EPA 560/5-75-004 A ., in_c
April 1975
LABORATORY TEST
METHODS TO ASSESS
THE EFFECTS OF CHEMICALS
ON TERRESTRIAL
ANIMAL SPECIES
FINAL REPORT
Office of Toxic Substances
U.S. Environmental Protection Agency
Washington, D.C. 20460
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April 1975
LABORATORY TEST METHODS TO ASSESS
THE EFFECTS OF CHEMICALS ON TERRESTRIAL
ANIMAL SPECIES
Prepared by
Ryckman/Edgerley/Tomlinson & Assoc. Inc.
12161 Lackland Rd.
St. Louis, Missouri 63141
Prepared for
Office of Toxic Substances
U.S. Environmental Protection Agency
Washington, D.C. 20"460
Contract/Order No. 68-01-1896
Project Director
Mr. Frank Kover
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This report has been reviewed by the Office of Toxic
Substances, EPA, and approved for publication. Approval
does not signify that the contents necessarily reflect the
views and policies of the the Environmental Protection Agency,
nor does mention of trade names or commercial products
constitute endorsement or recommendation for use.
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TABLE OF CONTENTS
Page No.
Letter of Transmittal i
Title Page ii
Table of Contents iii
List of Tables vii
List of Figures ix
Abstract xii
Acknowledgments xiii
SECTION
I CONCLUSIONS AND RECOMMENDATIONS 1
II INTRODUCTION 2
Toxic Substances and the Environment 2
Toxicity Testing: An Historical
Perspective 2
Pertinent Legislation 3
Methodology Used in this Study 4
III SPECIAL PROBLEMS 5
Problems in Toxicity Testing 5
Variations in Experimental Design 5
Homeothermic vs. Poikilothermic 5
Sex, Weight and Age 5
Random Sampling Variation 6
Variations in Breeding and Holding
Environs 7
Genetic Variations 8
Non-Genetic Variations 8
Genetic Drift 9
Resistance 9
Mechanisms 9
DDT Resistance - A Case Study 11
Natural Selection 11
Testing of Dynamic Ecological
Associations 12
Experimental Design of Test vs. Cost 13
Wild Native Species vs. Domestic
Laboratory Species 14
Chemical Problems in Testing:
Degradation 15
111
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TABLE OF CONTENTS
(continued)
Page No,
Nonbiological Oxidation, Reduction
and Hydrolysis 15
Nonbiological Miscellaneous Reactions 16
Biological Oxidation, Reduction and
Hydrolysis . 16
Dosages 17
Biostatistics in Toxicological Testing 17
Introduction 17
Randomization 17
Sample Size 18
The Normal Distribution 18
Tests of Hypotheses 19
Linear Regression and Correlation 19
IV ANIMAL SUITABILITY 21
Introduction 21
Establishment of Animal Groupings 21
Animal Suitability Tables 22
Invertebrates 26
Helminths 26
Mollusca 29
Arachnida 31
Orthoptera 3 5
Hemiptera-Homoptera 38
Coleoptera 40
Lepidoptera 42
Diptera 45
Hymenoptera 50
Miscellaneous Invertebrates 52
Vertebrates 53
Amphibians and Reptiles 53
Anurana 55
Chelonia 62
Squamata 64
Birds 69
Pelecanidae and Phalacrocoracidae 73
Gaviiformes and Podicipediformes 75
Anseriformes 76
Procellariiformes 86
Ardeidae and Threskiornithidae 88
Gruidae and Rallidae 90
Scolopacidae 91
Lari 93
Picidae 95
IV
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TABLE OF CONTENTS
(continued)
Page No.
Falconiformes and Strigiformes .96
Columbiformes 100
Colinus virginianus 104
Phasianus colchicus 107
Miscellaneous Galliformes 109
Passeriformes 111
Domestic Galliformes 121
Galliformes: Coturnix coturnix
japonica 124
Exotics 129
Mammals 132
Insectivora 135
Chiroptera 140
Lagomorpha 143
Rodentia 148
Carnivora 176
Ungulates 187
Primata 205
Marsupiala 210
Edentata 213
Exotic Species 216
Suggested Species 219
Final List 219
Selection of Animal Species for Testing 221
V. METHODOLOGIES 225
Introduction 225
General Scientific Procedure 225
Selection of Categories 226
Physical/Chemical Tables 226
Physical/Chemical Methods 239
Ciliary Transport 239
Stress 239
Respiration 240
Enzymes 241
Anesthesia 242
Telemetry 242
Observe/Monitor 242
Population Dynamics 243
Chromatography . 244
Residues 245
Bioassay 245
Metabolism 246
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TABLE OF CONTENTS
(continued)
Page No.
Genetics 246
Anticholinesterases 247
Hepatic Studies 247
Endocrinology 248
Anatomy 248
Physiology 249
Histology 249
Pathology 250
Pharmacology 251
Hematology 251
Oology 252
Embryology 252
Other 253
Behavioral Methods 254
Introduction 254
Repellents 254
Discrimination 262
Stress 266
Pheromones 267
Neurophysiology 267
Other 268
Instinct 270
Learning 276
Suggested Methodologies 279
Introduction 279
Suggested Approaches to Toxicity
Testing 279
General Studies 283
The Selection of Methods for
Toxicity Testing 283
State-of-the-Art 284
The Future 285
VI. APPENDICES
A. Contacts and Consultants
B. List of Suppliers
C. Suggested Species Synopsis Sheets
VII. BIBLIOGRAPHY
VI
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LIST OF TABLES
Table No. Page No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
Suitability of Invertebrates for
Toxicological Testing
Composition of an Artificial Medium Used
for Rearing Noctuid Species
Suitability - Herpeto fauna
Food Recommended for Amphibian Larvae
Young Animals and Adults
Natural Breeding Habits of Common North
American and European Anura
Suitability of Birds for Toxicological
Testing
Distribution of Indigenous Breeding
Waterfowl Species by Zoogeographic
Regions
Incubation Periods of Various Waterfowl
Fledging Periods of Various Waterfowl
Some Longevity Records for Waterfowl
Suitability of Mammals for Toxicology
Testing
Composition of an Experimental Creep-
Feed
Breeding and Rearing Characteristics
of the Wild Rats
Components of the Standard Laboratory
Feed Used for Maintaining Microtus
ochrogaster in the Laboratory
Reproduction of M. ochrogaster in the
Laboratory at DirTerent Temperatures
25
43
54
57
58
70
77
81
82
84
133
145
153
164
165
Vll
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LIST OF TABLES
(continued)
Table No. Page No.
16 Reproduction of M. ochrogaster in the
Laboratory on DifTerent Light Schedules 165
17 Reproduction in Mustelidae Under Natural
Conditions 181
18 Growth Ration 193
19 Gestation Ration 194
20 Maintenance Ration 195
21 Artificial Diets for Baby Pigs 196
22 Final List of Suggested Species 219
23 List of Suggested Species from Selected
Terrestrial Animal Groups 220
24 Ecological Roles of Suggested Species 223
25 Methodology: Physiological 228
26 Methodology: Behavioral 255
27 Selected Approaches to Toxicity Testing 280
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LIST OF FIGURES
igure No. Page No.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
Web Building Enclosure
Mallard Test Chamber
Raptor Cages Converted into Mallard Cages
Mallard Cages (Base 4 •' x: 4 ' )
Barn Owl Cages of Chicken Wire and Wood,
50' x 6" x 10', with Nest Boxes as Shown
Nest Box for Barn Owl Cages
Mourning Dove Cage
Food and Water Distribution System for
Mourning Dove Cages
Bobwhite Quail Outdoor Holding Pens
(Base approximately 4' x 8')
Starling and Wild Bird Cages
Wild Bird Traps - Birds Trapped Include
Starlings, Sparrows, Robins and Crows
Coturnix Laying Cages (12" x 10" x 16")
Coturnix Incubation Chamber
Coturnix and Mourning Dove Testing
Cages (10" x 10" x 12")
Miniature Swine Transport Apparatus
Miniature Swine Observation Enclosures
(20" x 30" Floor Area)
Miniature Swine in Holding Pen
(8' x 12' Floor Area)
Deer Holding Pen
Primate Cages - Squirrel Monkeys -
32
78
78
79
97
97
101
101
105
117
117
125
125
126
190
190
191
202
Metal - Commercially Available 206
IX
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LIST OF FIGURES
(continued)
Figure No. Page
20 Plexiglas and Metal - Fabricated at
Iowa State 206
21 Plexiglas Cage, Fabricated at Iowa
State University 207
22 Trophic Triangle 222
23 The Interrelationships of Testing
Methodologies 238
24 Diagram of Visual Acuity Chamber 263
25 Cutaway of a Visual Cliff Used as a
Test Apparatus 264
26 Discriminatory Device Used in Primate
Testing : 265
27 Inside of Visual Discrimination Chamber
for Primate Testing 265
28 Wiring Diagram of Traffic Counter.
Mountain Beaver Burrow with Pictorial
Drawing of Burrow Probe in Place 269
29(a) Runways with Frightening Device at End 271
29(b) Runway Mechanism for Operating
Frightening Device 271
29(c) Runways, Frightening Device, and Camera
Used to Record Chick's Response 271
30(a) Enclosure for 10 Runways 272
30(b) Runway with Speaker at Far End 272
30(c) Holding Area - Point of Release 273
31(a) Examination Table Prior to Photographing
Web 274
31(b) Examination Table Prior to Photographing
Web 274
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LIST OF FIGURES
(continued)
Figure No. Page No.
32 Apparatus for Photographing Web 275
33 An Automated Method for Studying
Aggression in Primates 277
34 Succession of Toxic Response 282
XI
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ABSTRACT
This report presents a review of test species and method-
ologies utilized in toxicological research on terrestrial
animals. It involved an extensive survey of available
literature, interviews with toxicologists, physiologists
and ecologists, and visits to research facilities around
the country. The report contains reviews of each major
animal group and individual species where specific infor-
mation was available, that have been used as test subjects
in past or present toxicological research programs.
Suitability for use in such testing, both actual and
potential, and general information relative to laboratory
maintenance and possible alternative species is presented
for each group. Brief synopses of general characteristics
for each group and/or species are also included. Existing
methodologies used in toxicological research are dis-
cussed and evaluated for their particular applicability
to testing programs and general recommendations for
approaches to toxicity testing, test animals, and methods
are made.
This report was submitted in fulfillment of Project
Number , Contract Number 68-01-1896, by Ryckman/
Edgerley/Tomlinson and Associates, Inc. under the sponsor-
ship of the Environmental Protection Agency. Work was
completed as of April 1975.
Xll
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ACKNOWLEDGMENTS
1
The investigation, data handling and report preparation
were performed by RETA and consultants. Primary contribu-
tors were R. Matter, Project Scientist, W. Zegel, Project
Manager, E. Edgerley, Project Principal, L. Hathaway,
T. Anderson, K. Cherryholmes, R. Bessent, K. Schaffer,
T. Weyenberg, M. Kane, C. Teig and B. Skwiot. C. Hamlin,
P. Braden, E. Ratz and M. McReynolds are given special
acknowledgment.
A special debt of gratitude is owed to those scientists and
technicians from numerous university research teams and
private industries who gave of their time, ideas and
information.
Keith Long, Director of the Institute of Agricultural
Medicine, University of Iowa, who reviewed the draft docu-
ment and whose comments and interests were very beneficial
to the RETA staff is acknowledged with sincere thanks.
RETA extends thanks to the EPA, Washington, D. C. personnel
including M. Prival and F. Kover for their interest and
schedule considerations.
Xlll
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SECTION I
CONCLUSIONS AND RECOMMENDATIONS
Selection of test organisms should depend primarily on the
following criteria:
1. The trophic level at which the test substance is known
to effect a reaction, either directly or indirectly.
2. The general habitat in which the test substance is
used.
3. The availability of the organism and level of back-
ground information on the prospective test species.
4. The degree to which the test species represents a
trophic level and accommodates the testing methodologies
of the program.
Optimally, multispecies test programs in which a number of
trophic levels are represented should prod-ice results of
greater ecological significance. Such programs, though
more difficult to design, fund and manage than traditional
single species tests, will hopefully illustrate composite
effects of test substances on ecosystems instead of indi-
vidual species.
Toxic substances can only be termed detrimental if they
adversely affect the reproductivity of a species. There-
fore, methodologies used in toxicity testing should be
selected for their ability to show short or long-term
effects on the survival of the test species (e.g., genera-
tion studies at subacute dosages). If this approach is
followed, morbidity and functional studies, biochemical
analyses and residue determinations are significant only
to the extent that they aid in projecting or precisely
determining the eventual effects of the test substance on
the species' survival (short and/or long-term). Ultimately,
toxicity testing results should provide an indication of
the relative health of the ecosystem.
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SECTION II
INTRODUCTION
TOXIC SUBSTANCES AND THE ENVIRONMENT
This report reviews terrestrial animal species and existing
testing methodologies to assess their suitability for use
in toxicological testing. A toxic substance has been de-
fined as a man-made chemical or man-caused concentration of
chemicals which may come in contact with living organisms
other than man, either accidentally or purposely, on a
small or large scale. This definition has been used to
avoid the problematic aspects of the concept of toxicity.
For example, many naturally occurring compounds (certain
plant fluids and animal secretions) are toxic if ingested
or touched.
Toxicity is incremental, depending on such factors as the
concentration of the substance in question, the contact
organism and its immune systems, sensitivities, and toler-
ance levels, and on routes of exposure. Some compounds,
which are essential to living systems, are beneficial at
low concentrations and lethal at high concentrations (e.g.,
salts and trace elements). The effects of toxic chemicals
can also be very slight and only discernible after long
periods of time, even several generations. In wild animals
the level could be so low as to be unmeasurable, but still
affect the ability of the creature to survive in a natural
system.
A more thorough and sophisticated testing system is required
to overcome these difficulties in assessing toxic substances,
This report provides the basic data on selection of test
species and methods necessary to develop this system. It
.presents the judgments of the scientific community on the
suitability of certain animals and methods for use in toxi-
cological testing and, in that light, attempts to determine
the adequacy of present testing procedures.
TOXICITY TESTING: AN HISTORICAL PERSPECTIVE
Modern toxicology is considered to have begun with the work
of M. J. B. Orfila (1787-1853) who investigated toxic chemi-
cals and their effect on physiology. His two-volume trea-
tise served as a basic reference for many years. Since
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Orfila's time, the science of toxicology has grown steadily
and incorporated discoveries and technology from chemical
and biological disciplines.
A rapid evolution in the science of toxicology has occurred
in the past few decades. In addition to the older LD5Q and
percutaneous tests, teratogenic, carcinogenic, biochemical,
and behavioral methods have been introduced as more effi-
cient and effective ways of quantifying and defining tox-
icity. This evolution has been provoked primarily by the
increasing amount of chemicals introduced by man into the
environment.
PERTINENT LEGISLATION
Toxic substance control legislation deals primarily with
pesticide useage. The Federal Food, Drug and Cosmetic Act
of 1938 is the cornerstone of all present pesticide legis-
lation in the U.S. This act deals primarily with the con-
trols placed on useage of economic poisons in the natural
environment. In 1955, an amendment to the hct defined an
orderly procedure for establishing pesticide tolerance
levels and provided penalties for violations (1).
The welfare of animals used in biological research is con-
trolled by Public Law 89-544, enacted in 1966 (2), and its
amendment, Public Law 91-579 of 1970 (3). The purpose of
the original Act was to establish the Secretary of Agri-
culture's authority to license and regulate suppliers of
laboratory animals. The Act as amended in 1970 states:
The Secretary shall promulgate standards to govern
the humane handling, care, treatment, end transpor-
tation of animals by dealers, research facilities,
and exhibitors. Such standards shall include mini-
mum requirements with respect to handling, housing,
feeding, watering, sanitation, ventilation, shelter
from extremes of weather and temperatures, adequate
veterinary care, including the appropriate use of
anesthetic, analgesic or tranquilizing drugs, when
such use would be proper in the opinion of the
attending veterinarian of such research facilities,
and separation by species when the Secretary finds
such separation necessary for the humane handling,
care, or treatment of animals.
As of 1971, legislation concerning toxic substances (cadmium,
mercury, miscellaneous heavy metals, PCB's, etc.,) had been
promulgated by a number of different federal agencies. Sec-
tion 12 of the Federal Water Pollution Control Act passed
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in 1970 is concerned with toxic materials discharged into
navigable waters. The Clean Air Amendments of 1970 contain
a section directly related to hazardous substances. The
Department of Transportation regulates transport of hazard-
ous materials with legislation such as the Transport of
Explosives Act and Hazardous Cargo Act (4). Current legis-
lation applicable to pesticides includes the Federal Envi-
ronmental Pesticide Control Act of 1972. Major concerns
of this act are registration of pesticides under EPA guide-
lines, statements of use, and unlawful acts in relation to
pesticide sale, distribution and use.
METHODOLOGY USED IN THIS STUDY
Data presented in this report were gathered from a variety
of sources by several means. First, a literature search was
conducted in over 120 scientific journals. Further relevant
literature sources were identified and obtained from bib-
liographies and from interviews with scientists conducted
by phone. In addition to phone contacts, a number of visits
were made to particularly interesting and relevant research
facilities where information was gathered using sketches,
photographs, further interviews and, in some cases, actual
observation of testing procedures. Finally, outside con-
sultants and correspondence with the Environmental Protec-
tion Agency offered direction and review throughout the
preparation of this document. (A list of contacts and
visits can be found in Section VI.)
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SECTION III
SPECIAL PROBLEMS
PROBLEMS IN TOXICITY TESTING
Special problems inherent in toxicological testing revolve
around the ramifications of experimental manipulations of
biological systems. Such deliberate interventions by
toxicologists are termed treatments. A direct result pro-
duced by a specific treatment is described as an effect.
The primary goal in designing and conducting a toxi-
cological experiment is to ensure, insofar as possible,
that no factor other than the treatment will contribute
to the observed effects. Unfortunately, this ideal
situation is almost impossible to attain due to extraneous
influences. Therefore, it is imperative to insure that all
influences except those of the treatment will act equally
on the test (treated) animals and the control animals
(those exposed to identical conditions excluding the
treatment).
VARIATIONS IN EXPERIMENTAL DESIGN
Homeothermic vs. Poikilothermic
The predominance of homeothermic animals in toxicity
testing is immediately apparent in a review of the
literature. There seems to be greater scientific
knowledge pertaining to the physiology, metabolism,
biochemistry and behaviorisms of homeotherms than there
is for these same parameters in poikilotherms. Poikilo-
therms, however, pose a special problem in toxicity
testing. They must be maintained in a carefully controlled
environment since their metabolic rate is &. function of
environmental temperatures. Environmental control is
important to all toxicity testing since inaclmissable
experimental error would be produced if the metabolic rate
of the test animal fluctuated from one exposure of the
toxin to the next.
Sex, Weight and Age
Sex, weight and age variations influence many toxicity
tests. Researchers using mammals and primates tend to
use males because of the problems encountered with the
estrous and menstrual cycle of the females. Not only
do these cycles arouse the male animals being kept in the
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same laboratory, but they also greatly affect the physiology
of the two groups. However, it is not scientifically sound
to extrapolate male toxicity data to females (5). The
fluctuations in hormone levels, coupled with the nervous
stress created by the estrous and menstrual cycles, may
greatly influence the results of a specific toxicity test.
Unless the toxicity test is aimed at postnatal toxicology,
age variation is often introduced since test animals are
generally selected from a stock of sexually mature
animals (6). The very old animal is almost always ex-
cluded from toxicity testing. Closely related to age is
the weight of the test animal. When the test animals
are sexually mature, their weights are comparable within
a certain deviation. Test animals that are procured
through animal supply houses ace supplied with a controlled,
well-balanced diet, and are relatively uniform in weight.
Undernourished or extremely obese animals are normally
excluded from laboratory supply house stocks.
Age, weight, and sex must be considered when interpreting
toxicity test results. The very young animal does not
respond (detoxify) to toxic materials nearly as fast as
the sexually mature, while the very old animal has even
less ability to detoxify poisonous materials. Weight can
be the key factor in determining a test animal's tolerance
of a certain poison. The undernourished as well as the
extremely obese animal is much more prone tc a high mor-
tality rate with a given toxin than a well nourished
animal of the same species (5).
Random Sampling Variation
Random sampling can be defined as selecting individuals
in such a manner that every animal in a population has
an equal and independent chance of being chosen. Unfor-
tunately, human errors and natural phenomena frequently
bias a "random" sample. When researchers are selecting
animals from within a species group, it is easy to introduce
variation into the sample by choosing potential test animals
that are less alert or slower to escape the collector.
The collector is also frequently prejudiced by a potential
test animal's well being (e.g., healthy looking fur, skin,
teeth, etc.). Thirdly, docile animals within a species
group are normally selected over pugnacious fellow-members.
These selection tendencies could all greatly bias a toxicity
test.
In addition to the problem of random selection within a
species group, there is a lack of true random selection
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between various animal groups. As stated previously,
homeothermic animals are generally selected for research
much more frequently than poikilothermic animals. Animals
that adapt well to laboratory environments are chosen
over those animal groups that do not. Size is another
factor which will bias the sampling of various animal
groups; small sized (rodents) to moderately sized (sheep)
animals are preferred over large animals (moose) due to
the limitations of laboratory space. Polyestrous animals
are more desirable than monestrotis animals such as toxicity
test animals, especially in chronic studies dealing with
a toxin's effect on fecundity and transplacental migration.
These kinds of biases affect the true random selection of
toxicity test animals. One must also consider the possi-
bility that the effects demonstrated in the laboratory by
a wild animal are not necessarily those which a toxin
would produce in the animal's natural environment.
Variations in Breeding and Holding Environs^
Variations are introduced into toxicological testing by
the ease of procurement of certain animals which are
potentially good test animals. Researchers have two main
avenues to obtain toxicological test animals: biological
supply houses, and actual field capture of test animals.
"Animals for Research" (7), a publication of the Institute
of Laboratory Animal Resources, National Research Council
of the National Academy of Sciences, lists 315 biological
supply houses throughout the United States that specialize
in selling various species of test animals. The animals
available through these supply houses fall into two
categories: (1) domestic laboratory animals and (2)
animals collected in the wild — 19 phyla of animals from
all over the world.
The major problems associated with animals collected from
the natural environment are: (1) the high cost per animal,
(2) the possibility of collecting diseased animals and
(3) the rarity of certain species. It is not uncommon for
large terrestrial animals, e.g., Artiodactyla, to cost
$200 to $1,500 per animal, depending on the species used.
Even the smaller animals, carnivores and rodents, are
generally very expensive compared to most domestic labora-
tory animals. Animals collected in the wild and then
brought into animal supply houses often suffer from disease
or stress. The animal may have been captured because it
was diseased and not as alert as its more healthy brothers.
In addition, the stress and fear involved in the capturing
can render these animals more sensitive to toxicants than
other members of their species in the wild. Thus, these
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animals may introduce significant bias into toxicity
testing. Many species sought as test animals also fall
into the rare or endangered species category. The En-
dangered Species Conservation Act of 1969, Public Law
91-135, sets very stringent regulations on the use of
endangered animal species as toxicological test subjects.
The best solution for obtaining an ample supply of native
North American terrestrial animals for toxicological
testing is to collect animals that are easily bred and
maintained in captivity. Collection of these potential
wild test species should be confined to geographical
areas where environmental contamination is minimaJ to
avoid collecting animals which have developed resistances.
If the animal does not lend itself to breeding in captivity
or if it is from an area of heavy environmental contami-
nation, it should be viewed with skepticism as a potential
test subject. Repeated collection of a certain species
from the wild should also be avoided since this may
seriously deplete species numbers. It must be kept in
mind that a breeding colony of."wild" animals in the
laboratory is very different from a breeding colony in
nature, and is subject to a loss of the genetic diversity
which enables wild populations in the natural environment
to adapt to change.
Genetic Variations
Genetic variations involve a chemical rearrangement of an
organism's DNA which produces a mutant form. This vari-
ation or change may be very subtle (alteration in a single
enzyme's activity) or the change may be very drastic such
as the critical shortening of the wings on a bird.
Mutation is the original source of variation within a
biological system, but it is only through many sexual re-
combinations that a mutation establishes itself in the
species. Mutations occur constantly in biological
organisms. The average spontaneous rate for a given gene
locus is 1-2 mutations per 100,000 genes per generation
(8). Of these mutations, the vast majority are of a
detrimental rather than of a useful nature. However, to
remain viable under continuous environmental changes, it
is extremely important for the population to carry this
potential for change.
Non-Genetic Variations
There are instances where non-inherited changes will occur
in .an animal, altering body structures, functions and
behavior, without being transmitted to the next gene-
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ration. For example, if certain muscles are used very
intensively in an animal they become much thicker and
stronger; if one organ is removed such as the kidney,
the other kidney will increase in size and degree of
function. Skin in areas that receive a lot of rubbing
or pressure become very thick and impervious. Animals
that are exposed intermittantly for short periods to
extremely cold weather or perhaps low oxygen concentrations
can learn to acclimate, whereas they would have died had
they been suddenly introduced to these environmental
conditions.
Genetic Drift
Biological variation in a population is greatly reduced if
the gene pool is small; some of the more uncommon vari-
ations of the pbpulation-at-large will not be represented
or will be lost. This phenomenon is commonly referred
to as genetic drift, which is simply the raudom loss or
gain of genes rather than loss or gain by natural selection
in large gene pools.
RESISTANCE
Resistance is a phenomenon resulting from a combination of
biological variations and genetic adaptation. In toxicity
testing, resistance is the developed ability of a species
to tolerate dosages of toxicants which would once have
been lethal to 50 percent (LD50) of the individuals
within a population.
Mechanisms
The vast majority of data collected on resistance deals
primarily with the various economic poisons. An animal's
resistance to an economic poison is observed as the pro-
gressive inability of a given compound applied at a
fixed rate to achieve control on its target organisms.
The mechanisms involved in developing resistance to
economic poisons are very similar to those used to avert
mortality and morbidity from other toxicants encountered
by animals in the natural environment.
It has been discovered that resistance can be induced in
laboratory test species by exposing the test animals to
toxic levels which kill 90 percent of the population, and
then breeding the survivors. Ozburn and Morrison (9) have
bred white laboratory mice (Mus musculus) in order to
develop a strain that after nine generations is 1.7 times
more tolerant to DDT than the controls. Dosages of DDT
to which these mice are exposed kill the control animals.
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Recent toxicological testing confirms that there are four
mechanisms working simultaneously or independently to
develop resistance: (1) physiological mechanisms that
reduce the penetration and transportation of a toxin to
the site of action{10); (2) unusual capacities for
storing toxins at inert sites coupled with rapid excretion
(10); (3) microsomal detoxification (10); and (4) genetic
resistance resulting from selective breeding within a
population (9). It has been found that reduced penetration
and transportation, rapid excretion and microsomal de-
toxification are manifested in populations of animals that
have never been exposed to toxic materials.
Reduced penetration is significant because even a small
reduction in the rate with which a toxin enters an animal
may enhance the activity of detoxification centers.
Certain animals have capacities for storing toxins in
inert areas of their body which prevent toxins from
entering metabolic pathways. Chlorinated hydrocarbon
pesticides can be assimilated in the adipose tissue of
animals having ample adipose reserve. Once isolated.in
the adipose tissue, chlorinated hydrocarbons exert little
if any effect on the animal until the animal is forced
to catabolize the tissue. This delayed reaction can be a
significant effect of a toxin in a time of stress. The
increased excretion of toxins has not been verified.
It is believed that certain enzymes convert foreign
substances into less toxic materials by making them more
water soluble. These enzymatic reactions are accompanied
by reduced tubular reabsorption in the kidney which in-
creases urine output.
Microsomal enzyme systems play a primary role in the
metabolism of foreign chemicals. Anything which tends to
change the effectiveness of this system in metabolizing
toxins alters the resistance of the animal. Microsomal
enzyme induction is another phenomenon related to
resistance. Chlorinated hydrocarbons are very good
microsomal enzyme inducers. Chlordane increases the
metabolism of hexabarbital aminopyrene and chloro-
promazine (11).
Research by Webb et al. (6) offers preliminary support
for the thesis that genetics play a role in specific
resistance processes. Pine mice (Pitymys pinetorum)
were exposed to endrin in apple orchards in Maryland and
Virginia and then live-trapped and brought into the
laboratory and bred. Offsprings of these field-trapped
pine mice were held between 60-90 days after birth and
then LD5Q values were established using endrin. It was
10
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the cohsensus of Webb et al. that the results of their
toxiclty studies on first generation offspring raised
under a pesticide-free environment offered preliminary
support for a genetic basis for the demonstrated endrin
resistance. Other genetic work by Cory et al. (12) supports
the thesis that exposure to environmental levels of DDT
induced chromosomal changes in Drosophila pseudoobscura.
DDT Resistance - A Case Study
The most celebrated case of resistance recorded in the
scientific literature is that which houseflies (Musea
domestica) have developed to DDT. The work of Sternburg
and Kearns (13) demonstrated that resistant houseflies
have a high titer of DDT - dehydrochlorinase which allows
them to dehydrochlorinate DDT to DDE. There is good
information to substantiate a correlation between dehydro-
chlororinase activity and DDT resistance.
It was first believed that only the cuticle of resistant
flies was the site of the enzymatic reduction (14);
however, later work has demonstrated that: most tissues of
the resistant flies are active (16) . Refined genetic
procedures conducted on the housefly by Lovell and Kearns
(17) reflected the fact that dechlorination and resistance
were genetically linked. It is also the belief of pesti-
cide toxicologists that the development of DDT de-
chlorinating ability in the housefly is preadaptive, that
is to say, it already existed prior to DDT application.
Resistance is important in toxicity testing because the
possibility always exists that researchers will select
test animals that possess one or all of the resistance
mechanisms for a particular toxin. These biased test
animals can produce erroneous dose/effect responses.
This is at best misleading and could be devastating if,
for example, the results are used to set threshold
exposure limits.
NATURAL SELECTION
Natural selection involves a combination of environmental
and interspecies pressures. Most species produce more
offspring than can possibly survive, necessitating a
competition for existence. Structural, functional, and
behavioral variations in some individuals provide them
with advantages over other individuals who lack these
distinctions. These variations must- be phenotypic before
natural selection can occur. If they are, they will
make a much greater contribution to the gene pool of
11
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the next generation. Accumulation of a desirable variation
within a population generally facilitates the species
adaption to its environment, enhancing its chances for
survival.
When the ratio of one characteristic to another changes
from 1:00 : 1:00 to 1:01 : 1:00 in each generation, it
is known that more individuals with this characteristic
are surviving than are individuals without it (8). With
a selective advantage of 1 in 100, it has been demon-
strated that a dominant characteristic will become
established in 99 percent of a population in about 1200
generations (18), a rapid evolution rate. When selection
has decreased to 1 in 100,000 or more, evolution is rela-
tively slow. The phenomenon of natural selection and its
effect on speciation can introduce a great amount of dis-
torted data in toxicological testing. Species collected
in geographical areas where natural selection and
resistance have influenced the population will be entirely
different from the same species collected in areas where
the parameters of selection and resistance are dissimilar.
Hence, the dynamic processes of resistance and natural
selection must be taken into careful consideration when
selecting species of toxicological test animals.
TESTING OF DYNAMIC ECOLOGICAL ASSOCIATIONS
An ecosystem may be generally thought of as the biotic
community and abiotic environment functioning together
as a system. From a structural standpoint there are four
constituents in the ecosystem: (1) abiotic substances
which are basic elements and compounds of the environment;
(2) producers, the autotrophic organisms that are com-
prised largely of green plants; (3) consumers, the
heterotrophic animals that ingest other organisms or par-
ticulate organic matter; and (4) the decomposers, the
heterotrophic organisms, chiefly bacteria, that
break down complex materials from dead biota.
The use of model ecosystems has recently been recognized
as being a valuable tool in toxicity testing. The
majority of the environmental toxicological testing to
date has involved the use of aquatic ecosystem models.
Kapoor et al. have constructed several aquatic model
ecosystems to assess the biodogradability of pesticides (19)
The use of terrestrial ecosystems in toxioity testing is
still in the developmental stages. Terrestrial model
ecosystems containing a predator require sizeable en-
closures and refinement (20).
12
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A food web is the system of dynamic transfer of food
energy from primary producers (plants) through various
trophic levels. It has been estimated by ecologists
that food webs allow approximately 10 percent of the
energy entering one trophic level to pass on to the next
higher level. The least complex version of this type
of system takes the form of a pyramid, with each suc-
cessive population receiving approximately a tenth of the
energy received at the level below it.
Unfortunately, nature seldom builds communities with such
a simple structure and the use of food webs as tools to
measure the biological magnification of environmental
toxins is not without problems.' Almost invariably, energy
is not passed along neatly ordered chains, but instead is
transferred to a great many organisms through a highly
complex sprawling web of pathways. It is this inherent
complexity that becomes one of the principal factors to
be considered in investigating how toxic materials are
disseminated and concentrated within food webs.
Another factor that becomes a problem in using food
webs as toxicity test devices is the nature of the various
metabolic processes of the members of a food web.
Normally less than 50 percent of the products of metabo-
lism go into the construction of new tissue, the remainder
is spent as respiration. The crux of the problem is that
some substances within a food web are not involved in
respiration, and consequently not excreted efficiently.
Therefore, these materials may be concentrated in the
tissues twofold or more when passed from one population
to another (21).
EXPERIMENTAL DESIGN OF TEST vs. COST
The experimental design of a toxicity test dictates the
cost of the research project. In toxicity testing, two
major considerations are reflected in costs: (1) retro-
spective versus prospective studies and (2) field investi-
gations. A retrospective study examines the distribution
of concentrations of toxins in the physical aspects of
an ecosystem and the biological aspects. From this residue
data, an attempt is made to assess the impact of the
various toxins on the animals colleqted. Retrospective
tests rely very heavily on data that has already been
collected and are usually of a shorter duration and are
much less costly than prospective studies.
Prospective studies are those that start with a thesis
which is to be experimentally tested, e.g., to ascertain
13
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if exposure to 200 ppb of polychlorinated biphenyls
(PCB's) in crayfish will have any chronic effect on the
fecundity of raccoons fed on these crayfish. In this
example, mature raccoons must be used that are known not
to have been exposed to PCB's (laboratory reared subjects,
perhaps) and they must be fed for one year or more on
dosed crayfish containing 200 ppb of PCB's. During this
study period the levels of PCB's in the crayfish and
raccoon must be closely monitored. Observations must
be made on the reproduction capabilities of these test
animals. Even if reproduction is accomplished in these
animals, it may be necessary as well to observe their
offspring for possible effects. It is clear from this
example that prospective testing can become highly complex
and in general is more costly than retrospective testing.
Field investigations are usually concerned with monitoring
toxic residues in terrestrial animals and comparing
these levels to those found in the air, water and soil of
the animal's ecosystem. Generally, the volume of bio-
chemical analyses, tissue residue level determinations
and histopathology examinations required by field studies
are fewer than in laboratory studies. Field investi-
gation studies will frequently require the services of a
commercial laboratory to conduct the biochemical, histo-
pathological and residue analyses. Although these two
characteristics sometimes make field studies less costly
than laboratory studies, it is generally the latter type
of study that is more expensive. There are unique
features which make laboratory studies more costly than
field studies: (1) animal maintenance, including
veterinary services; (2) original cost of the test
animals; (3) sophisticated laboratory instrumentation;
(4) large numbers of technically trained personnel; and
(5) the fact that they are generally prospective studies
(whereas field studies are generally retrospective).
WILD NATIVE SPECIES vs. DOMESTIC LABORATORY SPECIES
A unique group of domestic animals has been bred for use
in biological research throughout the world. This group
includes white mice and rats of various strains, crickets,
chickens, ducks, guinea pigs, hamsters, gerbils, dogs,
cats, etc. The majority of these animals reproduce at
relatively fast rates, are easily maintained in a labora-
tory environment, and have been inbred (especially the
white mice and rats) to produce a very homogenous,
well-defined animal. Inbreeding results in a loss of the
genetic diversity and plasticity which a natural population
of animals relies on to adapt to changing environmental
14
-------
conditions, and, in this sense, laboratory animals are
no longer representative of wild populations.
