Table Of ContentA SYMPOSIUM ON THE USE OF FISH AS AN
EXPERIMENTAL ANIMAL IN BASIC RESEARCH
SPONSORED BY THE UNIVERSITY OF SOUTH DAKOTA
VERMILLION, SOUTH DAKOTA
NOVEMBER 15-16, 1968
FISH
IN
RESEARCH
EDITED BY
OTTO W. IMEUHAUS
DEPARTMENT OF BIOCHEMISTRY
THE UNIVERSITY OF SOUTH DAKOTA
VERMILLION, SOUTH DAKOTA
JOHN E. HALVER
WESTERN FISH NUTRITION LABORATORY
U.S. BUREAU OF SPORT FISHERIES AND WILDLIFE
COOK. WASHINGTON
®
ACADEMIC PRESS
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INVITED SPEAKERS
Ashley, Laurence M., Western Fish Nutrition Laboratory, U. S. Fish and
Wildlife Service, Cook, Washington.
Bilinski, E., Vancouver Laboratory, Fisheries Research Board of Canada,
Vancouver, British Columbia, Canada.
Conte, Frank P., Department of Zoology, Oregon State University, Corvallis,
Oregon.
Gall, Graham A. E., Agriculture Experiment Station, College of Agriculture,
Department of Animal Science, University of California, Davis, California.
Goldberg, Erwin, Department of Biological Sciences, Northwestern
University, Evanston, Illinois.
Halver, John E., Western Fish Nutrition Laboratory, U. S. Bureau of Sport
Fisheries and Wildlife, Cook, Washington.
Hastings, W. H, Fish Farming Experimental Station, U. S. Fish and Wildlife
Service, Stuttgart, Arkansas.
Idler, D. R., Halifax Laboratory, Fisheries Research Board of Canada,
Halifax, Nova Scotia, Canada.
Mertz, Edwin T., Department of Biochemistry, Purdue University, Lafayette,
Indiana.
Ridgway, George /., Biological Laboratory, U. S. Fish and Wildlife Service,
West Boothbay Harbor, Maine.
Scarpelli, D. G., Department of Pathology and Oncology, University of
Kansas Medical Center, Kansas City, Kansas.
Sinnhuber, R. 0., Department of Food Science, Oregon State University,
Corvallis, Oregon.
Siperstein, M. D., Department of Internal Medicine, Southwestern Medical
School, University of Texas, Dallas, Texas.
v
INVITED SPEAKERS
Tarr, H. L. A., Vancouver Laboratory, Fisheries Research Board of Canada,
Vancouver, British Columbia, Canada.
Wellings, S. R. Department of Pathology, University of Oregon Medical
School, Portland, Oregon.
Zaugg, W. S., Western Fish Nutrition Laboratory, U. S. Fish and Wildlife
Service, Cook, Washington.
vi
PREFACE
This volume is comprised of papers presented at a symposium entitled
"Fish in Research" sponsored by the University of South Dakota in Vermil-
lion. The purpose of the symposium was to ask those directly involved in re
search on fish, "What unique information of biochemical and physiological
processes can be gained by using fish as experimental animals?" Discussions
at the meeting indicated this approach to be of great interest, but never
adequately considered. We sincerely hope that the unique question explored
will stimulate future meetings and other volumes of proceedings dealing
with the use of fish in research.
We wish to express thanks to the Bureau of Sport Fisheries and Wildlife
of the U. S. Department of the Interior for its interest and participation.
Special thanks are due Mr. John S. Gottschalk, Director, Bureau of Sport
Fisheries and Wildlife, for addressing the guests of the symposium at a
special banquet. His subject was "Basic Research Programs in the Bureau of
Sport Fisheries and Wildlife Using Fish as Experimental Animals." Also we
thank Clarence Johnson, Western Fish Nutrition Laboratory, Cook,
Washington, Bruno von Limbach, Fish Genetics Laboratory, Beulah,
Wyoming, and Dr. Norman Benson, North Central Reservoir Investigations,
Yankton, South Dakota, for their participation in the displays of fish.
