CN113215057B - 蛙源乳酸乳球菌乳亚种lcll1及其用途 - Google Patents
蛙源乳酸乳球菌乳亚种lcll1及其用途 Download PDFInfo
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Abstract
本发明公开了蛙源乳酸乳球菌乳亚种LCLL1及其用途。蛙源乳酸乳球菌乳亚种Lactococcus lactis subsp.lactis LCLL1的保藏号为CGMCC No.21817,将蛙源乳酸乳球菌乳亚种LCLL1添加到豆粕饲料中,能够提高牛蛙的产量;提高饲料利用率;增强牛蛙机体免疫力;提高牛蛙消化道抗炎能力,使消化道组织结构更完整、更健康;消化道黏膜微生物中有益菌的丰度增加,条件致病菌丰度降低。蛙源乳酸乳球菌乳亚种LCLL1对牛蛙表现出很好的益生效果作用。
Description
技术领域
本发明涉及微生物领域,尤其涉及一种蛙源乳酸乳球菌乳亚种LCLL1及其用途。
背景技术
牛蛙(Lithobates catesbeiana)属两栖纲(Amphibia)、蛙科(Ranidae)、牛蛙属(Lithobates),是大型可食用蛙类。近年来,牛蛙养殖在福建、浙江、四川、湖南、江西、广东、广西和海南等地发展迅速,已成为我国养殖量最大的经济蛙类。但由于盲目追求高密度养殖和降低养殖成本,且缺乏科学管理,养殖生产中牛蛙病害频发,导致抗生素等药物滥用,危害了食品安全和环境安全。
同时,为了降低水产饲料中鱼粉的用量,植物蛋白源已成为水产动物饲料中鱼粉的替代品。但植物蛋白中含有多种抗营养因子,且适口性差,因而,其在水产动物饲料的添加量受到限制。近年来,研究发现牛蛙饲料中豆粕蛋白替代50%鱼粉蛋白会对其生长和肠道健康造成负面影响。有益菌能有效提高养殖动物免疫力,降低养殖动物病害发生,并保证人类食品安全。因此,亟需开展牛蛙肠道有益菌的相关科学研究,以促进牛蛙产业的健康发展。
发明内容
本发明的目的在于提供一种蛙源乳酸乳球菌乳亚种LCLL1及其用途。
为实现上述目的,本发明提供一种蛙源乳酸乳球菌乳亚种Lactococcus lactissubsp.lactis LCLL1,其保藏号为CGMCC No.21817。
本发明还保护所述蛙源乳酸乳球菌乳亚种LCLL1的用途。
进一步,所述用途为对牛蛙具有益生效果。
进一步,所述用途为能提高牛蛙的产量。
进一步,所述用途为能提高饲料利用率;增强牛蛙机体免疫力。
进一步,所述用途为能提高牛蛙消化道抗炎能力。
进一步,所述用途为能使消化道黏膜微生物中有益菌的丰度增加,条件致病菌丰度降低。
进一步,所述用途为将蛙源乳酸乳球菌乳亚种LCLL1添加到豆粕饲料喂养牛蛙。
蛙源乳酸乳球菌乳亚种LCLL1添加到豆粕饲料中,能够提高牛蛙的产量;提高饲料利用率;增强牛蛙机体免疫力;提高牛蛙消化道抗炎能力,使消化道组织结构更完整、更健康;消化道黏膜微生物中有益菌的丰度增加,条件致病菌丰度降低。蛙源乳酸乳球菌乳亚种LCLL1 对牛蛙表现出很好的益生效果作用。
附图说明
图1是试验饲料对牛蛙空肠炎性因子基因和紧密连接蛋白基因表达量结果图。
图2是试验饲料对牛蛙空肠组织结构的影响结果图。
图3是NC组与LL组牛蛙消化道菌群结构比较热图(A为门水平,B为属水平)。
图4是用PICRUSt2预测牛蛙肠道菌群的功能概况图。
具体实施方式
下面详细描述本发明的实施例,所述实施例的示例在附图中示出,其中自始至终相同或类似的标号表示相同或类似的元件或具有相同或类似功能的元件。下面通过参考附图描述的实施例是示例性的,旨在用于解释本发明,而不能理解为对本发明的限制。实施例中未注明具体技术或条件者,按照本领域内的文献所描述的技术或条件或者按照产品说明书进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规产品。
实施例1:乳酸乳球菌乳亚种LCLL1的分离和验证
1.分离技术:
用双毁髓法处理摄食高豆粕饲料的牛蛙后,用75%的酒精棉球擦抹牛蛙腹部表面,无菌操作取其空肠和回肠,随后用灭菌的PBS缓冲溶液冲洗内腔,将肠组织与0.8%的无菌生理盐水按体积比1:9混合,低温均浆,以均浆液作10倍体积系列稀释,分别取稀释液10-2、10-3、 10-4等3梯度的稀释液各100μL涂到MRS固体培养基上,33℃培养48h。挑取单菌落经多次提纯得到纯化菌株。将纯化后的菌株与50%浓度甘油1:1等体积混合于冻存管中,最后放入-80℃低温冰箱保存,备用。
2. 16S rDNA的序列测定与分析
