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Applied and Environmental Microbiology, September 2003, p. 5492-5502, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5492-5502.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Development and Application of a Monoclonal-Antibody Technique for Counting Aureococcus anophagefferens, an Alga Causing Recurrent Brown Tides in the Mid-Atlantic United States

David A. Caron,^1* Mark R. Dennett,² Dawn M. Moran,² Rebecca A. Schaffner,¹ Darcy J. Lonsdale,³ Christopher J. Gobler,⁴ Robert Nuzzi,⁵ and Tim I. McLean¹

Department of Biological Sciences, University of Southern California, Los Angeles, California 90089-0371,¹ Biology Department, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543,² Marine Sciences Research Center, State University of New York at Stony Brook, Stony Brook, New York 11794-5000,³ Natural Science Division, Southampton College, Long Island University, Southampton, New York 11968,⁴ Bureau of Marine Resources, Suffolk County Department of Health, Riverhead, New York 11901⁵

Received 9 January 2003/ Accepted 24 June 2003

A method was developed for the rapid detection and enumeration of Aureococcus anophagefferens, the cause of harmful algal blooms called "brown tides" in estuaries of the Mid-Atlantic United States. The method employs a monoclonal antibody (MAb) and a colorimetric, enzyme-linked immunosorbent assay format. The MAb obtained exhibits high reactivity with A. anophagefferens and very low cross-reactivities with a phylogenetically diverse array of other protists and bacteria. Standard curves are constructed for each 96-well microtiter plate by using known amounts of a preserved culture of A. anophagefferens. This approach allows estimation of the abundance of the alga in natural samples. The MAb method was compared to an existing method that employs polyclonal antibodies and epifluorescence microscopy and to direct microscopic counts of A. anophagefferens in samples with high abundances of the alga. The MAb method provided increased quantitative accuracy and greatly reduced sample processing time. A spatial survey of several Long Island estuaries in May 2000 using this new approach documented a range of abundances of A. anophagefferens in these bays spanning nearly 3 orders of magnitude.

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* Corresponding author. Mailing address: Department of Biological Sciences, 3616 Trousdale Parkway, AHF 301, University of Southern California, Los Angeles, CA 90089-0371. Phone: (213) 740-0203. Fax: (213) 740-8123. E-mail: dcaron{at}usc.edu.

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Applied and Environmental Microbiology, September 2003, p. 5492-5502, Vol. 69, No. 9
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.9.5492-5502.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

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