Our sincere appreciation is extended to the president of the University of
South Dakota, first Dr. Edward Q. Moulton and now Dr. Richard Bowen,
and the Dean of the School of Medicine, Dr. George W. Knabe, Jr., for the
support and sponsorship of this venture. Special thanks are due to Mrs.
Dorothy E. Neuhaus for her extensive help in organization, public relations,
and undaunted efforts to ascertain that all went well. We wish to thank Mrs.
Myla Kampshoff for preparing the manuscripts and the South Dakota
Geological Survey for the use of the IBM Magnetic Tape Selectric Composer
system. Finally, we are grateful to the faculty of the Department of
Biochemistry and our many friends for their unstinting help.
Otto W. Neuhaus
John E. Halver
September, 1969
vn
WELCOMING ADDRESS
Otto W. Neuhaus
Sometimes scientists in the Middle West express the feeling that
nonmetropolitan areas are, scientifically speaking, hinterlands. I heard this
expressed at a major institution not long ago. In my opinion, it would be
better to speak of scientifically underdeveloped regions because then we are
challenged to do something about it. The present symposium is an effort on
our part to meet this challenge and to enhance the scientific climate in our
institution and the State of South Dakota. For once we are priviledged to
perform the duty of being host. We may not be expert at the job, but we
sincerely hope that you will find this weekend to be pleasant, informative,
and especially scientifically provocative.
Another purpose of this symposium is to challenge us, the faculty of the
Department of Biochemistry, to consider seriously the potentials of using
fish as experimental animals in anticipation of the possibility that research in
this area may lead to opportunities for expansion of the biological sciences
at this university.
I am deeply indebted to Dr. John E. Halver, director of the Western Fish
Nutrition Laboratory, and his staff, especially Clarence Johnson, for their
help in arranging part of this program and especially for bringing to
Vermillion fascinating displays of live fish as well as their movies. We hope
that you will take the time to visit these displays.
This symposium is purposely intended to be unique not only for us in the
field of biochemistry, but also for you who are active in the field of fish
research. We know that you have many meetings and symposia in which to
report your studies on fish; therefore, this in itself is not unique. The
challenge of the present symposium is not so much to review what is known
concerning fish, but to determine how studies on fish can yield unique
insights into biochemical and physiological phenomena.
In choosing this program we as a department are very much fish out of
water. Only one of our staff is actually using fish in his research activities. In
this instance, studies of collagen structure, formerly restricted to rat tail
tendon, are now being expanded to include fish skin. Otherwise, our research
activities center around the use of rats, frogs, Ascaris lumbricoides, and
algae. As biochemists, we are not really interested in the welfare of rats and
frogs or in learning more about algae and invertebrates. Indeed, each simply
serves as a model system for the study of specific biochemical mechanisms.
In other words, the primary motivation in biochemical research is to gain an
xi
WELCOMING ADDRESS
understanding of chemical mechanisms of life processes. The exact model
system or experimental organism used is often secondary.
I have asked myself many times why fish have not been used more
extensively for studies of biochemical mechanisms and, in fact, are we really
missing a good opportunity? The capacity of fish to adapt to their
environment suggests numerous opportunities to study metabolic control
mechanisms. For example, what is the sequence of events that allows the
normally ammonotelic lung fish to become ureotelic when cut off from
water? When the normal environment is restored, how do they become
ammonotelic again? What mechanisms control salt balance as a fresh water
adapted salmon enters its marine environment? How can trout perform
their metabolic activities in a 10°C environment whereas in warm blooded
animals metabolism functions best at 37°? Perhaps by using fish we could
learn more about metabolic control mechanisms and the mechanism of
action of hormones. Could we learn more about the mechanisms of enzyme
action and the protein structure required for specific catalytic activities by
using fish enzymes? Perhaps there are subtle differences in primary structure
or conformation of enzyme molecules from fish adapted to low
temperatures compared with the same enzymes of warm blooded animals.
