Biomonitoring of Environmental Status and Trends (BEST) Program: Selected Methods for Monitoring Chemical Contaminants and their Effects in Aquatic Ecosystems Cl Cl Cl O Cl Cl Cl Cl O Cl O (a) 2,3,7,8-Tetrachlorodibenzo- p-dioxin (b) 2,3,7,8-Tetrachlorodibenzofuran Cl Cl Cl Cl Cl (c) 3,3´,4,4´,5-Pentachlorobiphenyl (PCB 126) (d) Benzo[a]pyrene Information and Technology Report USGS/BRD/ITR--2000-0005 U.S. Department of the Interior U.S. Geological Survey Biological Resources Division Technical Report Series The Biological Resources Division publishes scientific and technical articles and reports resulting from the research per- formed by our scientists and partners. These articles appear in professional journals around the world. Reports are published in two report series: Biological Science Reports and Information and Technology Reports. Series Descriptions Biological Science Reports Information and Technology Reports ISSN 1081-292X ISSN 1081-2911 This series records the significant findings resulting from These reports are intended for publication of book-length sponsored and co-sponsored research programs. They monographs; synthesis documents; compilations of con- may include extensive data or theoretical analyses. ference and workshop papers; important planning and Papers in this series are held to the same peer-review and reference materials such as strategic plans, standard high-quality standards as their journal counterparts. operating procedures, protocols, handbooks, and manu- als; and data compilations such as tables and bibliogra- phies. Papers in this series are held to the same peer- review and high-quality standards as their journal coun- terparts. Copies of this publication are available from the National Technical Information Service, 5285 Port Royal Road, Springfield, Virginia 22161 (1-800-553-6847 or 703-487-4650). Copies also are available to registered users from the Defense Technical Information Center, Attn.: Help Desk, 8725 Kingman Road, Suite 0944, Fort Belvoir, Virginia 22060- 6218 (1-800-225-3842 or 703-767-9050). Printed on recycled paper Biomonitoring of Environmental Status and Trends (BEST) Program: Selected Methods for Monitoring Chemical Contaminants and their Effects in Aquatic Ecosystems Editors : 1 Christopher J. Schmitt 2 Gail M. Dethloff Contributors (in alphabetical order): 3 Vicki S. Blazer 4 Nancy D. Denslow 2 Gail M. Dethloff 5 Timothy S. Gross 5 Kelly K. McDonald 1 Christopher J. Schmitt 1 Donald E. Tillitt 6 Jeff J. Whyte 1 U.S. Geological Survey, Columbia Environmental Research Center, Columbia, MO 2 AScI, c/o U.S. Geological Survey, Columbia Environmental Research Center, Columbia, MO 3 U.S. Geological Survey, Leetown Science Center, National Fish Health Laboratory, Leetown, WV 4 University of Florida, Department of Biochemistry, Gainesville, FL 5 U.S. Geological Survey, Florida-Caribbean Science Center, Gainesville, FL 6 University of Missouri, Department of Fisheries and Wildlife, Columbia, MO Suggested citation: Schmitt, C. J. and G. M. Dethloff. editors. 2000. Biomonitoring of Environmental Status and Trends (BEST) Program: selected methods for monitoring chemical contaminants and their effects in aquatic ecosystems. U.S. Geological Survey, Biological Resources Division, Columbia, (MO): Information and Technology Report USGS/BRD-2000--0005. 81 pp. CONTENTS ABSTRACT...............................................................................................................................................................iv PREFACE.................................................................................................................................................................vi THE 1995 AND 1997 BEST PROJECTS. C.J. Schmitt, D.E. Tillitt, V.S. Blazer..........................................................1 SITE SELECTION..........................................................................................................................................1 ORGANISMS SELECTED FOR SAMPLING.........................................................................................................2 METHODS EVALUATED................................................................................................................................3 EROD ACTIVITY. J.J. Whyte, D.E. Tillitt...............................................................................................................5 Background ..............................................................................................................................................5 Performing the EROD Assay....................................................................................................................7 Factors That Can Afftect EROD Induction in Fish...................................................................................7 Value and Utility of EROD in the BEST Program....................................................................................8 H4IIE BIOASSAY. J.J. Whyte, D.E. Tillitt...............................................................................................................9 Background ..............................................................................................................................................9 Performing the H4IIE Bioassay...............................................................................................................10 Factors That Can Affect the H4IIE Bioassay...........................................................................................10 Value and Utility of the H4IIE Bioassay in the BEST Program ..............................................................12 CONDITION FACTOR AND ORGANO-SOMATIC INDICES. G.M. Dethloff, C.J. Schmitt............................................13 Background ............................................................................................................................................13 Measuring Condition Factor and Organo-somatic Indices......................................................................13 Factors That Can Affect Condition Factor and Organo-somatic Indices.................................................13 Value and Utility of the Condition Factor and Organo-somatic Indices in the BEST Program...............17 THE NECROPSY-BASED FISH HEALTH ASSESSMENT. V.S. Blazer.........................................................................18 Background.............................................................................................................................................18 Performing the Necropsy-Based Fish Health Assessment......................................................................18 Factors That Can Affect the Necropsy-Based Fish Health Assessment..................................................18 Value and Utility of the Necropsy-Based Fish Health Assessment in the BEST Program......................22 HISTOPATHOLOGIC ASSESSMENT. V.S. Blazer........................................................................................................22 Background ............................................................................................................................................22 Performing Histopathlogical Analyses...................................................................................................23 Factors That Can Affect Histopathological Biomarkers.........................................................................23 Value and Utility of Histopathological Biomarkers in the BEST Program.............................................25 IMMUNE SYSTEM INDICATORS. V.S. Blazer, G.M. Dethloff..................................................................................25 SERUM/PLASMA LYSOZYME ACTIVITY...................................................................................................................26 Background.............................................................................................................................................26 Performing the Lysozyme Assay.............................................................................................................26 Factors That Can Affect Serum/Plasma Lysozyme Activity...................................................................27 i Value and Utility of Serum/Plasma Lysozyme Activity in the BEST Program.......................................27 MACROPHAGE AGGREGATES.................................................................................................................................27 Background.............................................................................................................................................28 Measuring the Macrophage Aggregate Parameters................................................................................28 Factors That Can Affect Macrophage Aggregates..................................................................................29 Value and Utility of Macrophage Aggregates in the BEST Program......................................................30 REPRODUCTIVE INDICATORS. K.K. McDonald, T.S. Gross, N.D. Denslow, V.S.Blazer........................................30 SEX STEROIDS......................................................................................................................................................31 Background.............................................................................................................................................31 Performing Sex Steroid