Laboratory rats and mice -are excellent test animals where
the experimental design calls for consistent, genetically
similar animals in large quantities. However, using these
domestic animals in environmental toxicological testing
may bias the results and not really reflect the reactions
that a natural population might exhibit. For this reason,
new animals, either naturally or captively bred, should
be added to each captive colony, periodically introducing
new wild-type genes. The results of toxicity testing on
animals from populations whose ancestors have been sub-
jected to natural selection would seem to be less biased
and much more indicative of the real impact of a toxin
on the natural environment.
Wild animal species recommended for toxicity testing are
reviewed in detail in the Animal Suitability section of
this report.
CHEMICAL PROBLEMS IN TESTING: DEGRADATION
Once released into the environment, chemical toxicants
undergo a continual dynamic change with the rate of
change being governed by the chemical stability of the
toxin. Environmental toxicants, in general, go through
two degradation processes: (1) nonbiological transfor-
mations and (2) biological transformations.
Nonbiological Oxidation, Reduction and Hydrolysis
Except for a few locations (i.e., certain soil types),
oxygen is a ubiquitous force for environmental transfor-
mation of toxicants, especially pesticides. The usual
reactions of oxygen focus around the free-radical character
of its usual triplet electronic state. However, recent
research has revealed the significance of the more
reactive double-boned singlet oxygen produced by both
light and chemical means (22) . Photooxidation at aliphatic
carbons is a common reaction in some pesticides. However,
DDT is normally very resistant to oxidation - either
photochemical or otherwise - given normal temperatures
and light.
The removal of hydrogen atoms from organic solvents by
free radicals is well understood. Photochemical study of
pesticides has revealed that energetic irradiation in
methanol will lead to the replacement of aromatic ring
halides with hydrogens (22). Strictly chemical reductions
15
-------
by transitional metal ions are probably very widespread
throughout the environment, but have not been reported
in the literature. A good example of this type of
reaction is the reduction of DDT to DDD by iron
porphyrins.
Water, like air, is present to some degree almost every-
where in the natural environment. Water has unique
chemical properties: it has high heat capacity, polarity,
is a good solvent, and is transparent to uv light; thus
it is an extremely important medium for chemical re-
actions. Pesticide hydrolysis occurs very readily and
many researchers are surprised that the extent of these
environmental transformations has received so little
attention (22) .
Nonbiological Miscellaneous Reactions
A number of degradation reactions have been reported
under various other environmental conditions. It has been
demonstrated that dieldrin undergoes an internal cycli-
zation in the presence of light to form photodieldrin
(22). Many organophosphorus esters such as the phos-
phorothionates will isomerize under the influence of
heat and light.
Biological Oxidation, Reduction and Hydrolysis
Generally all biochemical oxygenations of pesticides and
other toxins are carried out by low-specificity, mixed-
function oxidases often localized in microsomes or
similar structures.
Reduction of toxins through metabolic pathways is much
less obvious than oxidative processes, perhaps because
life exists in and on an oxidizing system. The biological
reduction of C-C double bonds has been suggested to
explain the intermediates observed in the metabolism of
DDT and its chlorinated hydrocarbon relatives; however,
simple olefins to not display these reduction products.
It has been found that pesticides react with water within
living organisms. The hydrolysis of the epoxide ring of
dieldrin has provided one of the most important routes
for the metabolic degradation of dieldrin. Hydrolysis,
however, like many other types of environmental trans-
formations, is not always straightforward. The primary
route for the detoxification of DDT in animals is by
the hydrolysis of the trichloromethyl group to a
carboxyl group.
16
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DOSAGES
Dosages used on test animals in toxicity testing can be
critically important in obtaining reliable test results.
As discussed under Biological Variation, individuals from
a homologous population display variations. The real
nature of these variations is seldom obvious except when
the organisms are challenged, such as by exposure to a
toxic material. For example: if the toxic agent is
capable of producing an observable effect (such as death),
then the chemical dose can be selected to produce that
effect on each test animal in a series of tests using
the same species group. However, it has been demonstrated
that, if the effect could be measured by means other than
death, then the test would show that not all members of
the group responded in the same manner.
Another problem associated with dosages is the method
used to administer the toxin. Methods include filling
gelatin capsules with a prescribed dose, injecting marsh-
mallows with the toxin by using a microsyringe, and putting
the toxin dosage into the animals' normal food or water.
It is questionable whether the test animal really receives
the intended dosage since some of the dosed food may spill
or the loaded marshmallows may be unpalatable.
There are four major routes to administer toxicants:
(a) percutaneous, (b) inhalation, (c) oral, and (d)
parenteral. Within a species and from species to species,
these four routes of administration greatly influence the
toxicity exhibited. If not taken into careful consider-
ation, they can introduce variation into the testing.
BIOSTATISTICS IN TOXICOLOGICAL TESTING
Introduction
Toxicological studies should be well-designed to assure the
collection of unbiased and precise data which are tech-
nically defensible and amenable to biostatistical evalu-
ation. Some of the basic concepts and techniques of
sampling design and data evaluation by biostatistical
methods are discussed in the following paragraphs.
Randomization
In toxicity testing, the experimental animals must be
selected with known probability. As mentioned previously
in "Variations in Experimental Design,"" the manner in
which a random sample is obtained can introduce a great
17
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deal of variation into the experiment. The careful use
of random selection is the only feasible technique by
which to satisfy the "known probability" criterion.
For example, knowing the probability of selecting an ex-
perimental animal makes it possible to extrapolate from
the sample to the population in an objective way.
It becomes obvious that there is a fundamental distinction
between a "haphazardly-selected" experimental animal and
a "randomly-selected" animal. The distinction can be
made that a "haphazardly-selected" animal is one chosen
with no conscious bias, whereas a "randomly selected"
animal is one chosen when there is consciously no bias
(the rationale being that there is consciously no bias
since the randomization is planned, and therefore bias is
planned out of the selection).
Sample Size
In any toxicological study, the number of animals to be
exposed and the number of animals to be employed as con-
trols should be decided early. Sample size is important
since frequently too many animals are used which is
wasteful, or not enough experimental animals are used
which results in invalid data. There are two biostatistical
techniques which can be employed to ensure adequate random
sample size: estimation of a binominal proportion and
estimation of a population mean for measurement data.
The Normal Distribution
Due to unavoidable error in the measurement process, all
measurements in toxicity testing are uncertain to some
extent. There is, of course, only one correct measurement,
which researchers can only attempt to estimate by one
mere measurement or by the mean of a number of measure-
ments. If measurements of some treatments are repeatedly
redone in a specific test, it is found that most outcomes
tend to cluster about a central value while a few deviate.
A frequency distribution of the results forms a somewhat
irregular histogram with the density of measurements being
greatest in the center and diminishing to the right and
left. It has been observed that the accumulation of more
data tends to make the histogram appear less irregular.
This type of measurement distribution is approaching a
mathematically defined bell-shaped curve known as the
Gaussian curve of error, or simply the normal distribution.
The mean is a biostatistical technique that locates the
center of distribution of the x-axis of measurement.
18
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In other words, it is the long-run, arithmetical average
of all x-values.
The statistical variance is useful in describing the amount
of data variability or data dispersion which, in turn,
determines the distribution curve's breath. Variance is
the expected value of the squared deviation of a measure-
ment from the mean, or simply the long-run arithmetical
average of all such squared deviations.
The square root of the variance is known as the standard
deviation. The standard deviation is the distance measured
along the x-axis, from the center (mean) to the point of
inflection of the curve which is the steepest point.
Tests of Hypotheses
In toxicity testing, frequently some aspect of the study
is directed to answering a hypothetical question regarding
a population. If the hypothesis is quantifiable, then the
hypothesis can be tested statistically by either the T-test,
Chi-Square Test, or the F-test.
The T-test can be used to compare a sample statistic
(i.e., the mean) from a toxicity experiment with some value
for the purpose of making a judgment regarding the popu-
lation as indicated by the sample data.
If there are more than two alternative categories of
response known or assumed, the sample methods developed
for binomial populations no longer serve. A useful tool
that can be applied to various problems involving enumer-
ation data, without regard to sample size, is the Chi-
Square Test.
In biostatistics the F-test is used for testing the
equality of variance. The computation of the F-values
involves the ratio of two variances, each with associated
degrees of random.
Linear Regression and Correlation
In various toxicity testing it is often desirable to
investigate relationships between different variables,
i.e., rate of weight loss in a t^st animal and the concen-
tration of toxin in blood serum, or mortality of test
animals per unit of time and the concentration of poly-
chlorinated biphenyls in the test animals brain, etc.
Toxicologists appreciate the complexity of actual world
19
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relationships between variables and those mentioned
above; however, toxicologists may also wish to investigate
further approximating these relationships with a straight
line.
Such an approximation may prove invaluable if used
judiciously in toxicity testing within the limits of the
conditions where the relation holds. It is important to
recognize that regardless of how well the straight line
describes the data, a casual relationship between the
variables is never implied. Causality is much more dif-
ficult to establish than mere description by a statistical
relation.
When studying the relationship between two variables, the
data may be taken in one of two ways. One way is to
measure two variables, e.g., measure body weight and as-
sociated blood serum measurements. Where two variables
are measured, the data are termed bivariate. The al-
ternate way is to choose the level of one variable and
measure the associated magnitude of the other variable.
Straight line equations may be obtained for each of these
situations by the technique of linear regression analysis,
and if the object is to predict one variable from the
other, it is desirable to obtain such a relation. When
the degree of (linear) association is to be examined, no
straight line need be derived - only a measure of the
strength of the relationship. This measure is the cor-
relation coefficient, and the analysis is termed correlation
analysis.
Thus, linear regression analysis and linear correlation
analysis are two ways in which linear relationships
between two variables may be examined.
By judiciously employing the biostatistical techniques
described above, the conclusions drawn from toxicity
testing can be made much more valid. It must be remembered
that all biostatistical methods are based on an abstrac-
tion (math model) which is an approximation of some
actual world phenomenon. If the approximation is reliable,
researchers have a useful tool; if the approximation is
unreliable, researchers can easily draw erroneous
conclusions.
20
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SECTION IV
ANIMAL SUITABILITY
INTRODUCTION
After determining the suitability of animals or animal
groups for use in toxicological testing, species have been
recommended in this section as good potential test subjects.
The basic criteria for these recommendations are: (a) the
availability of the species; (b) the ability of the species
to breed under captive conditions; (c) the ease with which
it can be maintained; (d) the previous use of the animal in
toxicological testing; and (e) the degree to which a
species is representative of the group. Most of the species
considered are wild native North American forms. Exotic
or domestic animals were substituted only if suitable
organisms for a specific group could not be found (see
Section III - Wild Species Versus Domestic/Lab Species for
further discussion).
Establishment of Animal Groupings
The animal groupings reflect natural, taxoncmic associa-
tions. Certain mammalian taxa were divided or aggregated
because of different or similar food habits, or because of
size and/or habitat preference. Invertebrates have been
aggregated on the basis of phylogeriy due to the paucity of
research information.
Sample species for each group were used wherever possible
and are meant as general guides to the group. In all
cases, the text refers to North American members of the
animal groups unless otherwise specified and it should be
noted that the stated requirements may diffar widely from
those of non-temperate species. Indeed, animals of the
same species from different latitudes can require different
maintenance methods.
A format consisting of 12 major categories has been estab-
lished for the discussion of the animal groups. Where no
information was available in a category for a specific group,
that section was omitted from the presentation. Previous
research is briefly presented under the category of Toxi-
city Testing. The large amount of information accumulated
makes it difficult to describe each study in detail, but
they may be located for further information via the list of
references.
21
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An effort was made to identify the specific qualities of a
species that make it particularly suited for a certain type
of test or procedure. The inclusion of information on
habitat preference and ecological roles enables the reader
to further evaluate the suitability of an animal for a given
test or methodology.
Animal Suitability Tables
The suitability of each animal subgroup for use in toxico-
logical testing is described in a table found in each major
animal group section. These tables synopsize the char-
acteristics of the animal group discussed in the text, with
central consideration afforded the recommended test species
in each group. The final section of this chapter sum-
marizes pertinent information on each suggested form and
contains a final list of the most suitable species for
testing from each group (general use of the suggested
species is further discussed in Section V, Suggested
Methods).
22
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INVERTEBRATES
Invertebrates, a very large group, include all animals that
do not possess a backbone, from the tiniest single-celled
creature to highly developed forms like squids, snails,
insects, and crustaceans. Despite their enormous variety
and number, invertebrates have been scientifically studied
far less than vertebrates, partly because they are not as
conspicuous as vertebrates or as directly important to man,
and because they are considered by many to be "lower" forms
of life. Information is available for relatively few species
and these are largely pests and otherwise economically
important species.
By far the largest group of invertebrates both in numbers
of individuals and species is the insects. Insects belong
to the phylum Arthropoda and possess jointed appendages and
an exoskeleton. They are one of the three animal groups
that have attained true flight, a characteristic which sepa-
rates them from other Arthropods. Three pair of walking
legs, compound eyes, tracheal respiration and metamorphosis
also characterize insects.
The information on insects is very heavily skewed in favor
of pest species and honeybees, because both groups are of
major importance in agriculture. The seemingly unimportant
species have largely been neglected, but research on these
will increase as more importance is attached to evaluating
and assessing the effects of chemicals on whole ecosystems.
Invertebrates can be rewarding as well as difficult test
organisms. Their life cycles are complex, often involving
many distinctive, morphologically different stages which
can cause the researcher some difficult maintenance and
rearing problems. The limited knowledge of this group
hinders research, and the generally small size of its
members restricts testing methods. On the other hand, they
are important members of the world ecosystem as basic
sources of food for many animal species. Furthermore, most
are very instinctual and therefore routine and stereotyped
in their behavior so that differences or alterations in
behavioral norms can be easily measured. In statistical
studies, their characteristics make them potentially good
test organisms. Finally, the group offers the researcher
a vast array of ecological roles. The greatest amount of
research to-date is concentrated in genetics (Drosophila)
and bioassays, (Musca).
23
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General criteria for selection of a test species include
the position of the species in the food chain, its availa-
bility, breedability in captivity, ease of maintenance and
previous use in toxicity testing. These criteria are sum-
marized for each invertebrate species investigated in
Table 1.
24
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TABLE J. SUITABILITY OF INVERTEBRATES FOR TOXI COLOR 1CAt
INVERTEBRATES
Helminths
Molluscs
Arthropods
Arachnxds
Insecta
Orthoptera
Hemiptera
Homoptera
Coleoptera
Lepidoptera
Diptera
Bymenoptera
Misc.
Arthropods
^
•H
Availabi:
G
G
G
G
G
E
G
E
E
G
>i -U
4J -H
•H >
Breedabi:
in Capt:
G
G
P
G
G
E
G
E
E
F
Q) fl> 01
o 10 tr c
CDC 0
id -H -H
C M 4J *»
>w QJ 3d U
O 4-) O 0) -H
C -H M U
w id v c n
H S Ł "* «
E F
G P
G P No
previous
breeding
work
G G
E F
E G
E G
E E
E G
G F
>i
•H •-*
r-l Id
-rf U
*f\ -H
id id en
U 4J O O
OJ -H .-» rH
C 3 O O
o to u ^
U W
G E
G D
Ł.
F B2
G B- f D-
G D2
E B2,D2
G D,
E D2
E D2
E,
G B2
u
c
Habitat
Prefere
Fo
Gr
Fo
Gr
FO
Gr
Fo
Gr
Fo
Gr
Fo
Gr
Fo
Gr
Fo
Do
Fo
Gr
Do
Fo
Gr
•o
o in
4J CJ
W -H
a> o
tr 'Btri3
Helix naj^erso
Milax sp.
4ra^eus ^-aJ^.tMo
Gryllue penneylvanious
praying mantids
B lie eus leujci ~er^s
leafhoppers
Tribolium sp .
Tenebrio sp .
CQrpocapea per- 'veils
Aprotis ypsilci
Musca domestica
Drceophila me^ .i'io paste
Apis mel-ifera
Isopods
Centipedes
Legend:
General Ecological Role
A - Large carnivore
B - Small carnivore
1. General
2. Insectivore
C - Omnivore
D - Herbivore
1. Seed, fruit eater
2. Grass, foliage eater
E - Decomposers
Habitat Preference
Fo
Gr
De
Aq
Do
Forest
Grassland
Desert
Aquatic
Domestic
E = Excellent
G = Good
F = Fair
P = Poor
Sources: - literature cited in text
- interviews of Scientists
25
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Helminths
Introduction -
This invertebrate group is comprised of several phyla, the
number depending upon the classification scheme used. Gen-
erally, the flatworms (Platyhelminthes), round worms
(Aschelminthes), segmented worms (Annelida), and a few other
smaller groups (e.g., Onychophora), are recognized as the
major divisions (24).
Worms are distributed throughout the world and occupy many
types of ecological niches. Earthworms (class Oligochaeta)
are fossorial, and ingest organic materials found in the
soil. There are also a great number of parasitic worms
ranging from the externally attaching, blood sucking leeches
(class Hirudinea) to the basically internal, often highly
specialized groups (classes Trematoda, Cestoda, and Nema-
toda). Whether this last group can be classed as terres-
trial is questionable. However, they do infect terrestrial
organisms to a great extent. Because of bhe small amount
of data available on worms in toxicological testing and
the large extent of the group, most will be treated in
this section in a general manner.
The only worms used to any extent in toxicity testing are
members of the Oligochaeta because of their close associa-
tion with soils. Most of the following comments pertain
to earthworms.
Caging/Lab Conditions -
The common earthworm species, Lumbricus terrestris (family
Lumbricidae), is widely used in zoology courses and in re-
search. They are readily available in large quantities
from biological supply houses or from bait dealers. How-
ever, if greater quality control is desired, earthworms can be
easily reared. Three general methods are used: small box
cultures that are handled easily; large containers; and
open fields. Any size container will do, but for frequent
inspection or sorting, a depth of more than six inches is
not desirable. Containers should have drainage holes
covered with screen. The bottom should be layered with
one-half inch of gravel and, if kept outdoors, covers
should be provided (25,26). Temperatures of 50°F to 75°F
(depending on the species) should be maintained (25).
26
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Nutrition -
Culturing media should be a combination of organic matter
and soil. Any combination of peat moss, manure, leaf mold,
sod, decaying sawdust, straw, hay and leaves and one-third
soil of high organic content will be a sufficient substrate
for most earthworms. Other foods, like table scraps, chick
starter, or cornmeal should be added periodically. About
40 g of dried food for 300 worms is a sufficient food addi-
tion at any one time (26). The substrate must always be
moist, but not wet (25) with an approximate pH of 7 (26).
Breeding/Rearing -
If provisions are made for adequate space, temperature
(about 50°F for L._ terrestris, higher for some other species) ,
and moisture, earthworms will reproduce successfully in a
loose substrate of one-part dung, three parts soil, and five
parts peat moss (26). Though earthworms are hermaphroditic,
an exchange of sperm takes place during conjugation. The
fertilized eggs are deposited in the soil in small pea-sized
cocoons.
Special Needs -
Moisture and moderate temperatures are necessary. In
nature, worms will burrow until the desired conditions are
found. Since this may be impossible in small containers,
care should be taken to keep their environment satisfactory.
Ecological Role -
Earthworms occur in upper soil levels and feed on decaying
organic matter. They are particularly important as soil
mixers, aerators and drainers and serve ab food for many
insectivores (robins, woodcock, mice, shrews).
Problems with Mass Culture -
A bin 4x8x2 feet will support 50,000 worms, so crowding
is seldom a problem when sufficient moisture and food are
available (25).
Toxicity Testing -
Tissue residues in earthworms have been monitored extensively
in areas where toxic substances have entered the soil (27,
28). The effect of chemicals on earthworm population
levels is of great concern and has been investigated under
27
-------
various conditions (29, 30, 31, 32), including in the open
field (33, 34, 35, 36, 37, 38, 39). The U. S. Fish and
Wildlife Service has published a thorough review of the ef-
fects of chemicals on earthworms (see Reference 40).
General Suitability -
Worms occupy a variety of ecological niches and occur
throughout North America. Their diversity, wide distribu-
tion, and importance as a food source for many vertebrate
species make them desirable test species. Earthworms are
particularly valuable because of their role in soil eco-
systems, their past use and ease of maintenance.
Ecological Alternatives -
Isopods, snails.
28
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Molluscsa
Introduction -
Molluscs are mainly shelled invertebrates and most are aqua-
tic; however, the pulmonate snails (class Gastropoda) are
common terrestrial forms with a modified mantle for air
breathing. Certain slugs are also terrestrial and cause
crop damage in some sections of the U. S.
Cepaea nemoralis, a snail, Testacella sp. (carnivorous) and
Milax sp., both slugs, have been successfully used in lab
research (41). Helix aspersa, a nocturnal pulmonate snail
found throughout temperate areas of the world, has been suc-
cessfully bred and reared in captivity since the life cycles
of snails and slugs are similar, the comments in this sec-
tion, which are primarily concerned with Helix aspersa, can
be applied to many other species.
Caging/Lab Conditions -
Colonies of Helix aspersa have been kept in 55 x 40 x
28 cm wooden boxes with a 5 cm layer of damp soil. The
boxes were covered with aluminum screen. Room temperature
(avg. 23°C) is suggested (42). C. nemoralis has been bred
in 20 cm flower pots two-thirds filled with soil. Testa-
cella sp. has been bred in corked glass tubes and fed on
earthworms. Milax sp. seems*to reproduce better under
conditions of varying temperature (41).
Nutrition -
Snails in captivity thrive on lettuce and other fresh vege-
tables (43). Slugs will very probably do equally well on
similar foods. Carnivorous slugs do well or. a diet of
earthworms (41).
Breeding/Rearing -
Helix aspersa has been successfully bred and reared in quart
fruit jars containing 5 cm of damp soil (43). The best re-
sults occur when only two individuals are housed in each
jar (43). Copulation is extended (the average per pair of
62 pairs was recorded as 6.25 hours) compared with that of
other animals. The eggs, which'are white and about 3mm in
diameter, are oviposited in nests prepared in the soil. In
one study, the average number of eggs deposited was 53, of
which, on the average, 10 hatched. Sexual maturity occurred
within four months after birth (43).~ The photoperiod af-
fects egg-laying (45).
29
-------
Special Needs -
Constant moisture and suitable temperatures are required
for snails to remain active (43). If sufficient moisture
is not present, snails will withdraw into their shells and
excrete a seal over the opening.
Ecological Role -
Terrestrial snails and slugs are primary consumers and eat
a varied diet of plant materials. They are a food source
for larger insectivores.
Longevity -
The average lifespan of snails and slugs is unknown.
Toxicity Testing -
Helix sp. have been subjects in drug testing (46, 47, 48,
49)and in open field pesticide studies where general popu-
lation fluctuations were measured (50). Learning behavior
in Helix is described in reference 42.
General Suitability -
Helix sp. is a very widely distributed snail and abundant
in certain moist habitats. In terms of rearing, abundance,
and present use, it may be the only suitable terrestrial
mollusc for use in toxicity testing. However, other pul-
monate snails and slugs, with the exception of the carni-
vores , may prove to be equally suitable after preliminary
maintenance procedures are worked out.
Ecological Alternatives -
Earthworms, some herbivorous insects.
30
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Arachnida
Introduction -
Spiders, mites and ticks belong to the class Arachnida in
the phylum Arthropoda. Members of this class are pri-
marily terrestrial and possess two main body segments. The
spiders, mites and ticks have four pairs of legs on the
cephalothorax and no abdominal appendages. They have com-
pound eyes but lack antennae. Many Arachnids have poison
glands and some spiders and ticks are able to produce silk
from special glands in the abdomen. Other members of this
class are scorpions and harvestmen.
Caging/Lab Conditions -
These invertebrates can be very easily maintained in almost
any container from individual glass tubes to covered ter-
raria and fruit jars. Mites and ticks are parasitic and
can be kept with their host in its enclosure. However,
the life cycles of ticks vary, some requiring only one
host and some up to three different host species, with
maintenance conditions changing accordingly. Boophilus
sp. and Dermacentor sp. are examples of some species that
have been used in the laboratory ( 51 ).
Spiders are also relatively easy to keep in the laboratory.
Dr. Peter N. Witt and his colleagues from the North Caro-
lina Department of Mental Health have very successfully
used web patterns in orb-weaver spiders to test the effects
of drugs. Araneus diadematus is their best subject.
(see Toxicity Testing for -details). Special 50 x 50
10 cm aluminum enclosures (See Figure 1) with removable
glass windows and provision for air circulation were con-
structed for web-spinning. To encourage nightly spinning,
a photoperiod of 13L:11D has been used. A temperature
change (colder at night) is also important (52) .
Nutrition
The feeding of mites and ticks often requires a host. All
their nutritional requirements are met by the blood meals
they extract (51). Spiders are fed flies and mealworms.
They require water, which can be provided by spraying a
ball of cotton placed in the cage or, in lab reared in-
dividuals, administered by a syringe placed up to the
animal's mouth (53).
31
-------
Figure 1. Web building enclosure
Raleigh, NC
Source: (52,53)
32
-------
Breeding/Rearing -
Breeding mites and ticks is simple when the appropriate
host is provided. The instar stages vary in number and
length, but if their natural host is available, a con-
tinuous supply of organisms can be produced. Female ticks
usually fall from the host after engorgement and mating to
lay their eggs, necessitating some method of collection
and housing (such as glass vials) ( 51 ). Success at
breeding spiders in captivity has been minimal, probably
due to the lack of effort. Most supplies of spiders are
acquired by gathering cocoons in the fall and hatching
the young in the lab (53).
Special Needs -
The special needs of parasitic mites and ticks have been
described above under Caging/Lab Conditions. Spiders need
a cold-dark, warm-light rhythm in order to build webs
nightly. Water should also be given frequently ( 53 ) .
Ecological Role -
Mites and ticks are parasitic on plants and animals, de-
riving their sustenance directly from the fluids of their
hosts. Spiders are carnivorous invertebrates whose food
consists entirely of small animals, primarily insects.
All Arachnids are potential food sources for insectivores.
Longevity -
The lifespan of mites and ticks are relatively unknown.
Spiders of the genus Araneus can live up to 18 months
( 53 ) and Tarantula and wolf spider life spans are measur-
able in terms of years.
Problems with Mass Culture -
Spiders need more research on breeding before they can be
mass-bred under controlled conditions.
Toxicity Testing -
Because certain mites are serious crop pests, a significant
amount of toxicological research has been conducted using
these mites as test species ( 54, 55, 56 )• Spiders have
been used in drug testing to some extent. Since some
groups build very uniform webs, the researcher has a very
33
-------
discriminating parameter for measuring drug effects. The
application of this technique to the testing of toxic
substances seems rather simple ( 53 ) .
General Suitability -
Mites and ticks are easily maintained under controlled
conditions and because many derive their substenance from
the fluids of their hosts, they might be used in a variety
of ways to monitor the host.
Spiders are excellent test subjects because they are preda-
tors on many insects, they are relatively easy to maintain,
if not to breed, and their web-building provides a very
useful experiment tool. Araneus diadematus has been a
most successful web-weaver and quite common in the Eastern
U.S. Mr. Leonard Pankhurst, 204 Stroud Street, Canastota,
New York, 13032, supplies this species in limited
quantities (52).
Ecological Alternatives -
For spiders, centipedes.
34
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Orthoptera
Introduction -
This order includes many large and well-known insects:
crickets, grasshoppers, roaches, locusts, and praying
mantids. Most are plant feeders and can be very de-
structive to man's crops. A few species are predacious
( 57 ). The Orthopterans usually have wings (two pair)
and their mouthparts are of the chewing type. Auditory
stimuli are important for intraspecific communication and
oval eardrums, or tympana, are the receptor organs.
Stridulations (the songs) are the results of rubbing cer-
tain body-parts together (e.g., hind femora and wing).
Most past toxicological research has been done on wide-
ranging and easily obtainable species, the common cockroach
(Periplaneta americana), the field cricket (Gryllius
pennsylvanicus),the locusts (Schistocera gregaria and
Locusta migratoria), and the Mantids(Mantis sp., Stagmo-
mantis sp., Tenodera sp.).
Caging/Lab Conditions -
Crickets, G. pennsylvanicus, have been housed in a great
variety of containers. Successful breeding has occurred in
cages with a moist sand substrate to facilitate oviposition.
Stock cages 42 x 18 x 18 inches made of aluminum screen
will house 250 adults. Five hundred nymphs can be reared
together in an 11 x 7 x 8 inch glass aquarium. A 16-
hour photoperiod at a temperature of about 24°C and a rela-
tive humidity of 60 to 65 percent are appropriate conditions
for rearing crickets (58).
Roach colonies have been kept in larvae cages 40 cm high by
20 cm in diameter with plastic sides, metal bases and per-
forated lids. The ten hardboard platforms provided added
surface area and relative seclusion for the roaches. A 70
percent relative humidity and a temperature of 27.5°C were
maintained for the colonies. Any enclosure with layered
surfaces to increase the area should be adequate for roach
maintenance (59).
Locusts and grasshoppers have been housed in many types of
enclosures. Electric lights of 25 watts are suggested as a
heat source (60).
35
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Praying mantids can be kept at 75° to 88 °F and at a rela-
tive humidity of 50 to 70 percent, but after the fifth molt
they become cannabalistic and should be separated (61).
Nutrition -
The main diet of crickets in one lab consists of a mixture
of biscuit meal, dried milk, Bemax and dried yeast in a
ratio of 50:5:40:5.. Fresh greens are provided about once a
week. A constant supply of water is required (62).
Cockroach diets can have many components. Table scraps,
dog food, and rat food are suggested. Water should be
constantly available (59).
Fresh grass and dry wheat bran are the components of one
diet for locusts. Water is necessary at all times (60).
Praying mantids will eat almost any living insect of an
appropriate size (61).
Breeding/Rearing - •
Some crickets breed all year long, while others have but
one generation per year ( 58 ). For all types, moist sand
should be provided for oviposition and the eggs removed
to petri dishes for hatching. Incubation takes 15 days
at 24°C but hatchina can only take place after a three
month diapause period at 4 C ( 62 ).
Cockroaches oviposit their eggs in dried food mixtures,
from which they can be removed with a sieve and incubated
at a relative humidity of 70 per cent ( 59 ).
In the lab, locusts lay eggs in moist well-packed sand.
The pods can be removed and incubated at 32°C and will
hatch in 11 to 25 days, depending on the species (60).
All Orthopterans have simple metamorphosis: the nymphs
hatch from the egg as miniature adults except for the
wings and achieve adulthood by periodic molts.
Special Needs -
Most insects need high humidity and a constant supply of
water.
36
-------
Ecological Role -
Crickets and roaches are omnivorous insects and will feed
on many kinds of organic matter. Locusts and grasshoppers
are vegetarians and can occur in very large numbers, some-
times defoliating the countryside. Praying mantids are
predators and feed primarily on insects. All of these
species are possible food items for insectivorous inverte-
brates and vertebrates.
Longevity -
Few representatives of this order live longer than one
season as adults.
Problems with Mass Culture -
Adult praying mantids cannot be housed together for they
tend strongly toward cannabalism.
Toxicity Testing -
Cockroaches are extensively used in toxicity testing in a
great number of ways, e.g., effects of pesticides on enzymes
(63, 64, 65} and the nervous system (66). Orthopterans
have been used in studies of metabolic reactions (67, 68,
69, 70) and in open field studies (71).
General Suitability -
The insects in this order are easily maintained and very
abundant. Roaches, grasshoppers, crickets and praying man-
tids are available commercially from suppliers. Praying
mantids, being strictly carnivorous and relying heavily
on insects for food, might accumulate certain chemicals or
be more heavily exposed to target animals. This ecological
role might make them especially interesting test subjects.
Ecological Alternatives -
For crickets, leaf hoppers. For praying mantids, carni-
vorous beetles.
37,
-------
Hemiptera-Homoptera
Introduction -
Hemipterans are true bugs. Their upper half-wings, or
hemelytra, are proximally tough and leathery, and distally
membranous. Their mouth parts are the sucking-piercing
type, enabling them to extract vital juices from plants
and animals ( 57 ). The Homoptera are closely associated
with the bugs and share many characteristics with them.
A distinct difference is implied in their name: both pairs
of wings are similar and entirely membranous. However,
some members of this group are wingless. All Homopterans
are plant feeders (57). Both orders undergo simple meta-
morphis.
Caging/Lab Conditions -
There are no special requirements for rearing most of the
common species of Hemiptera and Homoptera, though the
life cycles of some groups may present difficulties
(e.g., cicadas). Milkweed bugs (Oncopeltus fasciatus) can
be reared in small glass tubes.
Nutrition -
In natural situations all of these insects utilize the
fluids of many kinds of plants and animals.
Ecological Role -
Hemipterans and Homopterans are feeders on organic fluids,
primarily plant juices. They can be destructive agri-
cultural pests. These insect groups are food for
insectivorous invertebrates and vertebrates.
Longevity -
It is doubtful that' most adult Hemipterans and Homopterans
will live longer than one year. Cicada larvae however,
care long-lived and could -provide a unique means for toxi-
city in soil ecosystems.
Toxicity Testing -
Chinch bugs.(Blissus) are serious pests of cereal crops.
The effects of chemical control measures on their numbers
have been studied ( 72 ). Assassin bugs (Triatoma) are
.blood-sucking insects which transmit a Trypanosome known
as Chagas disease in South America. The effects of chemi-
cals on this species have also been studied ( 73, 74 ).
38
-------
Aphids or plantlice are a large group of Homoperans and fre-
quent pests on vegetation. Toxicity testing with aphids have
been concerned with their control (75, 76).
General Suitability -
The herbivorous aphid species are good selections for use
in studies of environmental contaminants that may accumulate
or deposit on vegetation.
Ecological Alternatives -
For aphids , herbivorous invertebrates.
39
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Coleoptera
Introduction -
About 40 percent of all known insects are beetles. They
comprise the largest order of animals/ with well over
25,000 species in the U.S. alone. The sizes, habits, and
ecological niches of beetles vary greatly. Some are phyto-
phagus, some predacious, some scavengers, some saprophagus,
and others parasitic on plants and animals, Beetles are
found almost everywhere in freshwaters and on land ( 57 ).
The Coleopterous (sheath-wing) insects have four wings,
the top pair a thick, hard set (elytra) to protect the
membranous under set. Mouthparts are basically of the
chewing type though some are modified for sucking.
Metamorphosis is complete. The life cycles vary con-
siderably within the group (57).
Beetles that have been used in research are frequently
pest species, though not exclusively. Included are ground
beetles, [Harpalus, Agonum ( 77), Feronia ( 39)], lady
beetles, [Hippodamia, Coleomegi1la ( 7 8 , ~7 9 )] and flour
beetles, [Tribolium ( 80)]. Mealworms, Tenebrio sp. are
commonly reared as food for insectivorous animals ( 81 ).
Caging/Lab Conditions -
Depending on their size and habits, beetles are relatively
easy to maintain under controlled conditions. The proper
temperature, 25° to 30 °C, and humidity, 50 to 65 percent,
are essential to all insects. Insect breeding may depend
on the photoperiod. A list of species bred in Great Brit-
ain, their nutrition and temperature requirements is
found in reference 81.
Nutrition -
The food habits of beetles are so varied that a general
statement about them would be useless. A close matching
of natural foods has been the general system in animal
maintenance and sould be acceptable for beetles. For a
large number of pest species, artificial diets have been
determined and a keyed list of them can be found in
reference 81.
Breeding/Rearing -
Many species are easily bred in captivity, given the proper
medium e.g., grain weevils (Sitophilus) on cereal grains,
40
-------
or mustard beetles (Phaedon) on potted cabbage, turnips
or radish plants. Tenebrio sp. can be bred year round
with great ease ( 81 ).
Ecological Role -
Some beetles are pests on agricultural crops and other
are predacious ground dwelling species (e.g., Harpalus)
( 82). Others feed on fungi and carrion. All beetles
are potential food for insectivorous invertebrates and
vertebrates.
Longevity -
Like most other insects, the generation time of beetles
is seldom longer than a year and it is unusual when adults
live for more than one year.
Toxicity Testing -
Insects are the primary pests which afflict man and his
domestic stock and crops/ and millions of dollars are spent
yearly for insect control. By far, the majority of toxi-
city testing using beetles has been concerned with pest
species. Pesticide residues in many species have been
studies including Popillia, (83, 84, 85 ); Tribolium
(80 ); ground beetles ( 36, 39, 77, 82 ); and lady
beetles ( 78, 79 ). Large numbers of toxic chemicals
have been tested on beetles (78 ).