The central theme of this symposium is best expressed in the deliberate
choice of our title. Not "Research on Fish," but "Fish in Research." Let the
central challenging question be, "What special insights can be gained in
biochemical and physiological mechanisms using fish as an experimental
animal?"
xn
ENVIRONMENTAL ASPECTS OF NEOPLASIA IN FISHES
S. R. Wellings
I. Introduction 3
II. Hepatoma 4
III. Thyroid Hyperplasia and Neoplasia 5
IV. Epidermal Papillomas 6
V. Mesenchymal Tumors of Goldfish 8
VI. Spindle Cell Sarcomas of Pike-Perch 8
VII. Multiple Osteomas of the Red Tai 8
Vili. Lympho sarco mas of Esox 9
DC. Experimentally Induced Chordomas in Embryo Teleosts 9
X. Undifferentiated Sarcomas in Lebistes and Pristella 9
XI. Melanomas of Swordtail—Platyfìsh Hybrids 10
XII. Multiple Nerve Sheath Tumors 10
XIII. Teratomas Associated with Ichthyophonus Infestation 10
XIV. Discussion 11
XV. Conclusion 13
XVI. Acknowledgments 13
XVII. References 13
XVIII. Comments 16
INTRODUCTION
In the physical, chemical, and biological evolution of the earth, numerous
naturally occurring tumorigenic agents were initially present, or
subsequently appeared in soil and water. These natural agents include various
heavy metals, inorganic compounds, and hydrocarbons, in addition to
radioactivity, ultraviolet light, and tumorigenic parasites. The exponential
3
S. R. WELLINGS
acceleration of human cultural evolution, beginning perhaps with the
controlled use of fire, has introduced further carcinogenic agents: new and
unique chemicals, increased concentrations of radioactive substances, various
hydrocarbon and other atmospheric contaminants, and insecticides and
herbicides with carcinogenic properties. In addition, man has modified the
spatial distribution of various carcinogens by a variety of concentration and
transportation procedures. The future promises further contamination of the
environment by a multiplicity of agents, especially chemical ones, of
unknown potential. The general problem of environmental neoplasia has
been recently reviewed (1).
The study of environmental neoplasia in animals and plants becomes of
considerable importance as a means of recognizing the environmental
presence of unsuspected new carcinogens, or increased concentrations of
known carcinogens, before these agents damage the various populations of
living organisms, including man and his food sources.
In the interest of this conference, this review will point out some
environmental aspects of neoplasia in fishes. Some examples are clearly
related to human modifications of the environment, whereas other examples
are not. In this context it should be pointed out that naturally occurring
phenomena of any kind are best studied in detail before human-generated
modifications ensue. In the examples which follow, this ideal was generally
not attainable; the phenomena discussed are nonetheless of considerable
biological and practical interest.
HEPATOMA
The most remarkable example of neoplasia in fish resulting from human
activity is the appearance in 1960 of hepatomas in epidemic proportions in
hatchery-reared rainbow trout, Salmo gairdneri, at various localities in the
United States and Europe (2). Hepatomas were also observed in cutthroat
trout (S. clarki) and brown trout (S. trutta).
The appearance of the disease in epidemic proportions followed the
introduction of dry, pelleted commercial feeds as a primary dietary source in
private, state, and federal hatcheries. This and other circumstantial evidence
led to fractionation of known hepatomagenic diets, culminating in the
isolation of a group of four potent chemically related, lipid soluble
carcinogenic agents, named aflatoxins Bi, B2, Gi, and G2. These
compounds are synthesized by certain mutants of A spergillus flavus, growing
in vegetable dietary constituents, such as cottonseed or peanut meal
products, stored under damp conditions.
Experimental studies revealed that rainbow trout fed crude aflatoxin at
0.8 parts per million of diet developed classical multinodular hepatomas. The
feeding of 1 part per billion of purified aflatoxin Bi produced hepatomas in
a high percentage of fish. Careful attention to storage of dietary constituents
resulted in disappearance of the problem in affected hatcheries. The lesson
to be learned from this is obvious: modification of the content or methods
of preparation of human and animal foodstuffs without testing of the final
chemical constituents may result in the introduction into the diet of
unsuspected hazardous carcinogenic substances. In man in particular, the
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