1构成的中药制剂经植物乳杆菌(Lactobacillus plantarum)LBR-001发酵制成;所述植物乳杆菌(Lactobacillus plantarum)LBR-001从四川酸白菜中分离得到,保藏编号为CGMCC No.26774。
2.权利要求1所述的植物乳杆菌(Lactobacillus plantarum)LBR-001发酵中药制剂,其特征在于,所述的植物乳杆菌(Lactobacillus plantarum)LBR-001适宜生长温度为28-40℃;pH值为5.5-7.5;发酵时间24—72h。
3.权利要求2所述的植物乳杆菌(Lactobacillus plantarum)LBR-001发酵中药制剂,其特征在于,所述的植物乳杆菌(Lactobacillus plantarum)LBR-001适宜生长温度为37℃;pH值为6.5;发酵时间60h。
4.权利要求1所述的植物乳杆菌(Lactobacillus plantarum)LBR-001发酵中药制剂,其特征在于,其为抑制金黄色葡萄球菌、大肠杆菌、沙门氏菌的发酵中药制剂。
5.权利要求1所述的植物乳杆菌(Lactobacillus plantarum)LBR-001发酵中药制剂在制备饲料添加剂中的应用。
6.权利要求1所述的植物乳杆菌(Lactobacillus plantarum)LBR-001发酵中药制剂在制备发酵饲料中的应用。
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