DFO - Libra y MPO Biblio hèque III II I 11 1111 1204304311 1111 II Canadian Special Publication of Fisheries and Aquatic Sciences 44 einellme ---.211151m.yeee - 7,11r,r1 ft Perielin e—N Edited by E. Scherer QL 626 C314 no.44 c.2 1979 Government of Canada Gouvernement du Canada Fisheries and Oceans Pêches et Océans CANADIAN SPECIAL PUBLICATION OF FISHERIES AND AQUATIC SCIENCES 44 TOXICITY TESTS FOR FRESHWATER ORGANISMS Edited by E. SCHERER Department of Fisheries and Oceans Freshwater Institute Winnipeg, Manitoba R3T 2N6 DEPARTMENT OF FISHERIES AND OCEANS WINNIPEG, MANITOBA 1979 In addition to the Canadian Special Publications of Fisheries and Aquatic Sciences (formerly Miscellaneous Special Publications), the Department of Fisheries and Oceans publishes the Canadian Journal of Fisheries and Aquatic Sciences (formerly the Journal of the Fisheries Research Board of Canada) in annual volumes of monthly issues,and a Bulletin series. These publications are for sale from the Canadian Government Publishing Centre, Supply and Services Canada, Hull, Quebec, Canada KlA 059. Remittance must be made in advance, payable in Canadian funds, to the order of the Receiver General for Canada. THIS PUBLICATION IS AVAILABLE FREE FROM THE FRESHWATER INSTITUTE ED Minister of Supply and Services Canada 1979 Cat. No. Es 41-31/44 ISSN 0706-6481 Correct citation for this publication: Scherer, E. (ed.) 1979. Toxicity tests for freshwater organisms. Can. Spec. Publ. Fish. Aquat. Sci. 44: 194 p. Cover design by Don i A. Taite CONTENTS Page Preface Acknowledgements ix Abstract Résumé DE MARCH, B.G.E.: Reproduction and growth impairment tests with the amphipod Hyalella azteca . ..... 1 EVANS, R.E., and T.J. MARA: A histochemical test for olfactory function in fish 11 FLANNAGAN, J.F., and D.G. COBB: The use of the snail Helisoma trivolvis in a multigeneration mortality, growth and fecundity test 19 FRIESEN, M.K.: Use of eggs of the burrowing mayfly Hexagenia rigida in toxicity testing . .... 27 GILES, M.A., and D. KLAPRAT: The residual oxygen test: a rapid method for estimating the acute-lethal toxicity of aquatic contaminants . ..... 37 HARA, T.J.: An electrophysiological test for neuro- toxicity in fish ......... 46 HEALEY, F.P.: The use of growth rate for testing toxicant effects on algae 57 LAWRENCE, S.G., and M.H. HOLOKA: A microbial predator-prey test system 71 LEONHARD, S.L.: Tests for the crayfish Orconectes virilis 82 LEONHARD, S.L.: Effects on survival, growth, and reproduc- tion of Daphnia magna 91 LILLIE, W.R., S.E. HARRISON, W.A. MACDONALD, and J.F. KLAVERKAMP: The use of the zebrafish (Brachydanio rerio) in whole-life-cycle tests . .... 104 LOCKHART, W.L., and A.P. BLOUW: Phytotoxicity tests using the duckweed Lemna minor 112 iv LOCKHART, W.L., and D.A. METNER: Biochemical tests for fish 119 MACDONALD, W.A.: Testing embryonic and larval life stages of fish 131 MAJEWSKI, H.S., J.F. KLAVERKAMP, and M.A. GILES: Testing effects on cardiovascular and respiratory functions in fish 139 MALLEY, D.F., and L.J. TINKER: Calcium uptake: a sublethal test for crayfish 150 SCHERER, E.: Avoidance testing for fish and invertebrates 160 SCHERER, E.: Testing responsiveness to overhead-light stimulation 171 SCHERER, E., and S.E. HARRISON: The optomotor response test 179 SCHERER, E., S.H. NOWAK, S.E. HARRISON, and D.L. KRIPIAKEVICH: Testing locomotor activity responses with ultrasonic beams 186 PREFACE In Canada as elsewhere, hardly a decade has passed since ecolo- gists saw their young, if not obscure science suddenly pushed to center stage. Pollution was the cue, DDT and mercury emerged as villains, dramatically shaping and changing public consciousness. Identifying and describing the environmental actions of some of the 50,000 or so most common man-made substances (an estimated 1,000 or more being added every year) has been an ongoing effort ever since. Of course, the ultimate goal is not description and documentation of existing problems, but prevention or reduction of future environ- mental deterioration. To this end, new legal tools were devised in most industrialized countries. In Canada, the Environmental Contami- nants Act of 1976 is the general framework designed to restrict, and prevent from entering the environment, chemical substances of a kind and quantity that "cause biological change". Since 1977, amendments to Canada's Fisheries Act have been aimed specifically at protecting aquatic habitats. Enforcement of these well-intentioned laws, however, requires the capacity to evaluate and predict the kinds and extent of harmful changes a chemical might cause upon entering air, soil or water at various concentrations and under various biological, chemical and physical background conditions. Unfortunately, this capacity is still very limited. Already the early alerting experiences with chlorinated hydro- carbons and mercury had taught the lesson that conventional acute- vi lethal "bioassays" could not be relied upon to foresee and avert environmental danger. Sudden, easily recognizable kill-offs did not constitute the challenge, but