title: Sequencing of variable regions of the 16S rRNA gene for identification of lactic acid bacteria isolated from the intestinal microbiota of healthy salmonids.
abstract: The aim of this study was to identify lactic acid bacteria (LAB) using polymerase chain reaction (PCR) amplification of variable regions of the 16S rRNA gene. Thirteen LAB strains were isolated from the intestinal microbiota of healthy salmonids. A approximately 500-bp region of the highly conserved 16S rRNA gene was PCR-amplified and following this, a portion of the amplicon (272-bp) including the V1 and V2 variable regions was sequenced. The sequence containing both the V1 and V2 region provided strong evidence for the identification of LAB. The LAB strains were identified as Carnobacterium maltaromaticum, Lactobacillus curvatus, Lactobacillus sakei, Lactobacillus plantarum, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, and Leuconostoc mesenteroides. The method described was found to be a very simple, rapid, specific, and low-cost tool for the identification of unknown strains of LAB.
authors.affiliation: Laboratory of Fish Pathology, Faculty of Veterinary Sciences, University of Zaragoza, Zaragoza 50013, Spain. balcazar@unizar.es
authors.forename: José Luis
authors.initials: JL
authors.lastname: Balcázar
authors.forename: Ignacio
authors.initials: I
authors.lastname: de Blas
authors.forename: Imanol
authors.initials: I
authors.lastname: Ruiz-Zarzuela
authors.forename: Daniel
authors.initials: D
authors.lastname: Vendrell
authors.forename: Olivia
authors.initials: O
authors.lastname: Gironés
authors.forename: José Luis
authors.initials: JL
authors.lastname: Muzquiz
mesh_terms: D000818:Animals; D001483:Base Sequence; D005285:Fermentation; D005784:Gene Amplification; D006094:Gram-Positive Bacteria / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007135:Immunoglobulin Variable Region; D007422:Intestines / Q000382:microbiology*; D019344:Lactic Acid / Q000096:biosynthesis; D007778:Lactobacillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics*; D012487:Salmonidae / Q000382:microbiology*; D013045:Species Specificity
chemical_list: D007135:Immunoglobulin Variable Region; D012336:RNA, Ribosomal, 16S; D019344:Lactic Acid