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DEVELOPMENT OF A SANDWICH HYBRIDIZATION ASSAY FOR THE HARMFUL CYANOBACTERIUM MICROCYSTIS AERUGINOSA (E)

Cyanobacteria are responsible for the largest number of harmful algal blooms (HABs) worldwide. The genus Microcystis is a major contributor to these HABs, and species such as M. aeruginosa, produce the hepatotoxin microcystin that affects humans, pets, and wildlife through direct bloom contact, or consumption of contaminated drinking water. Stormwater detention ponds, such as those found along the South Carolina (SC) coast, often harbor M. aeruginosa blooms, and their close proximity to residential and recreational areas pose threats to public health. Molecular techniques enable rapid and accurate species identification and quantification of HABs and other plankton, thereby facilitating ‘early warnings’ of blooms and management responses that safeguard public health. Sandwich hybridization assay (SHA), the technique considered here, directly (e.g. no amplification of genetic material) identifies and quantifies planktonic species using ribosomal RNA (rRNA). SHA utilizes two DNA probes; a capture and signal probe that bind to the targeted rRNA sequence creating a “sandwich” wherein the results are measured by absorbance. SHA for M. aeruginosa was developed using 16S rRNA from a regional strain and validated using cultures and local (SC) isolates. Bloom samples spanned April 29-July 26, 2016, and multiple toxigenic, cyanobacteria species, including, M. aeruginosa were observed in succession. This presentation describes assay refinement, environmental sampling, and the potential influences of nutrient form on SHA responses.

Authors

Dearth , N., Belle W. Baruch Institute for Marine and Coastal Sciences, University of South Carolina, USA, njudy@email.sc.edu

Jones , W. J., Arnold School of Public Health, University of South Carolina , USA, w.joe.jones@gmail.com

Mortensen , R., Belle W. Baruch Institute for Marine and Coastal Sciences, University of South Carolina, USA, bec.mortensen@gmail.com

Doll , C., Marine Resources Research Institute, South Carolina Department of Natural Resources, USA, dollc@dnr.sc.gov

Pinckney , J. L., Department of Biological Sciences, University of South Carolina , USA, pinckney@sc.edu

Greenfield, D. I., Belle W. Baruch Institute for Marine and Coastal Sciences, University of South Carolina , USA, dgreenfield@belle.baruch.sc.edu

Details

Oral presentation

Session #:021
Date: 03/01/2017
Time: 12:30
Location: 323 B

Presentation is given by student: Yes