将分离并纯化后的菌株(命名为LCLL1)。保藏信息如下:
菌株名称:乳酸乳球菌乳亚种Lactococcus lactis subsp.lactis LCLL1
保藏日期:2021.2.7
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
保藏地址:中国北京
保藏编号:CGMCC No.21817。
以160000r/min离心30s,收集菌体。使用赛百盛硅胶膜型TM细菌基因组DNA提取试剂盒,按照说明书方法进行提取细菌的基因组DNA。1μL DNA模板(~50ng),1μL的正向引物(10μM),1μL的反向引物(10μM),25μL的Premix,22μL的去离子水作为反应体系进行PCR扩增。其中正向引物为用细菌16S rRNA扩增引物27F(5′ -AGAGTTTGATCCTGGCTCAG-3′,SEQ IDNO:1),反向引物为1492R(5′-ACGGCTACCTTGTTACGACT-3 ′,SEQ ID NO:2)。从基因组DNA中扩增出16S rDNA基因片段,PCR产物送往上海美吉生物医药科技有限公司进行上机测序。测序后的序列进行头尾低质量碱基去除、序列拼接后,在韩国ezbiocloud数据库(https://www.ezbiocloud.net/)进行blast比对。结果见表1,鉴定结果为乳酸乳球菌乳亚种LCLL1(Lactococcus lactis subsp.lactis)。鉴定序列如SEQ ID NO:3 所示。
表1序列比对结果表
菌株 | 登录号 | 最相似菌株及登录号 | 相似度 |
LCLL1 | JCM5805 | Lactococcus lactis subsp.lactis JCM5805 | 99.93% |
SEQ ID NO:3所示序列如下:
ATGCAAGTTGAGCGCTGAAaGTTGGTACTTGTACCGACTGGATGAGCAGCGAACGGGTGAGTAACGCGTGGGGAATCTG CCTTTGAGCGGGGGACAACATTTGGAAACGAATGCTAATACCGCATAAAAACTTTAAACACAAGTTTTAAGTTTGAAAG ATGCAATTGCATCACTCAAAGATGATCCCGCGTTGTATTAGCTAGTTGGTGAGGTAAAGGCTCACCAAGGCGATGATAC ATAGCCGACCTGAGAGGGTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAA TCTTCGGCAATGGACGAAAGTCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAACTCTGTTGGT AGAGAAGAACGTTGGTGAGAGTGGAAAGCTCATCAAGTGACGGTAACTACCCAGAAAGGGACGGCTAACTACGTGCCAG CAGCCGCGGTAATACGTAGGTCCCGAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGTGGTTTATTAAGTCT GGTGTAAAAGGCAGTGGCTCAACCATTGTATGCATTGGAAACTGGTAGACTTGAGTGCAGGAGAGGAGAGTGGAATTCC ATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCGGTGGCGAAAGCGGCTCTCTGGCCTGTAACTGACACTGA GGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAGATGTAGGGA GCTATAAGTTCTCTGTATCGCAGCTAACGCAATAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGG AATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACA TACTCGTGCTATTCCTAGAGATAGGAAGTTCCTTCGGGACACGGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGT CGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTGTTAGTTGCCATCATTAAGTTGGGCACTCTAACGA GACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGC TACAATGGATGGTACAACGAGTCGCGAGACAGTGATGTTTAGCTAATCTCTTAAAACCATTCTCAGTTCGGATTGTAGG CTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGGGAGTTGGGAGTACCCGAAGTAGGTTGCCTAACCGCAAGGAGGGCGCT。
