De Fabrizio S V, Parada J L, Ledford R A, Solari A, Brown J
Laboratorio de Microbiología de Alimentos, Facultad de Ciencias Exactas y naturales, Universidad de Buenos Aires.
Rev Argent Microbiol. 1989 Jan-Mar;21(1):1-7.
Streptococcus lactis and Streptococcus cremoris have an important role in the fermentation of milk during the manufacturing of lactic products. Bacteriophages are spread in the plant environment and they may infect the starters causing technical and economic problems to the dairy industry. It is now known that the usual methods of control are not completely safe against the proliferation of phages. It is necessary therefore to include resistant strains to the phages which infect the plant. This study introduces a simple and direct method to obtain spontaneous mutants which have performed clear resistance to a certain phage. The prolate (ARG) phages used in this work St11, St13 and St15 were isolated in Argentina, on S. lactis C2 while the prolate and isometric (AM) phages phi C2, D59-1, I16-1, and G72-1, F4-1, I37-1 respectively, were isolated in Cornell University. on the corresponding S. lactis strains except the phage D59-1 isolated on an S. cremoris strain. The propagation of phages was performed using M and M17 broth. The phage sensitivity was tested through spot test and overlay plaque plating method expressing the titre of phage suspension. as PFU/ml. In order to obtain spontaneous resistant mutants of S. lactis C2 to phage St15, and active culture of S. lactis C2 (sensitive) was spread on agar M plates, inoculated with drops of a suspension of phage St15 (10(8)PFU/ml) and incubated at 30 degrees C for 48 h. Colonies of mutants grown in the lytic areas caused by the phage were isolated and selected.(ABSTRACT TRUNCATED AT 250 WORDS)
乳酸链球菌和乳脂链球菌在乳制品生产过程中的牛奶发酵中起着重要作用。噬菌体在工厂环境中传播,它们可能感染发酵剂,给乳制品行业带来技术和经济问题。现在已知常规的控制方法并不能完全安全地抵御噬菌体的增殖。因此,有必要纳入对感染工厂的噬菌体具有抗性的菌株。本研究介绍了一种简单直接的方法来获得对特定噬菌体具有明显抗性的自发突变体。本研究中使用的长形(ARG)噬菌体St11、St13和St15是在阿根廷从乳酸链球菌C2中分离出来的,而长形和等轴(AM)噬菌体phi C2、D59 - 1、I16 - 1以及G72 - 1、F4 - 1、I37 - 1分别是在康奈尔大学从相应的乳酸链球菌菌株中分离出来的,除了噬菌体D59 - 1是从一株乳脂链球菌菌株中分离出来的。噬菌体的增殖使用M肉汤和M17肉汤进行。通过点试验和覆盖平板法测试噬菌体敏感性,以PFU/ml表示噬菌体悬浮液的滴度。为了获得乳酸链球菌C2对噬菌体St15的自发抗性突变体,将乳酸链球菌C2(敏感型)的活性培养物铺在M琼脂平板上,接种噬菌体St15悬浮液(10⁸PFU/ml)的液滴,在30℃下培养48小时。分离并选择在噬菌体引起的裂解区域生长的突变体菌落。(摘要截短至250字)