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SELECTIVITY OF LYSIS BY OCTANOL-WATER; APPLICATION TO HARMFUL ALGAE

Abstract

As reported previously, octanol-water extraction of chlorophyll and phycocyanine from the cyanobacterium Microcystis aeruginosoffers advantages in the methodology used for monitoring water bodies for harmful algal blooms (HABs). However, this methodology has not been extensively examined or reported to work for cyanobacterial species other than M. aeruginosa, nor have results been reported for other algal types. Here we show that octanol appears to lyse other species of cyanobacteria (evidenced by the release of chlorophyll into the upper octanol phase of the extraction mixture), as well as bacteria other than cyanobacteria (evidenced by the release of protein absorbance at 280 nm wavelength). We also show that nonbacterial algae, such as green algae, are not lysed by octanol, although they may be killed by it. The cyanobacterial species tested were (in addition to Microcystis aeruginosa) Cylindrospermopsum, Anabaena, Limnothrix, and Spirulina. Algal species Chlorella, Scenedesmus, and Chlamydomonas were not effectively lysed by this methodology. Euglena was quite effectively lysed. Other species are in the process of being tested. The selectivity for bacterial lysis shown here by octanol may potentially allow for more sensitive detection of the cyanobacterial component of algal blooms by eliminating the nonbacterial absorbance in the extract. Another strategy for reducing background is to collect the algae onto filters that allow the cells to be washed free of the filter, thus eliminating the homogenization of the filter along with the cells. We will show results of a Pall filter (Super 800 Pall 60110) that shows potential for achieving this result. We hope to develop a methodology for monitoring environmental waters for harmful algae that will lend itself to high throughput at lower expense. This work is supported by the East Georgia State College Baccalaureate Biology Program.

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