猪源益生性乳酸菌的分离鉴定及生物学特性研究  

Isolation,Identification and Biological Characteristics of Probiotic Lactic Acid Bacteria Derived from Porcine

作　　者：吴晓婷 王召路 朱莉莉 赵丽丽[1] 靳建军 史新娥[1] WU Xiaoting;WANG Zhaolu;ZHU Lili;ZHAO Lili;JIN Jianjun;SHI Xin’e(College of Animal Science and Technology,Northwest A&F University,Yangling 712100,China)

机构地区：[1]西北农林科技大学动物科技学院,杨凌712100

出　　处：《动物营养学报》2025年第4期2750-2762,共13页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：陕西省重点研发计划(2023-YBNY-109)。

摘　　要：本试验旨在从健康断奶仔猪粪便中筛选出产酸性能较好和具有益生特性的乳酸菌,为其作为饲料添加剂的应用奠定基础。首先用含碳酸钙(CaCO_(3))的MRS固体培养基分离纯化乳酸菌,然后通过菌株的产酸能力初步筛选出10株产酸能力较强的乳酸菌,对其进行分子生物学鉴定确定种属,并进一步评价其生长特性、耐受性、黏附能力、抗生素敏感性和抑菌能力,最后进行体内安全性评价。结果显示:筛选得到的产酸能力较强的10株乳酸菌(L2、L3、L4、L5、L7、L11、L13、L21、L22和L24)经16S rRNA测序鉴定均为唾液乳杆菌。在生长特性上,分离出的10株乳酸菌的生长情况相似,2~12 h为对数生长期;从耐受性上看,在pH为3.0的环境中培养3 h后,L4和L22表现出较高的耐酸性,存活率分别为36.24%和36.20%,在胆盐浓度为0.3%的胆盐环境中培养3 h后,L5的存活率最高,为41.14%;从黏附能力上看,分离出的10株乳酸菌的表面疏水率均在70%以上,其中L5最高,为87.76%,L7最低,为70.87%;L24表现出最强的自聚集能力,自聚集率为28.27%;抗生素药敏试验结果显示,分离出的10株乳酸菌对阿莫西林、呱拉西林、氟苯尼考、多西环素、头孢拉定、头孢呋辛和氧氟沙星均表现出敏感性,对苯唑西林和庆大霉素耐药;抑菌试验结果显示,分离出的10株乳酸菌均可以抑制目前常见的肠道病原菌的生长;选取L3和L5菌株进行小鼠体内安全性试验,结果显示灌胃乳酸菌(L3、L5和L3+L5)对小鼠体重和采食量均无负面影响,剖检后小鼠脏器无病理变化。综上所述,本试验从健康断奶仔猪粪便中筛选出的10株唾液乳杆菌具有较好的益生特性,尤其是唾液乳杆菌L3和L5,可作为饲料添加剂的候选菌株。The purpose of this experiment was to isolate and screen lactic acid bacteria with good acid production and probiotic properties from the feces of healthy weaned piglets,so as to lay a foundation for their future application as feed additives.In this study,lactic acid bacteria were isolated and purified using CaCO_(3)-MRS solid medium.Ten strains of lactic acid bacteria were initially selected based on their capacity for acid production,followed by species identification through molecular biological identification.Their growth characteristics,tolerance,adhesion ability,antibiotic susceptibility and antimicrobial activity were subsequently evaluated alongside an assessment of their in vivo safety.The results showed that all ten strains of lactic acid bacteria(L2,L3,L4,L5,L7,L11,L13,L21,L22 and L24)with strong acid-producing ability were identified as Lactobacillus salivarius by 16S rRNA sequencing.In terms of growth characteristics,these strains exhibited similar patterns with a logarithmic phase occurring between 2 hours to 12 hours post-inoculation.Regarding acidity tolerance,L4 and L22 demonstrated higher acidity tolerance with survival rate of 36.24%and 36.20%,respectively,after being cultured in an acidic environment(pH 3.0)for three hours;meanwhile,L5 achieved the highest survival rate at 41.14%when exposed to a choline salt concentration of 0.3%for three hours.Adhesion ability revealed that all ten strains of lactic acid bacteria had surface hydrophobic rate exceeding 70%,with L5 exhibiting the highest rate at 87.76%,while L7 showed the lowest rate at 70.87%.Among these strains,L24 displayed the most robust auto-aggregation ability with an auto-aggregation rate of 28.27%.Antibiotic susceptibility test revealed that all ten strains of lactic acid bacteria were sensitive to amoxicillin,clavulanic acid,florfenicol,doxycycline,cefadroxil,cefuroxime and oxifloxacin,but exhibited resistance against benzathine penicillin and gentamicin.Additionally,the antimicrobial test confirmed that all ten strains of lactic ac

关 键 词：乳酸菌 分离 鉴定 生物学特性 安全性 

分 类 号：S816.35[农业科学—饲料科学]