Colonial cell disaggregation and release of intracellular microcystin were evaluated following chlorine treatment of naturally occurring Microcystis. Microscopic observations of water samples collected from Lake Mead, Nevada, USA, confirmed the presence of colonial Microcystis with cells protected by an outer sheath up to 30 μm thick. During chlorination, two stages of cell decomposition were observed, stage 1: colonial cell disaggregation, and stage 2: unicellular degradation. Following a [Cl2]0:DOC0 ratio of 0.15 (t = 20 min, pH = 8.2-8.5) in unfiltered Lake Havasu samples, total particle count increased from (1.0 ± 0.11) × 10(5) to 4.2 × 10(5) particles/mL and fluorescent particle count increased from (1.2 ± 0.50) × 10(4) to 1.2 × 10(5) particles/mL, illustrating colonial cell disaggregation. Although total and fluorescent particles increased, the concentration of chlorophyll-a (Chl-a) decreased from 81 μg/L to 72 μg/L, and continued to decrease at higher [Cl2]0:DOC0 ratios. The preliminary second order rate constant for the reaction between Microcystis and chlorine in natural waters was estimated using either Chl-a (k = 15 M(-1) s(-1)) or fluorescence particle count (k = 38 M(-1) s(-1)) as an indicator of cell damage following colonial disaggregation (i.e., at [Cl2]0:DOC0 ratio ≥0.15). Complete release of intracellular microcystin-LR (MC-LR) was observed in both Lake Havasu and Lake Mead samples when applying a [Cl2]0:DOC0 ratio of 0.30 (t = 20 min), which was equivalent to a chlorine exposure of 8 min-mg/L for Lake Havasu samples. With chlorination, DOC increased by 3-18% indicating release of either colony-bound or cell-bound DOC. The results demonstrated the ability of chlorine to disaggregate/inactivate natural Microcystis colonies, and identified oxidation conditions resulting in complete release of intracellular MC-LR.
Keywords: Chlorine; Cyanobacteria; Digital flow cytometry; Microcystin-LR; Naturally occurring Microcystis; Sheath.
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