title: Evaluation of molecular methods to discriminate the closely related species Vibrio fluvialis and Vibrio furnissii.
abstract: Vibrio furnissii and Vibrio fluvialis are two closely related species which are regarded as emerging human pathogens. Human infections have been mainly associated with consumption of seafood or drinking of contaminated water. V. furnissii strains can be distinguished from V. fluvialis by their ability to produce gas from fermentation of carbohydrates. In this study, we compare two phenotypic (biochemical testing and matrix-assisted laser desorption/ionisation time of flight mass spectrometry, MALDI-TOF MS) and three genotypic techniques (rpoB sequencing, conventional PCR and real-time PCR) for determination of the two species. The methods were evaluated on a collection of 42 V. furnissii and 32 V. fluvialis strains, which were isolated from marine environments and from animals intended for food production. Four of the applied methods allowed the unambiguous discrimination of the two species, while the biochemical testing was the least reliable technique, due to a high variation in the phenotype of gas production from carbohydrates. In view of the One Health concept reliable diagnostic techniques are a prerequisite for preventive public health measurements, as pathogens isolated from animals can cross species borders and methods for detection of sources, reservoirs and ways of transmission of pathogenic bacteria are indispensable for the prevention of infectious diseases in humans and animals.
authors.affiliation: Bundesinstitut für Risikobewertung, Federal Institute for Risk Assessment, National Reference Laboratory for Monitoring Bacteriological Contamination of Bivalve Molluscs, Berlin, Germany.
authors.forename: Falko
authors.initials: F
authors.lastname: Schirmeister
authors.affiliation: Bundesinstitut für Risikobewertung, Federal Institute for Risk Assessment, National Reference Laboratory for Monitoring Bacteriological Contamination of Bivalve Molluscs, Berlin, Germany.
authors.forename: Angelina
authors.initials: A
authors.lastname: Wieczorek
authors.affiliation: Bundesinstitut für Risikobewertung, Federal Institute for Risk Assessment, National Reference Laboratory for Monitoring Bacteriological Contamination of Bivalve Molluscs, Berlin, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Dieckmann
authors.affiliation: Landesuntersuchungsanstalt für das Gesundheits - und Veterinärwesen Sachsen, Standort Dresden, Germany.
authors.forename: Karin
authors.initials: K
authors.lastname: Taureck
authors.affiliation: Bundesinstitut für Risikobewertung, Federal Institute for Risk Assessment, National Reference Laboratory for Monitoring Bacteriological Contamination of Bivalve Molluscs, Berlin, Germany. Electronic address: Eckhard.Strauch@bfr.bund.de.
authors.forename: Eckhard
authors.initials: E
authors.lastname: Strauch
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques / Q000379:methods*; D001431:Bacteriological Techniques / Q000379:methods*; D012321:DNA-Directed RNA Polymerases; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D008969:Molecular Sequence Data; D017747:Seafood / Q000382:microbiology; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014733:Vibrio / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000502:physiology; D014871:Water Microbiology
chemical_list: D012321:DNA-Directed RNA Polymerases; C105078:RNA polymerase beta subunit
keywords: MALDI–TOF mass spectrometry; Veterinary Vibrio isolates; qPCR; rpoB sequencing