Abstract
Vibrio fluvialis, an estuarine-based halophilic vibrio, causes opportunistic infections in humans resulting in bloody diarrhea. The organism produces an enterotoxin (ET), a cytotoxin (CT), and a cytolysin (CL). We created isogenic non-toxigenic mutants to assess the relative contribution of each toxin to virulence. High level streptomycin (Sm) resistant mutants of the prototype toxigenic strain 807-77 were used as conjugal recipients for TnphoA, which creates gene fusions between target genes and phoA, which encodes the alkaline phosphatase of Escherichia coli. Transposon insertions into genes for secreted or periplasmic proteins were selected on indicator plates. Putative CL- transconjugates (which appear as dark blue colonies) were recovered at a rate of ca. 10-6/recipient cell. Presumptive CL- mutants were screened on blood agar, egg yolk agar, and with a sensitive tube hemolysin assay. Twenty-two CL- mutants were tested vs. red cells from two different species. CL- production was attenuated in 17 and negative in 5 of the mutants. The verified CL- mutants were tested for pathogenicity in infant mice. The CL- mutants were unaltered in their ability to cause fluid accumulation in this model. Various CL- mutants responded differently to increased osmolarity in their growth media as measured by alkaline phosphatase activity of permeabilized cells.
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© 1990 Plenum Press, New York
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Richardson, S.H., Kaspar, J.V. (1990). Genetic Studies on Vibrio Fluvialis and its Enterotoxins. In: Pohland, A.E., et al. Microbial Toxins in Foods and Feeds. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-0663-4_16
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DOI: https://doi.org/10.1007/978-1-4613-0663-4_16
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