3.形态及生理生化指标
3.1形态特征
(1)菌落形态:将乳酸乳球菌乳亚种LCLL1菌提纯划线到MRS固体培养基平板上,经33℃培养48h后观察,菌落表面光滑凸出,边缘光滑呈圆形,乳白色,半透明或不透明。
(2)个体形态。乳酸乳球菌乳亚种LCLL1菌经革兰氏染色试验后呈阳性。经显微镜观察,细胞形态呈球形或卵圆形。
(3)运动性。乳酸乳球菌乳亚种LCLL1菌无运动性。
3.2生理生化指标
通过生理生化试验鉴定指标(见表2)。乳酸乳球菌乳亚种LCLL1菌的葡萄糖发酵试验、乳糖发酵试验呈阳性;柠檬酸盐利用试验、硫化氢试验、乳糖发酵试验、吲哚试验、甲基红试验、VP试验、明胶液化试验、淀粉水解试验呈阴性。
表2乳酸乳球菌乳亚种LCLL1菌的生理生化鉴定结果表
生化试验 | 乳酸乳球菌乳亚种LCLL1菌 |
葡萄糖发酵试验 | + |
乳糖发酵试验 | + |
甲基红试验 | - |
吲哚试验 | - |
硫化氢试验 | - |
明胶液化试验 | - |
柠檬酸盐利用试验 | - |
淀粉试验 | - |
VP试验 | - |
注:“+”表示鉴定为阳性;“-”表示鉴定为阴性。
4.最适宜温度和PH
4.1最适温度探索
分别制备8瓶100mL的MRS液体培养基,接种100uL的种子液置于摇床中180r/min,分别采用26℃、28℃、30℃、32℃、34℃、36℃、38℃、40℃培养18h,酶标仪设定波长为600nm,18h后取出菌液测定OD600值,计算最适的培养温度。结果显示,乳酸乳球菌乳亚种LCLL1 菌最适生长温度为36℃。
4.2最适pH探索
分别制备7瓶100mL的MRS培养基pH调节至:2、3、4、5、6、7、8。接种100μL的种子液置于恒温箱中180r/min培养18h,培养结束后测定菌液OD600值,计算最适的pH。结果显示,乳酸乳球菌乳亚种LCLL1菌最适生长pH为8。
5.对病原菌拮抗试验
本试验采用点种法。取浓度为106cfu/mL的100μL指示菌菌液均匀涂布于固体NA培养基上,再在培养基上加入10μL浓度为108cfu/mL待测菌液,33℃恒温箱培养24h后观察是否有抑菌圈。结果见表3。
表3乳酸乳球菌乳亚种对指示菌的拮抗作用结果表
菌株 | 乳酸乳球菌乳亚种LCLL1菌 |
霍乱弧菌LCUE_s | - |
维氏气单胞菌Aeromonas veronii | - |
嗜水气单胞菌嗜水亚种A.hydrophila | - |
注:“+”表示有抑菌圈;“-”表示无抑菌圈。
实施例2:饲料的制备
1材料与方法
1.1饲料制备
以豆粕为主要蛋白源,鱼油和豆油为主要脂肪源配制蛋白水平为41.10%,脂肪水平为 7.90%的基础饲料(表4)。饲料原料粉碎后全部通过60目筛网,所有饲料原料混合好后,加入蒸馏水,制成直径为3.0mm的膨化颗粒饲料,脂肪源暂时不添加,自然晾干后于-20℃保存。
表4试验饲料配方及营养组成表(干物质,WT%)
1.2乳酸乳球菌乳亚种LCLL1悬浊液准备
乳酸乳球菌乳亚种LCLL1菌接种于MRS液体培养基中于33℃培养24h。将液体培养物于4000r/min,4℃条件下离心10min,将菌体沉淀重悬于无菌生理盐水溶液中。无菌生理盐水洗涤2次后,将菌悬液稀释不同梯度,测定OD600值,同时用涂布平板计数法测定活细胞数。制作OD600条件下菌体吸光度与活细胞数量的线性关系。以均匀喷涂的方式添加到未加脂肪源的饲料中,添加量为1×107cfu/g,记为LL组,对照组喷相同量的无菌生理盐水 (NC组)。晾干后再按照饲料配方均匀喷涂鱼油和豆油,每两周进行一次饲料喷涂,然后将饲料密封于塑料袋中于-20℃冰箱保存。
1.3养殖试验
牛蛙是福建莆田某养殖场的同批次孵化蛙苗,暂养于集美大学水产试验场,期间投喂商品饲料。经三周暂养,停饲24h后,将78只牛蛙(29.61±0.07g)随机分配到6个养殖桶中 (π×30×90cm),共分为2组,每组3个重复,每个重复13只牛蛙。每天饱食投喂两次(8:00和18:00),养殖周期为58天,养殖试验的最后2周收集牛蛙粪便。养殖试验期间气温为28-32℃,水温为27-31℃,水深保持在4~6cm。
