Current Progress in Isolation and Characterization of Toxins Isolated from Pfiesteria piscicida

The isolation and partial purification of toxic substances derived from Pfiesteria piscicida Steidinger & Burkholder extracts is described. Four distinct bioassay systems were used to monitor bioactivity of the P. piscicida extracts, including a high throughput cell cytotoxicity assay and a repo...

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Bibliographic Details
Published in:	Environmental health perspectives 2001-10, Vol.109 (suppl 5), p.739-743
Main Authors:	Peter D. R. Moeller, Morton, Steve L., Mitchell, Brad A., Sivertsen, Scott K., Fairey, Elizabeth R., Mikulski, Tina M., Glasgow, Howard, Deamer-Melia, Nora J., Burkholder, JoAnn M., Ramsdell, John S.
Format:	Article
Language:	eng
Subjects:	2-ethylhexylphthalate Animals Anostraca Artemia Bioassay Biological Assay Brackish Brines Cytotoxicity Fishes Fractionation Freshwater Gene Expression Regulation Genes, Reporter Magnetic Resonance Spectroscopy Marine Mass Spectrometry Molecular Cell-Based and Bioassy Methods for Detection of Pfiesteria Species and Their Suspect Toxins Molecular Structure Oceans Pfiesteria piscicida Pfiesteria piscicida - pathogenicity Phthalates Pisces Reporter genes Sea water Solubility Toxicity Toxins Toxins, Biological - adverse effects Toxins, Biological - chemistry Toxins, Biological - isolation & purification
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Summary:	The isolation and partial purification of toxic substances derived from Pfiesteria piscicida Steidinger & Burkholder extracts is described. Four distinct bioassay systems were used to monitor bioactivity of the P. piscicida extracts, including a high throughput cell cytotoxicity assay and a reporter gene assay as well as assays using brine shrimp and fish. Using these bioassays to guide fractionation, we have isolated two distinct, active fractions from Pfiesteria culture medium and cell mass extracts on the basis of their solubility characteristics. We have identified and characterized a bioactive lipophilic substance from Pfiesteria-derived extracts as di(2-ethylhexyl)phthalate, a commonly used plasticizer. The source of this typically man-made substance has been identified as originating from Instant Ocean (Aquarium Systems, Mentor, OH, USA), a commercially available seawater salt mixture used to prepare our mass culture growth medium. We have developed chromatographic methodology to isolate a bioactive polar compound isolated from extracts of Pfiesteria culture and presently report the characterization of the activity of this substance. The molecular structural analysis of the polar active component(s) using mass spectrometry and nuclear magnetic resonance spectroscopy is currently under way.
ISSN:	0091-6765 1552-9924