General Suitability -
Beetles are an ecologically diverse group which generally
appears to be attractive for use in toxicity testing.
Ecological Alternatives^ -
For ground beetles, centipedes. For scarab beetles,
millipedes. For plant eaters, Hemiptera and slugs.
41
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Lepidoptera
Introduction -
Butterflies and moths are conspicuous and well-known in-
sects, among which are some of the most beautiful animals
we know. However, the adults that please the eye are often
metamorphosed from destructive phytophagus larvae that are
major insect pests of cultivated plants. The adult mouth
parts are the sucking type and metamorphosis is complete in
the species of this order. The larval forms can produce
silk for cocoons and shelter (57).
Many Lepidopterans have been reared under lab conditions.
Three general methods - two for pest species and one for
nocturnal moths (the largest family of Lepidoptera) will be
discussed.
Nutrition -
Some Lepidopteran larvae are general feeders and others are
very specific, feeding on one plant species exclusively.
Lab diets must reflect these natural habits and require-
ments.
Breeding/Rearing -
The black cutworm (Agrotis ypsilon) has been difficult to
rear due to cannabalism, but because of its economic impor-
tance, a method has been found by some Canadian researchers.
Pupae are obtained from nature and after emergence the adults
are held in 12 x 12 x 16 inch cages with sliding glass panels
to permit air circulation. Twenty moths per cage is the
normal capacity and ambient temperature is 80 + 2°F. Gallon
jars with moist paper are used to supply an egg laying
environment and a light-dark photoperiod is maintained.
Pyrex trays 12 x 8 x 2 inch with 1 inch moistened sand are
provided for larvae. At the initiation of the fifth instar,
each larva is separately placed in a small flint jar with a
1.5 inch layer of a 1:1:1 sand-sandy loam-vermiculate mix-
ture with about 6 percent moisture added (86).
The codling moth (Carpocapsa pomonella) is reared in 53.3
x 30.5 x 7.6 cm stainless steel trays with a 10 to 13 mm
layer of burrowing medium on the bottom. To prevent con-
tamination and dehydration, a thin layer of paraffin is
sprayed over the surface with a specially designed apparatus
(87). Larvae of Noctuid moths have been successfully reared
in the lab from eggs in 6 ounce waxed paper cups with
42
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standard cardboard lids. A diet, shown in Table 2, is
placed in each cup. The number of larvae per cup depends on
the species (88).
Adult black cutworm larvae are fed by means of dental cotton
plugs soaked in a 10 percent dextrose solution suspended
in the cages on strings. Larvae are fed red clover or
tobacco (86) .
Artificial diets for codling moths are based on ingredients
such as wheat germ, soybean meal, rice flour, pinto beans,
alfalfa meal, cottonweed meal, corn meal and carrot powder
(87).
Noctuid larvae have been successfully reared on the medium
described in Table 2 below.
Table 2. COMPOSITION OF AN ARTIFICIAL
MEDIUM USED FOR REARING NOCTUID SPECIES
Ingredient
Soaked lima beans
Soaked pinto beans
Dried brewers' yeast
Ascorbic acid
Methyl p-hydroxybenzoate
Sorbic acid
Formaldehyde (40%)
Agar
Water
Grams for 120-150 Cups
Present Method
2133
320
32
20
10
20
128
6400
Source: 88
43
-------
Special Needs -
Excessive humidity can cause fungi and bacteria growth on
insect growth media and infections of the animals them-
selves. On the other hand, a lack of moisture and the
resulting dehydration can also cause mortality in lab
colonies. A careful balance of temperature and humidity
is mandatory (87,88).
Special diets for the various life stages of Lepidopterans
may be expensive and time-consuming to prepare.
Ecological Role -
The larvae of butterflies and moths, often severe agricul-
tural pests, are economically much more important than the
adults, some of which never feed. They frequently supply
food for insectivorous predators.
Longevity -
Most Lepidopterans have an annual cycle. Eggs or pupae
overwinter. The larvae feed voraciously and pupate. Adults
often have the sole task and ability to mate and lay eggs.
Problems with Mass Culture -
Many larval forms are cannibalistic, especially in crowded
quarters. Isolation may be necessary during certain instar
stages (86,88).
Toxicity Testing -
Much of the research with butterflies and moths has been
with pheromones (89,90,91,92,93) often with the goal of
achieving population control. However, a few toxicity
studies are available (94,95,96).
Researchers have found a way of closely scrutinizing the
flight of insects and the comparison of certain measurements
between controls and test organisms gives a reliable, be-
havioral test (97) .
General Suitability -
Some Lepidopterous insects can be routinely bred and reared
in captivity and can make excellent test animals.
Ecological Alternatives -
Other phytophagous insects.
44
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Diptera !
Introduction -
The "flies" are a well-known group o'f insects and one of
the larger orders. They possess only one pair of wings -
the front pair. The hind set are reduced to balancing, knob-
like organs. Mouth parts are sucking and metamorphosis is
complete.
Most flies are small soft-bodied insects, many of which are
economically important. Mosquitoes, stable flies, house
flies, and blow flies are pests of man"and other animals.
Many diseases can be transmitted by Dipterans. Many other
flies are predacious on harmful insect species, some are
important as pollinators, and still others are useful
scavengers (57).
Most of the available maintenance procedures are concerned
with pest species and these will be reviewed. A group of
smaller insects (family Chironomidae) has also been suc-
cessfully reared in the lab and will also be discussed be-
low with mosquito (family Culicidae) rearing techniques.
Caging/Lab Conditions -
Larger flies (i.e., Muscidae, Calliphoridae) can be kept
in any escape-proof container at a temperature of 27.5 +
1°C and relative humidity of 50 to 55 percent. Windows are
not necessary but proper air circulation is important.
Wooden cages (45 x 45 x 45 cm) with screen sides that can
be dismantled for cleaning are used to keep adult house
flies (Musca domestica), and stable flies (Stomoxys calci-
trans) (98, 99).A minimum of 50 cm3 for each adult is a
good guide for space requirements (98).
Mosquito adults (Culex, Aedes, Anopheles) can be easily
housed in 18 x 18 x 18 inch plywood cages screened with
18 mesh screen. In most fly cages, a muslin sleeve is
attached (24 inches long, 9 inches circular opening) to
provide access while preventing the escape of the animals.
Temperatures between 24° and 27°C and relative humidity
around 80 percent are satisfactory ambient conditions. A
12L:12D photoperiod enables Anopheles quadrimaculatus to
carry out its normal life cycle "(100) .
Chironomids can be housed in cages similar to those of mos-
quitoes, but because they may need ample space to swarm in
order to reproduce, larger enclosures (5.25 x 4.5 x 7 feet)
45
-------
must be provided with a 75 gallon aquarium filled with mud
for the floor (see Breeding/Rearing for further discussion).
Chironomus plumosus will breed under such conditions with an
ambient temperature of about 25°C and a relative humidity of
70 percent (101).
Fruitflies (Drosophila), can be easily raised in the lab in
many sizes of cages and are widely used in many types of
research (102).
Nutrition -
Adult houseflies and blowflies are provided with a shallow
dish of sugar for feeding. Stable flies are fed blood
offered in soaked cotton, the live host, or in dishes on
the floor of the cage. Water soaked cotton pads should be
available for drinking in all fly. enclosures (98, 99, 103).
Larval houseflies and blowflies are reared on many media,
the general ingredients consisting of various combinations
of dog biscuits, fish, wheat bran, alfalfa meal, baker's
yeast, malt extract, manure, sugar, and water (98, 103).
Larval stable flies are more difficult to rear. Stoiaoxys
cultures should be fed the diet found in reference 99.
Only small amounts should be given at one time until larval
consumption increases. Shallow flat dishes are suitable
holding containers for larvae and medium. A pH of above
7 is desirable for the medium, and using only 24-hour old
medium assures that newly hatched larvae will not encounter
acid conditions (99).
Adult mosquitoes can be maintained on soaked seedless
raisins or a 10 percent sugar solution in saturated gauze padi
Females get a blood meal via a shaved guinea pig three
times a week ( 100). Adult female mosquitoes can also be
fed on shaved rabbits. Larval mosquitoes are fed a diet
of wheat flour, dry brewer's yeast, dried beef blood, and
non-fat dry milk. The finely ground mixture is sprinkled
sparingly on the surface of the water (100).
Breeding/Rearing -
The procedure for breeding houseflies is to transfer
adults to a cage made of gauze with a plywood bottom (see
Caging) (104). Cotton-wool pads soaked with a milk-water
solution (1:1) provide an oviposition site. The pads with
46
-------
eggs are then transferred to jars with rearing medium and
muslin caps. About eight days after the eggs are laid,
larvae will migrate to upper layer of medium and pupate.
This layer is removed to an adult cage where emergence
takes place (98) .
Ground horse meat is the medium most often used for ovi-
position by blowflies. Eggs remain in the meat and are
transferred to larvae medium in glass jars. They hatch in
24 hours. In eight days they pupate and emergence occurs
six days later. Sawdust is used as a sponge to soak up
excess moisture from the meat in the culture jars (103).
Oviposition cages for stable flies are of the same con-
struction as adult holding cages, but smaller in size
(25 x 25 x 25 cm). Oviposition occurs more readily in
confined spaces. About 6.5 days after emergence, well-fed
flies will produce the first eggs, but full production
doesn't occur until the 10th day, when the transfer of
gravid adults to Oviposition cages is suggested. Eggs are
laid through perforations in a zinc floor and caught on the
black cloth surrounding the cage. Flurorescent lighting
and adult food should be present. About 60 to 70 percent
of the eggs will hatch and placed on a culture medium, will
pupate in nine to ten days. Pupae are placed in adult
cages for emergence (99).
Mosquitoes (Anopheles guadrimaculatus) oviposit in 8.5 x
4.5 x 2 inch enamel dishes of aerated tap water two to
three days after a blood meal. Eggs are transferred to
rearing pans (9 x 15 .x 2.5 inch). Dechlorinated water
should be used. After hatching, larvae are separated to
500 per pan and fed the larval diet (see Nutrition). Pupae
are transferred to the holding cages for emergence. It
should be noted that different species of mosquitoes have
different specific requirements, though the rearing pattern
for A. quadrimaculatus is, in general, characteristic for
other species(100).
Midges (Chironomus plumpsus) have been bred and reared in
the laboratory. The Chironomid group swarm under specific
conditions, and successful breeding depends on suitable
ambient temperature and light. Cages are lit by fluores-
cent and incandescent lamps and natural conditions are simu-
lated by a gradual increasing and diminishing of intensities.
Swarming takes place in the "evening". Some successful
breeding has taken place in small cages (3.0 x 1.9 ~x 1.5 cm)
(105) . Larvae will develop to the fourth ?.nstar in the mud
substrate at the cage bottom. Trainer1s Dog rewards are
added as food after they are given thorough soaking. Pupa-
tion occurs in the substrate and adults emerge after nine
to 13 days (101).
47
-------
Special Needs -
Diets and maintenance requirements are different for each
species. There are many entomology laboratories throughout
the United States which can supply procedural data and
special needs for many Dipteran species.
Behaviora]^ Restrictions -
Life cycles can be very complex and difficult. Species-
specific behavioral idiosyncrosies abound in this animal
group. These facts can make lab breeding/rearing difficult
or at least, demand special provisions (e.g., swarming
cages for Chironomids).
Ecological Roles -
Many adult Dipterans are vectors of disease and nuisance
pests of other animals. However, they also can represent
staple foods for insectivorous predators (i.e., bats,
swallows, frogs). Aquatic larvae are frequently major food
sources for fish in quiet waters.
Longevity -
It is doubtful that adult Dipterans live for more than one
year; they probably live for a much shorter period of time.
Toxicity Testing -
Dipterans have been the subjects of conditioning experi-
ments, with mixed success (106, 107). They have been used
in a great number of experiments with toxic substances to
determine mode of entry (108, 109) and general effects (80,
110, 111). Sublethal effects on reproduction, behavior
under stress conditions (112) and physiology have also been
studied (113, 114, 115). Insect resistance to toxic sub-
stances (116, 117, 118) and even the effects of toxic sub-
stances on Dipteran eggs have been studied (119).
48
-------
General Suitability -
Many flies have been used in research and for a good number
of species the maintenance procedures are available. This
makes them worth consideration as test species. The double
association of some forms (e.g., mosquitoes) to aquatic and
terrestrial systems at different times during their life
cycles may make them particularly suitable subjects in ex-
periments in which land-water transfer of a substance is to
be studied.
Ecological Alternatives -
Blood sucking Hemipterans.
49
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Hymenoptera
Introduction -
The Hymenoptera (membrane-wing) contain what man regards as
the most beneficial group in the insect kingdom, the honey-
bees. There are also a good number of wasps that parasit-
ize other insects, many important pollinators and several
interesting social groups. Ants, sawflies, Ichneumons,
Chalcids, wasps and bees all belong to this order. They
have chewing and chewing-sucking mouthparts and undergo
complete metamorphosis. Bees are the only members that
have been maintained to any extent.
Caging/Lab Conditions -
Housing of honeybee (Apis) colonies is an old and well-known
practice and there are very extensive texts available on the
subject. Bumble bees (Bombus) are also important as polli-
nators of legume crops and have been bred and reared in the
lab. Wooden nest boxes contain 'two compartments: a feeding
and a brood chamber. Nesting material such as dried grass
or cotton is supplied, and, for feeding, a honey-and-pollen
mixture. The queens are introduced to the nest and their
natural nesting instinct usually takes over. Some reports
attribute success in colony establishment to the addition
of some workers to the nest with the queen. Queens are
caught in nature and fed on pollen and honey. After the
establishment of colonies in the lab, they have been set
outdoors to do their pollination job. Successes have been
reported for about 30 species. Some success at overwinter-
ing queens has also been reported (120).
The Agricultural Research Services, USDA, Tucson, Arizona,
has utilized honey bees (Apis mellifera) in toxicity testing
under lab and field conditions. Their lab cages are 6 x
6x2 inches with 100 bees per cage (121).
Nutrition -
Bumblebees have been kept in the lab on honfcy and pollen
(120). Honeybees have been maintained on 60 percent suc-
rose and pollen (121). The larvae require no special diet
because of the care given them by the adults.
50
-------
Breeding/Rearing -
Once a bee colony is established it will be self-perpetuating
for at least that season. Honeybee colonies remain intact
over winter and may last for many years, but whether
colonies which have been kept in the lab year-round are still
able to reproduce is uncertain, because uiost have been
initiated for research outdoors.
Special Needs -
Nest boxes are. necessary for the colony.
Ecological Role -
Many Hymenoptera are important as pollinators and as
parasites on other insects. They feed on pollen, plant
juices, and many other liquid foods.
Longevity -
i
The lifetime of most adult Hymenopterans can be measured
in months.
Toxicity Testing -
There have been numerous studies of the resistance of
Hymenopterans to the adverse effects of toxic substance
(122, 123, 124, 125, 126, 127, 128). General studies
both in the lab and field have also been completed (129).
General Suitability -
The social species of insects, e.g., ants and bees, appear
to be very interesting potential subjects because there
already exists a considerable body of knowledge on their
social organization and population dynamics and because
some species are relatively easy to keep and simple to work
with (121 ). Studies at the population level and of sub-
lethal effects on behavior could easily be performed and
the results reviewed in the light of known patterns.
51
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Miscellaneous Invertebrates
This section discusses a group of insects which have not
been extensively studied in the laboratory, but which may
prove to be useful subjects for toxicity testing. Included
here are the soil arthropods and those that are closely
associated with ground and other substrates, as well as
other miscellaneous groups. Centipedes (Chilopoda) and
millipedes (Diplopoda) are interesting animals and play im-
portant roles in the micro-systems in which they exist:
centipedes are predators, sucking the fluids from their prey
through their mandibles; millipedes feed mainly on dead or-
ganic matter. Both have been used as test animals (130).
Springtails (Collembola) are apterygote insects that loco-
mote by means of a forked structure which, extended, causes
the animal to jump. Springtails are inhabitants of leaf
litter, decaying logs and fungi. They are basically vege-
tarians .
Thrips (Thysanoptera) are also insects and plant feeders.
They occur on all types of vegetation and can be serious
pests of cultivated plants.
Another Arthropod group are the Isopods, or pill bugs. They
are frequently found around decaying matter and play an im-
portant role in the decomposition of organic solids. Iso-
pods are abundant throughout North America and because of
their saprophagous feeding habits and abundance, may prove
to be very good test animals. They have been used previous-
ly (131).
Scorpions (Arachnoidea) are rapacious arthropods that occur
primarily in subtropical and desert regions of the United
States. They nocturnally feed on insects and spiders, first
killing or immobilizing them with the sting apparatus on
the distal tip of the abdomen. Scorpions are relatively
easy to keep. Palamnaeus gravimanus have been kept in wood
cages 3 x 2.5 x 1 foot with a soft earth substrate for
burrowing. Wads of wet cotton kept them moist and cock-
roaches were the staple food. Some evidence indicates they
may be quite suitable for use in toxicity testing (132).
52
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VERTEBRATES
Amphibians and Reptiles
Amphibians and reptiles, the herpetofauna, are poikilother-
mic vertebrates that inhabit both terrestrial and aquatic
environments of all types. The herps are basically egg
laying animals. Amphibians usually require water for
successful reproduction. Reptiles on the other hand,
through evolution,have developed a land egg which can be
deposited in many types of terrestrial nests. Some rep-
tilian species are live-bearers and do not lay eggs.
Most reptiles and amphibians are carnivorous and feed on
insects, other invertebrates, ybung birds, eggs, and small
mammals. However, a few lizards and turtles are herbiv-
orous.
Amphibians and reptiles have been used in laboratory re-
search in a few ways that may prove applicable to toxicity
testing. For example, amphibian eggs are excellent for
embryological studies and are often used in college courses.
In toxicological research, however, there has been very
little attention paid to the poikilothermic vertebrates
probably because of their lack of economic importance.
The following text discusses species which are judged to be
potentially suitable for toxicity testing based on their
ecological importance, availability, ease of laboratory
maintenance, and reproductive success in captivity. Be-
cause of the paucity of information available in the liter-
ature on herpetofauna that might aid in such a selection, a
number of herpetologists were consulted for suggestions. A
summary table indicating the suitability of herpetofauna for
toxicology testing is provided in Table 3.
53
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TABLE 3. SUITABILITY - HERPf:P
HERPS
Amphibians
Turtles
Lizards
Snakes
>.
-U
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t-(
-H
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a
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A
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4J
-H
>
-H
C Q.
•a o
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G
F
01
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c
a
c
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c
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10 as
as
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G
o>
tn a>
a c
•H
tn -i*
3 01
O 01
j>
M *H
P*
F
P
w
C
o
-H
4J
O
•H
JJ
tn
Hi
K
Pure water
Housing
methods
Captive
breeding
>,
tJ
•H
i-t
-H
r-tŁ>
l-l 4J .
01 -H
c a
0) CO
G
F
iH
to
0
•H
Dl
O 01
r-J r-t
O O
o a
B2
B,
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(j
o
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10 O
-H OJ
CO 0.
X
All
Aq
Fo
•o
-p a
tn -H
01 0
O> 0
3 Ul
CO
Rana pi:
Amb'.fs tor
Terrapen
Cheliidrj.
ens
.1 ifxi -anu".
e sp.
8 erper.ti.na
FofGr AncLis earoZ inms
De Phrvnosorna eornut
F-G
All
Tha^ncphis sirtalie
Elarhe sp.
Legend:
General Ecological Role
A - Large carnivore
B - Small carnivore
1. General
2. Insectivore
C - Omnivore
D - Herbivore
1. Seed, fruit eater
2. Grass foliage eater
Habitat Preferences
Fo
Gr
De
Aq
Do
Forest
Grassland
Desert
Aquatic
Domestic
E = Excellent
G = Good
F = Fair
P = Poor
Sources: - literature cited in text
- interviews of Scientists
-------
Anurana
Introduction -
Amphibians are so named because they metamorphose from
aquatic forms into air-breathing and often terrestrial
tetrapods. The aquatic forms of frogs and toads, the tad-
poles, have provided scientists with unique opportunities
to study developmental biology.
There are six families of Anurans in North America: the
spadefoot toads, Pelobatidae, the narrow-mouthed toads,
Microhylidae, the other toads, Bufonidae, the leptodactylid
frogs, Leptodactylidae, tree frogs and their allies, Hylidae,
and the true frogs, Ranidae. Of the true frogs, the leopard
frog (Rana pipiens) has been utilized by researchers for
many types of investigations. It is a very abundant and
widespread species that occurs in many kinds of waterbodies
throughout the U. S., Mexico, and Southern Canada (133).
Many other species of toads (Bufo) and true frogs (Rana)
are locally abundant and also bred, reared and maintained
in labs throughout the U. S. However, since the leopard
frog is the major subject in amphibian research, the follow-
ing text is focused primarily on this specie..
Caging/Lab Conditions -
For adult frogs, just about any waterproof enclosure with
available water is sufficient to house large numbers (134).
Plastic tanks, large sinks (22 "x 14 x 14 inches) for about
25 adults (135), and plastic rat cages (22-3/4 x 11 x
6 inches) with perforated stainless steel lids (136) have
been suggested cages. Adults can also be maintained for
weeks under refrigeration (4°C) in water. No feeding is
necessary during this time. The water, however, should be
changed three times weekly (134). Further enclosure speci-
fications and handling techniques can be found under Breed-
ing/Rearing.
Water for use in amphibian cages should be unchlorinated and
unfluoridated, especially water in which larval forms are
kept (134, 137, 138, 139, 140).
Proper temperature regimes are quite variable for adults,
and should depend on the state of mobility desired and the
techniques to be performed. Certain anesthesia doses are
differentially effective at different temperatures (141).
For active adult animals, 18° to 20° C is suggested (134).
55
-------
Larval anurans are affected in their sexual development by
ambient water temperatures. For normal results, a 15° to
25°C range is sufficient (134) .
The Mexican axolotl, Ambystoma mexicanum, have been main-
tained in the laboratory in containers made of asbestos
cement at the time they reach 5 mm, for caging specifications
see reference 142. Water in the salamander containers
should be unchlorinated and circulated constantly (142).
Work in the Amphibian Facility at the University of Michi-
gan, Ann Arbor, is proving the feasibility of mass producing
Rana pipiens as well as other amphibian species (137).
Nutrition -
Cooked lettuce and various other greens have been most often
used as food for tadpoles (137, 138) , but one nutritional
study showed a 300 percent weight, increase at seven weeks in
larvae fed Purina No. 1 trout chow over those fed just let-
tuce (143). Natural pond plankton and bits of liver have
a.lso been successfully used to rear tadpoles (148) . Spinach,
however, has been shown to cause kidney stones in some
amphibians (144) and is not recommended (137).
For recently metamorphosed frogs, small mosquitoes are ex-
cellent food, and can be replaced by flies (137), crickets
and liver bits (134) as they mature. Adults have survived
in excellent condition by being fed twice weekly, crickets
(two to three) alternated with worms (134).
Suggested diets for the appropriate life stage for amphibi-
ans are presented in Table 4. Adult axolotls thrive on
beef heart with vitamins A, D, B complex and powdered CaCO0
added (142) . ^
Breeding/Rearing -
The breeding and rearing of most amphibious species are
more complicated than those of birds or mammals because
amphibians show no parental care for the young. In frogs
and toads, the eggs are laid in water and fertilized ex-
ternally by the males during amplexes. (Salamanders have
internal fertilization by means of spermatophores deposited
by the male and taken up by the females into her cloaca.)
The eggs are left to develop, hatch and mature on their own.
Therefore, rearing techniques must include care for the or-
ganisms from the zygote stage to the adult stage. Table 5
presents the natural breeding habits and numbers of eggs for
some North American and European Anurans.
56
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Table 4. FOOD RECOMMENDED FOR AMPHIBIAN LARVAE
YOUNG ANIMALS AND ADULTS
Stage In Life History
Anura
Newly hatched larvae
Older larvae
Boiled spinach or lettuce,
green part of Romaine let-
tuce; for Xenopus: dried
nettle or alfalfa powder,
dried yeast, egg or liver
powder
As above
Newly metamorphosed
animals (terrestrial)
Drosophila
Juveniles (terrestrial)
Larger Drosophila species,
mealworms, small earth-
worms, crickets
Metamorphosed animals
(more or less aquatic) and
large, neotenic larvae
(e.g., axolotl)
For Xenopus; Tubifex,
Enchytraeus, small pieces
of liver; later: earth-
worms and beef heart cut
into small blocks
Adults (aquatic)
For Xenopus: earthworms,
beef heart cut into small
blocks
Adults (terrestrial)
Larger species of Drosophila,
house flies, mealworms,
crickets, grasshoppers and
other insects.
Source (142)
57
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Table 5. NATURAL BREEDING HABITS OF COMMON
NORTH AMERICAN AND EUROPEAN ANURA
Scientific
Name
SPECIES
Common
Name
Breeding Season
(in northern
latitudes)
Approximate Number of Eggs
Laid by One Female, and
Mode of Deposition
I/I
CO
Rana catesbiana
Rana clamitans
Rana palustris
Rana pipiens
Rana sylvatica
Hyla grucifer
Hyla versicolor
Bufo americanus
Bufo Fowleri
Rana esculenta
Rana temperaria
Bufo bufo
Bullfrog
Green frog
Leopard frog
Woodfrog
Spring peeper
Treefrog
American toad
Fowler's toad
Edible frog
Common frog
Common toad
June-July
June-August
Pickerel frog April-May
April-May
March-April
April-June
May-July
May-June
May-June
May
February-April
March-April
10,000 or more, in large film on
surface, 60 cm across
1000-4000, in film on surface,
less than 30 cm across
2000-3000, in firm globular
masses attached to twigs
3000-5000, in flattened sphere
2000-3000, in globular mass
800-1000, laid singly among plants
1000-2000 in small packets of
30-40 eggs
4000-8000, in two long strings
Up to 8000, in two long strings
1600-1900, in clumps
1000-4000, in large clumps
3000-4000, in two long strings
-------
George W. Nace, et al. , began an amphibian maintenance pro-
gram in the mid-W60' s at Ann Arbor, Michigan and has very
successfully bred and reared a variety of frogs, and sala-
manders, with the purpose of developing genetically defined
strains. A general review of their techniques and inserts
from various other maintenance programs follows (for speci-
fic detail, see 137).
It has been discovered that breeding can be induced in
Anurans by injecting the animals with pituitary tissues
from other frogs, and progesterone (145). Females can be
injected and stripped of their eggs which are then insemin-
ated by sperm from excised testes, or males can also be in-
jected and natural breeding behavior allowed to occur. The
artificial method allows for unique egg-sperm distributions
or for other specific handling techniques. (The specific
procedure for inducing ovulation can be found in reference
138). Generation time (at Ann Arbor, Michigan) is 13 to 15
months.
Embryos are held in enamel pans and water (unchlorinated) is
changed every third day. When the tadpoles begin to swim
they are transferred into specially designed bottles which
hold 25 to 75 larvae, depending on the species and size of
individuals. These are one-gallon bottles with their bot-
toms removed. A one-hole cork is fitted in the neck, they
are positioned upside-down and a leveling-drain tube extends
from it to the desired water level. A wire screen keeps the
tadpoles from the narrow neck and from debris that collects
at the bottom of this aquarium. Water is slowly added to
the system and circulated through it. There is good evi-
dence that the average growth rate of tadpoles decreases as
the density rises (138).
Metamorphosed animals are transferred to larger plastic con-
tainers with appropriate water circulation. These are
tipped up on one side leaving a pool at the lower end and a
terrestrial area at the other.
Adults are housed in larger containers that can be easily
cleaned and that provide both aquatic and terrestrial areas.
Salamanders breed from January to June. After two to three
months some species can be bred again. Breeding is conveni-
ently induced in the Mexican axolotl by a sudden decline in
temperature (22°C to 12°C for two days). Males and females
are kept separately except to breed and put together into
the colder water. The breeding chamber should be-sheltered
from light and no food provided until after oviposition on
plastic tubes (13mm diameter) fitted to aquarium sides be-
low the water line. Eggs are reared in 14° to 18°C water
in shallow trays. During culture, crowding should be
59
-------
avoided. Larvae should be fed Artemia or small Daphnia ob-
tainable from supply houses. Mosquitoe larvae also can be
used. Containers should be cleaned of detritus daily.
Sexual maturity is achieved in males at about seven months
and in females in nine or ten months (142).
Special Needs -
Larval forms need fresh, unchlorinated water, which must be
changed frequently. Tap water can be treated in various
ways to remove the chlorine (134).
Fresh live insects must be continuously available for
adults (135, 137).
Embryos and tadpoles need special handling (137).
Ecological Role -
Anurans are carnivorous animals that feed on a great variety
of invertebrate species, particularly insects. Many larger
predators utilize them as a food source. Birds, snakes,
turtles and mammals feed on the adults, and the tadpoles
provide food for predators associated with aquatic habitats
(133).
Longevity -
Rana pipiens has achieved life spans of three years in the
laboratory (134).
Problems with Mass Culture -
Mass culture of anurans requires a large supply of unchlo-
rinated water (134,137) and sufficient lab personnel to
feed and maintain the various life stages.
The susceptibility of frogs and toads to parasites is well
known and health maintenance programs including penicilin
inoculations, etc., may be required (137,146).
Toxicity Testing -
Larval and adult frogs and toads have been used in studies
of the effects of pesticides on survival and reproductive
behavior in outdoor enclosures '147,148,149,150 151) and
under lab conditions (152,153,154,155,156).' Heavy metal
(157 ) and irritant vapor (158,159) studies have also been
performed. Their use in cancer research is very extensive
(160,161,162,163,164).
60
-------
General Suitability -
In toxicity tests, the accessibility of the early life
stages of anurans and the sensitivity of frog skin makes
them very suitable subjects. An increase in breeding
amphibians in recent years has made them more available than
in the past.
Rana pipiens is the most likely subject due to its wide use
in many research areas, its availability and the large body
of basic information that has been gathered. Ambystoma sp.
are salamanders that have received much previous attention
and are being bred at the University of Michigan Amphibian
Facility (137).
Ecological Alternatives -
Turtles, Lizards.
61
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Chelonia
Introduction -
Turtles are reptiles with a very long history on earth.
They are shelled organisms that are mostly associated with
aquatic environments, but wholly terrestrial species are
not uncommon. The feeding habits of turtles are quite
variable and most species seem to be omnivorous, feeding
on a great variety of plants and animal materials. They
are found in many natural habitats, particularly around
ponds, streams and rivers.
Caging/Lab Conditions -
Aquatic turtles have been adequately housed in concrete
tanks (130 x 72 cm) lined with paraffin through which
unchlorinated water was constantly circulated and main-
tained at a depth of 25 cm. Wooden platforms were pro-
vided onto which the turtles could crawl. Terrestrial
species were kept in dry tanks and provided with water
for drinking (165).
Terrariums are also excellent enclosures for keeping tur-
tles, and are easily transported (133).
Nutrition -
Ground horsemeat and lettuce is a suitable diet for many
species of turtles (165) and raw hamburger and table scraps
are also recommended ( 133). In the wild they consume
insects, crayfish and other invertebrates.
Breeding/Rearing -
No information is available on breeding turtles in captiv-
ity. Since they are egg layers, the collecting of eggs
from natural nest sites is a possibility. Most species
produce large amounts of eggs yearly.
Ecological Role -
Turtles are basically omnivorous reptiles and are important
elements in the aquatic systems. They are predators on all
types of invertebrates and some species are avid consumers
of aquatic vegetation.
62
-------
Problems with Mass Culture -
Turtles are susceptible to a great variety of parasites and
diseases including tuberculosis and many types of Salmonella
(166).
Toxicity Testing -
Turtles have been used in terrestrial DDT studies (167) and
parathion distribution studies in aquatic ecosystems (152).
General Suitability -
Turtles might prove very suitable for testing. Their court-
ship rituals are unique and may supply a means by which
behavioral modifications, etc., might be studied. However,
their external dermal skeleton makes minor surgical tech-
niques difficult.
Their morphology and taxonomy are known and sexes are easily
distinguished (165). Terrapene sp. has a number of terres-
trial species and is recommended for use in the toxicity
testing of terrestrial forms. Young snapping turtles
(Chelydra serpentina)are frequently used in laboratory ex-
periments, and though primarily an aquatic species, they are
a likely second choice.
. 63
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Squamata ~ Lizards
Introduction -
The lizards are represented in North America by five fami-
lies: the iguanids (Iguanidae), the skinks (Scincidae), the
glass lizards and alligator lizards (Anguidae), the whiptails
(Teiidae), and the geckos (Gekkonidae).
Caging/Lab Conditions -
Lizards can be housed in terrariums (133) , wooden cages
with glass doors (168), or nearly any escape proof enclos-
ure. Many lizard species are desert or tropical forms and,
being poikilothermic, require rather warm ambient conditions.
Some recommended temperatures are 85° to 90°F for iguanas
(169) (this can be achieved locally by infrared lamps) and
75° to 85°F for gila monsters (168). Arboreal species should
have appropriate props (170).
Photoperiod is important for the regulation of the endocrine
system in some lizards and should be regulated if animals
are kept indoors (171). Light might also influence feeding
in certain lizard species and it has beer reported that re-
luctant feeders can often be induced to eat by putting them
under a wide spectrum artificial light (170).
Nutrition -
Most lizards are carnivorous and can be maintained on a
large variety of foods. However, there are some native
herbivorous species, such as chuckwalla, in the southwest.
Iguana iguana, a common South American lizard seen frequently
in pet stores is also herbivorous and can be satisfactorily
maintained on fresh green vegetables (169).
Carnivorous species like Sceloporus, a native iguanid,
have been fed larval wax moths (Galleria mellonella) (172)
and other easily obtainable insects. Some desert lizards
will not eat if their body temperature is below 35 to 40°C.
Gila monsters, Heloderma, have been maintained for years on
a diet of hens eggs(one every two weeks). Ground whole
rat was substituted for a time with no significant effect
on weight (168).
Water should be provided ad libitum, though certain desert
species will make little use of it. Some Anolis may not
64
-------
drink at pools, but will only take water in the form of dew '
off of leaves. Spraying the cage may be the only means of
watering this lizard group (173).
Breeding/Rearing -
There is no information concerning captive breeding in
lizards. They are egg layers and usually breed in the
spring in nature.
Special Needs -
Infrared lamps or some other local heat source may be re-
quired for lizard cages.
Live food may be necessary because lizards respond to
movement and under natural conditions attack only moving
prey.
Behavioral Restrictions -
Many desert lizards are highly territorial {e.g. Crotaphytus)
and there may be some difficulty in caging individuals of
this species together in any numbers. However, Anolis can
be housed in rather large quantities in cages with suffi-
cient vertical twig surface for all individuals (173).
Ecological Role -
Lizards are both primary consumers, insectivorous, and
carnivorous (Gila monsters eat eggs, young oirds and
mammals). They all, in turn, are consumed by predatory
snakes, birds and mammals.
Problems with Mass Culture -
Lizards, like most reptiles, are relatively unprolific.
Their breedability in captivity has not been thoroughly
investigated and their potential as laboratory colonial
animals does not appear to be very good.
Toxicity Testing -
Lizards have not been extensively used as subjects for
toxicity testing. However, field investigations of DDT
effects on faunal elements have included lizards (174).
General Suitability -
Their poor availability and the lack of knowledge of their
65
-------
reproductive behavior in laboratory situations makes lizards
rather poor candidates for wide use as toxicity experiments.
Anolis carolinensis has been successfully bred in captivity
and would be the most likely candidate (175).
Ecological Alternatives -
Toads, snakes.
66
-------
Squamata-Snakes
Introduction -
Snakes are more closely related to lizards than to other
reptiles. They are legless poikilothermic vertebrates that
are significant predators on smaller animals. North America
has three families: the constrictors, Colubridae, the
rattlesnakes, Viperidae, and the coral snake, Elapidae.
Snakes of the last two groups are venomous (133).
Reptiles differ from amphibians in their ability to breed
without returning to an aquatic environment and in pos-
sessing internal fertilization. Their eggs are laid in a
terrestrial nest. There is no metamorphosis.
