title: Occurrence and distribution of Vibrio vulnificus and Vibrio parahaemolyticus--potential roles for fish, oyster, sediment and water.
abstract: Vibrio vulnificus and Vibrio parahaemolyticus are Gram-negative marine bacteria naturally found in estuaries such as the Gulf of Mexico and can be pathogenic to humans. We quantified both of these organisms in fish, oyster, sediment, and water using culture-independent (quantitative PCR; qPCR) and culture-dependent (direct plating-colony hybridization; DP-CH) techniques during the transition period between winter and spring. We correlated these levels to environmental conditions and to abundance of total bacteria and total vibrio. By qPCR, fish intestine samples had the highest V. vulnificus densities and were 2·7, 3·5, and 4·2 logs greater than oyster, sediment and water samples, respectively. Densities of V. parahaemolyticus in fish samples by qPCR were 1·0, 2·1, and 3·1 logs greater than oyster, sediment and water samples, respectively. Similar differences between sample types were also observed by DP-CH. The difference between the more favourable and less favourable environmental conditions identified in this study was small (mean salinity 4·3 vs. 13 ppt). However, V. vulnificus and V. parahaemolyticus were consistently detected in fish intestines, but infrequently detected and at lower levels in oysters and during the less favourable period. This trend was observed by qPCR and DP-CH, indicating fish intestines are a significant source of pathogenic vibrios in the environment.
  
  
  CONCLUSIONS
  This is the first study to report the densities of Bacteria 16S rRNA, Vibrio 16S rRNA, Vibrio vulnificus, and V. parahaemolyticus in fish intestine, oyster, sediment and water samples, as well as compare these values through culture-dependent and culture-independent methodology. Vibrio vulnificus and V. parahaemolyticus were detected in samples of fish intestines by qPCR and colony hybridization when conditions were less favourable for their occurrence in the environment. In contrast, V. vulnificus and V. parahaemolyticus were infrequently detected and at lower levels in other niches examined. These results indicate that fish intestinal tracts are a significant source of these pathogens.
authors.affiliation: Department of Marine Science, University of Georgia, Athens, GA, USA.
authors.forename: C E
authors.initials: CE
authors.lastname: Givens
authors.forename: J C
authors.initials: JC
authors.lastname: Bowers
authors.forename: A
authors.initials: A
authors.lastname: DePaola
authors.forename: J T
authors.initials: JT
authors.lastname: Hollibaugh
authors.forename: J L
authors.initials: JL
authors.lastname: Jones
mesh_terms: D000818:Animals; D005399:Fishes / Q000382:microbiology*; D041981:Gastrointestinal Tract / Q000382:microbiology; D019015:Geologic Sediments / Q000382:microbiology; D060851:Gulf of America; D006801:Humans; D058889:Molecular Typing; D010125:Ostreidae / Q000382:microbiology*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D045483:Rivers; D054712:Salinity; D012621:Seasons; D012758:Shellfish / Q000382:microbiology*; D014736:Vibrio parahaemolyticus / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D041261:Vibrio vulnificus / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D014774:Virulence; D014871:Water Microbiology
chemical_list: D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S
keywords: environmental; fish (live); sediment; shellfish; water