inconspicuous, slow, gradual derangements of functions at all levels of biological organization, from biochemistry to behaviour of a species, and from bacterium to vertebrate predator. In addition to basic information on physical and chemical properties of existing and new compounds (e.g. solubility, adsorption/desorption, persistency), a new, much more comprehensive kind of toxicity testing was needed - and still is. In 1971, Dr. R.D. Hamilton of the Freshwater Institute chaired a federal task force that investigated the state of toxicity testing vis à vis the new environmental laws being planned at that time. Their conclusion, valid then and now: enforcement would be limited due to lack of sound testing procedures. While no single country has surged significantly ahead in eco- toxicological methodology, some international organizations such as the International Organization for Standardization (ISO), the Food and Agriculture Organization of the United Nations (FAO), and the Organization for Economic Co-operation and Development (OECD), have begun to work jointly with member nations towards incorporating indi- vidual tests into standardized, tiered (step-sequence) testing proce- dures ("protocols"). There are strong political and economical reasons for standardized tests, and for cost-saving tiered testing. The problem is - and may well remain so for a number of years - to put the right kind and number of tests into the right kind of tiered system to predict hazard at an acceptable level of certainty, at least for a given class vi i of chemicals in a given environmental "compartment". In 1972, the Freshwater Institute, housing a large group of aquatic biologists with a wide range of expertise, added toxicity test development to its research activities. This publication sum- marizes what has been achieved so far. Considering the state of the art, we chose not to present our 20 tests in some form of tiered system. Rather, they are described as discrete elements, to be used independently or in any desired com- bination. The sections called "Examples of Application" and "Discus- sion" with literature references will give the reader an indication of the test's state of development; "Costs and Staff Requirements" contain estimates for specifically needed items, in 1979 Canadian dollars, with basic laboratory supplies and equipment including toxi- cant delivery systems usually assumed to be available. As the reference sections will show, some of the tests described are based on previously published material, reviewed and rewritten to fit a user-oriented common format. Others are proposed for the first time. All should be considered guidelines or proposals rather than cut-and-dry standard recipes. The authors are only too aware of the gap between test development at the research stage and the testing of tests in routine use by regulatory agencies. In fact, trying to bridge this gap has been a major motive for this publication. Accor- dingly, readers and potential users are invited to contact the editor and individual authors for changes and improvements, expanding on procedural details, etc. This publication can only be a start. Any future editions would benefit, I believe, not only from critical outside viii comments, but also from external authorship under an enlarged editorial panel, thus pooling and focusing the somewhat scattered Canadian testing expertise. Tests dealing with invertebrates frequently refer to a "Manual for the Culture of Selected Freshwater Invertebrates" edited by S.G. Lawrence, planned to be published shortly in the same "Special Publi- cation" series as this volume. Further, while compiling and editing material for these toxicity tests, a comprehensive bibliography on testing methods was assembled; this will also be available (probably in this series) within the next few months. E. Scherer Winnipeg, November 1979 ix ACKNOWLEDGEMENTS First and foremost, this publication was only possible through the non-abating cooperation of 24 authors, going diligently through the various stages, from collating and reviewing material over adopting a common format to several revisions and final drafts. Drs. R.D. Hamilton and J.F. Klaverkamp, former heads of the F.W.I. Toxicology Section, provided much encouragement and helpful discussions, from pre- ceding research to write-up. Miss M.P. McLean, later joined by Mr. R.E. McNicol, assisted most commendably in many editorial aspects including proofreading and reference and symbol standardization, with technical advice by Mrs. M. Layton and Mr. K.E. Marshall. Reviews by Drs. G.B. Ayles, W.G. Franzin, J.A. Mathias and Mr. D.G. Wright led to many sub- stantial improvements. Drawings, except those previously published, were done by Mr. and Mrs. L.D. Taite. Certainly not least: Mrs. G.M. Decterow transcribed some of the early, at times not very legible, handwritten outlines, notes and comments; Miss M.D. Ziprick typed in a most reliable and efficient manner all the seemingly never-ending drafts without, miraculously, ever losing her patience or friendliness.