1.4样品采集
养殖试验结束后,停止喂食24h后进行采样,用毁髓法处死牛蛙,每一只牛蛙称重记录。每桶随机取3只牛蛙留作全体组成分析样本。每桶再随机取9只牛蛙,先取心脏动脉血管血液,于离心管中,低温静置24h,低温离心10min(3000rpm/min),取上清,-80℃保存;然后逐一称量每只牛蛙的肝脏和腿重;随机挑选其中3只蛙,测定胃体部、空肠中部以及大腿肌肉的pH(Hanna HI1083B,意大利);取3只蛙的空肠肠段1cm浸于波恩氏液中,用于制作切片;剩余6只蛙的空肠两两混合于冻存管,于-80℃冰箱保存,用于测定消化酶活、基因表达和肠道微生物多样性等。
2结果
2.1生长性能和饲料利用率
经过58d的养殖试验,结果见表5。可以看出,LL组的终末均重、增重率和蛋白质效率显著高于NC组(P<0.05)。LL组的饲料系数显著低于NC组(P<0.05)。两组之间的摄食率、存活率、肝体比和腿体比未呈现显著差异(P>0.05)。由以上数据可以得出,牛蛙全豆粕饲料中添加乳酸乳球菌乳亚种LCLL1可以促进牛蛙的生长和提高牛蛙对饲料利用率。
表5试验饲料对牛蛙生长及生物学性状的影响结果表
NC | LL | |
终末均重(FBW,g) | 160.27±2.01<sup>a</sup> | 180.09±2.06<sup>b</sup> |
增重率(WG,%) | 443.87±9.54<sup>a</sup> | 503.55±6.14<sup>b</sup> |
摄食率(FI,%/d) | 2.44±0.06 | 2.40±0.12 |
饲料系数(FCR) | 1.03±0.01<sup>b</sup> | 0.97±0.01<sup>a</sup> |
蛋白质效率(PER) | 2.18±0.01<sup>a</sup> | 2.24±0.01<sup>b</sup> |
存活率(%) | 100±0.00 | 100±0.00 |
肝体比(HIS,%) | 4.73±0.54 | 5.09±0.74 |
腿体比(HLI,%) | 33.90±3.97 | 40.65±0.41 |
注:同一行内不同上标的均值有显著性差异(P<0.05),下表同。
2.2牛蛙对饲料营养物质表观消化率
牛蛙对试验饲料表观消化率见下表6,LL组的饲料干物质消化率和粗蛋白消化率显著高于NC组(P<0.05)。Ca和P的表观消化率各试验组间的差异不显著(P>0.05)。因此,乳酸乳球菌乳亚种LCLL1可以提高牛蛙对全豆粕饲料的消化率,促进豆粕蛋白的利用。
表6试验饲料对牛蛙饲料中营养物质表观消化率的影响(%)结果表
NC | LL | |
干物质 | 72.79±0.17<sup>a</sup> | 75.08±0.37<sup>b</sup> |
粗蛋白 | 86.31±0.30a | 88.11±0.13<sup>b</sup> |
钙 | 56.10±1.00 | 58.28±0.88 |
磷 | 56.83±0.79 | 57.11±0.38 |
2.3牛蛙消化道pH值和消化酶活性
结果见表7。LL组的胃部pH值相较NC组显著降低(P<0.05)。LL组的蛋白酶和淀粉酶活性显著高于NC组(P<0.05)。由此可知,乳酸乳球菌乳亚种LCLL1降低了牛蛙消化道内的酸碱度,提高蛋白酶和淀粉酶活性。因此,牛蛙豆粕饲料中添加LCLL1可以降低胃内pH,促进了牛蛙对饲料的消化和利用。
表7试验饲料对牛蛙空肠消化酶活性的影响结果表
NC | LL | |
胃内容物pH | 4.03±0.42<sup>b</sup> | 3.44±0.13<sup>a</sup> |
空肠pH | 7.70±0.02 | 7.57±0.08 |
蛋白酶(U/mg prot) | 3.04±0.23<sup>a</sup> | 5.00±0.30<sup>b</sup> |
脂肪酶(U/g prot) | 2.45±0.95 | 2.71±0.23 |
淀粉酶(U/g prot) | 33.49±1.21<sup>a</sup> | 41.87±1.69<sup>bc</sup> |
2.4牛蛙血清生化指标
结果见表8。LL组血清LZM活性、血清IgM含量、血清补体C3和C4含量,LL组均显著高于NC组(P<0.05),说明乳酸乳球菌乳亚种LCLL1可提高牛蛙机体的免疫力。
表8试验饲料对牛蛙血清生化指标的影响结果表
2.5牛蛙空肠炎性因子和紧密连接蛋白基因表达量
