Date of Award

Spring 5-14-2016

Degree Type

Thesis

Degree Name

MS Microbiology

Department

Biology

Advisor

Tin-Chun Chu, Ph.D

Committee Member

Daniel B. Nichols, Ph.D

Committee Member

Constantine Bitsaktsis, Ph.D

Committee Member

Angela V. Klaus, Ph.D

Committee Member

Jane L. Ko, Ph.D

Keywords

flow cytometry, cyanobacteria, algal bloom, freshwater, cyanotoxin

Abstract

Eutrophication, a process that occurs due to excessive accumulation of nutrients such as nitrogen and phosphorous is primarily from anthropogenic nitrogen and natural occurrences. This phenomenon causes cyanobacterial overgrowth which can potentially lead to toxic algal blooms that affect public drinking water sources and recreational usage. An immediate need to detect bloom forming cyanobacteria in freshwater bodies early hand is critical to implement prevention strategies. These microorganisms contain phycobiliproteins such as phycoerytrhin, and allophycocyanin as part of the phycobillisome that allow autofluorescence. In this study, 36 freshwater bodies from 14 New Jersey counties were collected and processed for flow cytometric analysis for forward- scatter, phcyoerthrin and allophycocyanin parameters. Pure cultures of Synechococcus sp. IU 625 (S. IU 625), Cylindrospermum spp. and Microcystis aeruginosa (M. aeruginosa) were utilized as references. Polymerase chain reaction (PCR)-based assay was performed for the validation of the flow cytometric analysis. The results revealed 17 out of the 36 sites showed all three reference species and their related. 6 waterbodies showed Cylindrospermum like species, 7 waterbodies showed Microcystis and Cylindrospermum like species, 4 waterbodies showed Microcystis and Cylindrospermum like species and 2 sites showed Microcystis like species. PCR results validated these results by showing positive results for phytoplankton, cyanobacteria and Synechococcus. Flow cytometry has high potential for the rapid detection of cyanobacteria in live form due to their autofluorescence properties from the phycobilisome.

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