title: The evaluation of four recent culture-based methods for the isolation and enumeration of Vibrio vulnificus bacteria from oyster meat.
abstract: The most common cause of seafood-borne death in the United States is the bacterium Vibrio vulnificus which can be concentrated into high numbers in the tissues of oysters or other shellfish. The ability to quickly, accurately, and inexpensively isolate living strains of this organism from oyster tissues is crucial for effective research on this pathogen. In this report, we evaluate four methods for isolating and quantifying V. vulnificus from oyster tissues, the solid media CPC+ (a refined version of cellobiose-polymyxin B-colistin medium), CHROMagar Vibrio, VVX (Vibrio vulnificus X-gal), and a method termed "Triple plating". Up to 1225 presumptive isolates were detected by each method, and 335 were subjected to molecular typing. The selectivity and sensitivity of each method was examined and VVX was found to be the most accurate method, with each of the other methods being recommended for task-specific uses. CHROMagar Vibrio is recommended for ease of use and relative accuracy, CPC+ is best used to differentiate between clinically associated and environmental strains.
authors.affiliation: UNC-Chapel Hill, Institute of Marine Sciences, 3431 Arendell Street, Morehead City, NC 28557, USA. Electronic address: bafroeli@unc.edu.
authors.forename: Brett A
authors.initials: BA
authors.lastname: Froelich
authors.affiliation: UNC-Chapel Hill, Institute of Marine Sciences, 3431 Arendell Street, Morehead City, NC 28557, USA.
authors.forename: Mary Jo
authors.initials: MJ
authors.lastname: Weiss
authors.affiliation: UNC-Chapel Hill, Institute of Marine Sciences, 3431 Arendell Street, Morehead City, NC 28557, USA.
authors.forename: Rachel T
authors.initials: RT
authors.lastname: Noble
mesh_terms: D000818:Animals; D058491:Bacterial Load / Q000379:methods*; D003470:Culture Media; D005516:Food Microbiology / Q000379:methods*; D005838:Genotype; D010125:Ostreidae / Q000382:microbiology*; D015203:Reproducibility of Results; D041261:Vibrio vulnificus / Q000302:isolation & purification* / Q000502:physiology*
chemical_list: D003470:Culture Media
keywords: CPC; CPC+; Oysters; Pathogen; Shellfish; TCBS; VVX; Vibrio; Vibrio vulnificus X-gal medium; cellobiose–polymyxin B-colistin medium; refined cellobiose–polymyxin B-colistin medium; thiosulfate–citrate–bile salts–sucrose; vulnificus