结果见图1。抗炎因子白细胞介素10(IL-10)的基因相对表达量LL组显著高于NC组(P<0.05);抗炎因子白细胞介素4(IL-4)的基因相对表达量LL组与NC组差异不显著 (P>0.05),但LL组有高于NC组的趋势。促炎因子白细胞介素17(IL-17)和肿瘤坏死因子α(TNF-α)基因相对表达量LL组显著低于NC组(P<0.05)。由此推测,豆粕饲料中添加乳酸乳球菌乳亚种LCLL1可以提高牛蛙消化道的抗炎能力。
图1中,紧密连接蛋白ZO-1基因的相对表达量LL组显著高于NC组(P<0.05)。紧密连接蛋白Occludin基因的相对表达量LL组高于NC组,但差异不显著(P>0.05)。表8 中血清DAO含量显著高于NC组(P<0.05)。血清D-乳酸含量各组间无显著差异(P>0.05)。由此推测,豆粕饲料中添加乳酸乳球菌乳亚种LCLL1可以减少肠道黏膜损伤,降低肠道黏膜的通透性。
2.6牛蛙空肠组织结构
结果见图2和表9。其中A是指NC组、B是LL组的空肠切片结果图。从图2中可知, NC组可观察到空肠固有层与黏膜分离(如图中箭头所示)、黏膜下层与肌层分离(如图中三角形所示);LL组空肠结构相对完整,肌层更厚,肠黏膜排布更紧凑。通过对切片拍照测量,可以看出,LL组的黏膜皱襞数目和肌层厚度显著高于其NC组(P<0.05)。两组黏膜皱襞宽度、黏膜皱襞高度和固有层厚度没有显著性差异(P>0.05)。因此,豆粕饲料中添加乳酸乳球菌乳亚种LCLL1可以使牛蛙消化道结构更完整。
表9试验饲料对牛蛙空肠组织结构的影响结果表
NC | LL | |
黏膜皱襞数目 | 28.83±1.45<sup>a</sup> | 36.17±1.01<sup>b</sup> |
黏膜皱襞高度(μm) | 526.00±21.34 | 619.00±22.39 |
黏膜皱襞宽度(μm) | 157.00±8.25 | 169.17±9.18 |
肌层厚度(μm) | 34.83±3.46<sup>a</sup> | 84.83±7.42<sup>b</sup> |
固有层厚度(μm) | 21.95±1.76 | 20.88±1.88 |
2.7牛蛙肠道菌群
豆粕饲料中添加乳酸乳球菌乳亚种LCLL1增加了牛蛙消化道内的梭杆菌门比例,降低了变形菌门和厚壁菌门的比例(见图3的A)。在属水平上(见图3的B),LL组的鲸杆菌属、乳球菌属、邻单胞菌属和梭菌属的比例增加,降低了肠杆菌属、爱德华菌属、不动杆菌属、埃希氏杆菌/志贺氏杆菌、芽孢菌属、Lelliottia和Lysinibacittus的比例。乳酸乳球菌乳亚种LCLL1降低了牛蛙消化道内条件致病菌的丰度,增加了有益菌的丰度。
利用KEGG Orthology(KO)数据库比对各组样品肠道菌群的OTU对应的基因,进行PICRUSt基因功能预测,比较对照组与LL组富集功能信息之间的差异(基于welch’s T检验,P<0.05),可以看出,LL组主要有8个功能通路富集显著高于NC组(见图4),包括萜类化合物和聚酮类化合物代谢、复制和修复、聚糖生物合成与代谢、折叠、分类和降解、翻译、其他次生代谢产物的生物合成、以及细胞生长与凋亡等。
尽管上面已经示出和描述了本发明的实施例,可以理解的是,上述实施例是示例性的,不能理解为对本发明的限制,本领域的普通技术人员在不脱离本发明的原理和宗旨的情况下在本发明的范围内可以对上述实施例进行变化、修改、替换和变型。
SEQUENCE LISTING
<110> 集美大学
<120> 蛙源乳酸乳球菌乳亚种LCLL1及其用途
<130> JMDXL-21021-CNI
<160> 3
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> DNA
<213> 人工序列
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 20
<212> DNA
<213> 人工序列
<400> 2
acggctacct tgttacgact 20
<210> 3
<211> 1414
<212> DNA
<213> 牛蛙
<400> 3
atgcaagttg agcgctgaaa gttggtactt gtaccgactg gatgagcagc gaacgggtga 60
gtaacgcgtg gggaatctgc ctttgagcgg gggacaacat ttggaaacga atgctaatac 120
