Flow Cytometric Analysis of Freshwater Cyanobacteria: A Case Study

Abstract

Eutrophication is a process that occurs due to the excessive accumulation of nutrients, primarily nitrogen and phosphorus, from natural and anthropogenic sources. This phenomenon causes cyanobacterial overgrowth, which over time leads to cyanobacterial harmful algal blooms (CHABs) that affect public drinking water sources and water sites with recreational usage. The rapid detection of bloom-forming cyanobacteria in freshwater bodies is critical in order to implement prevention strategies. Cyanobacteria contain phycobiliproteins such as phycoerythrin and allophycocyanin as part of the phycobilisome that allows autofluorescence. In this study, samples from 36 freshwater bodies in 14 New Jersey counties were collected and analyzed using flow cytometry with forward-scatter phycoerythrin and allophycocyanin parameters. Pure cultures of Synechococcus sp. IU 625, Cylindrospermum spp. and Microcystis aeruginosa were used as references. The results revealed that 17 out of the 36 analyzed sites contained all three references and related species. Seven sites showed Microcystis and Cylindrospermum-like species, while four sites indicated Microcystis and Cylindrospermum-like species. Six water bodies showed Cylindrospermum-like species, and two sites showed Microcystis-like species. Polymerase chain reaction (PCR)-based assays further confirmed the flow cytometric results. The findings from this study suggest that flow cytometry could potentially serve as a rapid method for freshwater cyanobacteria detection and screening.

Keywords

• cyanobacteria
• flow cytometry
• cyanobacterial harmful algal blooms (CHABs)