Snakes inhabit many types of habitat and are carnivorous
in their natural feeding habits.
Caging/Lab Conditions -
Snakes are easily housed in terrariums with tightly sealed
lids (133 ), but can be kept in various types of cages.
Pea gravel or artificial grass are recommended substrates.
Fibrous substrates have been found unsuitable. Rocks and/or
branches should be available to all snakes as aids in
ecdysis (171,173).
Preferred ambient temperatures range from 26° to 36°C
depending on the species. Local heat (infrared lamps) if
supplied, will allow the snakes to position themselves in
order to achieve a desired body temperature.
Humidity can be critical for some snakes and a range from
33 to 60 percent is recommended. At very high humidities
snakes succumb easily to bacterial infections and at low
humidity levels the danger of dehydration is always present
(171).
Proper lighting is important for feeding responses in some
reptiles and sufficient light is most important (l7l)«
Anesthesias for snakes are variable (141,176>111/178»179 )•
Nutrition -
Carnivory is the rule with snakes. Some species may need a
visual stimulus to attack and eat (173).
67
-------
Water should be supplied in open bowls or other containers
ad libitum.
Breeding/Rearing -
Very little information is available on the breeding and
rearing of snakes in captivity.
Special Needs -
A source of heat is necessary for snakes. Live food may be
an additional requirement for some species.
Behavioral Restrictions -
Some species are prone to biting each other and their
keepers.
Ecological Role -
Snakes are predators on various small mammals and other
animals. In turn, they are a food source for larger preda-
tors, especially birds and mammals.
Problems with Mass Culture -
Reproductive potential seems to be rather low for snakes.
Toxicity Testing -
Snakes (Opheodrys) have been analyzed for DDT content in
open areas where spraying took place ( 174 ) . In toxicity
testing their use has been very limited, but the effects of
radiation on some species is known (180 ).
General Suitability -
It would be difficult to obtain a large number of snakes
for testing. Thamnophis sirtalis has been successfully
bred in captivity and is suggested for that reason. Elaphe
obsoleta is a common snake and a likely second choice.
Ecological Alternatives -
Lizards, turtles.
68
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Birds
Birds are a fairly large vertebrate group (about 1,780
species in North America) that is unique because all of its
members are feathered. With the exception of such birds as
emus, ostriches, penguins and a few South American waterfowl,
all birds are capable of flight. They are as diverse in
their ecological roles as they ate in physical size. Des-
pite these facts, far fewer scientific endeavors use birds
as subjects than mammals for a number of reasons. First,
most birds are protected by federal and state laws. Second,
birds have a high metabolic rate which means they need a
constant supply of food, and migration is an innate urge
which makes the maintenance of many species very difficult.
Lastly, even when the birds can be maintained, they still
may not breed in captivity (this is true of other animal
groups as well). Because of relaxed statutes, easier main-
tenance and breedability, exotic species have received the
most attention in labs. Wild North American species have
been used only infrequently, so many gaps occur in the data
presented here.
The life cycles of birds are complicated and controlled by
their endocrine glands which are stimulated by the photo-
period of their surrounding environment. Establishment and
maintenance of appropriate photoperiods is a major problem
in breeding birds.
In spite of these difficulties in using birds in the labora-
tory, they are potentially very valuable subjects. First,
birds are very important in the world ecosystem. They are
primary and secondary consumers, feeding on plants, inverte-
brates and vertebrates alike. They in turn are food for
mammalian predators (including man), a few amphibians and
reptiles, and a few species of birds. Second, most birds
are highly instinctual and therefore are routine and stereo-
typical in certain aspects of their behavior. This means
that differences or alterations in some behavioral norms
can easily be measured. Last, a vast array of ecological
roles can be seen in this group of animals.
While limited background information hinders in-depth testing
of most birds, it is evident, nevertheless, that many avian
species are good indicators of environmental quality and
good potential test subjects. A summary table indicating the
suitability of birds for toxicology testing is.provided in
/ Table 6.
69
-------
TABLE 6, SUITABILITY OF BIRDS FOR TOXICOLOG1CAL TESTING
•tl -H >.
!o c >
a -H -H
i-( T) -P
•H (UP.
B 0) 10
> l-l U
5 M
Pelicans and F P
Cormorants
Loons and F P
Grebes
Waterfowl E E-G
Shearwaters P P
and Petrels
Herons and Ibises G P
Cranes, Rails E P
and Coots
Woodcock E P
Gulls and E P
Terns
Woodpeckers E P
Raptors F P
Doves and E E
Pigeons
Bobwhitc Quai] E E
Pheasant E E
S S2 § 2 -3
S -3 2 -H S
*w 0) D W U >— 1 A4 •'-'
O-U OOJ 'H T]fl CF1
11 -H > *> " 'H "-1 73
MB 01 Ci 01 C 3 P.P.
BE H -^ «! /" " «
U & K 0 U
F P *Large size, F DZ
slow repro-
ducers, expen-
sive to house
F P *Don't live well P ' Di'D2
in captivity
E-G G-F * E A««B,
Al
J
P P 'Don't live P D. ,D.
well in
captivity
F P *Don't live well P D, ,D,
in captivity
F P *Hard to house, F D,
don't breed in
captivity
P P "Very high P D,
strung, hard
to teed
P F *Don't breed F-P Di'D?
in captivity
F P *Doii't breed in F-P A, ,C.
captivity . i
F F **Require large F C,E
cages
E E E A3,A4
E E 'Cannibalistic E A3'A4
if not "de-
be aked"
E E 'Cannibalistic E A,, A,
if not "de- J
beak od "
01
u
c
01
tf O
•f-l CJ
Is
Aq
Aq
Aq
Aq
Aq
Aq-Ed
Fo-Ed
Aq
• Fo •' .
Fo-Gr-Do
Gr-Do
Fo-Gr
Go-Gr
OJ W
4-* 0
01 -H
(U O
en (U
Cn CL
3 Ul
Phalacrocorax curitus
Podilymbus podiceps
Anas platyrhynchos
Branta canadensis"
Puffins griseus
Puffinus griseus
Fu] ica. americana
Ph^lohela minor
Larus argentatus
Helanerpes erythrocephalus
/
Falco sparverius
Tvto alba
Columba livia
Colinus virqinianus
Phasianus coJchicus
-------
TABLE 6 (CONL), SUITABILITY OF BIRDS FOR TOXICOLOGICAL TESTING
Miscellaneous
Galliforms
Perching
Birds -
Granivores
Perching Birds -
Insectivores
Perching Birds -
Onmivores
Domestic
Ga.1. liforms
Japanese Quail
Exotic;-,
Availability
E
E
E
E
E
E
E
Breeding
in Captivity
E
P
P
F-P
E
. E
F
Ease of
Maintenance
E
G
F
E-G
E
E
G
Previous Use
in Testing
F
P
F
G
E
E
P
y>
c
o
•H
4J
U
• H
M
JJ
V)
0
K
*Cannibalistic
if not "de-
beaked"
*Territoriality
*Territoriality
**u-starlings ,
weaver finches,
territorial ity
Cannibal istic
if not "de-
beaked"
Cannibalistic
if not "de-
beaked"
**Many species
>~
4J
•H rt
r-* Id
•H U
f-H 43 -H
tj 4J O Q)
0) -H --, ^
C 3 O O
O W O (S
0 W
E A ,A
3 4
P A3'A4
F A2/C
G C,A,,
•E '. A3,A4
E A3,A4
G A,CI,
IU
-P l-l
ra m
4-* ^-1
•H QJ
43 H
(tj ft
Fo-De
Fo-Gr
Fo-Gr
Fo-Gr-Do
Do
Do
•a
o en
4-) C)
Dendragupus obscurus
Melospiza melodia
Richmondena cardinali s
Turdus migratorius
Sturnus vu3garis
Passer domesticus
Callus qallus
Coturnix coturnix
lielopsittacus undulatus
in captivity and
unavailable in
large numbers
-------
TABLE 6 (CONT.), SUITABILITY OF BIRDS FOR TOXICOLOGICAL TESTING
LEGEND
ECOLOGICAL ROLE
A. Herbivorous - Terrestrial
1. Nector Feeding-
2. Fruit Eating
3. Seed Eating
4. Granivores
B. Herbivorous - Aquatic
C. Carnivorous - Terrestrial
1. Insectivorous
2. Other Invertebrates
3. Vertebrates
D. Carnivorous - Aquatic
1. Invertebrates
2. Vertebrates
E. Carrion Foragers
C = Nestling Diet
HABITAT PREFERENCE
Fo
Gr
De
Aq
Ed
Do
Forest
Grassland
Desert
Aquatic
Edge
Domestic
E = Excellent
G = Good
F = Fair
P = Poor
RESTRICTIONS
*A11 species protected
by legislation
**Some species protected
by legislation
Unprotected
Sources: Literature cited in text, interviews of scientists
-------
Pelecanidae and Phalacrocoracidae
Introduction -
These two families are represented by the pelicans and cor-
morants. Pelicans are large, bulky water-birds with long
flat bills and tremendous throatpouches. Cormorants are
dark water-birds the size of loons and geese. Both are pri-
marily coastal birds, are active during the day, and feed on
aquatic animal life. They are colonial nesters and migrate
to their northern breeding grounds each year where most nest
on the ground along the rocky shores. They make their nests
from pebbles and other debris they find on the nesting site.
Like most other birds, they are protected by legislation.
Caging Conditions -
A cage 6: x 6 feet made of wood, cement and chicken wir^,
painted, and fitted with indoor-outdoor carpeting will
house three pelicans or six cormorants. This cage was
equipped with tree trunks and large tanks for bathing and
drinking (181).
nutrition -
In captivity, pelicans and cormorants can be maintained on
a diet of fish ( 182) . Food and water should be given ad
libitum.
Breeding/Rearing - _
Pelicans and cormorants usually lay two eggs per clutch and
one clutch per year ( 183 ) . Their young are altricial and
born with their eyes closed. They are also nidicolous
(remain in the nest until fledged) (183)- These birds are
colonial nesters (182).
Behavioral Restrictions -
These birds are large and have voracious appetites (182).
Ecological Role -
t
Pelicans and cormorants are at the top of their food chain.
They are carnivores and feed mainly on marine .vertebrates.
73
-------
Problems with Mass Culture -
Pelicans and cormorants are difficult to maintain in captivit
for long periods of time and are even harder to breed in
captivity (184) . They do not mature rapidly as smaller
birds do and they lay small clutches (183). They require
a good deal of space and are expensive to feed (182).
Toxicity Testing -
Because of maintenance problems most of the work on these
birds has been done in the field. DDT and toxaphene levels
have been checked in birds found dead in the field (185).
DDT and its metabolities were studied in penned cormorants
(186) . The largest body of knowledge comes from oological
studies done in the field (184, 187, 188, 189, 190).
General Suitability ~
Pelicans and cormorants make good subjects for short-term
and field studies because they are large and easily marked
for identification. Their eggs are also easily obtainable.
They feed on a wide variety of marine organisms thus making
them good indices for environmental contamination levels.
The difficulty encountered in maintaining and breeding them
makes them poor candidates for chronic toxicity testing.
The cormorant was chosen as a selected species because it
has been used more frequently in laboratory tests.
Ecological Alternatives -
Gulls, terns, herons, shearwaters.
74
-------
Gaviiformes and Podicipediformes
Introduction -
These two orders are represented by loons and grebes. Loons
are swimming birds that are larger than ducks but have
shorter necks than geese. Grebes are duck-size. Both loons
and grebes are expert divers. They have elaborate courtship
displays, like those of ducks and geese. Both groups are
ground nesters and migrate to their breeding grounds each
year. Loons are basically northern birds ranging from the
north edge of the United States to Labrador and Newfoundland.
Some populations winter in the Gulf of Mexico. Grebes are
more widespread in their distribution and are found through-
out the United States. Grebes, like most birds, are di-
urnal, while loons are crepuscular. Both groups are pro-
tected by legislation.
Breeding/Rearing -
Loons and grebes usually lay two eggs per clutch and one
clutch per year. Their'young are precocial and hatch with
their eyes open. Like most precocial young, they are
nidifugous (183) .
t
Ecological Role -
Loons and grebes are aquatic carnivores arid feed on a variety
of vertebrate and invertebrate species.
Problems with Mass Culture -
These birds are poor breeders in captivity (182).
Toxicity Testing -
DDT and toxaphene have been applied to the habitat of loons
and grebes. They have also been fed in capsule form to
these birds (185) .
General Suitability -
Loons and grebes are fairly available, but they are poor
breeders in captivity. The pied-billed grebe was chosen as
a selected species on the basis of its-availability.
Ecological Alternatives -
Diving Ducks.
75
-------
Anseriformes
Introduction -
There are 43 species of waterfowl in the U.S. (see Table 7),
nine species of swans and geese, and 34 species of ducks.
All species of waterfowl have webbed or at least partially
webbed feet to increase swimming efficiency. They have
down-covered precocial young that are able to swim soon after
hatching. The individual species exhibit a remarkable diver-
sity in size, form, behavior, and environmental requirements,
They range in size from the pygmy geese (Nettapus) to the
trumpeter swan (Cygnus cygnus) and have a variety of bills
which are modified to suit the individual species' feeding
habits.
Some species such as the mallard (Anas platyrhynchos) are
practically omnivorous in their diet. Others, such as the
common shoveler (Anas clypeata) are purely strainers of
aquatic life. A flock of geese can cause significant damage
to an agricultural grain crop.
Waterfowl are migratory and the largest concentration of
breeding is in Manitoba and the potholes of the Dakotas. A
less impressive number of them breed along the coasts of
North America. A few refuges have established flocks of
year 'round residents, especially in the case of the Canada
goose (Branta canadensis) (191).
Caging/Lab Conditions -
A recommended pen for ducks is 30 x 15 x 6 feet. This will
house five adult birds. The birds must be provided with
access to water 10 inches or more deep (192). Pens of this
size with nest boxes, 250 gallon water troughs and feeders,
were used to breed mallards (193). A 14L:10D photoperiod
is recommended (194). See Figure 2 for the mallard test
chamber used at the Denver Wildlife Research Center and
Figure 3 for the cages used. See Figure 4 for the outdoor
mallard cages used at Patuxent Wildlife Research Center.
A more elaborate structure is a 12 x 7 foot room with one-
way glass windows so that the birds can be observed without
being disturbed (194) .
Nutrition -
Commercial game bird breeder ration is available, but many
labs supplement this with millet, cracked corn, wheat, milo,
and barley, all fed ad libitum (195). An alternate diet
76
-------
Table 7. DISTRIBUTION OF INDIGENOUS BREEDING
WATERFOWL SPECIES BY ZOOGEOGRAPHIC REGIONS
Palearctic^-Nearctic Neotropicals Australian Ethiopian Oriental
Magpie Goose (1 sp.) 1
Whistling Ducks (9spp.) 24 233
Swans and Geese 15 9 2 2
(21 spp.)
Freckled Duck (1 sp.) 1
Sheldgeese and Shelducks 4 623
(15 spp.)
Steamer Ducks (3 spp.) 3
Perching Ducks (13 spp.) 214 3 43
Dabbling Ducks 11 9 10 8 8 - 3
(39 spp.)
Pochards (16 spp.) 652 2 21
Sea Ducks (20 spp.) 14 16 1 1
Stiff-tailed Ducks 113 2 1
(8 spp.)
Europe, Asia north of the Himalayas, and northern Africa
i
2
North America, Greenland, northern Mexico, and Hawaiian Islands
South and Central America (except Mexican uplands), and West Indies
Australia, New Zealand, New Guinea, Melanesia, and Polynesia
Africa (except northern part), southern Arabia, and Madagascar
India, Southeast Asia, Sumatra, Java, Borneo, and Philippine Islands
Source: (196)
-------
Figure 2. Mallard test chamber. Denver Wildlife Research
Center, Denver, Colorado
Figure 3. Raptor cages converted into mallard cages
78
-------
gj
Figure 4. Mallard cages (base 4* x 4').
Patuxent Wildlife Research Center
79
-------
consists of cracked corn with quartz and oyster shell grit
fed aid libitum (192). In the wild, grazers, such as B. can-
adensis feed on all sorts of grains, while aquatic strainers
such as A. clypeata feed primarily on spaeriids, gastropods,
mayflies, and caddisflies. Waterfowl consume an average of
230 gm of food per day (191). Water should be available at
all times.
Breeding/Rearing -
Waterfowl are territorial during the breeding season. Some,
such as the gadwall (Anas strepera), Canada goose (B. cana-
densis), and greater scaup (Aythya marila) are colonial
nesters. Waterfowl seek nesting sites that suit their indi-
vidual ecological niches. Hence, they are found nesting on
land, on improvised platforms, in tree holes, on limbs, and
over water in marshy areas.
Hole nesters such as the wood duck (Aix sponsa) have larger
clutches and longer incubation periods than ground nesters
(see Table 8). Clutch size in all waterfowl varies with
environmental and physiological stimuli and with the
breeding abilities of the adults. In general, a large clutch
size means small eggs and vice versa ( 191). Some ducks
such as the ruddy duck (Oxyura jamaicensis) lay their own
weight in eggs (191 ).
There is a great variety in average clutch size between
species (5.1 to 9.3 eggs per clutch) and also in the time it
takes to lay the clutch (191).
Renesting, dump nests, parasitic layers, and indeterminate
layers are some phenomena which are characteristic of the
duck nesting period. The best known parasite layer is the
redhead (Ay thy a americana). Mallard (Anas platyrhynchos}
and pintail (Anas acuta) are the most persistent renesters.
Gadwall (Anas sjtrepera) are indeterminant layers (191).
Geese and swans become mature in their second or third year.
Ducks mature in their first, but are usually not successful
breeders until their second year. Waterfowl young are all
precocial. For an idea of fledging periods see Table 9.
Imprinting is a behavioral phenomena found only in precocial
young. During a relatively brief period shortly after hatch-
ing, the young will develop a permanent attachment for any
moving object presented to it whether it be a human, a dog
or a matchbox on a string Q.91).
-------
Table 8. INCUBATION PERIODS OF VARIOUS WATERFOWL
Species or Group Days
Magpie Goose 28-30
Whistling Ducks 26-30(White-backed Duck 31)
Swans 34-40
True Geese 24-30(Ross1 Goose 21-23)
Sheldgeese and Shelducks 28-30
Perching Ducks 28-32(Brazilian Teal 25;
Muscovy Duck 35)
Dabbling Ducks 21-28(Hottentot Teal 18-20;
1 Crested Duck 30)
Pochards 23-29
Sea Ducks 25-30(Bufflehead 22; Goosander
M and Harlequin Duck 30-32)
Stiff-ta.'.led Ducks 21-27 (Musk Duck and Black-
headed Duck unknown)
Source: (196).
-------
Table 9. FLEDGING PERIODS OF VARIOUS WATERFOWL
Species
Magpie Goose
Fulvous Whistling Duck
Mute and Trumpeter Swans
Black and Bewick's Swans
Pink-footed Goose
Graylag Goose
Snow Goose
Hawaiian Goose
Canada Goose (large races)
Cackling Canada Goose
Ross1 Goose
Freckled Duck
Shelducks
Egyptian Goose
Gadwall
Mallard
Northern Shoveler
Pintail
Blue-winged Teal
Redhead
Canvasback
European Pochard
Ring-necked Duck
White-winged Scoter
Common Goldeneye
Ruddy Duck
Days
Source: (196).
82
-------
Behavioral Restrictions -
Most waterfowl are very territorial during the breeding seas-
on. It is necessary to minimize crowding during this time
if reproductive studies are being carried on.
Ecological Role -
Waterfowl are primarily herbivores, utilizing many plant
foods but preferring grains and aquatic vegetation. In late
spring and summer they become insectivorous, scouring the
lakes and rivers for all forms of insect life. Ducklings,
goslings and cygnets thrive on this high protein diet. Dur-
ing the nesting period waterfowl prefer secluded sites near
small bodies of water. Once the young are off the nest the
families head for open, bodies of water whare the young are
preyed upon by turtles, hawks and larger carnivores (191).
Longevity -
Waterfowl are rather long-lived, but between species there
is a wide range of life spans. For a more specific list see
Table 10.
Problems with Mass Culture -
Waterfowl are protected by legislation and so might be hard
to procure.
Ducks, geese and swans can be heavily parasitized but the
effects of this are not known. Juveniles are more prone to
parasitism, because of their feeding habits, than adults are.
Bacteria pose the greatest threat to waterfowl although fowl
cholera, avian tuberculosis and malaria also occur. Infesta-
tions with trematodes, mites and leeches are common (191).
Toxicity Testing -
Because of their large size, ecological role, and importance
in hunting, much testing has been done with waterfowl, al-
though work with captive birds has been limited because of
the stringent migratory bird code. Most of the penned
studies have used the oral route of exposure for pesticide
testing (193,195,197,198,199,200,201). Disease studies have
been another area where oral administration was used (202).
Heavy metals and pesticides are also introduced into wild
populations by the oral route (203,204,205,206,207). This
has been the major route of exposure for heavy metals too
(192,208). Spraying enclosed fields and ponds has been
another field study method-(209,210,211).
83
-------
Table 10. SOME LONGEVITY RECORDS TOR WATERFOWL
Species
Magpie Goose
Wandering Whistling Duck
Trumpeter Swan
Whistling Swan
Graylag Goose
Canada Goose
Egyptian Goose
Northern Green-winged
Teal
Common Mallard
Canvasback
European Pochard
Redhead
Common Goldeneye
Maximum Age in Years
26 (captivity)
15 1/4 (captivity)
32 1/2 (captivity)
19 (captivity)
26 (captivity)
33 (captivity);
23 (wild)
25 (captivity)
20 (wild)
20 (captivity);
16 (wild)
19 (captivity)
20 (captivity; fertile
entire period)
16 1/2 , (captivity)
17 (wild)
Source: (196).
84
-------
Post mortem metabolism studies are another area of study (113).
Yolk injection is a common method of pesticide and drug study.
The stomach tube has been used in toxicological test-
ing (212). Tests for the presence of certain metals have
also been developed (213). Reference 214 has extensive tests
of pesticides on waterfowl.
General Suitability -
Waterfowl are large enough to be easily handled. Many of
them will breed readily in captivity, especially the mallard
(Anas p1atyrhynchos) and black duck (Anas rubripes). Some
waterfowl species are indeterminate layers(e.g., A. strepera)
and will continue to renest if their eggs are destroyed or
removed. This makes them good candidates for oological
studies as they offer an almost unlimited supply of eggs dur-
ing the breeding season. Because of their sensitivity, they
are good models for ecological indices. Finally, most species
adapt readily to laboratory conditions. A major drawback to
their use in toxicological research is the migratory water-
fowl code. The Canada goose and mallard duck were chosen as
selected species because of their abundance and adaptability
to laboratory conditions.
Ecological Alternatives -
Other waterfowl of the world.
85
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Procellariiformes
Introduction -
This order is commonly called the tubenosed swimmers because
of the peculiarly shaped openings of the birds' nasal pass-
ages. These sea birds have the ability to convert sea water
to usable drinking water by means of their nasal salt glands.
Shearwaters and petrels are usually found far out at sea.
They are gull-like birds but their flight is distinctive.
They are migratory and have diurnal habits. Their popula-
tions are small compared to most other birds and they are
protected by legislation.
Caging Conditions -
Sea birds do not keep well in the lab, and to date, no suc-
cessful arrangement has been made for long term laboratory
studies of them.
Nutrition -
In nature, sea birds eat fish, Crustacea, cephalopods, and
macroplankton (215,216). It is not necessary to give them
freshwater as they can convert sea water with their nasal
salt glands. However, food and water should be supplied
ad libitum.
Breeding/Rearing -
Shearwaters and petrels are colonial nesters (215). They
have altricial young which are nidicolous. Most petrels are
born with their eyes closed, while other sea birds are born
with their eyes opened (183) . The Bermuda petrel is wholly
pelagic and visits land only to breed (216). Most lay one
egg per year (183,215). If their nest is destroyed, they
will not lay again that year (215). Incubation varies from
six weeks to 11 weeks, depending on the species. Young
petrels fledge at 9 1/2 weeks while shearwaters take 13 to
14 weeks (215). Young sea birds never breed their first
year (215). The incubation period broadly correlates with
the fledging period but apparently is not affected by egg
size. Fledging period does not correlate .with body size.
The young have fat deposits (subcutaneous and visceral)
which probably act as reserves- when food is scarce (215).
86
-------
Ecological Role -
Sea birds are aquatic carnivores feeding on both vertebrates
and invertebrates.
Problems with Mass Culture -
Sea birds, although not hard to obtain, are not easily main-
tained in captivity and are hard to breed (215).
Toxicity Testing -
Because of their limited number and poor adaptation to lab-
oratory conditions, little testing has been done on sea
birds. What little has been done, has been conducted in the
field. The effects of DDT and organochlcrines on population
dynamics have been studied (216,217,218/219). Gas liquid
chromatography has been used on PCB concentrations (220).
General Suitability -
Sea birds range widely so they are not suitable for local-
ized studies. They are slow reproducers (183). Their
existing populations are too small for extensive testing to
be done (216). Sea birds have not yet been successfully
maintained for long periods under laboratory conditions.
The sooty shearwater was chosen as a selected species on the
basis of its relative abundance.
Ecological Alternatives -
Gulls and terns.
87
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Ardeidae and Threskiornithidae
Introduction -
These two families are commonly known as herons and egrets
(Ardeidae) and ibises (Threskiornithidae). They are all
stork-like wading birds with long necks, long legs and
pointed, herons and egrets, or decurved, ibises, bills.
They are semi-aquatic birds associated with coasts, marshes
and rivers. They are large birds with body lengths ranging
from 11 inches, least bittern, to 52 inches, Great blue
heron (221). They are primarily colonial tree nesters and
their colonies are called rookeries. Herons, egrets and
ibises are diurnal and migratory. There were never large
numbers of them, but man's encroachment has made many of
them endangered species. For this reason, they are all pro-
tected by legislation.
Caging Conditions -
An outdoor cage, made of chickenwire with a wooden frame and
measuring 50 x 10 x 6 feet has been successfully used (20).
Breeding/Rearing -
Herons and their allies usually lay a clutch of three eggs
once a year (222). Their young are altricial but have their
eyes open at hatching. As with other altricial young, they
are nidicolous (183). Young ibis fledge at about five weeks
of age (222) .
Ecological Role -
Herons, ibises and egrets are aquatic carnivore. They feed
on both vertebrates and invertebrates.
Problems with Mass Culture -
Herons and their allies are slow breeders. They require a
good deal of space because of their size and habits (182).
Toxicity Testing -
These birds have been used very little in toxicological
studies. However, DDT and toxaphene have been fed to them
in capsule form as well as sprayed in their natural environ-
ment (185). General pesticide studies have been done on
population dynamics in the field (222).
88
-------
General Suitability -
Size is a somewhat limiting factor in using some herons.
As with most other birds, Ciconiids are protected by law.
They are fairly easy to maintain in captivity; however,
they are not easy to breed. The American egret was chosen
as a selected species because it typifies this group of
birds.
Ecological Alternatives -
Cranes, rails, coots.
89
-------
Gruidae and Rallidae
Introduction -
The first family is reoresented by the cranes, the
second family by the rails and coots. Cranes are found on
the prairie and, like rails and coots are semi-aquatic.
Cranes are large stork-like birds wnilo rails and'coots
are plump somewhat chicken-like birds. Both families have
secretive habitats, are shy and much more often heard than
seen. Cranes, rails and coots have diurnal habits, are
migratory and are protected by legislation.
Breeding/Rearing -
Gruiiforms have precocial young that are born with open eyes.
Like most precocial birds, they are nidifugous (183)'.
Ecological Role -
Cranes, rails and coots are primarily aquatic carnivores
and feed on invertebrate populations.
Problems with Mass Culture -
Cranes are slow reproducers (182).
Toxicity Testing -
Not much testing has been done on this group, however, tissue
residual studies have been done using dieldrin and organo-
chlorides (223,224). Hatchability studies have also been
done on a limited scale (223).
General Suitability -
Rails and coots are readily available for study, but cranes
are not nearly as numerous. The American coot was chosen
to represent this group because of its abundance.
Ecological Alternatives -
Herons, ibises, ducks.
90
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Scolgpacidae
i
Introduction -
Shorebirds have not been used very extensively in laboratory
research. The most widely used member of this group is the
woodcock (Philohela minor) and for this reason it will be
discussed here. The woodcock is a little larger than the
bobwhite quail and has an extremely long bill. It inhabits
woodland swamps and leafy thickets throughout the United
States. At dusk, during the breeding season, it makes
aerial displays, calling loudly, and beating its wings. The
woodcock is migratory, territorial and protected by legis-
lation.
Caging Conditions -
Various types of cages have been employed to house woodcock,
*-'' ranging from a 20 x 40 x 8 feet field enclosure and a
./ 4 x 4 x 5 feet cage resting on the ground, to a 22 x 29
x 30 inch stainless steel cage in a laboratory. All cages
had water and food pans (225). Hemmed burlap was placed
over the top and front of the cages to prevent the birds
from flushing and injuring themselves at the least disturb-
ance. The burlap was hemmed to keep the woodcocks from be-
coming ensnared in its ends and injuring themselves. Injur-
ies of birds attempting to flush were also reduced by clip-
ping feathers from one wing, making the cage walls opaque
and using high cages or false ceilings of fabric (225). The
smaller cages were found to be more satisfactory because
they allowed the birds to find their food more readily.
Smaller steel cages also helped eliminate the sanitation
problem common in the maintenance of woodcock.
Nutrition -
Woodcocks were fed a diet of earthworms, mealworms and fly
larvae placed in peat moss in food pans (225). Because the
bird has a high metabolism rate, food and water should be
supplied ad libitum (225).
Breeding/Rearing -
Woodcocks have precocial young that are born with their eyes
open (183). They are nidifugous (183).
91
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Special Needs -
Woodcock require a constant supply of fresh, live earth-
worms (38,225).
Behavioral Restrictions -
Woodcock are high strung and therefore prone to injuries ir
caged conditions. Their cages are impossible to keep clear
and being nonperching birds they live on the contaminated
floors, causing themselves to become soiled (225) .
Ecological
Woodcocks are carnivorous, feeding on invertebrates they
find near the shores of ponds, lakes and streams.
Problems with Mass Culture -
The major problem with maintaining woodcocks is keeping an
adequate supply of fresh, live earthworms (38,225). This
is particularly hard to do in winter. Another important
problem arises from the unsanitary conditions under which
the birds are caged; foot infections are a particular prob-
lem (225) . Cages must be built to maximize cleaning effici-
ency and minimize self-inflicted injuries of the birds.
Toxicity Testing -
Because of the effort it requires to keep woodcocks, not
much work has been done with them. DDT studies (226,227,
228) and Heptachlor studies (228,229) have been conducted
however, in addition to general pesticide studies (27,230).
General Suitability -
Woodcocks are fairly available and socially tolerant of one
another. They are relatively hardy in captivity, withstand-
ing heat and cold well and fighting off a wide range of
minor injuries and infections. However, they are costly and
troublesome to feed and create a difficult cage sanitation
problem. Cages must be built to minimize injuries as wood-
cocks are far more prone than other birds to injure them-
selves (225) .
Ecological Alternatives -
Snipe.
92
-------
Lari
Introduct ion -
The suborder Lari is composed of two groups of birds: gulls
and terns. They are all colonial nesters. Gulls and terns
are primarily coastal birds, but some species are also
found along river banks. In any case, they are associated
with the aquatic environment. Gulls range in size from the
great black-backed gull (Larus marinas) (28-31 inches long)
to the Bonaparte's gull (Larus Philadelphia) (12-14 inches
long) ( 221). The most widely known gull is the herring or
"sea" gull. Terns are the size of smaller gulls and are
more streamlined than a gull. Members of the suborder Lari
are diurnal, migratory and protected by legislation.
Nutrition -
Gulls and terns eat a variety of aquatic life, both verte-
brate and invertebrate. In captivity, food and water
should be supplied ad libitum.
Breeding/Rearing -
Gulls and terns usually lay four eggs to a clutch and one
clutch per year (183 ). They are colonial nesters and will
renest if their nest is destroyed early enough in the
season (184) . The young are precocial and have their eyes
open when they hatch, but they are nidicolous ( 183) ; the
average incubation time is 27 days, and the young are able
to fly at 43 days (231).
Ecological Role -
Gulls and terns are carnivorous in the wild, feeding on
vertebrates and invertebrates alike. They have become
scavengers of refuse dumps.
Problems with Mass Culture -
They do not live well in captivity (185 ). Like other
birds they are hosts for fleas, mites, ticks, and lice(182)
93
-------
Toxicitv Testing -
Most testing has been done in the field because long term
studies are not possible at present .in the laboratory.
The primary route of exposure has been oral and the birds
have been tested for DDT (135, 231 )» PCB's (220,232,233,
234, 235), organochlorines (210)/ chlorinated hydrocarbons
(236,237), general pesticide (231 ) and pollution effects
(184) as well as eggshell changes (ics) •
General Suitability -
Gulls and terns are hard to maintain in captivity (185).
For short term studies and ecological studies they make
good subjects because they are colonial nesters and it is
relatively easy to capture large quantities of birds in
a short time. Their eggs are also easily obtainable,
Herring gulls were chosen as a selected species because 01
their abundance and previous use in the laboratory.
Ecological Alternatives -
Pelicans, cormorants.
-------
Picidae^
Introduction -
This is the family of chisel-billed tree-climbers known as
woodpeckers. These birds have stiff, spiny tails which act
as props as they hitch their way up the trees. The males
of most species have some amount of red on the head. They
are represented throughout the U. S. in woodlands. They are
hole nesters, usually excavating their homes in tree trunks.
Woodpeckers are diurnal and migratory. Like most birds,
they are protected by legislation. Very little is known
concerning their adaptability to laboratory conditions.
Breeding/Rearing -
Woodpeckers lay two to six eggs per clutch. There is a one-
day laying interval between eggs and a 12-day incubation
period (215). The young are altricial and are born with
their eyes closed. They are nidicolous (183) and usually
fledge in about 24 days (215). Sexes are readily disting-
uishable in breeding adults (182).
Ecological Role -
Woodpeckers primarily eat insect larvae which inhabit the
interior of tree trunks and limbs but also eat seeds. They
are a major check on insects that attack trees.
Problems with Mass Culture -
All birds are hosts for fleas, mites, lice and ticks and
can become heavily infested with them (132).
Toxicity Testing -
Very little work has been done with woodpeckers in this
area. Field spraying has been done, however, and the popu-
lation dynamics that result have been studied (238).
General Suitability -
The red-headed woodpecker has been chosen as a representa-
tive of this group because it is abundant. However, no
member of this group of birds is judged to be very suitable
for testing.
Ecological Alternatives -
Brown creeper, nuthatch.
95
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Falconiformes and Striqiformes
Introduction -
The Falconiformes are eagles, hawks, arid falcons. Strigi-
formes are owls. Most of these birds are impressive in sist
and perform spectacular aerial displays. Of all the birds,
they are probably the best knov/n in American folk stories.
Their feathers are especially adapted for soaring, sudden
aerial dives and silent flight. Their beaks and claws are
modified into sharp cutting and tearing implements with
which they can grasp, kill.and eat their prey. Members of
the order Falconiformes are diurnal, while Svrigiformes are
nocturnal. Like most other North American species of birds,
raptors are migratory. Some of them, such as the bald eagle
(Halioeetus leucocephalus) are on the Federal endangered
species list. Although some species are very restricted in
their distribution, most are represented at least by a sub-
species throughout the United States. With the exception of
eagles and fish hawks, which prey on fish, all raptors are
associated with terrestrial environments. Most raptors
build their nest platforms in high tree tops or fairly inac-
cessible nooks and crags. The sparrov; hawk (Falco sparveri-
us) is an exception as it is commensal with humans. It can
be found nesting in attics, and other nooks and crannys of
barns, factories, school buildings, churches, etc. (182).
Caging Conditions -
A recommended outdoor pen for a pair of sparrow hawks is
8 x 12 x 7 feet with a roof to keep predators out. The
cage consists of a wooden framework with 1 x 1-1/2 inch
mesh partially sunk in the ground. Each cage has a nest box
10 x 10 x 15 inches with a 3 inch diameter opening and a
slab perch below the opening. Larger scaled (50 x 20 x
6 feet) versions of this cage were used to house multiple
pairs of hawks (239) . An alternate cage for a pair cf
sparrow hawks is 6 x 6 x 8 feet solid wire (240). See
Figure 5 (Cage) and Figure 6 (Nest Box) for the outdoor
housing used for barn owls at the Patuxent Wildlife
Research Center.
