Graduate Thesis Or Dissertation
 

Human health significance of vibrios from the aquatic environment

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  • For the enrichment and enumeration of Vibrio fluvialis, a broth medium was designed by modifying alkaline peptone (AP) medium. This new V. fluvialis enrichment medium (FEM) was shown to be more effective than AP medium in field samplings where a total of 177 samples (estuarine waters and sediment, sewage, and crabs) were processed over a 14 month period. FEM was particularly superior to AP for water and sewage with low salinities (<6%0). V. vulnificus was shown to have a species-specific antigen by analyzing sonicated whole-cell antigens with two-dimensional immunoelectrophoresis. The antigen (designated as VVA) was purified by various protein chemistry techniques, and specific antiserum to VVA was prepared. Using anti-VVA serum, a simple and rapid microimmunodiffusion method was designed which allowed the specific identification of V. vulnificus in as early as 10 hrs after preparing a bacterial cell lysate from a single colony. suckling mouse assay was utilized to examine enteropathogenicity, i.e., potential to cause diarrhea, of environmental and clinical isolates of 0-1 and non 0-1 V. cholerae, V. mimicus, and V. fluvialis. Cultures from both environmental and clinical origins induced intestinal fluid accumulation (FA) in 3-day-old mice at 4 hr postinoculation followed by diarrheal feces and high mortality. The virulence-associated factor(s) that caused FA were different from cholera toxin (CT) in terms of kinetics of FA and serological behavior, but were correlated with extracellular cytotoxic factors active against Y-1 mouse adrenal tissue cultures. Furthermore, all clinical isolates of non 0-1 V. cholerae, V. mimicus, and V. fluvialis, when grown in brain heart infusion broth supplemented with 0.5% sodium chloride, were found to produce a new extracellular heat-labile enterotoxin which induced significant FA in 3-day-old mice and was distinct from previously reported enterotoxins including CT. Most of the environmental isolates, however, did not produce the newly discovered enterotoxin.
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