title: Comparative analysis of Edwardsiella isolates from fish in the eastern United States identifies two distinct genetic taxa amongst organisms phenotypically classified as E. tarda.
abstract: Edwardsiella tarda, a Gram-negative member of the family Enterobacteriaceae, has been implicated in significant losses in aquaculture facilities worldwide. Here, we assessed the intra-specific variability of E. tarda isolates from 4 different fish species in the eastern United States. Repetitive sequence mediated PCR (rep-PCR) using 4 different primer sets (ERIC I & II, ERIC II, BOX, and GTG5) and multi-locus sequence analysis of 16S SSU rDNA, groEl, gyrA, gyrB, pho, pgi, pgm, and rpoA gene fragments identified two distinct genotypes of E. tarda (DNA group I; DNA group II). Isolates that fell into DNA group II demonstrated more similarity to E. ictaluri than DNA group I, which contained the reference E. tarda strain (ATCC #15947). Conventional PCR analysis using published E. tarda-specific primer sets yielded variable results, with several primer sets producing no observable amplification of target DNA from some isolates. Fluorometric determination of G+C content demonstrated 56.4% G+C content for DNA group I, 60.2% for DNA group II, and 58.4% for E. ictaluri. Surprisingly, these isolates were indistinguishable using conventional biochemical techniques, with all isolates demonstrating phenotypic characteristics consistent with E. tarda. Analysis using two commercial test kits identified multiple phenotypes, although no single metabolic characteristic could reliably discriminate between genetic groups. Additionally, anti-microbial susceptibility and fatty acid profiles did not demonstrate remarkable differences between groups. The significant genetic variation (<90% similarity at gyrA, gyrB, pho, phi and pgm; <40% similarity by rep-PCR) between these groups suggests organisms from DNA group II may represent an unrecognized, genetically distinct taxa of Edwardsiella that is phenotypically indistinguishable from E. tarda.
authors.affiliation: Thad Cochran National Warmwater Aquaculture Center, College of Veterinary Medicine, Mississippi State University, Stoneville, MS 38776, USA. griffin@cvm.msstate.edu
authors.forename: Matt J
authors.initials: MJ
authors.lastname: Griffin
authors.forename: Sylvie M
authors.initials: SM
authors.lastname: Quiniou
authors.forename: Theresa
authors.initials: T
authors.lastname: Cody
authors.forename: Maki
authors.initials: M
authors.lastname: Tabuchi
authors.forename: Cynthia
authors.initials: C
authors.lastname: Ware
authors.forename: Rocco C
authors.initials: RC
authors.lastname: Cipriano
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Mauel
authors.forename: Esteban
authors.initials: E
authors.lastname: Soto
mesh_terms: D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology; D001482:Base Composition; D020609:Edwardsiella tarda / Q000145:classification* / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification; D004756:Enterobacteriaceae Infections / Q000382:microbiology / Q000662:veterinary*; D005393:Fish Diseases / Q000382:microbiology*; D005399:Fishes; D005798:Genes, Bacterial / Q000235:genetics; D014644:Genetic Variation; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D014481:United States
chemical_list: D000890:Anti-Infective Agents