Nutrition -
Sparrow hawks have been maintained on day-old dead cockerels
(240), but a more rounded diet consists of ground beef
(mostly muscle, liver, tongue, jowls and heart), vitamins,
minerals and turkey breeder crumbs (239).
96.
-------
:
•••' •;
. X
.•*»
•>
•
r
^
Figure 5. Barn owl cages of chicken wire and wood,
50* x 6' x 10', with nest boxes as shown.
Patuxent Wildlife Research Center (Dr. I. Klaus)
.
-
33ŁL'
rat
Figure 6. Nest box for barn owl cages.
Patuxent Wildlife Research Center
(Dr. I. Klaus)
97
-------
A combination of whole white rats, white mice or hamsters,
chicken heads and skinned chicken necks can be substituted
for the ground beef (239). This diet will sustain most
other hawks and owls also, but eagles and. fish hawks must h
given fish (182). Food and water should be available ad
libitum.
Breeding/Rearing -
Sparrow hawks lay three to seven eggs per clutch and will
breed their first year. Pairs need not be housed separate
(239,241). Peregrine falcons also lay three to seven egg:;
per clutch, and lay one clutch per year (239). Prairie
falcons lay two to five eggs per clutch per year (242).
Peregrine and prairie falcons cannot be housed in multiple
pairs. There is usually a two to three day laying interval
between eggs in raptors and incubation generally takes 30
days. The usual fledging period is also 30 days (215).
Raptors have altricial young which are born with closed eyt
They are nidicolous (183). Most raptors do not breed until
they are two to three years old (182).
Spec i a 1 Ne eds -
Raptors need some types of roughage in their diet to help
them form fecal pellets.. This can be in the fo:rm of
feathers, fur or crumbs (239). They need shelters to esc.n,
the wind if they are kept outdoors (239). They also need
smaller mesh around the cage by the 'nest box to keep mammal
ian predators away from the eggs and young (239).
Behavioral Restrictions -
Wild adult raptors never tame and even hand-reared young
must constantly be worked with to keep them gentle. Even
the smaller hawks are capable of inflicting deep lacera-
tions.
Ecological Role -
With the exception of eagles and fish hawks, raptors are
carnivorous mammalian predators and constitute a major chec
on small mammal populations. Eagles and fish hawks are prŁ
dators on fish populations.
Longevity -
Next to members of the parrot family (Psittacidae), raptors
are probably the most long lived birds on earth.
98
-------
Problems with Mass Culture -
Raptors are relatively slow reproducers and require large
pens for breeding and rearing (182). These birds are hosts
for external parasites also, but not to the extent that most
birds are (182).
Toxicity Testing -
Most testing has been done on wild trapped birds. The birds
were usually fed the toxic substances in foods. Whole bird
analysis of DDT has been done on sparrow hawks (243,244),
PCB analysis with most raptors (240,242,245,246), and
Methyl mercury has been studied in the red-tailed hawk (247).
Finally, egg shell thinning as the result of organchlorines
has been studied in nearly all raptors (138,184,188,210,237,
248), while DDT residue studies have been done with bald
eagles (249).
General Suitability -
Raptors are fairly available and fairly easy to maintain but
with the exception of sparrow hawks they are hard to breed
in captivity and they are slow reproducers. The larger
species are somewhat hard to handle. However, they can be
easily marked for identification and are good indicators of
pesticide levels in their environment. The barn owl and
sparrow hawk were chosen from this group because of their
abundance and previous use in testing.
Ecological Alternatives -
Kites, shrikes.
99
-------
Columbiformes
Introduction -
This group is unique in North America because it. feeds j.-;
young crop (or pigeon) "milk", a pasty substance produce-
in the bird's crop. Both male and female birds are capar
of producing this substance. The domestic pigeon is the
most common and widespread member of this group. Most >-:
of this group are diurnal, migratory, and territorial.
domestic piegon is an exception to the last two of thes>}
classifications. Except for the domestic piegon, these
birds are protected by legislation.
Caging Conditions -
Cage specifications differ as the number of birds to be
caged varies. For a cage designed to house one bird, a 2i
x 29 x 19 cm steel rat cage (250), and a 40 x 40 x 64 cm
cage made of adequate materials (251) have been suggested.
Pairs should be kept in 7.3 x 2.5 cm2, welded wire^cages,
having an area of 45.7 cm2. A perch, food can, automatic
watering device and a nest box (15.2 cm2, 5.1 Cm deep) sK
be provided (252). A cage having the dimensions 115 x 13-
x 229 cm has also been suggested for a pair of birds (253)
A 14L:10D photoperiod is recommended for breeding pairs
(251, 252, 253). Cages should be kept in an environment ha
ing a temperature of 20°C + 1°C (253). See Figure 7 for tfc
caging system used at the Denver Wildlife Research Center,
Nutrition -
Various diets have been suggested for this group of birds.
One includes a strictly commercial pellet diet while the
other incorporates a mixture of wheat, maize and tick beatf
(251) . Grit should be available to the birds at all times
as a digestive aid and mineral supplement (182). See
Figure 8 for the feeding and watering system used at the
Denver Wildlife Research Center.
Breeding/Rearing -
In the wild, this group of birds breeds all year. A reduce
rate of breeding is generally noticed between October and
December (254) . Domestic pigeons breed readily in captivi-
and are available from commercial breeders (255). Under
tropical conditions, two eggs make up a clutch. The usual
laving interval between eggs is 1.5 to two days, the
100
-------
fi:
Figure 7.
Mourning dove cage. Denver Wild
life Research Center, Federal
Center, Denver, Colorado
Figure 8.
Food and water distribution system
T;°^f^rning dove ca
-------
incubation period is 15 days, and the fledging period is
17 days (215) . Adult birds stay sexually active for years
(256). Fertility in these birds ranges from 83.3 percent
to 92.9 percent, while hatching occurs in 81.9 to 90.5 p--;•:-
cent of eggs produced (252). In general, experienced pa; ••<•
prove to be more successful propagators than Inexperience.:
pairs (252).
At birth, these birds have their eyes closed and they are
both altricial and nidicolous (183).
Ecological Role -
Pigeons and their allies eat seeds and grains. They are
preyed upon by domestic cats, other carnivorous mammals,
and birds of prey.
Toxicity Testing -
Much toxicity testing has been done using pigeons, ring
doves and mourning doves as test specimens in the laboratc:
PCB's have been administered to these birds to examine ef-
fects on eggshells (257) and to measure residue (253).
Methods of PCB administration include oral injection (253,
257), and intraperitoneal injections (257) . Lead residues
have been analyzed at laboratories in birds wounded by
hunter shotgun pellets (258). The synergistic effects of
dieldrin and DDE on the thyroid gland (251) have been stuJi
along with steroid breakdown due to dieldrin and DDT admin-
istered through the diet. The reproductive effects of DDi:
(250), the effect of DDT on the thyroid gland, and metabol:
(256,259)have been observed after oral administration. Res
dues have been analyzed in eggs and tissues after aldrine
and dieldrin have been orally administered through a bird's
diet. Acute toxicity has also been measured (260). Resict
studies have been performed using dieldrin which was admin-
istered orally (255). Breeding and reproductive effects oJ
mestranol when added to synthetic grit have been studied
(252,261). Parathion metabolism and liver activation have
been investigated (152). Acute toxicity of a wide variety
of pesticides has been investigated (199).
Anesthetic effects of drugs and other materials on this
group of birds have been evaluated. Pentobarbital and
ketamine, injected into the pectoral muscle, have been
used to evaluate these effects. Anesthesia chamber experi-
ments have also been documented (262).
102
-------
Various behavior studies have been performed on this bird
group. The behavioral effects of pentobarbital (263) ,
chloropromazine (264), and promazine (264) , orally adminis-
tered, have been evaluated. The reversal of discriminatory
performance due to orally administered scopolamine has baen
observed (265). SC-12937 has been placed on food to evalu-
ate a bird's taste discrimination (266). General drug tests
and their relation to behavior have been documented (267),
and numerous Skinner-type behavior experiments have been
performed (268,269,270,271,272).
Field studies on these birds include an evaluation of breed-
ing effects after exposure to SC-12937 via wheat, and of
lead residue in tissues of birds to determine air pollution
levels (273).
General Suitability -
This group of birds has been found to be fairly suitable for
testing because they are large enough for good samples, yet
they are easily housed and marked. Pigeons, ring doves, ana
mourning doves also have good temperaments (182) . Easy con-
ditioning and training are other advantages of this bird
group (267,272). The rock dove (domestic pigeon) has been
chosen to represent this group on the basis of availability
and previous use in testing.
Ecological Alternatives -
Seed-eating passerines.
103
-------
Colinus virginianus
Introduction -
This is the bobwhite quail, one of th
-------
"J'lfSp
• ^,
*- o
V •
~ «.
J
f
I
*
••
->a "»>
•
m . ^.,
•^^:r%- -
;
Figure 9.
Bobwhite quail outdoor holding pens
(base approx. 4' x 8"). Patuxent
Wildlife Research Center
105
-------
Ecological Role -
Quail eat seeds and grains. They are pray for larger mam-
malian carnivores. Their young are preyed upon by raptor;
as well as mammalian carnivores.
Problems with Mass Culture -
Birds can become heavily infested with a number of fleas.
mites, ticks and lice (182).
Toxicity Testing -
In all penned studies the toxic substances were fed in
dietary form. Blood parameters (274), behavioral (280) an:
residue studies (278) have been done with DDT. Population
dynamics were studied using chlorinated hydrocarbons (231,
282) and PCB's (283). Assays were done with, aroclor (27"
and chlorinated hydrocarbons (284). General observation
studies were done with fenuron (279) . Residue studies were
done with mosquitoe larvicides (275) and mirex (276) . 'Thy-
roid studies were done using calcium deficient diets (2u< )
Studies on bait acceptability were done with chemical repel
lants (285). Population dynamics were also studied in
fields sprayed with heptachlor (286).
General Suitability -
The bobwhite is a prolific breeder and does not require rau;
housing space. It is readily available and easy to maints:
For these reasons it is a recommended species.
Ecological Alternatives -
Coturnix, pheasant, grouse.
106
-------
Phasianus colchicus
Introduction -
This is the ring-necked pheasant, an introduced bird in the
Americas. It has become established in farming country,
mainly north of the Mason and Dixon Line. A long pointed
tail readily distinguishes it from other upland game birds.
The pheasant is the largest of the wild gallinaceous birds
and, like all members of this, it is a diurnal creature.
The pheasant is a denizen of open country and an excellent
bird for sportsmen. It is protected by legislation. Al-
though the pheasant is not as mobile as passeriforms, it is
still considered to be migratory.
Caging Conditions -
A pen complex 22 x 6.1 . x 1.7 m was portioned into 27 in-
dividual cages. Of these, nine pens were 2.4 x 2.4 x 1.7m
with dirt runways and 1.2x1.2 x 1.7 m plywood shelters.
The other 18 pens were only half as wide but otherwise ident-
ical (287).
Nutrition -
As with all birds, food and water should be fed ad libitum
(282,287). A basal diet was that recommended for~~nine to
eighteen week old pheasants in the Poultry Formula Guide (288)
Breeding/Rearing _
Like other gallinaceous birds, pheasant young are precocial
and hatch with their eyes open (183). They are nidifugous
(183) . Sexes are distinguishable in adults (182).
Behavioral Restrictions -
Pheasants are cannibalistic in close quarters. To prevent
this, young chicks should be "debeaked" (289).
Ecological Role -
t
Pheasants are primary consumers, feeding on seeds and grain.
Young pheasants, however, are primarily insectivorous for
the first weeks of their life.
107
-------
Problems with Mass Culture^ -
Birds can become heavily infested with ticks, lice, mites
and fleas (182) .
Toxicity Testing -
Numerous studies have been done on the pheasant. The
two major routes of exposure to toxic substances are
mixing the substance with chow and force feeding gel at i:.
capsules. The former method was used to study populati :..
dynamics with dieldrin (290,291) chlorinated instctici...
(282)i chlorinated hydrocarbons (281), DDT and toxaphene
(291) and ethyl mercury p-toluene sulfonanilide (292).
It was also used in monitoring studies with pesticides
(293 ), metabolism studies with PCB (294), residue studi,-.
with heptachlor and 14c-lindane (295 ), behavior studies
with PCB's and general physiology studies with DDT (296).
The latter method was used to study dieldrin residues (29
PCB residues (298 ), eggshell thinning caused by dieldrin
(299)/ pharmacodynamics of dieldrin (300) population
dynamics caused by dieldrin (301)and stress due to PCB's
(302 )• Behavior due to dieldrin (303).LD50s of pesti-
cides (199), monitoring of aldrin (304 ), residue studies
on organochlorine insecticides (248, 305 ), aldrin (306),
and organochlorines in general (210) were also done in
the field.
Genera!! Suitability -
Pheasants breed readily in captivity. They are easily
maintained although they require larger cages than most
other gallinaceous birds.
Ecological Alternatives -
Grouse, turkey.
108
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Miscellaneous Galliformes
Introduction -
This group includes several species of grouse, prairie
chicken and partridge. The birds on the whole are chicken-
size or slightly smaller. Of the galliforms, this group
has the most spectacular courtship displays. The booming
grounds of the prairie chicken and drumming logs of the
grouse attract many amateur and professional ornithologists
each year. These displays are used to establish breeding
territories and also to attract mates. Grouse designate
certain trees in their territories as roosts and use these
trees habitually. Galliforms are classed as upland game
birds because of their topographical distribution and the
sport they provide for hunters. Like other gallinaceous
birds, they are diurnal creatures. They are migratory and
are protected by legislation.
Caging Conditions -
A number of different cages have been used for housing
gallinaceous birds. A 24-1/2 x 2-1/2 x 2-1/4 inch concrete
tank with wire mesh top is recommended (307). An alternate
type of housing is 20 x 10 x 6 feet pens constructed of
fish netting on steel posts (307). A third method of hous-
ing consists of pens 8.1 ' x" 16.2 feet with turf floors and
mesh roofs (308) .
Nutrition -
Commercial game bird chow in pellet form has been fed these
birds (308). Food and water were given ad libitum.
Breeding/Rearing -
Gallinaceous birds have precocial young that hatch with
their eyes open. They are nidifugous (183) . Grouse reach
sexual maturity in approximately 112 days (309).
Behavioral Restrictions -
Gallinaceous birds aire cannibalistic and need to be "de-
beaked" (289).
109
-------
Ecological Role -
Galliforms are primarily seed and grain eaters. Their
young, however, eat insects as well for the first weeks of
their life. All these birds are preyed upon by mammalian
carnivores.
Problems with Mass Culture -
Birds are hosts for a number of ticks, mites, fleas and
worms (310).
Toxicity Testing -
Zectran was fed to birds in capsule form followed by gross
observation and monitoring (307). Dieldrin and malathion
were mixed with game chow and general behavior disruptions;
in the peck order were studied (311). Chlorinated hydro-
carbons and organophosphate were mixed with chow and fed tc
the birds. Various parameters of population dynamics were
then studied (308). Field studies include work with phos-
phamidon (312), DDT (313), and zinc phosphide baits (314).
Telemetric studies were also done with dosed birds (307, 31
316) .
General Suitability -
Gallinaceous birds are easy to maintain and breed in capti/\
ity. They are particularly useful in field and laboratory
telemetric studies because of their physical size and high
fidelity to their territories (307). The blue grouse has
been chosen to represent this group because of its previous
use in testing.
Ecological Alternatives -
Coturnix , bobwhite, pheasant.
110
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Passeriformes-Granivores
Introduction -
This group includes the cardinal, song sparrow and other
finches. These are mostly small, sparrow-size birds and
are distributed throughout North America. They are in-
hibitants of wooded areas and fields and are associated
therefore with a terrestrial environment. Members of this
group are diurnal, migratory, highly territorial and
protected by legislation.
Caging Conditions -
Different investigators have used different sized cages
to house this bird group. One cage used measures
120 x 60 x 60 cm; the floor, iroof, and back of this cage
are wood, while the front and sides consist of 12 mm
mesh (317). Food and water must be available at all times.
A nest box should be provided in the cage, or materials for
building nests may be put at the birds' disposal (318).
Another investigator specifies a cage 61 x"41 x 38 cm for
a single bird (319 )• Large communal aviaries are recom-
mended for long term experiments (320).
Nutrition -
In the lab, these birds are usually fed either pigeon chow
(275), canary or millet seed (319), or a mixture of
biscuit meal, dried milk, soya bean meal, and maw seed.
The mixture diet is considered good for breeding (319).
Cuttle bone should be available and grit should be pro-
vided as a digestive aid and mineral supplement (182,319).
Breeding/Rearing -
This bird group commonly has four nestings a season with
two to five eggs per nesting. The female usually incubates
the eggs, and incubation lasts 13 to 14 days. Hatchlings
usually leave the nest after nine to 11 days, and are con-
sidered fleged at 45 days. A complete molt occurs, in both
young and adults, during August or late summer (288).
Behavioral Restrictions -
These birds are highly territorial and so require separate
cages for each breeding pair (182).
Ill
-------
Ecological Role -
Cardinals, sparrows and finches serve as granivores in th=
food chain. Small mammals commonly eat their eggs, and
cowbirds parasitize them (321).
Longevity -
A song sparrow's life span in the wild is approximately r-,.
years (327). A cardinal lives between three to six years
in a natural environment (321).
Problems with Mass Culture -
The main problem encountered with mass culture is parasite
Common parasites on this bird group are lice, flies, mite?
and ticks (321).
Toxicity Testing -
Limited toxicity testing has been performed on this bird
group. Acute toxicity of mosquitoe larvicides has been
evaluated after ingestion (275). Reproductive effects of
orally administered DDT (319) and residue analysis (323)
have been documented.
Various non-toxicological investigations have been done in-
cluding prehatching and hatching behavior (324) , flocking
behavior (317), hostile, sexual, and social behavior, and
adaptive behavior (325).
General Suitability -
This group, although readily available in the wild, does
not do well under laboratory conditions. They are relativ;
ly difficult to breed in captivity, although there has bee:
some success with house finches (320). Cardinals, song-
sparrows and house finches were recommended as test specie:
because of their numbers and somewhat successful breeding
in captivity; the house finch was also chosen because it i-
representative of western birds.
Ecological Alternatives -
Blackbirds.
112
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Passeriformes-Insectivores
Introduction -
This category is represented in laboratory testing by the
families Turdidae (thrushes), Troglodytidae (wrens),
Alaudidae (larks), and Hirudinidae (swallows). Most species
are highly territorial with monogamous mating systems the
rule. This presents inherent problems for attempting to
rear them under controlled laboratory conditions.
Most passeriformes are diurnal. The swallows are crepuscul-
ar. All are migratory and most winter in Central America.
Although they are associated with streams and other bodies
of water, insectivorous passerines are purely terrestrial
denizens. With the exception of the horned lark which
nests on the ground, and the swallows, which nest where
their common names suggest (i.e., banks, cliffs, barns,
chimneys), these birds are tree and shrub nesters.
Virtually all species of passerines are protected by legis-
lation which makes them generally unavailable from the wild.
Caging Conditions -
A recommended communal cage is 9 x 12 x 7 feet. This will
house 20 to 70 individuals (326). Ten birds can be housed
in a cage 20 x 15 x 22 inches (327). Individual metabo-
lism cages for these birds are 9-3/4 x 7 x 8 inches (327).
To stimulate spermatogenesis, birds were maintained on a
15L-.9D photoperiod (328) at 70° to 80°F (329).
Nutrition -
These birds feed on a variety of insect life including
beetles, true bugs, spiders, caterpillars, ants and other
small Hymenoptera. Grasshoppers, crickets, craneflies,
moths, millipedes and snails are minor food items while
dragonflies, mites, sowbugs, daddy-longlegs and pseudo-
scorpions are merely tasted (330).
Breeding/Rearing -
Passerines usually lay five to six eggs per clutch with a
one-day laying interval between eggs (330). The incubation
113
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period is usually 13 to 16 days (215, 330). The young are
altricial and are born with eyes closed (183) . Temperature
is an important factor in determining the length of incuba-
tion (330). They are nidicolous (183) and fledging on the
average takes 13 days (215). Sometimes 3 broods are reared
per season (330). Both parents usually care for the young
after hatching (330) . Swallow eggs have a 20 day incubation
period and young are fledged 40 days after hatching (215).
Adult passerines undergo a complete postnuptial molt in
August and young birds undergo a partial juvenal molt at
this time also (330).
Behavioral Restrictions -
Passerines hare highly territorial birds and so require
individual cages for each breeding pair (182).
Ecological Role -
Wrens, swallows, thrushes and horned larks are all insec-
tivores. Wrens are forest foragers, swallows and horned
larks are denizens of open country, and thrushes occupy
both habitats. All are preyed upon by carnivorous mammals
and predatory birds.
Longevity -
The average life span for wild birds is one to three years,
but passerines have been known to live 12 years (331).
Problems with Mass Culture -
Passerines are territorial so each pair must be kept in
separate cages if breeding studies are undertaken (182).
Passerines can become heavily infected with mites and
ticks (330). Swallows are poor breeders in captivity and
cannot be easily maintained for long periods in the lab
(182).
Toxicity Testing -
Birds were dosed by mixing the toxic substances with their
food or by loading the food with the substance. DDT was
fed and residual studies were done (326, 332). DDT was
also used in field studies of populations (333, 334) and
residue (37, 335, 336, 337). Loaded food was used to
study the residue and pharmacodynamics of dieldrin (38).
Methoxychlor was fed in field and lab residual studies.
114
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General Suitability •-
Because they require a constant and abundant supply of
insects, they are poor candidates for testing subjects.
The robin is the best candidate among them because of its
distribution and abundance.
Ecological Alternatives -
Vireos, warblers, goatsuckers.
115
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Passeriformes-Omnivores
Introduction -
This group includes four North American families: Stur^id
(starling) , Corvidae (jays and crows) , Icteridae (blacr.-
birds) and Ploceidae (weaver birds) . Ecologically the;;o
families range from prairie and open field denizens (inea-
dowlarks) to early successional and edge species (grac;- ••••[
cowbirds, redwings) to late successional woodland (jay3
and crows). With the exception of the house sparrow
(Passer domesticus) and the European tree 5;p,irrcw (P. u.vŁ
tanus), which are both permanent residents, these birds
are all migratory. Most species are highly territorial
and monogamous, breeding in loose but discrete colonies
and foraging and roosting in flocks at other times of I:...-
year. Many of these birds are not protected by the Mi-
gratory Bird Treaty Act and so they are readily available
from the wild.
Crackles and redwings are usually found around water,
while the other members of this group are usually associ-
ated with purely terrestrial environments. Blackbirds
generally nest in swampy or marshy areas or wet meadows,
Jays and crows usually nest in tree tops and weaver birds
and starlings, who are commensal with man, nest in almost
any available nook or cranny.
Caging/Lab Conditions -
Because these passerines are so diverse in their physical
sizes, a number of different cage dimensions are in use.
For the sparrow-sized members of the group, a cage 7 :x 7
x 7 feet will hold 50 to 100 birds indoors for a four to
six week period (285). A cage 2 x 2*2 feet will hole
seven to ten of these birds for three days (285) . Indivi-
dual test cages are 6 'x 9 x_ 6 inches. One inch mesh
poultry netting is used in ail cases. A community cage
for grackles, red-wing blackbirds and other blackbirds is
9 x 12 x- 7 feet; this will house 20 to 70 individuals
(326). Ten birds of this size can be housed in a cage
20 x 15 x 22 inches (327). Individual metabolism cages
for these birds are six inches cubed (339, 340) or 9-3/4
!x 7 " x8 inches (327). Caging requirements for crow-
sized passerines were not available. See Figure 10 and
Figure 11 for cages and traps used at the Denver Wildlife
Research Center.
116
-------
•
"aa»- .'*«*»**,,
'•; v - ,$*••
1 ^1"$*
\ -
,
I
Figure 10. Starling and wild bird cages.
Denver Wildlife Research Center,
Denver, Colorado
«*
•
.
Figure 11.
Wild bird traps - birds trapped
include starlings, sparrows, robins and
crows. Denver Wild]ifo Research Center,
Denver, Colorado
117
-------
To stimulate spermatogenesis, birds were maintained on a
15 L: 9D cycle (328) at 70° to 80°F (329).
Nutrition -
Pigeon chow checkers (275) and chick starter mash (335)
were used to feed house sparrows. Hulled rice (285, 3^/!
turkey starter crumbs (326, 340, 341), chicken mash (342
and a cracked corn and oats mixture (339) were used to
the smaller blackbirds. Crows, bluejays and ravens were
fed pigeon chow checkers (275). A suggested diec for
breeding smaller omnivores is a seed mixture (75:25:1 -
canary - rape - thistle) supplemented with patainine,
collard greens, lettuce and mealworms (343). Water and
food should be given ad libitum.
Nesting food consists of the following: five pounds pou.
cage bird nesting food, two pounds yellow cornmeal, 10 ':,.
ounces Kretschmer wheatgerm with sugar and honey, two po::.
ten ounces quick Quaker oats and one teaspoon iodized sal;
These ingredients are mixed thoroughly and refrigerated
until used. To this is added dried crushed wholewheat
bread and hard boiled eggs as follows. Equal volumes of
the first mixture and wheat bread are mixed, then to ever.
1 1/2 cup of this mixture, one finely ground hard boiled
egg and shell is added. All this is mixed thoroughly and
enough water is added to make the mixture crumbly (343).
Breeding and Rearing -
Like other passerines, blackbirds and their allies have
altricial young that hatch with their eyes closed (183,
344). They are nidicolous (183). In general, passerines
lay two to six eggs per clutch, have a one day laying
interval between eggs, a 13 day incubation period and a
13 day fledging period (215). The incubation and fledgin
period is longer for jays and crows. Passerines usually
rear two clutches per breeding season, and the first clut
is generally larger. Both male and female partners incu-
bate the eggs and feed the young. Starlings (Sturnus
vulgaris) have a clutch size of four to six eggs, incuba-
takes 12 days and there is a 21 day fledging period after
hatching (344). House sparrows (Passer domesticus) havf
clutch size of three to six eggs, an 11 day incubation
period and a 14 day fledging period (345). They rear two
to four clutches per breeding season (March to September)
(345, 346).
118
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Redwings (Agelaius phoeniceus) usually have four eggs per
clutch and fledge 11 days after hatching. Two clutches
per season is standard (310). Cowbirds (Molothrus ater
ater) are completely parasitic, but an egg laying potential
of four to five eggs has been suggested. Incubation is
believed to take 12 days (310). Crackles (Quiscalus
quiscula) also lay four to five eggs. The "Incubation
period is 14 days and the young are fledged 18 days after
hatching (310).
All junenile passerines have a partial postjuvenal molt in
early August and adults have a complete postnuptial molt
a few weeks later (310). Omnivorous passerines as a whole
are sexually mature and breed at one year of age (310).
Behavioral Restrictions -
Most of these species are highly.territorial and so re-
quire separate cages for each breeding pair (182).
Ecological Role -
Blackbirds and their allies are omnivores, feeding primarily
on grains, seeds and insects. All these birds are consi-
dered to be agricultural pests, but cowbirds are really
more beneficial than harmful. Passerines are food for
mink, weasel, owls, hawks, and falcons. In addition their
young are often attacked by other species of omnivorous
passerines.
Longevity -
Life expectancy in nature for smaller members of this group
of passerines is three years, but they have been known to
live up to 12 years (331). The larger members, such as
the crow (Corvus brachyrhynchos) can live past 20 years of
age (182).
Problems with Mass Culture -
The house sparrow (Passer domesticus) is a carrier of St.
Louis encephalitis (346). All birds are parasitized by
lice, flies, mites and ticks (310).
Toxicity Testing -
Pesticide studies were undertaken using a stomach tube and
LDso's were done (347). Field applications of pesticides
were used for tissue analysis (335), eggshell thinning
119
-------
studies (188), and residue (210). DDT was given in loaded
food and necropsy was done (342). LD^Q'g were done on
birds fed mosquito larvicides (275) and DRC-1339 bait (3451
LDgs's were conducted on birds fed strichnine bait (339).
SC-12937 was fed and physiological and histological studie;
were undertaken (328). A field study on the repellant
power of 4-Aminopyridine was also undertaken (349).
General Suitability -
\
Bluejays are particularly sensitive to toxic substances
(275) . This group of passerines is fairly easily raised
in captivity (182). The starling and house sparrow were
chosen to represent this group because they are numerous
and not protected by legislation.
Ecological Alternatives -
Most non-grain eating seed-eaters.
120
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Domestic Galliformes
Introduction -
This group is comprised of two species: the chicken(Callus)
and the turkey(Meleagris). Both of them have down-covered,
precocial young. Many different strains of these domesti-
cated birds have been developed for commercial purposes
but the leghorn is probably the best known. For this
reason, the text that follows will concern itself only
with the leghorn unless otherwise stated. Chickens, like
all gallinaceous birds, are grain and seed eaters and are
strictly terrestrial in their habits. They are ground
nesters, but they roost in trees.
Caging Conditions -
A recommended cage for domestic fowl is 30 x 20 x 8 inches
will house ten to 50 chicks. For animals used in experi-
mentation, a pen 15 x 10 x 8 inches will house ten to 50
chicks. Individual cage units and brooder facilities are
available commercially. Shavings, peat moss or commercial
litter should be used on the floor of the cage (289). A
14L:10D photoperiod should be observed. Relative humidity
for day-old chicks should be 78 percent, for five-week old
chicks 65 percent, and for adults 45 to 55 percent (289,
350). Adult domestic fowl should be kept at a temperature
of 23.9°C (350). A restrainer for blood collection has
been described in 351.
A commercial brand of chow was fed, food and water were
available ad libitum ,(289) .
Breeding/Rearing -
The chicken has been domesticated for so long that it no
longer has a clutch size; a hen lays for a few months con-
tinually and then ceases for a few weeks (352). Chickens
have an incubation period of 21 to 23 days. They mature
sexually in 20 to 22 weeks. At four weeks, secondary sex
characteristics appear and sexing becomes a relatively
unskilled job. Before this time vent sexing and feather
sexing are used. There are two types of feather sexing -
growth rate of primaries and color of feathers. Both of
these traits are sex linked (352). One cock can service
several hens (352).
121
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Special Needs -
A supply of calcium must be available for good egg laying
results (353). Coarse textured food is preferred over a
food with fine texture (354). Drafts should be avoided
and noise and excitement should be kept to a minimum. Th<:
pens should be kept dry and sudden changes in temperature
should also be avoided (289). For the first four to six
weeks, temperature control is most important. Chicks
should be started at 90 to 95 F and gradually decreased
(approximately 5 per week) to 60 to 65°F (352).
Behavioral Restrictions -
Fowl have cannibalistic tendencies. To help eliminate
these tendencies they can be "debeaked" as young chicks
(289, 352) and the light intensity can be diminished
(352).
Ecological Role -
Because they are domesticated and have been for centuries,
it is doubtful that these birds can be considered to
have an ecological role in the wild.
Longevity -
Domestic fowl, like other larger birds, are fairly long-
lived. Age limits have not been definitely established,
however, as the adults are usually sacrificed after a
few years (352).
Problems with Mass Culture -
Fowl are.prone to many parasites and diseases. Some, such
as Newcastle's disease, bronchitis, fowl pox and coccidio-
sis can occur in epidemic proportions unless controlled
by vaccines (352). Other diseases and parasites are
common but can usually be eliminated without much effort
or loss to the flock (352).
Toxicitv Testing -
Because they can be readily supplied and their biology and
anatomy are well-known, chickens are frequently used in
toxicity testing. Almost every conceivable route of ex-
posure has been used on adults and juveniles. Injecting
chemicals into yolk sacs (355, 356) and studying the
122
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development of embryos (357) are some of the experiments
done with the eggs themselves. Ciliary transport studies
(358, 359) and teratological studies have been done (360,
361, 362, 363) along with disease studies (350), enzyme
studies (364) drug studies (363, 365, 366, 367), population
dynamics studies (368, 369, 370, 371), hjstological studies
(372), x-ray morphology studies (373) and stress studies
(374).
Methods of tissue culture (375, 376) and devices for collec-
tion of excretions (377) have been developed. The piezoe-
lectric transducer has also been used in domestic fowl
study (378).
General Suitability -
Chickens are large yet easily handled. They can readily be
marked for later identification (289). They are obtainable
in large numbers from commercial breeders on short notice.
They are continuous nesters, so ecological studies could be
carried out year-round. Chicks can be housed in limited
space fairly cheaply.
Ecological Alternatives -
Jungle fowl, wild turkey.
123
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Galliformes: Coturnix coturnix japonica
Introduction -
The Japanese quail was introduced into this country by the
Missouri Department of Conservation in an effort to supple-
ment existing game bird species. Coturnix are now found in
several states. It is slightly smaller than the bobwhite
quail in size and far more amenable to laboratory condi-
tions. Because it is an upland game bird, it is associated
with a terrestrial environment. Like other gallinaceous
birds, it is diurnal in habits and migratory.
Caging Condi tions -
Several cages have been used in the laboratory testing of
Coturnix. A frame of 3 x 6 x 1.5 feet covered with 1/2
inch mesh hardware cloth is suitable to house 20 adult
birds (379). Cages to house 25 brooding birds have a floor
surface area of 69 x 100 cm and a height of 24 cm; three
sides are mesh and a fourth side is sheet metal. This caae
also serves as a test chamber in behavior studies of
Coturnix (380). Breeding cages for Coturnix having vari-
able dimensions may be purchased from GOF Mfg. Co., P.O.
Box 8152, Savannah, GA 31402 (381). Laying, incubation,
and testing cages used at the Denver Wildlife Research
Center are shown in Figures 12, 13, and 14 respectively.
Diurnal light schedules differ. Some workers recommend a
14L:10D photoperiod (379, 382, 383, 384); others recommend
an 18L:6D photoperiod (385). Cages should be kept in an
environment of 60 to 80 percent humidity (379) . Temperature
in brooding units should range between 29.3°C to 37.8°C
(380).
Nutrition -
It is common practice to feed Coturnix commercially pre-
pared game bird feed in experimental laboratories. In some
cases, the birds are given turkey starter ration which is
approximately 22 percent protein (380, 386, 387). Some
labs place Coturnix on feeding regimes. An example is
Game Bird Startena (Purina) for two weeks; then Game Bird
Growena is increasingly mixed with Game Bird Startena until
Growena becomes the exclusive diet of the bird. After
weeks the young bird's diet consists exclusively of Game
Bird Layena (379, 381). An oyster shell should be avail-
able to the birds at all times to provide calcium for
124
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Figure 12.
Coturnix laying cages (l2" x 10" x 16").
Denver Wildlife Research Center,
Federal Center, Denver, Colorado
Figure 13. Coturnix incubation chamber.
Denver Wildlife Research Center,
Denver, Colorado
125
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--
. ?
1_
... 1 •• ' »«»%'"
*Ł- «i^. »'^i*^M*- 7/ * -»•>»' .v '. - <• .tiiS^
Figure 14
• :
Coturnix and mourning dove testing
cages (10" x 10" x 12"). Denver
Wildlife Research Center, Denver,
Colorado
126
-------
general good health and for adequate egg development in
females (382, 386).
Food and water should be available at all times because of
the high rate of metabolism of Coturnix (379, 380).
Breeding/Rearing^ -
The natural breeding season for Coturnix is April to Sep-
tember. A year-round breeding season can be induced in
the lab by providing the birds with the correct lighting
and photoperiod. Eggs laid by Coturnix required an
incubation period of 16 days + eight hours at a temperature
of 100°F. It has been found tTha't temperatures above 102°F
are harmful to developing embryos. For purposes of embryo-
logical study, eggs may be held at 25°C for as long as two
weeks before incubation without damage (388).
Coturnix are born with their eyes open, they are precocial
and nidifugous (183).
Behavioral Restrictions -
Migratory restlessness occurs from April to September and
birds may damage themselves when they fly against their
cage (379).
