被膜态与浮游态猪源乳杆菌上清液抑菌活性比较分析  

Comparative Analysis of the Antimicrobial Activity of Lactobacillus Supernatants from Swine in Biofilm and Plankton

作　　者：靳胜男 舒慧萍 张冬星[1] 赵泽林 康元环[1] 张蕾 单晓枫[1] 钱爱东[1] JIN Shengnan;SHU Huiping;ZHANG Dongxing;ZHAO Zelin;KANG Yuanhuan;ZHANG Lei;SHAN Xiaofeng;QIAN Aidong(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学动物科学技术学院

出　　处：《中国畜牧兽医》2019年第11期3358-3369,共12页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金项目(31201927);吉林省教育厅“十三五”科学技术项目(JJKH20180693KJ、JJKH20190910KJ);吉林省科技厅自然科学基金学科布局项目(20170101016JC);吉林农业大学博士启动基金(201801)

摘　　要：试验旨在比较分析猪源约氏乳杆菌L-76和嗜淀粉乳杆菌L-102被膜态和浮游态下其上清液对病原菌的抑菌活性。采用结晶紫染色法测定L-76和L-102的生物被膜形成能力,研究其被膜态和浮游态乳杆菌上清液及上清液与不同影响因素作用后的抑菌活性,并利用扫描电镜观察其两种状态下的上清液对指示菌形态的影响。结果显示,L-76、L-102乳酸菌具有较强的生物被膜形成能力,被膜态和浮游态乳杆菌L-76、L-102具有良好的抑菌活性,其上清液经蛋白酶作用后,被膜态L-76和L-102上清液的抑菌圈直径比浮游态明显变小;经过氧化氢酶和不同温度作用后,两种状态的乳杆菌上清液的抑菌活性无明显变化(P>0.05),但在pH 3.0作用下,两种状态的乳杆菌上清液的抑菌效果最好;扫描电镜结果显示,被膜态乳杆菌上清液对金黄色葡萄球菌的形态影响略大。综上所述,两种状态下乳酸杆菌上清液中均含有抑菌物质,且被膜态乳酸杆菌上清液中的抑菌物质含量略多或活性略高。The aim of this study was to compare and analyze the bacteriostatic activity of the supernatants of Lactobacillus johnsonii L-76 and Lactobacillus amylovorus L-102 from swine in biofilm and plankton to pathogenic bacteria.The biofilm formation ability of L-76 and L-102 were measured by crystal violet staining assay.Then the supernatants of Lactobacillus in biofilm and plankton were treated with different influencing factors to study the inhibitory effect of Lactobacillus supernatants on pathogenic bacteria.Scanning electron microscopy was used to observe the effect of supernatant on the morphology of indicator bacteria.The results showed that L-76 and L-102 Lactobacillus had strong ability of biofilm formation and that the growth of pathogenic bacteria was inhibited after co-culture with the supernatant of Lactobacillus in biofilm and plankton.After the action of protease,compared with plankton,the inhibitory zone diameters of biofilm L-76 and L-102 supernatant on the indicator bacteria were significantly reduced.After the action of catalase and different temperatures,there was no significant difference in the antimicrobial activity between L76 and L-102 Lactobacillus supernatants in both conditions(P>0.05).However,they had the best inhibitory effect when pH was 3.0.We observed that the supernatant of biofilm Lactobacillus had a slightly greater effect on the morphology of Staphylococcus aureus by scanning electron microscopy.In conclusion,the supernatant of Lactobacillus contained antimicrobial substances in both conditions,and the activity of bacteriostatic substances in the supernatant of biofilm Lactobacillus was slightly higher or the content was slightly higher.

关 键 词：猪 乳杆菌 生物被膜 约氏乳杆菌 嗜淀粉乳杆菌 抑菌性 

分 类 号：S858.28[农业科学—临床兽医学]