Sequencing of variable regions of the 16S rRNA gene for identification of lactic acid bacteria isolated from the intestinal microbiota of healthy salmonids.
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.
Balcázar
José Luis
JL
Laboratory of Fish Pathology, Faculty of Veterinary Sciences, University of Zaragoza, Zaragoza 50013, Spain. balcazar@unizar.es
de Blas
Ignacio
I
Ruiz-Zarzuela
Imanol
I
Vendrell
Daniel
D
Gironés
Olivia
O
Muzquiz
José Luis
JL
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
D007135:Immunoglobulin Variable Region; D012336:RNA, Ribosomal, 16S; D019344:Lactic Acid