Ecological Role -
Japanese quail are seed and granivores primarily although
newly hatched young feed on insects. They are prey for
predatory birds and carnivorous mammals,
Toxicity Testing -
A wide variety of toxicity testing has been performed on
Coturnix with numerous test substances. Enzyme assays (386,
389, 390, 391), egg production and shell characteristic
studies (382, 392), physiological effect studies (326, 393),
acute toxicity studies (249), stress studies (394) and
residue studies ( 392, 395) have been conducted after the
oral administration of DDT and DDE to these birds. The
effects of oral administration of dieldrin and PCB's have
been evaluated through egg studies (396), acute studies
(249), and tissue residue studies (255, 396, 397). Sleep
studies have been documented after the oral administration
of PCB's and terphenyls (398). Reproductive effects have
been evaluated upon oral administration of Hexachloroben-
zene (399), and physiological and endocrinologic effects
127
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of Kepon have been evaluated after it was administered in
the birds dret (385) . Enzymatic effects have been examined
upon dietary application of Bidrin and Azodrin (400), and
parathion and guthion (401). Coturnix sterility has been
studied after egg application of Mestranol (402). Pesti-
cides have had their effects evaluated, after oral applicr.-
tion, in physiological testing (403), sleep testing (364V
synergistic testing (293) , behavior testing (380) , and res:
due testing (404). A crop-tube method for administering
pesticides for acute toxicity tests has been documented
(199) . Solitary PCB testing has been performed to evaluate
liver residue, biological effects, and behavior after oral
administration (380, 405). The development of different
pesticides has dictated acute, subacute, and chronic testn.:
after oral administration of a particular material (406).
General Suitability -
The Japanese quail is considered a good laboratory aniiv.ul
because it is hardy, grows rapidly, reaches sexual maturity
early, and is a prolific egg producer under lab conditions
(379, 383, 400, 406, 407). It also has simple housing
requirements (379, 383, 406, 407), and is considered repre-
sentative of upland game birds (199, 406). Research with
these quail has become sophisticated to the point that
strain numbers have been assigned.
Ecological Alternatives -
Bobwhite quail, ring-necked pheasant.
128
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Exotics
Introduction -
This group encompasses a wide variety of foreign species
including members of the parrot family, mynahs, and finches.
The most popular and well-known member of the group is the
parakeet or budgerigar. Along with canaries, mvnahs and cock-
atiels, it is sold by the thousands as a household pet.
The physical size of these exotic birds is as varied as their
distribution and ecological niches, ranging from the large,
brilliant colored macaws to the almost hummingbird-size
finches. The bills and claws of these birds reflect the
peculiar habits each has acquired from evolution. Many
members of this group are considered to be the most intell-
igent birds in the world. Parrots, parakeets and mynahs
can be taught to talk, whistle, do tricks and solve problems.
Most of the exotic birds are colonial nesters and are tro-
pical inhabitants. Many of them are native to Australia,
Africa, Central and South America.
Caging/Lab Conditions -
Standard breeding cages, nest boxes, and next bowls are
available commercially for the smaller members of this
group such as the finches and parakeets. Lovebirds can use
parakeet cages, but they need larger nest boxes with bigger
holes (408). Nest material for canaries and finches can be
bought commercially (408). Cages for small finches should
be 24 x 12 x 12 inches and at least four feet long for
larger finches. Perches should be made from twigs or
branches rather than doweling (408). A suitable nest box
for cockatiels is 9 x 9 x* 15 inches with two to three
inches of coarse sawdust or woodchips on the floor and a
ladder of hardware cloth stapled inside the box from the
entrance hole to the bottom of the box (408) . Birds ac-
customed to flying should be housed accordingly (354).
An alternate method of housing is the aviary. This should
be eight feet high with four foot high doors, a small
entrance chamber to prevent birds from escaping, and a
shelter of some sort for the birds at one end (354).
Planted aviaries are best (408).
Nutrition
Commercial chows are available for all these birds. Sun-
flower seeds should be used to supplement this diet in seed
eating birds. Mealworms, flies, maggots, crickets and
cockroaches have been used to feed the insectivorous
129
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members of this group (409) . Water should be kept availe
ble at all times with these birds (410).
Breeding and Rearing -
Lovebirds are sexually distinguishable only by surgery.
They lay three to eight eggs per clutch and have a 23 day
incubation period. The young are fledged at seven weeks
(408). Parakeets average three to four eggs per clutch
and can have as many as six eggs per clutch. They have ?.;
18 to 23 day incubation period and the young are f ledger.
at six to nine weeks of age. Parakeets become sexually
mature at six months and at this time the sexes are fairly
easy to distinguish (408). Parakeets will breed year-
round, starting a new clutch of eggs before the old cluth
is fledged. In order to insure quality stock, a pair
should only be allowed to raise three overlapping broods
before they are given a rest of a month or so (408).
Canaries show no sexual dimorphism, except that mature (r^l
sing while females do not. They lay three to five eggs pe
clutch and breed year-round (408) . Some finches are sex.oa
dimorphic. They have the shortest incubation time of exot
birds — only 12 days, and their young are fledged at two
to three weeks (408) . Sexes are distinguishable in cocka-
tiels but their breeding habits are not well-known. Mynai
and parrots show no sexual dimorphism and many species do
not breed readily in captivity (408).
Special Needs -
Routine care and definite scheduling are a must for these
birds (213) . Social birds should not be kept in individual
cages if they can still hear and see members of their own
species (409) . Adequate flying space is a must (354, 409).
Ground foragers should be fed on the ground and tree
foragers should have their feeders placed higher (409).
Grit is essential for all exotic birds along with vitamins
(410). Parakeets need iodine in their diet (410). Parrots
and parakeets need pieces of tree branch to chew on and
keep their bills trimmed (408). Lovebirds need a supply
of green twigs for moisture for hatching eggs (408)."
Behavioral Restrictions -
Larger birds such as parrots and mynahs can give nasty
bites. Lovebirds can only be caged with members of their
own species as they are vicious in interspecific encounter:
(408).
130
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Ecological Role -
These birds feed primarily on seeds, grain or fruit. When
not kept in check by their natural predators (snakes ancl
carnivorous mammals), they can fnflict serious damage on
agricultural crops.
Longevity -
Larger birds such as mynahs and parrots live 20 years or
longer. Parakeets have been known to live 12 to 15 years
and the small finches probably live three or four years in
captivity (182).
Problems with Mass Culture -
With the exception of finches, these birds are all very
noisy (407). They are extremely messy eaters and sani-
tation can become a problem (182). The insectivorous
species require a constant supply of live insects (408) .
All are hosts for fleas, ticks, lice and mites (182).
General Suitability -
The smaller birds are easily bred and prolific. All of
these birds are relatively easy to maintain. The Aus-
tralian parakeet, or budgerigar, is recommended as a test
species because its initial cost is low and its life
history and diseases are best known in this group. It is
also the most prolific breeder of the group.
131
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MAMMALS
Mammals are a homeothermic group of vertebrates character-
ized by the presence of hair and milk producing glands.
They are considered by many to be the dominant group of
animals on the earth today. Man and most of his domestic
animals belong to the class Mammalia and probably for this
reason our biological knowledge of this class far exceeds
what is known about the rest of the animal world. This 1"
particularly true in the medically-oriented scientific
disciplines such as physiology.
While most research has been concerned with the economical-
ly important mammals, other species are valuable to their
ecosystems and have recently received increased attention.
The following text attempts to consider all mammal groups
in light of present knowledge and to select from each, one
or two species suited for laboratory testing of the effect-
of chemicals on terrestrial species. A summary table
indicating the suitability of various mammal species for
toxicology testing is provided in Table 11..
132
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TABLE 1].. SUITABILITY OF WM.S FOR TOXICOLOGY TESTING
MAMMALS
Shrews
Bats
Vesperti-
lionidae
Phyllosto-
matid
Rabbits and
Hares
Desert
Rodents
Wild rats
Ground
squirrels
and allies
Chipmunks
and allies
Voles and
allies
Deermice and
allies
Lab rats
and mice
Large
herbivores
Large
carnivores
Small
carnivores
Domestic
carnivores
Horse and
allies
Swine and
allies
Sheep and
goats
Deer and
allies
Primates
Opossum
Armadillo
Exotic
species
•fi
r-\
_f)
flj
•H
nj
*
G
F
Ea
E
G
G
G
G
E
E
E
F
P
G
E
E
E
E
G
PC
G
F
E
C
-H
tP-P
C -H
•H >
O 4J
0) a
^ ft
03 U
P
F
F
E
F
E
F
F
E
E
E
G
G
G
E
E
E
E
G
G
G
F
E
ance
U-4 C
C G
.p
G C
in -H
W Ł
F
G
G
E
G
E
G
G
E
E
E
G
G
G
E
G
G
G
G
G
G
F
E
U]
tfl -H
3 -P
O W
•H 0
t»- ^
1)
Vj C
F
P
P
E
P
G
G
F
E
E
E
G
P
G
E
G
E
E
F
G
F
P
E
C
o
-rH
-P
U
-H
4J
tn
a
K
Difficult to
house
Breedability
in captivity
poor
Territoriality
Not social
animals
Not social
animals
Unavailable
in large
numbers
Some very
hard to
keep
large size
large size
long
generation
time
very
susceptahle
to disease
difficult
to house
j?
.-t -.-(
« a
n r3
0) P
C'H
a> 3
O w
F
F
F
E
•F
G
G
F
E
E
Eb
G
P
G
E
G
E
E
G
G
G
F
E
o
•H
tj"
o
o
U
w
B2
B2
D!
D2
DI (C)
C
C
C
D2
DI (C)'
C
Dl-2
A
B-C
C
D2
C
D2
D2
C
c
c
c
O •Ł
C OJ
.p cj .P in
(0 W tr. G
P 0) O -H
•r4 *±4 CJ1 U
.Q iu
De Dipodo-ni/s jrdi
Fo-Gr-Aq A'aotcna li^ijs
Gr Sperr.cpki Ins
beethey I
Fo-Cr FutaTias ^ini'tiis
Fo-Gr Micro tun
per.nsy lyan-izue
.'.'. of Jircfjaat f r
Fo-Gr-De P. rsntV:. .'ur UP
DO Mli? '•lUSu'U t U.S
Sat f us r.crvenic-jf
Fo-Gr '/litre." ;". "r.;«
Fo-Gr- Musi^;.[ "'e . >
Aq-De
Do C.ir.is ''-vi? i .'/'.'s
Do
Do-Gr Miniature swine f^u
Do
OV'.'-p ŁP-
Fo-Gr -'«: j'i<'ua virfii*::-'
0, h;.^ic~.us
Fo-Gr .'.'jinj'rt .;<7!i,.^.?i..>
Fo-Gr i' •:>;.. ;': > .• .:
r ..-iri.ri^;; .-
Fo-Gr ii^if.vr iii- f
no'.'t:r^i j".?i«s
133
-------
TABLE 11 (CONTINUED). SUITABILITY OF MAIWL:-- FOP: TOXlCOLfl'
Legend:
General Ecological Role
A - Large carnivore
B - Small carnivore
1. General
2. Insectivore
C - Omnivore
D - Herbivore
1.
2.
Seed, fruit eater
Grass foliage eater
E = Excellent
G = Good
F = Fair
P = Poor
a - Tropical species
availability is good in tropi..
b - The suitability of long
inbred ' (horaozygous) popu-
Jacions in toxicologies 1
testing is suspect.
c - No native non-human species
Habitat Preference
Fo
Gr
De
Aq
Do
Forest
Grassland
Desert
Aquatic
Domestic
Sources:
- literature cited in text
- interviews of Scientists
134
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Insectivora
Introduction -
Two of eight Insectivora families are represented in North
America: Soricidae (shrews) and Talpidae (moles). There
is little literature on moles and they will not be treated
in detail in this report. Crandall (411) gives one ac-
count of a captive Condylura cristata, and two accounts
from other authors for Scalopus aquaticus . Most of the
seven U.S. mole species are restricted to a few states.
The eastern mole,(Scalopus aquaticus), however, has a large
distribution (34 states); it is found generally east of
the 100th meridian and south of 43° latitude (412) . Of the
27 U.S. species of shrews, three have wide distributions.
Cryptotis parva [the least shrew (or lesser short-tail
shrew], and Blarina brevicauda (the short-tail shrew) , are
generally found east of the 100th meridian. C. parva occurs
in 31 states, mostly south of 43° latitude. B. brevicauda
occurs in 37 states. Sorex cinereus is a northern species,
distributed roughly from 40° latitude to the Arctic Ocean.
Identification of shrew species is often difficult and
should be verified (412).
B. brevicauda, the principal subject of this section, lives
Tn "well wooded. . . low ground" (413) , mature beech-maple
forest with abundant leaf litter (414) , and sometimes in
grasslands (415). Like other shrews, it is fossorial, but
not so completely as moles. The short-tail shrew main-
tains two nests, a breeding nest about 2 x 2-3/4 inches
which is lined with grass, whole or broken leaves, and
sometimes mouse fur and a resting nest described as about
the size of a large apple (413). Hamilton (413) found a
dung pile about four inches from a breeding nest and, com-
monly, snail shells in the runways. Scats are usually
piled on one side of the runway near the nest.
Caging/Lab Conditions -
Shrews do well in cages made in a variety of materials and
sizes, plastic (14 x 18 inches) and glass have been used,
with cages made as small as 20 x 2 inches (314, 411, 416).
There is little danger of shrews jumping out of their cage
as mice do. For example, Blus (417) kept shrews in
2 x 1. x 1 foot (288 square inches) open-topped aluminum
boxes. In designing cages, it should be remembered that
Rood (416) observed the animals frequently licking screen
sides and tops of cages (without speculating on the cause
135
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of this behavior) and that Blus (417) rejected wire cages
because of the inadequacy of the substrate.
Shrews are frequently provided with a tin can (411, 413, 418)
for a nest, but have been forced to sleep in the open, liv-
ing months with no cover and very little soil (416) . When
cover, such as sod, is provided, the shrews use it (416)
Cotton (418), leaves, and grass (411, 416, 417) have been
provided as nest material. Shrews treat nest materials
in a variety of Ways: the softer may be placed inside the
rougher; they may be burrowed into (417) , or dragged into
a corner, and they may be ignored when there is a good
layer of soil in the cage where the animal can enlarge one
tunnel for a sleeping chamber (416). Two nests are pro-
vided a pair of shrews by Blus (417), who also placed a
plywood platform supported by dowel legs above the nest
chamber to support the weight of a water dish and to pro-
vide additional cover. Usually, captive shrews are sup-
plied with soil - from just enough to cover the bottom of
their cage, to several inches (411, 416) - or sphagnum
(413, 417). Shrews immediately start to dig in either
substrate, honeycombing it with tunnels (see 416 for de-
scription of digging behavior) (417). Other beddings are
wood shavings (418 genus unspecified), pine chips (417),
corn cob litter (417), peat (417), grass [(413 genus and
condition unspecified)]. Blus (417) identifies milled
sphagnum peat moss, three inches deep, as the best substrate.
Rood (416) and Blus (417) kept the animal room at 70°F.
Doremus (419) kept the room at an "air-conditioned" 17°C
(referring to temperature, not quality or humidity) and
controlled lighting to "approximate outside conditions"
(probably off-on roughly with sunset-sunrise, ignoring
brightness and spectrum). Blus (417) kept a regimen of 14
hours continuous fluorescent light followed by ten hours
continuous darkness.
Only Hamilton (413) mentions odor, which he found most pro-
nounced in April, lingering two days after a pair of shrews
had been removed from the room. No authors have reported
on the method and frequency of cleaning cages. Some shrews
dropped scats in the sphagnum and the water, and never in
the nest box, and a female with a litter left scats in the
corner of the nest box, rarely venturing from the nest (413).
Blus1 animals usually defecated and urinated in one corner
of the next box, either on or below the surface (417).
136
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Nutrition -
Shrews are very sensitive to shortages of water and food
but the amounts required by Blarina have probably been con-
fused by experiences with Sorex '(418) . Blarina consumes
less than one-half their body weight in food every 24 hours
(416) . Blus (417) , however, provided 15 to 20g food per day
per individual. In nature, soricids consume earthworms,
slugs, snails, insects, mice, fungi, berries, seeds, and
greens (411, 420). In captivity, they have survived on
an array of foodstuffs: ground raw horse meat, bone meal
with cod liver oil, chopped hard-boiled egg, earthworms,
mealworms, mice, diced fruits, greens, and cottage cheese
(416). Additional items are dead shrews, dead birds,
chicken, canned dog food, peanuts, crackers, whole wheat
•bread, cheese (416). There is much individuality in food
preferences (416). Pearson (421) fed a mixture of ham-
burger, ground liver, and dry commercial dog food; Mann
(418) substituted commercial dog food for the meat portion.
Mann kept shrews up to 16 months solely on canned dog food,
starting the animals on it immediately upon their arrival.
Doremus (419) kept animals on a basic diet of canned dog
food, occasionally supplementing with mealworms and raw
beef. A mixture of "ground carcasses of laboratory rodents
(61.3 percent), chicken heads (15.4 percent), turkey
breeder ration (7.1 percent), Vionate, a vitamin supplement
(0.5 percent}, and bone meal (0.3 percent)" was fed by
Blus (417). Rood (416) found that most shrews readily
ate dead shrews. A shortage of food may induce cannabilism
in a group (418) .
Breeding/Rearing -
The most complete description of breeding captive Blarina
is that of Blus (417). The average number of young is
five to seven (413, 417) with a range of four to ten (413,
422) . The peak of breeding activity in nature is in April
(413, 414, Ithaca, New York) when there is maximum testes
development and captured females have embryos. In May and
early June, females are nursing. Breeding is minimal in
late August and early September (413, 415) . Dapson (414)
concludes that breeding may occur throughout the year.
There are probably spring and late summer litters, and
occasionally, a vigorous female produces a third litter
(413, 415). Crandall (411) reports that E. P. Walker had
a captive Cryptotis garva female which produced 66 young
in one year. Some individuals breed one or two months after
birth (414, 423). As a rule, males with testes greater than
5 by 7 mm have sperm in the cauda epididymis if the acces-
sory reproductive glands are developed (414). The
137
-------
gestation period is21 to 28 days (413). Hamilton (422) be-
lieves post-partum estrus is normal for wild Blarina, as
well as for Sorex fumeus and S_. cinereus.
The development of the newborn shrew has been described in
detail by Hamilton (413). At birth, the shrews are dark
pink and the size of a honeybee. A wild female was ob-
served moving each of her young (about nine days old) by
the belly when the nest was disturbed in the wild. By 13
days, shrews weigh about 9 g; mammae are visible on the
females; the ears are open; the side glands are obscured;
the mid-ventral bare patch is prominent. Litters have been
kept together up to one month following weaning (417).
Special Needs -
Shrews are very sensitive to food and water shortages and
require continuous supplies of both.
Behavioral Restrictions -
The greatest concerns in maintaining shrews in captivity
are their voracious appetites and pugnacious behavior, both
over-rated. Although Blair (415) found that Blarina are no'
territorial, most researchers keep shrews in individual
cages except when pair are mated (417). Shrews pair for
very long times in the wild, perhaps for life, and parents
may stay together until the young are born (413). Rood
(416) describes the behavior of captive shrews in various
combinations and as strange animals are introduced. Indi-
viduals are marked with #1 bird bands on the hind legs (416)
or by toe-clipping (415, 424). Playful pushing and snug-
gling occurs among a few individuals living together with
a minimum of quarrel (416) . The important factor is the
disposition of the individuals involved, with age and sex
only secondarily important. Rood (416) doubts that Blarina
are "as solitary as is generally believed."
Ecological Role -
Shrews are energetic predators in the ecosystem, consuming
quantities of earthworms, snails, and slugs, various in-
sects, and mice. The role of shrews in controlling mice
has been disputed and probably depends on the local popula-
tion as well as individual differences (416).
138
-------
Longevity -
The longevity record for Blarina brevicauda may well be that
established at the National Zoological P'ark of two years,
three months, and one day (411). Life expectancy in nature
is 20 months (414). The age to toothwear relationship is
curvilinear (414).
Problems with Mass Culture -
No diseases have been identified for Blarina (413).
Toxicity Testing -
Shrews have not been used much as subjects for toxicity test-
ina except in field studies where lead content was analyzed
(425).
General Suitability -
Blarina has had previous maintenance work completed (416,
417) , and is of a suitable size to be easily housed in
relatively small cages. Its voracious appetite and
abundance in nature makes Blarina a good candidate for use
in toxicological testing.
Ecological Alternatives -
Other shrews, moles.
139
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Chiroptera
Introduction -
Bats (Chiroptera) are the only mammals capable of true
flight. Almost all North American bats (about: 30 species)
are members of the family Vespertilionidae, and are
nocturnal, insectivorous, and weigh no nore than a fraction
of an ounce. During the day they roost in various types
of shelters including caves, attics, trees, and old mine
tunnels (412). Some species are migratory, changing
their range with the seasons and availability of insects.
Other bats hibernate in caves and buildings during the win-
ter. Four Phyllostomidae are found in the southwestern
United States and feed on pollen and nectar. Six species
of Molossidae occur in the United States and are primarily
insectivorous.
Caging/Lab Conditions -
Because of the small size of bats, many different cage en-
closures have been successfully used. A 20 "x ^J x 25 cm
cage was used to house 12 individuals. A wood-wire mesh
structure 80 x 92 x 128 cm, with a roostinq box 80 x 92 x-
51 cm was used to house a bat colony of unknown size (426).
Bats have been individually housed in pint ice cream con-
tainers with wire mesh lids (427), and in battery jars
15 cm wide and 20 cm high (428). All types of materials -
wood, metals, etc. can be used for cages.
Sanitation of cage facilities seems to be the major dif-
ficulty in the maintenance of bats. Overcrowding leads to
sanitation problems and much time is spent cleaning the
cages C428, 429, 430). Paper on the bottom of the cages
should be changed daily (431).
Because bats are nocturnal, they require dim light for feed-
ing. Other special needs include room to exercise or to
fly (431) , chitin in their diets (428) , and water made
available in such a way that bats can readily find it.
High relative humidity (71-81 percent) and high ambient
temperatures (78-84°F) increase their activity and facili-
tate the feeding process (427, 428, 432) . The use of
light bulbs to provide local heat is a common practice
(427).
140
-------
A photoperiod of 13L:11D, a 21-28°C temperature range, and
a relative humidity of 55-92 percent, have proven effective
in maintaining Phyllostomid colonies (426).
Nutrition -
Eats eat a great variety of foods in captivity. One elabo-
rate diet consists of hard-boiled egg yolk, dry cottage
cheese, ripe banana, live mealworms, six drops of liver
extract with iron, six drops of wheat germ oil, and three
gms of milti-vitamin preparation, all mixed in a blender.
If desired, it can be frozen. This preparation is given
ad_ libitum (428) . Insects are a necessary component of tne
diet of insectivores because the chitin aids the formation
of fecal pellets (428).
Successful diets for Phyllostomids include young birds, in-
sects, raw beef, banana (433), cereal, wheat germ, milk,
powder, protein and mineral supplements, corn oil mixture
in a fruit base (426, 429). Mealworms are used widely
as a staple food for bats in captivity (434).
Breeding/Rearing -
North American bats do not usually breed in captivity (431).
In nature copulation takes place in late autumn and birth
in spring. The sperm is stored for several months in the
body of the female (429, 433).
Gestation lasts 56-100 days in Corynorhinus (435) and is
rather variable in many species. Usually only one or two
young are produced each year (429). There have been a
number of tropical bats maintained in captivity with mixed
breeding success (436).
Behavioral Restrictions -
One drawback to keeping bats, since bats usually forage on
the wing, is that they must be conditioned to feed in cages
from the food source (see 427 for a thorough description
of techniques). North American bats are hibernators and at
roost they assume the temperature of their immediate en-
vironment. If temperatures are low enough they will enter
into a semi-torporous state every day. The seasonal hiber-
nation pattern conflicts with long-term physiological
experiments and many researchers have resorted to tropical
species which do not hibernate (434).
141
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Ecological Role -
The bats of North America are primarily insectivores feed-
ing entirely on airborne species. Bats are known to loca:e
their prey by means of ultra-sound and navigate with ease ir
the absence of light. Their ecological role is not fully
understood.
The leaf-nose bats, Phyllostomidae, feed in part on pollen
and nectar, acting as pollinators of certain angiosperms
specifically adapted to their visitations.
Longevity -
For Mexican free-tailed bats,(Tadarida brasiliensis), the
life span is greater than 4.5 years in captivity. Pacific
pallid bats have lived to eight years , three months and cer-
tain Desmodontidae average five to six years (433).
Toxicity Testing -
Bats are rather unusual members of the laboratory world.
Myotis lucifugus has been used for taste reception work
'(437) , and the Mexican free-tailed bat (Tadarida brasilien-
sis) (427) for metabolic versus temperature studies.
The little brown bat fay otis lucifugus), has been shown to bn
highly sensitive to DDT (438) and studies have been used
also for the effects of ethyl alcohol on subcutaneous
microcirculation (439).
General Suitability -
Most species of North American bats are not plentiful
enough to withstand large harvests to stock lab colonies.
The possibilities for breeding in captivity are relatively
unknown. This, added to the seasonal activity changes of
temperate-region species, makes them poor subjects for wide
use in toxicity testing. Tropical species have a wide
range of ecological niches and may be better test subjects
(434).
If a North American species is chosen to begin lab breeding/
rearing work, Eptesicus fuscus, because of its large size,
is recommended (434).
Ecological Alternatives -
Other bats, insectivorous animals, (wrens, shrews).
142
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Lagomorpha * "
In troduc t i o n -
The Lagomorphs are composed of tv/o families, the rabbits and
hares (Leporidae) and the pikas (Ochotor.idae) . Though they
resemble the rodents, there is good evidence that they have
been separately evolving for a very long time in terms of
mammalian history.
Rabbits (Sylvilagus) and hares (Lepus) have long ears
and long hind legs modified for saltatorial locomotion. In
warmer climates, they are carriers of tularemia. There are
about 15 native species that are basically openland-edge
animals. The largest hares are about 26 inches long and
weiqh up to 12 pounds. Leporids are basically herbivorous
(412).
Pikas, Qchotona, are inhabitants of talus slopes in the
Northern and Central Rockies. There are only two native
species of this smaller, rat-sized, tail-less animal. They
store food in hay piles and feed almost entirely on grasses
and herbs (412) ..
In research, the domestic New Zealand white rabbit,
Oryctolagus cuniculus, is utilized almost exclusively, and
the following comments refer to this species unless other-
wise stated. They weigh from nine to 14 pounds and are
very gentle lab animals (289).
Caging/Lab Conditions -
Recommended cages for New Zealand white males are 30 x 18 x
18 inches, which are also sufficient for a breeding aiea.
Does with litters need larger enclosures (30 x~ 36 x 18
inches) and a nest box, 18! x 9. xr 7 inches, with fresh straw
bedding. Temperatures from 65-75°F are suitable and rela-
tive humidity of 50 percent is ideal, but not a necessity.
A 14L:10D photoperiod is recommended. Drafts should be
avoided (289).
A breeding pair of European hares, (Lepus europaeus) were
successfully housed in an 8 x 6 x 8 foot "brick"", outdoor
enclosure with a concrete floor and a wire front which was
covered in winter. One-half of the floor was covered with
straw (440) .
143
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Snowshoe hares have been kept in captivity. One-half inch
wire cloth flooring is suggested as a deterrent to the
spread of coccidiosis, a perennial problem with this species,
This cage alteration increased survival of new born rabbits
from 11.6 percent (when they were born on the ground) to
63.3 percent (441).
For handling lab rabbits the restraint device described in
reference 442 has proven very effective.
Nutrition -
Commercial rabbit pellets are available, but many labs
supplement the diet with greens, raw vegetables, hay,
and oats (441). Adults should receive four to six ounces
of food daily, pregnant females six to eight, and lactating
does eight to 16 ounces. Growing juveniles need from one
to two pounds per diem (289) . . Pikas have been maintained
on lettuce, cabbage, raw potatoes, dandelions, grass, oat-
meal, and bread (441). Lagomorphs are coprophagus and
apparently obtain some nutritional benefit from this habit
(443). Water should be supplied ad libitum to all species.
Growth of suckling rabbits has been shown to increase faster
in creep-fed individuals than in those reared naturally
(444). Creep-feeding is the practice of self-feeding
concentrates to young suckling animals in a separate en-
closure away from mothers. The composition of one experi-
mental creep-feed diet is shown on Table 12.
Breeding/Rearing -
Domestic rabbits and many wild Lagomorphs are polyestrous.
The gestation period for New Zealand whites is 30-32 days,
litter size varies from one to 18 with an average of eight,
the young are weaned in eight weeks and both males and
females can breed approximately six months from birth.
Females can be rebred about 35 days after parturition. The
breeding life of females and males is one to three years.
One buck can service six to ten females in a breeding
colony. Pair mating is recommended in the buck's cage (289) •
European hares have a 42-day gestation period, snowshoe
hares 37 days, and pikas 32 days. There has been some suc-
cess breeding pikas in captivity (441).
In the wild, cottontails (Sylvilagus floridanus) gestate
for 27 to 30 days, and produce three to five litters per
annum. The number of young per litter is four to seven
(412).
144
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Table 12. COMPOSITION OF AN EXPERIMENTAL CREEP-FEED*
Protein, 22 percent; fat, 5 percent; fiber, 13.4 percent
Ingredients: Dehydrated alfalfa meal, soybean meal, oats, red wheat bran, distillers
dried corn solubles, skimmed milk, salt, anise oil powder, vitamin A,
balance of trace minerals, vitamin pre-mix, and vegetable fat or oil.
Mixing Formula;
Dehydrated Alfalfa Meal
Soybean Meal (expeller)
Oats
Red Wheat Bran
Distillers Dried Corn Solubles
Skimmed Milk
Vegetable Fat or Oil
Salt (NaCl)
Vitamin Pre-mix
Anise Oil Powder
Trace Minerals
Vitamin A
Percent
Of Total
37.5%
10.0
10.0
15.0
13.5
11.5
1.25
0.5
100%
Pounds
Per Ton
750
200
200
300
270
230
25
10
7.5
4.5
2
1
2,000 pounds
*Feed was a crumble in Trial 1 and pelleted at 3/32" diameter in Trial 2. Also, in
Trial 2, the formula was modified slightly by Albers Milling Company. Essentially,
the modification was to increase the level of protein and reduce the level of fat.
SOURCE: (444).
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Behavioral Restrictions -
The restrictions associated with maintaining domestic rab-
bits are very minor. Wild species may have some difficulty
adjusting to caging. Frequently the cage elicits an escape
response which sends the animal nose first into the wall of
the cage, often with sufficient force to cause self-injury
(441).
Pikas are very territorial creatures in their natural en-
vironment. This behavior may restrict attempts to cage a
number of-them.
Ecological Role -
Rabbits and hares are primarily herbivorous, utilizing many
plant foods, but preferring grasses and herbaceous materials
(412). They are typically openland-edge inhabitants and
remain active year round (443). Pikas have similar food
habits and are easily detectable on the talus slopes where
they live because of the "hay" piles they build in the fall
as storage heaps (412).
All Lagomorphs are desirable prey for larger predaceous
birds and mammals.
Longevity -
In captivity, eastern cottontails have lived to five years
and European hares to 11 years of age (441).
Problems with Mass Culture -
Rabbits and hares are quite susceptible to a variety of ills,
particularly coccidiosis which is highly communicable and
causes high infant mortality when it occurs (441) .
Toxicity Testing -
Domestic rabbits are a rather popular subject in American
laboratories because of their gentleness and in some cases/
their relatively large size. They have been used in many
types of toxicity experiments, particularly those testing
dermal reaction where the substance is directly applied to
the shaved skin (445, 446, 447, 448, 449, 450, 451). Other
routes of exposure are also utilized with a great variety
of substances. Work with wild species is limited to a few
pesticide studies of free roaming subjects (452).
146
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General Suitability -
Domestic rabbits are readily available and easy to use in
toxicity testing. The applicability of such work to natural
populations is difficult to judge.
Wild species, particularly the eastern cottontail (Syl-
vilagus floridanus) and a western species (S.'auduboni)
are available and should prove adaptable to laboratory
conditions.
Ecological Alternatives -
Woodchucks, nutria.
147
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Rodentia: Heteromyidae
Introduction -
The Heteromyidae, the family of kangaroo rats,and pocket
mice, are found in the southwestern United States and in
Mexico. They are specially adapted to desert habitats.
They have fur-lined cheek pouches. The large forms have
modified rear limbs for leaping and very long tails used
for balance. All Heteromyids are nocturnal. There are
more than 20 species of pocket mice, one kangaroo mouse,
and 14 species of kangaroo rat in North America. The rats
are the largest, with four to eight inch bodies and weigh-
ing from two to five ounces. Pocket mice are similar to
laboratory mice in size.
Caging/Lab Conditions -
For the kangaroo rats (Dipodomys) glass front metal cages,
2x2x3 feet with dried leaves for a floor and a small
shelter, have been used. A quantity of sand was supplied
and used by the rats for cleaning their pelage (453).
Other cages, 10 x 11 x 16 inches ( 3 x 4 m) are also
adequate, but for breeding a larger enclosure, 5x7x2
feet, is recommended. Shelters, such as small boxes or
mailing tubes and a sand substrate are also desirable
(454).
Pocket mice (Perognathus) were routinely housed in 12 * 12
x 12 cm cages with sand bottoms (455). It was found that
the pocket mouse (P. flavus) tolerates physiological stresses
very well such as sound, heat, vibration, and G forces (456)-
The Heteromyids/ though desert forms, are greatly affected
by exceedingly high or low ambient temperatures (457) and
should be protected from them.
Nutrition -
Kangaroo rats were fed a diet of 26 percent rolled oats, 25
percent rolled barley, 20 percent meat meal, 10 percent
powdered milk, 11 percent whole wheat kernel, 2 percent
cod liver oil, 2 percent sunflower seeds, and traces of
birdseed, salt, powdered calcium carbonate. Lettuce was
provided every other day. They seemed to do equally well
on just barley, sunflower seeds, and lettuce (454).
148
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Pocket mice have been successfully nourished for seven
years on mixed seeds alone (453). A more varied diet in-
cluded dog food, greens, and vegetables (453). Desert
rodents may need green vegetation to induce breeding in the
spring.
The water requirement of heteromyids is negligible in the
laboratory; even when water is made available, they do not
drink it (453) .
Breeding/Rearing -'
Heteromyids are very territorial and will usually destroy
each other when two or more are caged together. Increased
cage size may improve the success of co-habitation (455,
458). In one case, individual kangaroo rats were .kept
separate until estrous and then a pair were placed together
in a neutral cage, 30 x 30 x 10 inches. This resulted in
pregnancy in six of ten females (459). Successful lab
breeding is recorded for D_. ordii, D. merriami, and
D_._ deserti in large rooms (453) . Some success at group liv-
ing has been reported for the, Fresno kangaroo rat, D_._
nitratoides, but only in large cages with rather violent
initial skirmishes (453).
All heteromyid rodents are seasonally polyestrous. The ges-
tation period in captivity for Dipodomys is 29 to 32 days
and varies little from one species to another (453).
Young of D. merriami and p_._ deserti were weaned at 22 to 25
days and were fully mature at 90 days. Sex of the young can
be determined after 11 days (454). The number of young 'per
litter for Dipodomys is two to four (459).
Behavioral Restrictions^ -
Though heteromyids are nocturnal, some specimens have over-
come this habit in the lab. Under red light, these rodents
have been quite active (453).
Kangaroo rats are very easy to handle and are essentially
very gentle creatures (453). However, because of their
territorial instinct, communal caging may not be possible,
with the exception of a few species.
Ecological Role -
Desert heteromyid rodents are primarily seed eaters and
serve as food for many predators. They are particularly
important in snake diets.
149
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Longevity -
In captivity, the life span of various pocket mice
(Perognathus) differs from four years (P . pqryus) to seven
years (P. longimembris) to eight years TP. fallax) (453).
Their mortality in nature is much higher as would be ex-
pected.
Toxicity Testing -
Because of the unique fashion in which these desert rodenc;
metabolize water, their physiology has been rather well
studied. Some blood work has been done for kangaroo rats
(454).