cgcataaaaa ctttaaacac aagttttaag tttgaaagat gcaattgcat cactcaaaga 180
tgatcccgcg ttgtattagc tagttggtga ggtaaaggct caccaaggcg atgatacata 240
gccgacctga gagggtgatc ggccacattg ggactgagac acggcccaaa ctcctacggg 300
aggcagcagt agggaatctt cggcaatgga cgaaagtctg accgagcaac gccgcgtgag 360
tgaagaaggt tttcggatcg taaaactctg ttggtagaga agaacgttgg tgagagtgga 420
aagctcatca agtgacggta actacccaga aagggacggc taactacgtg ccagcagccg 480
cggtaatacg taggtcccga gcgttgtccg gatttattgg gcgtaaagcg agcgcaggtg 540
gtttattaag tctggtgtaa aaggcagtgg ctcaaccatt gtatgcattg gaaactggta 600
gacttgagtg caggagagga gagtggaatt ccatgtgtag cggtgaaatg cgtagatata 660
tggaggaaca ccggtggcga aagcggctct ctggcctgta actgacactg aggctcgaaa 720
gcgtggggag caaacaggat tagataccct ggtagtccac gccgtaaacg atgagtgcta 780
gatgtaggga gctataagtt ctctgtatcg cagctaacgc aataagcact ccgcctgggg 840
agtacgaccg caaggttgaa actcaaagga attgacgggg gcccgcacaa gcggtggagc 900
atgtggttta attcgaagca acgcgaagaa ccttaccagg tcttgacata ctcgtgctat 960
tcctagagat aggaagttcc ttcgggacac gggatacagg tggtgcatgg ttgtcgtcag 1020
ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccctat tgttagttgc 1080
catcattaag ttgggcactc taacgagact gccggtgata aaccggagga aggtggggat 1140
gacgtcaaat catcatgccc cttatgacct gggctacaca cgtgctacaa tggatggtac 1200
aacgagtcgc gagacagtga tgtttagcta atctcttaaa accattctca gttcggattg 1260
taggctgcaa ctcgcctaca tgaagtcgga atcgctagta atcgcggatc agcacgccgc 1320
ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac accacgggag ttgggagtac 1380
ccgaagtagg ttgcctaacc gcaaggaggg cgct 1414
Claims (8)
1.一种蛙源乳酸乳球菌乳亚种Lactococcus lactis subsp. Lactis LCLL1,其保藏号为CGMCC No.21817。
2.权利要求1所述蛙源乳酸乳球菌乳亚种LCLL1用于制备养殖牛蛙饲料的用途。
3.如权利要求2所述用途,其特征在于,所述用途为对牛蛙具有益生效果。
4.如权利要求2所述用途,其特征在于,所述用途为能提高牛蛙的产量。
5.如权利要求2所述用途,其特征在于,所述用途为能提高饲料利用率;增强牛蛙机体免疫力。
6.如权利要求2所述用途,其特征在于,所述用途为能提高牛蛙消化道抗炎能力。
7.如权利要求2所述用途,其特征在于,所述用途为能使消化道黏膜微生物中有益菌的丰度增加,条件致病菌丰度降低。
8.如权利要求2所述用途,其特征在于,所述用途为将蛙源乳酸乳球菌乳亚种LCLL1添加到豆粕饲料喂养牛蛙。
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