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Craig A. Harms,¹ Christopher A. Ottinger,² Vicki S. Blazer,² Christine L. Densmore,² Laurence H. Pieper,³ and Suzanne Kennedy-Stoskopf⁴

¹Environmental Medicine Consortium and Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, North Carolina, USA
²U.S. Geological Survey, Biological Resources Division, National Fish Health Research Laboratory, Kearneysville, West Virginia, USA
³Maryland Department of Natural Resources, Stevensville, Maryland, USA
⁴Environmental Medicine Consortium and Department of Microbiology, Pathology and Parasitology, North Carolina State University, Raleigh, North Carolina, USA

Abstract

Fish morbidity and mortality events in Chesapeake Bay tributaries have aroused concern over the health of this important aquatic ecosystem. We applied a recently described method for quantifying mRNA of an immunosuppressive cytokine, transforming growth factor-ß (TGF-ß) , by reverse transcription quantitative-competitive polymerase chain reaction to a field study of fish health in the Chesapeake Basin, and compared the results to those of a traditional cellular immunoassay macrophage bactericidal activity. We selected the white perch (Morone americana) as the sentinel fish species because of its abundance at all of the collection sites. White perch were sampled from Chesapeake Bay tributaries in June, August, and October 1998. Splenic mononuclear cell TGF-ß mRNA levels increased and anterior kidney macrophage bactericidal activity decreased, particularly in eastern shore tributaries, from June to August and October. The results of the two assays correlated inversely (Kendall's b = -0.600 ; p = 0.0102) . The results indicated both temporal and spatial modulation of white perch immune systems in the Chesapeake Basin, and demonstrated the utility of quantitative PCR for TGF-ß as a molecular biomarker for field assessment of teleost fish immune status. Key words: Chesapeake Bay, field study, macrophage bactericidal activity, Morone americana, quantitative PCR, transforming growth factor-ß, white perch. Environ Health Perspect 108:447-452 (2000) . [Online 28 March 2000]

http://ehpnet1.niehs.nih.gov/docs/2000/108p447-452harms/ abstract.html

Address correspondence to C.A. Harms, Depart-ment of Clinical Sciences, North Carolina State University, 4700 Hillsborough Street, Raleigh, NC 27606 USA. Telephone: (919) 515-8112. Fax: (919) 515-4237. E-mail: craig_harms@ncsu.edu

We thank T. Childers for technical assistance, and C. Weedon and the Maryland Department of Natural Resources for fish collection and site characterization.

Funding sources include a U.S. EPA Science to Achieve Results Fellowship (U-915209-01-0) , a NCSU College of Veterinary Medicine State Research grant, and the USGS Place-Based Studies Program.

Received 26 July 1999 ; accepted 7 December 1999.