The effects of psychomimetics like LSD-25 and psilocybin
have been studied as has the uptake of l!31 in the thyroid
of D. merriami (454). Kangaroo rats have also been used
in a~field study on the effect of a rodenticide (Gophacide)
(460). They seem very suitable for these experiments.
General Suitability -
Kangaroo rats and pocket mice are gentle laboratory sub-
jects and would make good species for use in experiments.
However, the difficulty of breeding them in captivity de-
tracts from their suitability.
Ecological Alternatives -
Other rats, mice.
150
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Rodentia: Cricetidae, Muridae - Wild Rats
Introduction -'
The wild rats of North America are from two rodent families,
the Cricetidae and the Muridae, and four genera, Rattus,
Neotoma, Sigmodon, and Oryzomys. Members of all four have
been used in some way in the laboratory.
Neotoma, the woodrats or packrats, is a widespread genus
ranging from the east coast forests to the deserts of the
west. There are eight species, all easily distinguished by
their hairy tails.
Sigmodon, the cotton rats, are represented by S. hispidus,
which is abundant in grassland habitats throughout the
southern U. S. Two minor species are found in the foothills
of northern Mexico, southern Arizona, and New Mexico.
Oryzomys palustris, the rice rat, occurs in marshy areas in
the southeastern U. S. It is a semi-aquatic species that
usually nests under debris above high-water level.
The Rattus species have been introduced by man and are
adapted to living in and around man-made structures, parti-
cularly where food is stored.
Rattus and Neotoma weigh about six to 12 ounces and their
body length is from seven to nine inches. Sigmodon and
Oryzomys are smaller weighing between two and six ounces
and measuring between four to seven inches in body length
(412). All have long tails, are typically rodent-like in
appearance, nocturnal, and omnivorous. They are relatively
abundant locally within their ranges.
The multimammate mouse (Rattus natalensis) , an African wild
rodent, has been introduced to the U. S. for laboratory
testing and is laboratory-adapted (461).
Caging/Lab Conditions -
Wire mesh, glass, and sheet metal are acceptable materials
for cages (453, 462). Wooden enclosures should be avoided.
The dimensions of cages in use range from 27 x 37 x 34
inches for a mesh cage with a galvanized removab]e floor
tray, to 16 x 15 x 15 inches for a sheet metal cage with
a wire mesh door (463) . A 3 x 8 x 35 foot enclosure with
151
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28 compartments has been used to house a colony of wild t
(462). Nesting materials such as cotton, excelsior, and
twigs are necessary (464, 465) as are nest boxes (453,
463) .
Conditions under which Central American cotton rats
Sigmodon, were successfully maintained in a lab are 15 to
16 C, 75 to 80 percent relative humidity, and 24 hours of
light per day (466) .
Male rice rats were kept in a common enclosure while ferna]-
were housed individually and no serious confrontations or""
fights were reported (467).
Nutrition -
The rice rat has been successfully maintained on a diet
of small grains, whole or rolled oats, greens, fruit and re-
vegetables, plus dog food fortified with cod liver oil (453
468) .
Recommended diets for Neotoma floridana include carrots
sunflower seeds, peanuts, stale bread, tomatoes, oatflakes,
berries, hard and fresh corn (463, 464).
Cotton rats have been fed combination diets of sunflower
seeds, whole wheat, oatmeal, cod liver oil, corn, and pow-
dered protein (meat scrap) with periodic greens and apple
or tomato supplements (463).
Diets for Rattus are not available, but it is likely that
combinations of the above would prove suitable because the
feeding habits of all the wild rats are comparable.
Water should be provided ad libitum for all wild rats.
Breeding/Rearing -
The breeding and rearing characteristics of the wild rats
are summarized in Table 13.
Behavioral Restrictions -
Researchers who have kept Sigmodon hispidus, the cotton rat,
report that cage mates are likely to fight, and that the
cotton rats do not tame easily or lend themselves to han-
dling (453, 463). Rice rats are docile and handle easily
(469). The eastern woodrat (Neotoma floridana) fight when
caged in numbers, but usually becomes habituated to others
and ceases hostilities (464). Wild populations are
152
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Table 13. BREEDING AND REARING CHARACTERISTICS OF THE WILD RATS
Group
Siqmodon
(463, 470)
Litters/
Years
3-6
Gestation
27 days
Young/ Age at
Time of Estrous* Litter Maturity
Feb. to Nov.
3-6 50 days
Rattus
(412, 467)
12
21-22 days Year-round
8-10 90 days
U1
U)
Neotoma
(464, 465, 471,
472, 473)
30-36 days March to Nov. 3-4
60-90
days
Oryzomys
(467, 468)
25 days
Feb. to Oct.
3-4 50 days
*The wild rats are polyestrous in captivity,
-------
somewhat colonial and should adapt well to lab conditions.
Species of Rattus which have been bred for laboratory
animals, will be treated elsewhere.
Ecological Role -
All wild rats are ominivorous. The woodrats (Neotoma) and
Old World rats (Rattus) feed more on animals than the cot-
ton rats (Sigmodon) and rice rats (Oryzomys) which favor
green vegetation (412).
All wild rats are potential and often valuable sources of
food for predatory snakes, birds, and mammals (412).
Sigmodon hispidus affects grassland bird populations by
eating eggs and are agricultural pests in certain areas
(463).
Longevi ty -
Desert woodrats, Neotoma lepida, have lived over five years
in captivity and the other species of wild rats probably
are capable of rather long lives in captivity (412).
Problems with Mass Culture -
Woodrats have a musky odor that may become rather unpleasant
to lab workers (465) and are very suitable hosts for fleas,
mites, etc. Care should be taken to eliminate parasites
from new individuals received from natural populations
(471, 474).
Toxicity Testing -
The genus Rattus has been used extensively to determine the
feasibility of using U-5897, a male chemosterilant, to
control pest populations of wild rats. Single oral doses
ranging from 0 to 300 mg/kg were administered to males of
four species of Rattus with mixed results (475, 476).
Other rodenticide studies have employed Norway and Black
rats with a variety of administration methods, primarily
forced ingestion of food additives (477, 478, 479).
General Suitability -
The suitability of wild rats for lab research is good due
to their ability to reproduce in large numbers, their
small size, and their relatively inexpensive diets.
Oryzomys palustris is recommended by labs that use it (469)-
154
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Ecological Alternatives -
Cotton rat alternatives include voles, Microtus, and rice
rat (412).
The eastern woodrat could be replaced by other woodrats and
by the Norway and Black rat (412).
The rice rat alternatives are the Norway rat, packrat, and
cotton rat (412) .
The genera of the wild rats are somewhat ecologically inter-
changeable .
155
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Rodentia: Sciuridae - Ground Squirrels
Introduction -
Ground squirrels (Spermophilus, Ammospermophilus) , are ro-
dents that inhabit openlands of many types and are limited
to the midwestern and plains region of North America. The
are diurnal, burrow dwellers, and hibernate during adverse
seasons. Their average body length is five to ten inches,
and they possess long furry tails. Adult body weights in
this group range from six to 38 ounces (144).
Caging/Lab Conditions -
Wooden cages will be gnawed through by ground squirrels
(453). However, 12 cubic feet packing crates surrounded
with wire mesh with a sand floor have served as suitable
housing (480). Metal suspension cages from four to ten
cubic feet have been used (481) . Litter composed of pine
shavings and burlap sacking has been used (481).
The California ground squirrel, Spermophilus beecheyi ,
has been housed and bred in 4 x 8 x 2 feet enclosures of
1/2 x 1 foot welded wire. A nest box, 12 x 12 x 12 inches
should be included (481).
Nutrition -
Ground squirrels are omnivorous and have been successfully
sustained on combinations of seeds, greens, fresh vege-
tables, dog food, raw meat, bone meal, and cod-liver oil
(853). Purina lab show has proven adequate for Spermophilus
(=Citellus) parryi (480). Ground squirrels like to
hoard food in nest boxes even if it is supplied a<5 lib it urn
and the nest boxes should be cleaned daily (482).
Breeding/Rearing -
The gestation period for the Columbian ground squirrel
(Spermophilus columbianus) is 24 days (453). The young of
the thirteen-lined ground squirrel ( S. tridecemlineatus)
are weaned at 30 days, but those of the spotted ground
squirrel (s . spilosoma) are not weaned until about 48 days
after birth (483).
156
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Various ground squirrels have been bred successfully in
captivity, including the golden-mantled ground squirrel,
ftmmospermophilus lateralis) the antelope squirrel ( A_._
leucurus)for three generations, the Columbian ground
squirrel (S. columbianus) the thirteen-lined ground squirrel,
S. tridecemlineatus (453) , and S_._ beechayii (481) . Green
vegetable material (i.e., lettuce) given before spring
breeding (60 to 70 g per day) may influence the reproductive
performance in ground squirrels (481).
Behavioral Restrictions -
Ground squirrels are solitary in nature and present problems
when confined in small areas. When housed together they
fight, except for a few weeks during the breeding season
(484). The arctic ground squirrel S^ parryi is canniba-
listic in captivity (480) and there is evidence that other
species will eat their young when caged (484) . While
single individuals adapt well to cage living, they do not
become docile and cannot be handled easily. Ammospermo-
philus from the Mojave Desert makes a good cage squirrel
and becomes fairly domesticated (485).
Ecological Role -
Ground squirrels are primarily herbivores though certain
species rely heavily on insects during dry seasons in arid
habitats. They are a food source for raptors and large
carnivorous mammals.
Longevity -
The life span of ground squirrels in nature is quite vari-
able, but in captivity they have been known to live up to
eight years (453) .
Toxicological Testing -
Bishydroxycoumarin has been successfully administered to the
California ground squirrel via an oral administration tube
and later assayed from liver and fecal samples. A dose of
20 mg/kg body weight was given every 24 hours to adult
squirrels. Differences in plasma dicoumarol were reported,
the concentration depending on differences in diet (486) .
157
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General Suitability -
There are problems with breeding wild caught ground squir-
rels. Successes with S. beecheyi are encouraging and makes
this species a prime choice for future use.
Ecological Alternatives -
Chipmunks , prairie dogs.
158
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Rodentia; Scuridae - Chipmunks and Squirrels
Introduction -
The chipmunks are rodents of the squirrel family and closely
related to ground dwelling squirrels. There is one species
common throughout eastern North America, Tamias, and six-
teen western species, Eutamias. They are from three to
seven inches in length, excluding their long, furry tails.
They weigh from two to five ounces, the eastern chipmunk
being larger than the western forms.
Chipmunks occur in all types of habitats from woodlands in
eastern North America to rocky slopes, high deserts and
conifer scrubland in the Rockies. They are diurnal, pri-
marily ground dwellers, and omnivorous although they can
exist for long periods on seeds alone.
These small squirrel-like creatures are very active and
alert. They require facilities with adequate room for
exercise.
Tree squirrels (Sciu.rus , Tamia sciurus) and flying
squirrels (Glaucomys) are well studied in the wildlife
literature, but there appears to be very little maintenance
information available.
Caging/Lab Conditions -
Glass or metal cages 2x2 feet with tops of fine wire
mesh have been successfully used to house chipmunks in the
lab. A substrate of dead leaves or another adequate
substance should be used (453). An exercise wheel would
probably increase the acceptability of the enclosure.
Nutrition -
Chipmunks are omnivorous, but feed mainly on fruits, nuts,
seeds, and invertebrates. Their natural diets conform to
the seasonal availability of foods. In the laboratory,
mixtures of seeds (canary and sunflower) with fresh greens,
fruits, chopped raw meat and cod liver oil have been
successful diets (453). The average daily water intake for
Tamias is about 33 ml. (487), and it has been found that
Eutamias need less water per gm of body weight than its
eastern cousin (488) .
159
-------
Tree squirrels are primarily consumers of nuts and these
dietary items are suggested as food for captive indi-
viduals (489).
Breeding/Rearing -
Chipmunks are not readily bred in captivity, but the long-
eared chipmunk (Eutamias quadrimaculatus) has been success-
fully bred in a large, screened, outdoor cage in Californis
(453). Pregnant females brought into a lab may eat their
young (487) or refuse to care for them (490).
The gestation period is 31 .days for all chipmunks. Tamias
striatus produces two litters each year of four to six
young each (491), while Eutamias breed but once in the
spring and average five young per litter (490) .
The gestation period in Sciurus is about 44 days, in
Tamiasciurus about 38 days, and for Glaucomys about 40
days.They all breed once a ye at and give birth in late
spring or early summer. The number of young varies from
two to eight (412). There is little known about captive
breeding in these genera.
Behavioral Restrictions -
Because chipmunks and other squirrels are solitary in
natural situations, there may be difficulty in keeping
more than one individual in each cage. Their social
intolerance is well documented (453, 492). Various
species of Eutamias have been housed in groups with no
fighting; however, most chipmunks and squirrels are non-
social and territorial in nature and would probably be
difficult to house in numbers.
Longevity -
Chipmunks have been known to live up to eight years in
captivity. Because they adapt well to conditions of
captivity, their mortality is minimal in laboratory
situations (453) .
Squirrels will live from 10 to 15 years in captivity (&17.)
Toxicity Testing -
Other than for simple residue analysis, squirrels and
chipmunks have not been used in toxicological testing.
160
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General Suitability -
Chipmunks and squirrels are difficult to. breed and
maintain in a laboratory. Suggested species from this
group are the eastern chipmunk (T. striatus) because of
its abundance in the eastern United States and its size,
and the least chipmunk (E. minimus) for similar reasons,
though its range is northern and western. These species,
and Glaucomys are available from some dealers.
Ecological Alternatives -
Mice, ground squirrels.
161
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Rodentia; Cricetidae - Voles and Allies
Introduction - '
The microtines include the lemmings (Dicrostonyx, Lemmus)
the red-backed voles (Clethrionomys) and other basic grass-
land dwellers (Phenacomys, Microtus).
This group includes both woodland, grassland and tundra
species. All occupy similar niches in these habitats, being
consumers of green vegetation with dentition specifically
adapted for crushing grasses, sedges, seeds, bark, etc.
They are mouse-like in form and size.
Because species of Microtus for example, M. pennsylvanicus,
M. ochrogaster, are the most utilized in laboratory work,
and most of the available information refers to these
forms, the following comments are generally restricted to
them.
Caging/Lab Conditions -
Two basic sizes of enclosures have been used to house these
small rodents. Smaller cages of various sizes and materials
have been used to house breeding pairs; 12 x 8 x 5 inch metal
cages with lift off lids, (493 , 494) 12 x 11 x 6 inch
plastic cages with wire mesh tops 12 x 18 x 11 inch
stainless steel, 18 x 7 x 4 inch fiberglass (496); 29 x
19 x 13 inch polypropylene cages (496). A 36 x 30 cm
cage for mother and young has been used (496). Substrates
of peat moss (493, 494) , and sand (497) have been used.
It has been reported that wire mesh flooring is unsatis-
factory (495) .
Large enclosures for colonies have been used by some labs.
A 6 x 25 x 2 foot galvanized metal cage with 48, 2x3x6
inch nest boxes, has been used for studies on adrenocortical
activity in confined populations (498).
Feeders have been used of 6 x 2-1/4 x 9/16 inch dimensions
with a 2 x 2-1/4 x 1 inch catch basket for overspill (398).
Bedding materials are necessary, and some that have been
used include cotton, corn husks (499), dried grass, hay and
wood shavings (496).
162
-------
Nutrition -
Corn, hay, wheat, oats, mixed vitamized olive oil and
excess hay (493,500) are some foods administered to
Microtus in the lab. Others include Rockland mouse diet
supplemented with fresh greens (lettuce and cabbage) (498)
though one report states green vegetable material is not
necessary and when given causes diarrhea and undue mess
(495). One lab, whose feed is given in Table 14, reports
that M. ochrogaster is quite sensitive to deficiencies cf
any sort (495). Other feeds can be found in references
499, 501 and 496. All sources state that water should
be available ad libitum.
Breeding/Rearing -
Microtus breeds well under laboratory conditions. The
gestation period is 21 days (453, 493, 502)and M.
agrestis averages 3.7 pups per litter. M. oregoni averages
four or five litters each year under natural conditions.
Gestation is about 25 days in this species (503) . M.
ochrogaster pups are weaned at .21 days, females reach
maturity at 35 to 40 days and males at 42 to 45 days (495) .
Grasshopper mice, Onychomys, are seasonally polyestrous
in the wild with single females producing three to six
litters each year for two or three years (497).
Female pine voles, M. pinetorum, usually have two to four
litters each year, but could produce eight (504).
The effect of ambient temperature and light conditions
on the reproductive ability of M. ochrogaster is shown in
Tables 15 and 16.
Special Needs -
Overcrowding Microtus impairs their reproductive potential
and sociability(498). Acceptable population optimum
densities are not known. Small cages are sufficient for
one breeding pair and a single litter (495).
Grasshopper mice (Ocychomys; should be supplied with a
substrate suitable for dust baths (497).
Pine voles (M. pinetorum) are inhabitants of the cooler
woodland floor and are sensitive to high temperature (504).
163
-------
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TABLE 15. REPRODUCTION OF U. Of.HROGASTER IN THE LABORATORY
AT DIFFERENT TEMPERATURES
Temperature*
Night Day
25
20
10
5
0
30
24
15
9
4
Number of
Days Pairs
160
120
102
60
48
16
15
12
16
20
Reproductive
Period
38
34
31
24
27.5
No.
Born
3.6
3.7
4.0
4.6
4.4
Young
Weaned
3.3
3.4
3.5
3.5
3.0
Percent
Survival
91
92
88
82
67
•There was no consistent shift from one temperature to another, with some animals spending
several months at the same temperature. Light was maintained above 20 foot candles for
14 hours.
TABLE 1G. REPRODUCTION OF ft. OCHROGASTER IN THE LABORATORY
ON DIFFERENT LIGHT SCHEDULES
Hours*
Light
14
12
10
11
12
13
Number of
Days Pairs
60
35
32
37
38
32
16
25
26
28
30
31
Average re-
productive Period
24
36
48
59
28
23.5
% Bred 3 Days
Postpartum
86
43
39
40
83
88
Average
No . Young
4.57
(..2
3.7
3.7
4.55
4.6
Repr oduc tive
Efficiency
93
73
46
38
84
94
*The colony was stabilized at 14 hours light, 10 hours dark, then shifted down to 10 hours
light by the steps and periods listed below, with approximately 50 cycles accumulated in
each stage. Temperature was maintained at 9 C day and 5 C night, on 12-12 hour changes.
Source:
495
165
-------
Behavioral Restrictions -
Fighting is frequently reported among the microtine
rodents (412) and aggressiveness, litter mortality and
infectious disease have been shown -to increase as the
density increases (498).
Ecological Role -
The small herbivorous rodents are important components of
nearly all natural terrestrial communities. They are a
primary food source of smaller predacious reptiles, birds,
and mammals.
Longevity -
Members of this group have a life-time of about one year
(503).
Problems with Mass Culture -
Parasites can be a problem (497, 504).
Toxicity Testing -
The use of Microtus in behavioral studies of overcrowding
is well documented (498, 505) and their reactions to
certain pesticides (e.g., endrin, dieldrin) have also been
studied. The substances were administered orally in a corn
mixture and the subjects individually caged (6) . The effect
of endrin on an enclosed outdoor population was studied
by periodic trapping following administration of the
chemical by spraying. The result was an immediate decline
in numbers and no recovery in numbers for years after
(506). Similar studies with endrin administered
orally to pine voles were conducted (507). Rodenticide
studies have also utilized voles (508, 509).
General Suitability -
They are small animals, easily and inexpensively kept. M.
pennsylvanicus, for example, is plentiful, has a large
range, and is easily bred in captivity. These are very
suitable animals, in general.
Ecological Alternatives -
Other voles (412), Onychomys (grasshopper mice), Clethriono'
(red-backed voles).
166
-------
Rodentia: Cricetidae - Wild Mice
Introduction -
The mice of the genus Peromyscus and allies, Onychomys,
Reithrodontomys, Zapus, are by far the most abundant and
widespread native mammal group in North America. The
white-footed mice are medium-sized mice with grey to brown
backs and sides and a very light underbody. There are
about 16 species in the United States and representatives
of the genus can be found in all types of habitat. The
most widespread species, P. maniculatus, ranges through
North America with the exception of the southeastern U.S. ,
the tundra of northern Canada, and the hot deserts of
Mexico.
All members of the genus are nocturnal; they are generally
ground dwellers, though some species nest in trees. In
nature, they rely heavily on seeds for their food, but
eat invertebrates readily. Spedies that have been used
for laboratory research include the white-footed mouse
(P. leucopus) , the deer mouse (P_. maniculatus) , the golden
mouse (Peromyscus = Ochrotomys nuttali), and "the cotton
mouse (P_. gossypinusTT
Caging/Lab Conditions -
Lab enclosures that have proved satisfactory for housing
wild mice include a 15.5 x 14.2 x 12 inch cage of 1-1/4
inch wire mesh with baking pans for the tops and bottoms,
quart fruit jars with cotton for nesting and a sawdust
floor (510); a 9 x 7 x 5 inch wire mesh cage with re-
movable wood top and a metal pan for the floor and cotton
for nesting (511); large aquariums or wire cages with
solid bottoms covered with a layer of sand or wood chips
and nest material in small nest boxes (512). For numbering
systems and data card see reference 494.
Nutrition -
Successful diets include frisky dog crackers and lettuce
(510); rolled oats, dry meat scraps, dry skim milk, whole
wheat, wheat germ, cod liver oil, sunflower seeds, hemp
seeds, canary seeds, millet, iodine salt and lettuce
(494, 513); and commercial mouse chow supplemented with
rolled oats, whole corn, sunflower seeds, nuts, greens,
and fruits (512). Food and water are always supplied
ad libitum.
167
-------
Breeding/Rearing -
The members of this group that have been used in labs, for
the most part, breed year round and have three to six young
per litter. Gestation last from 23 to 30 days, the young
are weaned at about 25 days and achieve sexual maturity at
about 55 days (510, 511, 514). P. maniculatus females
mature faster than males (514).
P^ gossypinus is known to hybridize with P._ leucopus in
captivity and produce fertile offspring. Cotton mice
(P_. gossypinus) , are somewhat colonial in captivity and a; -
parently reach sexual maturity later than some other wild
mice (approximately 70 days) (510). This species usually
nests off the ground in nature (511).
Golden mice (O. nuttali) share communal nest boxes in the
lab. In nature, they do not breed from November to
February.
Special Needs -
Nest boxes are better for litter rearing than open cage
floors.
Ecological Role -
Mice are small omnivores that are an important food source
for small carnivorous reptiles, birds, and mammals. They
are found in almost all habitats throughout the world and a
very plentiful in many areas. In some cases they are sig-
nificant dispersers of seeds.
Longevity -
It is very unlikely that the average life span of wild mice
in nature exceeds two years, but in captivity, a life span
of five to eight years is not unusual (412) .
Problems with Mass Culture -
There are many known parasites of mice, e.g., nematodes
(510, 515), fur mites, and fleas (516), lice (494), and
these should be controlled. Otherwise, wild mice
(Peromyscus) seem quite suitable for mass culture.
168
-------
Toxicity Testing -
Wild mice, (Peromyscus maniculatus, 1?. leucopus) , have been
used in studies of the effects of DDT (167) and Endrin
(348, 506, 517) both in the lab and under natural condi-
tions. Mouse populations declined after the ground appli-
cation of endrin by spraying (eight ounces per acre) and
when fed endrin with their food in the laboratory, showed
adverse effects (348).
Experiments on the effects of sodium flouracetate - the
rodenticide was fed with bait in a natural environment -
have also been performed (518).
General Suitability -
This group in general is very easily bred and maintained
in the lab. P^ maniculatus is the most abundant and wide-
spread species, but others are just as suitable for re-
search.
Ecological Alternatives -
Other mice (412).
169
-------
Rorientia: Muridas - Lab Rats and Mice
Introduction -
The choice of research animals in the past has been based
on availability and ease of maintenance. No group of
animals was more available than the old world rats (Rattus
norvegicus) and mice (Mus musculus) which have adapted so
well to living with man. The white lab rats and mice of
today are albino strains of these species that have beer.
selectively bred for research purposes.
Caging/Lab Conditions -
There are numerous suitable cages available commercially
for housing laboratory animals, largely designed for lab
mice and rats. Unique apparatuses for restraining, anesi':
tizing, bleeding, and feeding them are available, and are
specially designed for these gentle, docile, easily mani-
pulated species.
Lab rats and mice can be housed in nearly any enclosure
including fruit jars (519), screened cages (520), metal or
plastic cages (521, 522). Overcrowding should be avoided.
Typical mice cages are 18 x 12 x 6 inches and typical rat
cages are 24 x 20 x 10 inches (289).
The type of bedding seems to be more important than the
cage. Pregnant mice produced larger litters, and a
greater percentage of their progeny was successfully
weaned on sawdust than on a commercial deoderized cellu-
lose (523). Mice housed on sawdust bedding also out-
produced those on corn cob bedding (524). Other bedding
suggestions can be found in references 525, 526 and 527.
Other environmental factors found to affect the performance
of lab mice are ambient light conditions (528, 529), noise
(530) , feeding schedules (531) , isolation and stress (532),
NH3, dust, and relative humidity (521). A thorough mainten-
ance procedure for mice is presented in reference 533.
Sawdust bedding for rats is considered inadequate due to
the intestinal obstruction it causes. When it is screened
of small particles it provides a better substrate (534).
Ventilation of the lab facilities is a proven necessity
(535).
170
-------
Nutrition -
Commercially prepared lab diets are used exclusively by
many institutions and together with water and a few minor
additions in specific cases, is supplied ad libitum (519,
522, 536, 537, 538, 539, 540, 541). There are food tube
dispensers available for accurate dispensing of certain
diets. Feeding frequencies have been shown to affect growth
in young rats (543). There are controls for diarrheal
disease available (544).
The suggested ration for adult mice is four to five gins per
day and for adult rats, 12 to 15 gms per day (289).
Breeding/Rearing -
The breeding of lab rats and mice is very easily accom-
plished if they are allowed space enough. The mouse
gestation period lasts an average of 19 days; the litter
size averages 10 to 12 young and they are weaned at 16 to
21 days of age. They are sexually mature at 60 days.
Females can produce up to ten litters in a lifetime and the
male breeding life lasts about 18 months. One male to
three females is a suggested ratio for colonies (289).
Albino rats are sexually active for about one year. Females
gestate for 20 to 22 days and eight to 12 young per litter
can be expected. They are weaned at about 21 days and
sexually mature at 100 days (289) .
Behavioral Restrictions -
Overcrowding should be avoided.
Ecological Role -
Lab rats and mice are bred for man's uses only and have no
ecological role in the wild.
Toxicity Testing -
Albino rats and mice have been the most used subjects to
date in toxicity testing. LDso's for large numbers of
compounds are available (545). (Testing methodologies, it
should be noted, vary greatly). Behavioral tests have
also been developed. Reviews of these methods and their
validity may be found in references 546 and 547.
171
-------
General Suitability -
Laboratory bred rats and mice are readily available and
easily maintained. However, because they have been selec-
tively bred for laboratory work and long removed from the
genetic influences of wild-types, their relationship to
natural fauna is indefinable, and the results of studies
using them may not be extrapolable to populations in the
wild.
Ecological Alternatives -
Wild strains of rats and mice.
172
-------
Rodentia: Castoridae, Cricetidae, Erethizontidae, Sciuridae,
Capromyidae - Large Herbivores
Introduction -
The beaver (family Castoridae), the muskrat (family
Cricetidae), the porcupine (family Erethizontidae), the
yellow-bellied marmot, the woodchuck (family Sciuridae), and
the nutria (family Capromyidae) are large herbivores not
often used in laboratories, In nature the beaver (Castor
canadensis) is distributed locally throughout North America,
utilizing streams and ponds with nearby cottonwoods, alders,
aspens, and so on (412). Both the muskrat (Ondatra
zibethica) and the nutria (Myocastor coypus)are inhab-
itants of marshes, swamps, and bayous. The muskrat ranges
throughout most of North America. The nutria occurs only
in the southern U.S. and Mexico, having been originally
a South American form. The porcupine (Erethizon dorsatuir.)
is a heavy bodied inhabitant of forested areas of the
western mountains and Canada. The woodchuck (Marmota monax)
occurs throughout the boreal Canadian forests and down into
the eastern U.S. It prefers open woods, and brushy ,
rocky ravines (412). The yellow-bellied marmot, (Marmota
flaviventris) is an inhabitant of talus slopes, valleys,
and foothills throughout the Rocky Mountains (412).
These large herbivores measure from 20 inches in total
length (muskrat) to 45 inches (beaver). They feed on all
types of vegetation - leaves, bark, twigs, and roots.
Caging/Lab Conditions -
Woodchucks were caged outdoors in 5 x 20 x 5 feet cages
made of wire mesh which was extended into the ground to
prevent escape by burrowing (548). Porcupines (2-3)
have been housed in 3 x 6 x 6 foot wire mesh (549), and a
pair of nutria in 3 x 2 x 1 foot metal cages with sawdust
bedding (550).
Nutrition -
Captive woodchucks were fed on ear corn and smoked herring
(548), porcupines on lawn grass, hay, fruits, oats, and
lab chow (549), and nutria on sugar beets and grasses (550).
173
-------
Breeding/Rearing -
Woodchucks breed in February and March and average 3.5
young per litter (551,552). Marmots breed about the same
time as woodchucks and produce from three to six young per
litter. The young emerge from the burrow after about 30
days.
The colonial beaver has one litter each year which averages
three young. They are born between April and July. Female
beavers do not breed until they are 2 1/2 years old (412).
The gestation period for porcupines is seven months and
usually there is one young each time. Breeding takes place
in the fall and birth in spring. There has been some suc-
cess breeding porcupines in captivity (549).
Nutria gestate for 120 to 139 days (the average is 128 days
and average five young per litter. There is post-partem
estrous and two litters a year seems to be the norm. In
captivity the sexual life of nutria lasts to five years.
Large cages are needed for breeding (550).
Special Needs -
Fresh food for the beaver, if needed, would probably be
difficult to provide, and its enclosure would have to have
a large water supply, a necessity for the grooming and
cleaning of their oily fur.
Fresh food for the woodchuck and marmot, which should in-
clude fresh greens, might also prove difficult and expen-
sive to gather.
Outdoor cages might be required for some species in this
groups.
Behavioral Restrictions -
Woodchucks and marmots are solitary, territorial creatures
which do not readily accept other individuals of their
species in their cages (548). They also hibernate for
months during the winter.
None of these species can be maintained in wooded enclosure'
for all of them would find no difficulty in gnawing their
way out.
174
-------
Ecological Role -
The beaver is a furbearer; its dams create lakes and ponds.
Like the marmot and woodchuck, it is a primary consuir.er.
Nutria and muskrats are also classified as furbearers.
The porcupine feeds on bark and twigs in forested regions
of western U.S. and Canada. All provide food resources
for the large carnivores.
Longevity -
Beaver have been known to life up to 11 years in the wild
and to 19 years in captivity. The woodchuck's life span is
four to five years (412). The life span of marmots is
unknown, but is probably very similar to that of the wood-
chuck. Nutria will live 12 years in captivity (412) .
Problems with Mass Culture -
Space will be a limiting factor for these large rodents
(551). The Sciurid forms will not readily accept confine-
ment in colonies. The large rodents are hosts of the
usual external parasites including ticks, fleas, etc.(551).
Porcupines are covered with quills and may prove to be
difficult to handle.
Toxicity Testing -
Though no references were located dealing with the use of
the beaver, marmot, or woodchuck in toxicity testing, mar-
mots and woodchucks have been used frequently for physio-
logical studies of hibernation.
General Suitability -
These rodents are probably too large to handle easily. In
addition, they require rather large enclosures. Nutria
are used by some researchers who consider them quite suit-
able subjects (550).
Ecological Alternatives -
Rice rat, voles.
175
-------
Carnivora: Canidae - Large Species
Introduction -
Large carnivores have been decreasing in number as their
habitats have decreased in size. Wolves (Canis lupus, ar.d
C. niger) have lost large amounts of habitable area in the
U.S. and are practically extirpated from this country ex-
cept from Alaska (Canada still has many wolf packs).
The common fox species are the red fox (Vulpes vulpes) and
the gray fox (Urocyon cinereoargenteus). Both species are
locally common throughout North America, the red fox oc-
curring in open woodlands north of Mexico and the gray
fox in woodlands south of Canada (412).
The coyote.(Canis latrans) is an inhabitant of open brush-
land prairies and deserts throughout central and southern
North America.
The wolf and coyote are dog sized and the foxes generally
smaller (412) .
Large cats are too rare to consider for mass culture and
no lab work on bears (ursidae) is known.
Caging/Lab Conditions -
Most cages for large carnivores are outdoors and large.
Nutrition -
Foxes have been maintained on fish and cooked cereals (553)
In the wild the large carnivores are predators and eat
most animal matter, as well as some berries and grasses.
Breeding/Rearing -
Wolves are generally social creatures and live in closely
knit packs. Their breeding is tied to their social struc-
ture; they probably produce only the number of offspring
needed to keep their pack viable. Gestation is nine weeks
long (554).
Foxes breed from February to May, depending on latitude and
local conditions, and produce from one to seven young per
litter (555,556,557). Their gestation period is 52 days
and the young breed at one year (558,559).
176
-------
Coyotes have a gestation period of 64 days (554). Five to
ten young are born in April or May and mature in one year.
Data on the breeding of large carnivores in captivity is
negligible, but in large enough enclosures there should be
no problem.
Special Needs -
Large carnivores prefer a den to sleep (556) . They prob-
ably need large areas for exercise and high protein diets.
Ecological Role -
The smaller forms are predators on rabbits and rodents.
The wolves generally prey on larger animals. Infrequently,
livestock is taken by all of these species. Of economic
importance to man is the fur of these species (412,443).
Longevity -
Fox have a ten to fifteen year life span in captivity (555).
Problems with Mass Culture -
Canids are very susceptible to external parasites such as
ticks, lice, etc., as well as tapeworms and roundworms (443)
Toxicity Testing -
In general, there is very little data concerning wild
Candis and toxicity testing. The effect of diethylstil-
bestrol, a synthetic estrogen, on red foxes has been
studied with good success. The foxes were force-fed the
substance (560).
Coyotes have been subjected to the management of their
numbers by the administration of diethylstibestrol in
bait (561).
General Suitability -
Due to their size, the large carnivores are probably less
suitable than smaller ones for laboratory research. How-
ever, fox have been commercially reared by the fur indus-
try and are breedable in captivity.
177
-------
Due to their dwindling population and large size, wolves
are considered very unsuitable for lab testing.
Coyotes are more plentiful than wolves and probably more
available to researchers.
Ecological Alternatives -
Domestic dog.
178
-------
Carnivora; Procyonidae, Mustelidae
Introduction -
The small carnivores include the family Procyonidae which
has one representative in North America, the raccoon
(Procyon lotor) which occurs throughout the U.S. with the
exception of the high Rockies. The species of Mustelidae
are numerous: the marten (Martes americanaV the long-
tailed weasel (Mustela frenataV the mink (Mustela vison),
the badger (Taxidea taxus),the striped skunk(Mephitis
mephitis), and the European ferret (Mustela putorius).
The ring-tail (Bassariscus astutus), the only North Amer-
ican member of the Bassariscidae, has not been extensively
used in lab work, but could prove very suitable. The
following information refers to those species of small
carnivores which have been selected because of previous
laboratory use.
Caging/Lab Conditions -
Mink, weasels, and ferrets have been maintained under lab
conditions. A good enclosure for weasels is an 11 x 9 x
8 inch wooden nest box connected by a doorway at one end of
the box with a runway of 1/2 inch galvanized wire mesh
(18 x 9 x 8 inches) where feeding and watering take place.
The nest box is provided with a removable plywood lid held
in place by a fastening device. The lid must be water
right if the box is used outdoors. A false top is placed
underneath the lid, consisting of a frame filled with 1/2 inch
galvanized soldered wire, resting on two cleats and
held in position by two wire finishing nails pushed through
holes in the sides of the box. Shavings and excelsior are
adequate nesting materials. Great care must be taken to
secure all edges and joints because weasels are superb
escape artists (562).
The proper housing of mink can be readily learned from
mink ranches. European ferrets have been successfully
housed in rabbit cages with a 12 x 6 x 6 inch nest box (563)
and in double compartment cages similar to those
described for weasels (564). They have been domesticated
for at least 2,000 years and their exact genetic ancestry
is quite nebulous. A natural photoperiod is nscessary to
effect breeding in this species (565).
