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Details for the preparation and partial purification of culture supernatant fluids of Vibrio cholerae (V. comma) 569B which retain rabbit ileal loop fluid-accumulating activity are presented. These preparations were fractionated on Sephadex G-200 and on diethylaminoethyl-Sephadex. On the latter, two fractions were obtained by elution with a linear sodium chloride gradient. The fraction designated "fraction I" retains the toxic activity as demonstrated in the rabbit ileal loop model. Chemical and immunological properties of this active fraction are described.

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