title: A multilocus sequence analysis scheme for characterization of Flavobacterium columnare isolates.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Centre of Excellence in Biological Interactions, Department of Biological and Environmental Science, University of Jyvaskyla, P.O. Box 35, Jyväskylän yliopisto, FI-40014, Finland. roghaieh.ashrafi@jyu.fi.
authors.forename: Roghaieh
authors.initials: R
authors.lastname: Ashrafi
authors.affiliation: Department of Biological and Environmental Science, University of Jyvaskyla, P.O. Box 35, Jyväskylän yliopisto, FI-40014, Finland. katja.pulkkinen@jyu.fi.
authors.forename: Katja
authors.initials: K
authors.lastname: Pulkkinen
authors.affiliation: Centre of Excellence in Biological Interactions, Department of Biological and Environmental Science, University of Jyvaskyla, P.O. Box 35, Jyväskylän yliopisto, FI-40014, Finland. lotta-riina.sundberg@jyu.fi.
authors.forename: Lotta-Riina
authors.initials: LR
authors.lastname: Sundberg
authors.affiliation: Department of Biological and Environmental Science, University of Jyvaskyla, P.O. Box 35, Jyväskylän yliopisto, FI-40014, Finland. nina.a.pekkala@jyu.fi.
authors.forename: Nina
authors.initials: N
authors.lastname: Pekkala
authors.affiliation: Centre of Excellence in Biological Interactions, Department of Biological and Environmental Science, University of Jyvaskyla, P.O. Box 35, Jyväskylän yliopisto, FI-40014, Finland. tarmo.t.ketola@jyu.fi.
authors.forename: Tarmo
authors.initials: T
authors.lastname: Ketola
mesh_terms: D000818:Animals; D017756:Aquaculture; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005387:Finland / Q000453:epidemiology; D005393:Fish Diseases / Q000453:epidemiology* / Q000382:microbiology*; D045826:Flavobacteriaceae Infections / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D005417:Flavobacterium / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D020043:Genes, Essential; D014644:Genetic Variation; D005838:Genotype; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D058885:Multilocus Sequence Typing / Q000379:methods*; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
chemical_list: D004269:DNA, Bacterial