A multilocus sequence analysis scheme for characterization of Flavobacterium columnare isolates

BMC Microbiol. 2015 Oct 30:15:243. doi: 10.1186/s12866-015-0576-4.

Abstract

Background: Columnaris disease caused by Flavobacterium columnare is a serious problem in aquaculture, annually causing large economic losses around the world. Despite considerable research, the molecular epidemiology of F. columnare remains poorly understood.

Methods: We investigated the population structure and spatiotemporal changes in the genetic diversity of F. columnare population in Finland by using a multilocus sequence typing (MLST) and analysis (MLSA) based on DNA sequence variation within six housekeeping genes. A total of 83 strains of F. columnare were collected from eight different areas located across the country between 2003 and 2012.

Results: Partial sequencing of six housekeeping genes (trpB, tuf, atpA, rpoD, gyrB and dnaK) revealed eight sequence types and a moderate level of genetic diversity (H=0.460). Phylogenetic analysis of the concatenated protein-encoding gene sequence data (ca. 3,509 nucleotides) formed two lineages, which could be further divided into five clusters. All analysed F. columnare strains appeared to have a genetic origin distinct from that of another important fish pathogen form the genus Flavobacterium, F. psychrophilum. Although the value of the index of association between alleles, 0.292 (P<0.001), supports some degree of clonality for this species in Finland, recombination has introduced molecular diversity to the population almost three times more than mutation.

Conclusion: The results suggest that Finnish F. columnare strains have an epidemic population structure followed by clonal expansion of successful genotypes. Our study with reproducible methodology and comparable results establishes a robust framework for the discrimination and phylogenetic analysis of F. columnare isolates, which will help to improve our understanding about geographic distribution and epidemiology of columnaris disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aquaculture
  • Cluster Analysis
  • DNA, Bacterial / chemistry
  • DNA, Bacterial / genetics
  • Finland / epidemiology
  • Fish Diseases / epidemiology*
  • Fish Diseases / microbiology*
  • Flavobacteriaceae Infections / epidemiology
  • Flavobacteriaceae Infections / microbiology
  • Flavobacteriaceae Infections / veterinary*
  • Flavobacterium / classification*
  • Flavobacterium / genetics*
  • Flavobacterium / isolation & purification
  • Genes, Essential
  • Genetic Variation*
  • Genotype
  • Molecular Epidemiology
  • Molecular Sequence Data
  • Multilocus Sequence Typing / methods*
  • Phylogeny
  • Sequence Analysis, DNA
  • Sequence Homology

Substances

  • DNA, Bacterial

Associated data

  • GENBANK/LN624115
  • GENBANK/LN624116
  • GENBANK/LN624117
  • GENBANK/LN624118
  • GENBANK/LN624119
  • GENBANK/LN624120
  • GENBANK/LN624121
  • GENBANK/LN624122
  • GENBANK/LN624123
  • GENBANK/LN624124
  • GENBANK/LN624125
  • GENBANK/LN624126
  • GENBANK/LN624127
  • GENBANK/LN624128
  • GENBANK/LN624129
  • GENBANK/LN624130
  • GENBANK/LN624131
  • GENBANK/LN624132
  • GENBANK/LN624133
  • GENBANK/LN624134
  • GENBANK/LN624135