一株安庆六白猪源乳酸菌的分离鉴定及生物特性研究  

Isolation,Identification and Biological Characteristics of a Lactic Acid Bacteria from Anqing Liubai Pigs

作　　者：吴雨晗 吴婷 涂健[1] 李哲 盛婷婷 李峰 姚皓洲 WU Yuhan;WU Ting;TU Jian;LI Zhe;SHENG Tingting;LI Feng;YAO Haozhou(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230000,China)

机构地区：[1]安徽农业大学动物科技学院,合肥230000

出　　处：《动物营养学报》2022年第8期5415-5425,共11页CHINESE JOURNAL OF ANIMAL NUTRITION

基　　金：安徽省中央引导地方科技扶贫专项(201907d06020016);高校协同创新项目(GXXT-2019-035);2020年度高校优秀人才支持计划项目(gxyqZD2020009)。

摘　　要：本试验旨在筛选优质猪源乳酸菌,为开发微生态制剂奠定基础。通过加入碳酸钙的MRS固体培养基分离纯化出6株疑似乳酸菌,对上述菌株进行耐受试验、抑菌试验,挑选出1株优秀菌株,再进行产酸试验、黏附性试验,测定其生长曲线和产酸曲线。结果表明:从六白猪粪样中分离筛选出1株抗逆性好且抑菌活性强的菌株,经过16S rRNA测序、生化鉴定、革兰氏染色鉴定,该菌株为约翰氏乳杆菌(Lactobacillus johnsonii),命名为Tβ。Tβ可以耐受高温与低pH,在pH为2.0时的存活率为12.1%,90℃时的存活率为14.02%。Tβ具有优秀的产酸能力,最低可达到pH 3.4。Tβ对大肠杆菌、沙门氏菌、金黄色葡萄球菌皆具有良好的抑菌效果。体外黏附试验证明Tβ具有较低的自聚合能力,较高的疏水能力,与金黄色葡萄球菌的共凝集性为52.11%,与沙门氏菌的共凝集性为75.64%,与大肠杆菌的共凝集性为73.95%。肠道内定植观察可以明显观察到异硫氰酸荧光素(FITC)标记的Tβ荧光,说明其黏附能力较为优秀。由此可见,Tβ是一株抗逆性、抑菌效果、黏附效果都十分优秀的猪源乳酸菌微生态制剂备用菌株。The aim of this paper was to screen a high-quality pig-derived lactic acid bacteria,to lay a foundation for the development of microecological preparations.Six strains of suspected lactic acid bacteria were isolated and purified by MRS solid medium with the addition of calcium carbonate,the above strains were subjected to tolerance test and inhibition test to select an excellent strain,then subjected to acid production test and adhesion test,and measured its growth curve and acid production curve.The results showed that a strain with good resistance and strong antibacterial activity was isolated and screened from Liubai pig manure samples,and identified as Lactobacillus johnsonii after 16S rRNA sequencing,biochemical identification and Gram staining,which named Tβ.Tβcan tolerate high temperature and low pH,the survival rate was 12.1%at pH 2.0,and the survival rate was 14.02%at 90°C.Tβhas excellent acid production capacity down to pH 3.4.Tβhas good antibacterial effects on Escherichia coli,Salmonella and Staphylococcus aureus.The in vitro adhesion test proved that Tβhas low self-agglomeration ability and high hydrophobic ability,with co-agglutination of52.11%with Staphylococcus aureus,75.64%with Salmonella and 73.95%with Escherichia coli.The fluorescin isothiocyanate(FITC)labeled Tβfluorescence could be clearly observed in the intestinal colonization observation,which indicated its superior adhesion ability.In conclusion,Tβis an alternate strain of pig-derived lactic acid bacteria microecological preparation with excellent resistance,bacterial inhibition and adhesion effect.

关 键 词：安庆六白猪 乳酸菌 分离 鉴定 

分 类 号：S816.7[农业科学—饲料科学]