179
-------
Nutrition -
In the wild, the small carnivores feed on crayfish, small
birds, insects, chipmunks, rabbits, mice, and shrews (566).
Diets for weasels in captivity include hamburger, mice,
fish, bread, milk, and ground liver (567) . Ferrets eat
similar foods as well as dry dog food and Gerber's Meat
Base for Babies (563,564). Bottles containing milk may
be placed in the cage to facilitate serving that item (562;
Mink are fed cotton rats, raw meat (568), mice, fish, musp-
rats , and frogs (569). For the fur trade, a special mink
food is prepared and sold commercially.
Breeding/Rearing -
The breeding and rearing characteristics of the Mustelidae
are summarized in Table 17.
Raccoons produce one litter each year of two to seven young
which are born in April or May about 65 days after concep-
tion. They are weaned at about two months (412, 554).
The ringtail also produces one litter each year of three to
four young that are born in May or June and develop at a
rate comparable to that of the raccoon (412).
Captive weasels breed readily but about 58 percent of
females bred in captivity fail to produce young and the
number whelped in captivity is often less than the four to
nine average whelped in the wild (560) . The family Musteii-
dae has delayed implantation which makes gestation times
difficult to determine (570).
Special Needs -
Weasels need dark sleeping chambers (562,564) and escape-
proof cages (562). A natural photoperiod is essential for
successful ferret breeding (565). The diets of the small
carnivores should include a substantial proportion of meat
(563).
Behavioral Restrictions -
All mustelids have musk glands which are responsible for
their peculiar odors. The best known example is the skunk
(571, 572). These glands can be removed. The weasels,
ferrets, and mink are nervous and frequently vicious ani-
mals (443, 474, 563). Skunk nest boxes should be left
undisturbed while young are present (573).
180
-------
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181
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Glass, wood, or any other material that can be gnawed or
broken may be hazardous (562).
Ecological Role -
The small carnivores are predators on many smaller verte-
brates and invertebrates. Many also eat seeds, fruits, an:
certain herbaceous material. Skunks and raccoons are often
scavengers and will readily feed on carrion and human gar-
bage.
Longevity -
Ferrets have lived to nine years (563) and the life spans
of other mustelids are probably comparable; smaller forms
will have shorter life spans.
Problems with Mass Culture -
Skunk parasites are numerous and many of them are easily
transmitted (515, 576, 577, 578). Influenza, canine
distemper, and tuberculosis are a few diseases reported in
captive ferrets (563); however, distemper can be treated.
The small carnivores are usually solitary in nature and
there are reports of individuals caged together displaying
great animosity towards their cage mates (574).
All of these species are nocturnal by habit; scheduling,
and so on, must be made to fit their cycle (571, 572).
Toxicity Testing -
Mink, which are farmed by the pelt industry and readily
available, have been used in testing of psychoactive com-
pounds, These were administered orally to adult males in
capsule form by means of a hollow wooden rod. Direct
observations of the males' behavior followed. Their innate
aggressiveness made them appropriate models for this work
(579). Wild mink have been trapped, killed and tested for
DDT residues in body tissues (580) and used in lab feeding
experiments with heavy metals, PCB's, etc. (581, 582, 583).
Wild trapped raccoons were examined for incidence of DDT
DDD, and dieldrin in a Missouri study (584).
182
-------
In Great Britain a study was conducted of pesticide levels
in the European badger (Meles meles), found dead along road-
ways from 1964 to 1968 (585).
General Suitability -
Small carnivores are relatively unavailable in large numbers
with the exception of mink that have been used in psycho-
logical behavior testing and judged quite suitable for such
research (579). Culls are available from some ranchers.
Raccoon and skunks are locally abundant, but large-scale
trapping could decimate populations. European ferrets are
an exotic domestic species and their suitability in
toxicity testing of wild species is questionable.
Ecological Alternatives -
Domestic cats.
183
-------
Carnivora: Canidae, Felidae - Domestic Species
Introduction -
This subgroup of carnivores includes the dog (Canis
familiaris) and the cat (Felis domestica), both familiar
animals in the laboratory. They are adapted to living wit
man, being handled, etc., and are very suitable for lab
work.
Caging/Lab Conditions -
A 36 x 16 x 18 inch cage of any easily cleaned and struc-
turally sound material is adequate for the housing of cats
Breeding cages are larger, 36 x 24 x 30 inches and both
require litter (289).
Dogs have been housed in 4 x 4 foot pens v/ith water and i'c
containers and resting platforms. These are connected
to outside running pens ranging in length from 12 to 20 fr
Inside pens 6x8 feet wil] house five adults (289) . Con-
crete floors in dog kennels provide easy cleaning (289).
Nutrition -
Commercially available diets with cooked horsemeat or brot
and additional vitamins are used for dogs. Cats are also
maintained on commercial foods. Water should be provided
ad libitium for both (586, 587).
Suggested dog rations are one-half ounce dry dog chow per
one pound of body weight. Adult cats should be fed four
percent of their body weight each day. Twice this ration
should be given to growing animals (289).
Breeding/Rearing -
Dogs vary somewhat in their breeding schedule, so the
beagle is chosen as the pattern subject because of its
size, availability, and adaptability to lab conditions.
Beagle females undergo estrous every 150 to 200 days and
apparently can produce only one litter each year. Gesta-
tion lasts 63 days and four to eight young are born per
litter. They are weaned in six to eight weeks and are
sexually mature in one year. The breeding life of female
beagles is six to ten years and that of males a little
longer. One male can service about 60 females in a
breeding colony though hand-mating may be required•(289) •
184
-------
Domestic cats can breed a number of times each year. The
estrous cycle is intermittent and irregular. Gestation
lasts an average of 62 days and litter size is from one to
six with an average of four. Weaning takes place in six to
eight weeks and the young are sexually mature in six months.
The breeding life of females is four to five years and males
generally can breed a few years longer. In a breeding
colony, one male for six females is suggested (289).
'Special Needs -
Dogs require no special bedding but a sleeping platform
raised from the floor. Cats should be provided bedding and
litter of wood shavings, etc.
Pregnant female dogs near term should be provided with
whelping boxes lined with shredded paper. Pregnant female
cats near term should be provided with nest boxes. They
should always be separated from other animals (289).
Behavioral Restrictions -
Both dogs and cats are easily handled and should present no
behavioral problems in the lab.
Ecological Role -
The ecological role of carnivores similar to the dog and
cat is discussed in the section on large carnivores.
Longevity -
Domestic dogs are productive for six to 14 years, cats for
four to seven years (289) .
Problems with Mass Culture -
There are no apparent problems associated with mass culture
of domestic dogs and cats.
Toxicity Testing -
The use of domestic cats and dogs in research is quite
extensive. Tests and methods from many disciplines includ-
ing physiological, immunological, biomedical, and toxico-
logical areas are available for both species.
-------
General Suitability -
This group is available in large numbers and adapted to
laboratory life. The animals are handled with ease and
are..most suitable subjects for experimental work. How suit
able they are for toxicity testing is difficult to say be-
cause their gene pools have been manipulated by man and,
in some important ways, have been free for many years fred
natural selection pressures.
Ecological Alternatives -
Mink, European ferret.
186
-------
Ungulates: Equidae, Bovidae
Introduction -
The domestic ungulates that have been used for scientific
testing other than the swine and allies are the donkey,
mule, horse, pony, and cattle. They are all large herbi-
vores, eating a variety of plant foods.
Caging/Lab Conditions -
The enclosures to contain domesticated ungulates must be of
suitable dimensions to accommodate their large size. Access
to an outdoor lot is desirable. No better description of
suitable enclosures is available, probably because the
maintenance of these domestic animals is so routinely
accomplished.
Special Needs -
Salt licks are necessary for all these animals.
Behavioral Restrictions -
All these animals may be difficult to handle and control
because of their large size. The biting and kicking of
donkeys and mules is well known.
Ecological Role -
The ungulates are grazing and browsing animals which rely
entirely on plant material for their food.
Problems with Mass Culture -
The space needed to house these animals is considerable.
Likewise, the food expense is much greater than for other
groups.
Toxicity Testing -
Because these animals are of economic importance, they have
been used quite extensively in many types of experimental
work, particularly in the study of the effects of pesti-
cides on their bodily functions. Chemicals tested on
cattle include heptachlor (588), DDT, BHC, lindane, methory-
chlor, chlordane, toxaphene, aldrin, dieldrin, endrin
(589, 590) and others (591, 592). These chemicals are
187
-------
administered in foods and through capsules. The testing
was conducted primarily to assay chemical residues in animal
products consumed by man. Substances that have been studiec
for their effects on cattle are nicotinic acid (593),
dimethyformamide (594) , arsenic, flourine, lead, salt,
selenium (595), iodine (596), and molybdenum (597).
Calves have been used in studies of taste reception; the
peripheral nerves were successfully exposed and impulses
monitored (598, 599).
Horses, donkeys, and mules have also been subjects in
studies of such pesticides as organophosphorous and carbo-
mate pesticides (591), DDT, toxaphene, and chlordane (592).
General Suitability -
The large size and domestic status of this group reduces
its suitability for the laboratory. Smaller ungulates
(sheep, miniature swine) may be better subjects.
Ecological Alternatives -
Deer, sheep.
188
-------
Ungulates: Suidae, Tayassuidae
Introduction -
Because of their smaller size, the swine (Sus scrofa) and
javelina (Tayassu tajacu) are used more in research than
other domestic ungulates (600). The special breeding of
miniature swine in the last decade or so has been very ex-
tensive and their availability is increasing. (For a re-
view of breeding stock and strains developed, see reference
600). While miniatures possess all the characteristics
of standard swine, they are far smaller and weigh from 70
to 90 kg, about as much as the average man. Their size
and availability have led to their increasing use in bio-
medical research (600). The javelina, or collared peccary,
is a native North American animal found commonly in Texas,
New Mexico, Arizona, and Northern Mexico. It belongs to
the family Tayassuidae. Another closely related species,
the white-lipped peccary (T. Pecari), occurs from Central
America to Paraguay. This swine-like animal weighs between
16 and 20 kg, and averages about 90 cm in length with a
shoulder height of approximately 54 cm. The ears are
small and erect, and the hair is long, bristly and dark in
color (601).
The following discussion is restricted to the miniature
swine unless otherwise indicated.
Caging/Lab Conditions -
Housing for miniature swine is generally a large community
cage of pipe and fencing. From one to ten pigs, depending
on age, are housed in 4 x 4 x 8 feet indoor cages with con-
crete floors and adequate ventilation. Outdoor cages 8 x
16 feet with earthen floors and a shelter roof with sawdust
bedding at one end of the pen housed an unknown number of
animals (602). Old dog kennels are apparently quite suit-
able enclosures (603). The transport apparatus, obser-
vation and holding pens used at the Sinclair Research Farm
are shown in Figures 15, 16, and 17 respectively.
Techniques for handling miniature swine for laboratory pro-
cedures are presented, with diagrams, in reference 604.
Pentobarbital sodium is a recommended anesthesia at 50 mg/4
kg body weight administered via the anterior vena cava.
Blood collecting has been accomplished from the same vessel
using 19 or 20 guage needles. Tranimul (600) and a spinal
two percent procaine (603) have proven acceptable tran-
quilizers.
189
-------
Figure 15.
Miniature swine transport apparatus,
Sinclair Comparative Research Farm,
University of Missouri, Columbia,
Mo. 65201
Figure 16.
Miniature swine observation enclo-
sures (20" x 30" floor area).
Sinclair Comparative Research Farm
190
-------
Figure 17.
Miniature swine in holding pen
(81 x 12' floor area). Sinclair
Comparative Research Farms
191
-------
Nutrition -
Commercial pelleted hog feed given twice daily is a suitable
diet (602). Water should be supplied ad libitum. A small
container for the water should be used to prevent the swine
from playing in it (600). The Hormel Institute has utilized
four different pig diets: pig starter, growth ration, ges-
tation rations, and maintenance ration, containing about
19, 15, 16, and 13 percent protein, respectively. The
Institute has also developed an artificial diet for baby
pigs. These four diets are given in Tables 18 through 21.
Breeding/Rearing -
With a regimen of full feeding, an average of 5.5 pigs per
litter are obtained by the Minnesota Hormel Institute. A
piglet weighs 750 to 800 gms at birth, grows to 18 kg at
140 days, and is full grown and about 90 kg at one year
(600). The Battelle Northwest Biology Laboratory uses
Pittman-Moore miniature swine, but limits the feeding
regimen of adult swine to 900 gms of feed per day. Their
swine average six young per litter but do not reach adult
weight until they are two to three years old. Gestation
ranges 112 to 115 days (603) for the miniature swine. For
the javelina which averages about three young per litter,
the period ranges from 142 to 148 days. Some females
have two litters each year (601).
The Oregon Dental School, Portland, has maintained Pittman-
Moore swine for some years, and reports that young were
weaned at 42 days and all members of the same age group
housed together for four months, segregated according to
sex; this communal housing effort tends to increase the
sociability of the swine (602).
Special Needs - , . •
Pen-to-pen contact between individuals should be limited to
prevent transmission of disease (603). New arrivals
should be temporarily quarantined; vaccinations for certain
diseases are available (603).
Swine housed outdoors need protection from the weather and
should always be provided shelter from direct sunlight.
Concrete floors are recommended to control parasites (603).
Exercise areas are needed if swine are housed indoors (603).
192
-------
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Table 21. ARTIFICIAL DIETS FOR BABY PIGS
Per Quart Homogenized Cow's Milk
One Whole Egg Plus
5 ml Salt Mixture Consisting of:
49.80 g FeSO. 7 H_0
*± ^
3.90 g CuSO. 5 H_0
3.60 g MnCl2 4 HO
0.26 g KI
200 ppm Aureomycin on Solid Basis plus water to make one
liter solution.
SOURCE: (605).
196
-------
Pregnant females should be separated from other swine on
the 109th day of gestation, washed and cleaned of parasites,
and housed in a disinfected cage with clean straw bedding
(see 603 for a plan for farrowing pens).
Behavioral Restrictions -
Sexes should be separated and sows with a new litter should
be handled with care because they can be quite dangerous
(603) .
Ecological Role -
In the wild, pigs are omnivorous, feeding on nuts, fruits,
leaves, cacti, grubs, etc. (412). Javelina inhabit areas
of rather dense vegetation (chaparal, mesquite, brushlands)
and are usually localized around a waterhole (412).
Longevity -
Miniature swine are known to live for more than six years
(600).
Problems with Mass Culture -
Although miniature, these swine are still large as lab
animals (600, 602). Maintenance is expensive and the
food costs may be limiting (606). Swine are subject to a
host of diseases, some of which are curable (603).
The javelina is rather new to the laboratory world and
little is known about its diseases and parasites (601).
Toxicity Testing -
Miniature swine are used in biomedical research (607, 608,
609, 610, 611, 612)and other fields as well. The Sinclair
Farms, University of Missouri, uses them extensively as
test animals (613).
Toxicity testing has been done for a variety of substances,
including manganese chloride (614) and selenium (615).
Pesticide effects have been investigated by scores of work-
ers (152, 590, 591, 592, 616, 617, 618, 619, 620, 621).
Javelinas have been used in cardiovascular research (601).
197
-------
General Suitability -
Swine are domestic animals. Their availability and the vast
amount of knowledge we possess about their breeding and
maintenance are their greatest advantages as lab animals.
As test animals they have been used successfully in re-
search in immunology, nutrition, dental, radiocardiology,
cardiovascular physiology, and pathology (600, 606, 622).
Their suitability as representatives of a complex, natural
ecosystem is unknown. The javelina, however, is a wild
form and would be quite suitable for toxicity testing, if
available in the future in large numbers.
Ecological.Alternatives -
Sheep, cattle.
198
-------
Ungulates: Bovidae
Introduction -
Sheep and goats are grazing and browsing herbivores. There
are.both domestic and wild species but the native members
of this group - Dall's sheep (Ovis dalli) the bighorn
sheep (Ovis canadensis), the mountain goat (Qreamnos
americanus), the Bison (Bison bison) and the musk ox (Ovibos
moschatus) are rare species and not suitable for use as
test animals.
Caging/Lab Conditions -
Cages should be dry and well ventilated and of sufficient
size tc allow free movement, with access to sizable runs.
Both sheep and goats can be communally housed, though the
uncastrated males should be limited in their access to
other animals during the rutting season, which occurs in
the fall, because they become aggressive and produce a
great odor (623).
The layout of an operating suite for sheep (at the
Agricultural Research Council Institute of Animal
Physiology) is detailed in reference 624.
Nutrition -
Mature goats do well on sun cured 15 percent protein alfalfa
pellets and block salt with the addition of one pound of an
inexpensive roughage such as hay or straw. Does used for
reproductive purposes should receive from one to 1.5 pounds
per day of a mixture of choice legume hay and 14 to 16 per-
cent quality grain. Clean water should be available at all
times (623) ..
Breeding/Rearing -
Goats ordinarily breed in the fall of the year and give
birth from February to June. The kids need milk for six
weeks and then should be weaned on a high protein dry food
diet. They reach maturity at three years and weigh 120 to
150 pounds as adults (623).
Ecological Role -
i
Goats and sheep are herbivores and feed on a great variety
of plant materials. The wild forms are high mountain
animals.
199
-------
Problems with Mass Culture -
t
The long generation time and large size of these animals
makes mass culture expensive.
Toxicity Testing - "
Pesticide residues in free-roving mountain goats have been
determined (625) by standard tissue analysis. Domestic
stock have been utilized for many types of research includ-
ing toxicity studies of various substances (590, 592, 616,
517, 618, 619, 620, 626, 627). Noteworthy is the Veteri-
nary Diagnostic Laboratory at the Iowa State University
which have been working with long-term effects of pesticide-
on domestic sheep (628, 629).
3eneral Suitability -
The suitability of all domestic animals as subjects in
toxicity testing is suspect because of their close associa-
tion with man's world, and because they have been so long
removed from the selection pressures of their natural en-
vironment. However, the sheep is probably more suitable in
this respect than other domestic species (613, 630).
Handling of these domestic creatures is not difficult.
Their suitability for surgical methods, etc., is discussed
at length by Fletcher, et al., 1964
Ecological Alternatives -
Other ruminants,
200
-------
Ungulates: Cervidae, Antilocapridae
Introduction -
The deer and allies are grazing and browsing animals from
two families. The Cervidae are the deer-like ungulates:
elk, moose, mule deer, white-tailed deer. The Antilocap-
ridae have a single representative in the U.S., the prong-
horn, which inhabits open prairies and sagebrush plains of
the west central U.S. Unlike Cervids, in which only males
have deciduous antlers, both sexes of pronghorn possess
true horns that are never shed.
The weight of the adult male elk (Cervus canadensis) and
the moose•(Alces alcesj is from 800 to 900 pounds; female
weights are about 200 pounds less. They stand five to six
feet high and the males of both, species have magnificent
racks. The moose antlers are broadly palmate and elk, ant-
lers are morphologically similar to those of native deer,
but considerably larger.
The mule deer (Odocoileus hemionus) and the white-tailed
deer (0. virginianus)are smaller than elk, standing be-
tween three and four feet high and weighing between 150 and
400 pounds. The mule deer ranges throughout western North
America in many habitat types. The white-tail is an
eastern species inhabiting all types of forest and brush-
land (412) .
Pronghorn (Antilocapra americana) stand about three feet
high and weigh between 75 and 130 pounds (412) .
Caging/Lab Comditions -
Any large outdoor enclosure of adequate areal extent and
height is suitable, but cover is essential both from the
sun and the cold in northern climates. An outdoor holding
pen used at the Denver Wildlife Research Center is shown
in Figure 18.
In the wild, deer eat twigs, shrubs, acorns, grass farm
crops, and certain fungi (412, 443). Elk and pronghorn
have similar diets. The moose inhabits swampy, marshy
areas and feeds particularly on aquatic vegetation, but
also on shrubs and saplings. White-tailed deer were
maintained on Ralston Purina Dairy chow for three years
during a pesticide study (631).
201
-------
Figure 18. Deer holding pen. Denver Wild-
life Research Center, Federal
Center, Denver, Colorado
202
-------
Water in large quantities is essential to all of these
animals (412, 443, 631).
Breeding/Rearing -
White-tail deer have been bred and reared in captivity, pro-
ducing one or two fawns each year. Females mature in their
second year (631). In nature, all the ungulates breed once
a year and produce one or two fawns (412). Gestation for
the pronghorn, moose, and elk is about eight months and for
the deer species from six to seven months (412).
Special Needs -
Moose prefer aquatic habitats for feeding and wading during
the summer months and may require water facilities for such
activity in captivity. All ungulates require mineral (salt)
licks (443).
Behavioral Restrictions -
Many species have a migratory tendency and may become quite
restless at certain times during the year (412, 443).
Ecological Role -
The ungulates are rather strict herbivores and graze on
many plant foods (412). They are prey for large predators,
particularly wolves. They also constitute the major big-
game animal group in North America. In some agricultural
localities, they can be serious pests of crops (412).
Longevity -
Elk have lived to 25 years in captivity. In the wild their
life span is probably less. Deer live to 16 years in the
wild, moose to 20 years, and pronghorn have been known to
reach 14 years (412).
Problems with Mass Culture -
The cost of maintaining a heard of large animals is high and
the reproductive rates are rather slow. A deer, for
instance, gives birth to one or two fawns each year.
In enclosed situations, deer chew each others tails and
ears. Mortality due to the "white muscle disease" can be
significant (631).
203
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Toxicity Testing -
DDT residues have been assayed from samples of free-running
pronghorn, deer, elk, and moose (625). Radioiodine has
been sampled from thyroid tissues of free running deer and
in Colorado following underground explosions in Nevada (632),
Strontium-90 has been assayed from the mandibles of free
running mule deer (633) .
One study of the effects of dieldrin on reproduction, mor-
tality, etc., in white-tail deer in Missouri was conducted
under controlled conditions on captive subjects (631).
General Suitability -
Limited reproduction and large size problems probably ex-
clude the large ungulates from mass culture and extensive
use in toxicity testing. The white-tail deer or mule deer
would be the most likely candidates from this group for
use in toxicity testing.
Ecological Alternatives -
Sheep.
204
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Primata
Introduction -
Because of their close relationship to man, non-human pri-
mates have been used in many fields of research. Primates
are omnivorous animals that have acquired the use of their
hands and often their feet as effective grasping organs
through the development of opposable digits. This makes
them particularly adapted for climbing, and most primate
species are arboreal.
They are found in the Old and New World tropical and sub-
tropical latitudes. The species used in research in North
America come from South America, (Platyrrhines), Africa
(Papio, Cercopithecus, Erythrocebus, Theropithecus) and
AsiarMacaca),(634). Recent restrictions on the capture
and shipment of primates may affect availability in the
future.
The most studied primate species other than man is the
Rhesus monkey (Macaca mulatta). Adults weigh 20 to 25
pounds and their life spans are much longer than those of
the other mammalian groups. The comments below refer to
this species, but many are applicable to other species as
well (i.e., squirrel monkeys, Saimiri).
Caging/Lab Conditions -
Cages for adult Rhesus monkeys should be at least 24 x 24
x 28 inches and house no more than a pair while breeding
or a mother and her infant. Smaller juveniles can be kept
in 14 x 12 x 18 inch cages (289). Cages of various dimen-
sions can be used to house large numbers of primates. They
are social animals and intra-specific contact is desirable.
Enclosures should be provided with suitable props for use
in climbing and exercise. Cages used at Iowa State Univer-
sity are shown in Figures 19, 20 and 21.
Nutrition -
Primates cannot metabolize ascorbic acid, so a daily ration
must be provided. After weaning, all ages of monkeys can
be sustained on commercially prepared monkey chow (289).
In nature they are omnivorous, feeding primarily on fruits.
Supplements to the ration of chow can include all types of
fruit and vegetable foods. Daily rations should be four
percent of body weight, and in communal cages a number of
205
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Figure 19. Primate cages - squirrel monkeys
metal - commercially available.
Iowa State
Figure 20.
Plexiglas and metal - fabricated
at Iowa State. (For dimensions
see Figure 21.)
206
-------
Figure 21. Plexiglas cage, fabricated at Iowa State
University.
207
-------
feeding areas should be established because dominant indi-
viduals will monopolize food sources. Water should be sup-
plied ad libitum (289).
Breeding/Rearing - ,
Gestation for Rhesus monkeys averages 165 days, and there
is usually only one young born at a time. They are fre-
quently removed from the mother soon after birth and reared
in a separate nursery. This procedure decreases infant mor-
tality. After three to six months, the young will eat solid
foods. Sexual maturity is achieved at five years for female;
and six years for males. The breeding life of Rhesus mon-
keys is 12 to 15 years ^nd females are receptive periodically
throughout the year. The breeding colony sex ratio can be
one male for up to ten females.
Special Needs -
Vitamin C is a diet requirement for primates (289).
Behavioral Restrictions -
Monkeys are often difficult to handle and unpredictable.
Special training of prospective handlers should be con-
sidered for their safety (289). A review of physical and
chemical restraints is presented in reference535.
Ecological Role -
Non-human primates are omnivorous animals of tropical and
sub-tropical regions of the world. Their staple foods are
fruits, leaves, roots, insects; they frequently consume
larger animals. They are prey for large predators.
Longevity -
Life spans of 15-20 years are not uncommon for Rhesus mon-
keys (289).
Problems with Mass Culture -
The generation time for primates is probably too long to
allow extensive use of this order of mammals when large
numbers are required.
Toxicity Testing -
Because of the long life spans of primates, they are invalu-
able in long-term research. The close relationship of
208
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monkeys to man is also an inducement to their use in research.
Experimental work in which primates have been subjects in-
clude pesticide toxicity work (272, 620, 636, 637), inhala-
tion of various gases (638). and other toxicity studies (104,
639, 640, 641).
General Suitability -
Non-human primates are desirable as experimental subjects be-
cause of their close relationship to man. Their long life
spans and specialized behavior also make them very suitable
for certain types of studies (630). However, extensive use
of primates in research is impossible because of their slow
rate of reproduction. Recent export restrictions may reduce
their availability in the future and increase cost.
Ecological Alternatives -
Fruit-eating bats, man.
209
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Marsupiala
Introduction -
The only marsupial native to North America is the opossum
(Didelphis marsupialis) which exists throughout North Amer-
ica except for the hzgher Rockies and the plains of the
Dakotas. It is probably limited in the north by the low
winter temperatures.
The opossum is an unspecialized mammal with a prehensible
tail. Its pelt is of poor quality. It is primarily noc-
turnal and omnivorous. Farming areas and disturbed wood-
lands are preferred habitats (412).
Adult opossums are about 18 inches in length with a tail of
equal length and weigh between nine and 13 Ibs. (412).
The pouchless opossum (Marmosa mitis) has only recently
been used in the laboratory.
Caging/Lab Conditions -
Cages for breeding pairs are 20 x 22 x 15 inches of gal-
vanized metal and 32 x 26 x 24 inches of galvanized metal
for individuals (642).
Large outdoor cages seem to better the reproductive perfor-
mance in opossums. Large wooded areas (150 x 300 feet)
were closed off by chain 1 ink fence eight feet high with an
eighteen inch wide metal strip around the top. Nest boxes
5x5x3 feet were provided (642). A 36 x 16 feet en-
closure of poultry netting surmounded by tin strips with
walls extending two feet in the ground has been used (643).
Breeding pens 20 x 40 feet are large enough to accommodate
ten adults (644). Other suitable nest boxes include dog-
house type structures with removable roofs (645) and
barrels with straw bedding (644).
In one lab, relative humidity under 50 percent resulted in
swollen and cracked tails (646) .
Marmosa have been kept in 8 x 8 x 11 inch cages with a
4 inch metal can secured to a cage top for a nest facility.
Colonial metal enclosures of 2 x 2 x 3 feet have also
been used (647).
210
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Nutrition
Opossums do well on simple diets such as dog pellets sup-
plemented twice a week with canned dog food and some fresh
fruit or vegetables (645).
4
Other supplements include raw meat scraps, bone meal (644)(
cat chow, and mealworms (642).
Water is supplied ad libitum in most labs.
Breeding/Rearing -
Marsupial reproduction differs from that of other mammals
in one unique way: the young are born after a very short
gestation period (12-13 days for the opossum [643]) and
crawl to a pouch where further development takes place.
There are usually 13 teats in the mothers pouch. This can
be a limiting factor in litter size. The litter average
for 17 females in one colony was 6.7 (644). There are
usually two litters each year (648).
Breeding in captivity is apparently best achieved in large
outdoor pens (642, 645). In some cases, in smaller con-
fines the nest boxes were removed during the mating period.
One lab team removed the young from the mothers pouch and,
in one case, the female produced a second litter of 12
only 16 days later (644).
Special Needs -
Nest boxes are considered necessary (647) as is suitable
bedding changed weekly—straw, shredded paper (642, 643,
644, 645, 649). Litter pans should be changed daily (642)
Behavioral Restrictions -
Some reports of pair incompatibility (647, 649) suggest
social problems in keeping opossums.
Another lab reported a very high mortality rate (to 75
percent) in colonial housing, although a great decrease in
this rate was shown when individuals were caged separately
and sanitary practices strictly enforced. (The high mor-
tality was due mainly to infectious diseases) (642). The
nocturnal habits of opossums may pose problems in their
handling and use.
211
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Ecological Role
The opossum will eat any food, plant or animal. Insects
make up a large portion of their diet during certain seasons
of the year. Other food items include bird eggs, birds,
snails, earthworms and carrion (413). The opossum is a
food source for large raptors and carnivorous mammals (412),
Longevity -
The opossum has been known to live up to four and one-half
years in captivity (646) . Mortality rates in captured in-
dividuals are very high in some cases (642).
Problems with Mass Culture -
Parasites are probably the greatest deterrent to the main-
tenance of large colonies (647, 649, 650, 651, 652, 653,
654). The parasite load is very great in captive indivi-
duals, and the colony must be cleaned up every six months
(645).
There is evidence that litter size decreases in captivity
and that breeding takes place more frequently (644) .
The opossum does best in outdoor enclosures because it need
large spaces. This kind of caging also eliminates the need
to change litter pans daily (642, 643, 644, 655).
Toxicity Testing -
Though little toxicological testing has been done with opos
sums, the ready accessibility of relatively undeveloped
young offers a unique possibility for work with developing
organisms. (See reference 656 for a successful technique
for the removal of "embryos" from the teat and subsequent
handling procedures.)
General Suitability -
Because of the accessibility of its young and its ability
to breed in captivity, the opossum seems well suited for
toxicological testing.
Ecological Alternatives -
Other omnivores.
212
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Edentata
Introduction -
The armadillo is a member of a tropical family (Dasypodidae)
that have degenerate teeth and a horny protective body cov-
ering resembling a plated coat of armor. One species
(Dasypus novemcinctus) has established itself in the central
southern U.S. and is commonly seen at night within its range.
Armadillos feed on the plant and animal material which they
unearth with their specially adapted, well-clawed forefeet.
The average adult is about 16-inches in length and has a
15-inch tail. They weigh from eight to 17 pounds (432).
Caging/Lab Conditions -
A single L-shaped unite consisting of two pens separated by
a removable plywood divider placed diagonally through the
center of a wading pool is described as suitable for arma-
dillos. The first pen in the L-shaped structure is 9 x 4
x 4 feet. The second is 18 x 4 x 4 feet. Fir boards
and plywood were used for the walls and floor and lined with
two or three layers of 44 inch wide fiberglass cloth and
treated with a resin compound and surface curing agent.
(Enclosures of metal or wood proved inadequate because of
injury to the animals or maintenance costs). In the pool,
the bottom was covered with gravel. Cedar shavings were
used as floor bedding (657). Smooth two-foot metal plates
covered with wire netting (658) and tile-covered brick
walls four feet high in an enclosure 26 x 15 x 6 feet
(659) are other cages that have been used to house armadillos,
Nutrition -
Armadillo diets vary, but a combination of animal and vege-
table material is the rule. Canned dog food, raw beef,
milk, eggs, liver and fruits are all desirable components.
One diet consisted of 1 1/2 cups water, 1/2 cup milk, three
hard-boiled eggs, two to three chopped bananas, one chopped
apple, one cup cat chow, one pound ground meat, and three
T multivitamins (657). Other possible additions to the
general diet are tomatoes, cod-liver oil, mink Developer
Chow, molasses, honey and potatoes (660).
Water should always be available.
213
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Care should be taken to provide sufficient vitamin K. K-Sol
powder from Vet-a-mix, Inc., Shenandoah, Indiana is one
possible source (660) .
Breeding/Rearing -
The gestation period of armadillos is difficult to deter-
mine because the implantation of the blastocyst is delayed
for a variable length of time. Polyembryony has also been
reported in armadillos. One to twelve young are born,
usually in the spring (658, 661).
Only a few armadillos have been reared under zoological gar-
den conditions, but the Apelt Armadillo Farm at Comfort,
Texas has bred the nine-handed armadillo in some numbers
(658).
Special Needs -
Armadillos need an abundant floor covering of materials like
wood shavings, straw and sand, or an earth floor (657, §58,
659) Logs and large rocks allow for scratching and diggu;
which alleviates localized pyogenic infection of claws and
footpads (657). Concrete floors are not desirable (658).
Vitamin K is necessary to prevent major blood loss from
minor, superficial lacerations and can be supplied by the
inclusion of K-Sol powder in the diet (see Nutrition).
Armadillos are tropical animals and seem to be adversely
affected by ambient temperatures of less than 50°F (659) ••
A large source of water is recommended (657, 658, 659, 660).
Behavioral Restrictions -
Small wooden cages cause undue stress, and metal enclosures
are inadequate because of injury caused when the armadillo
digs at the sharp corners (657).
Armadillos feed at night in nature and remain nocturnal in
captivity (659). Red lights may be needed in the lab.
#
Ecological Role -
Armadillos are secondary consumers and scavengers. They
consume many insects and other terrestrial invertebrates
(412). Their meat is edible, but predators other than
men have a difficult time overcoming their armor.
214
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Longevity -
Nine-banded armadillos have lived over six years in captiv-
ity. Other species have life spans of 10 to 15 years (658).
Problems with Mass Culture -
Armadillos require special caging and a lot of space (657).
They often injure themselves during shipping and are very
susceptible to pyogenic infection of claws and footpads.
It is difficult to cleanse them thoroughly because of their
dermal plates and they are therefore more susceptible to
parasites.
Toxicity Testing -
Although toxicity studies with armadillos as subjects are
unknown, physiological research has been conducted which
may indicate the suitability of armadillos for such testing.
This research includes general hematological studies (662,
663, 664, 665, 666).
General Suitability -
Depending on the caging facilities available, the armadillo
is more or less suitable for toxicological research. Sur-
gical techniques are seriously hindered by the dermal skele-
ton (659) .
Ecological Alternatives -
Moles
Six-banded armadillo: Euphractus sexcinctus
La Plata three-banded armadillo: Tolypeutes matacus
Bolivian hairy armadillo: Chaetophractus nationi
Small hairy armadillo: Chaetophractus vellerosus
Pichiy: Zaedyus pichiy
215
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Exotic Species
Introduction -
There are a number of exotic species used in research in
the U.S. Some are recent introductions and some have been
used for years. The guinea pig (Cavia) has been used in
medical research because of its immune responses to foreign
proteins and other somewhat unique metabolic characteristics.
Adults weigh between 850 to 1200 gms and are gentle animals
that are very easy to handle. Golden hamsters (M.esocricetusi
and gerbils (Meriones) are closely related to rats and mice
(family Cricetidae) and somewhat similar in size and appear-
ance to smaller rodents, with a body length of about five
inches. The lab rats and mice are of European origin and
are treated elsewhere. Hamsters are natives of temperate
Europe and Asia. There are three varieties used in re-
search, the Golden, European and Chinese hamsters, but the
Golden is the most prominent in research. Gerbils are of
the same size as hamsters and of two varities, African and
Mongolian. The latter is more prominent in research.
Other exotic species that have been utilized as test ani-
mals are the hedgehog, the chinchilla, and the sand rat
(Psammomys obesus). Because of the more extensive use of
hamsters, gerbils and guinea pigs, the following text will
concern itself primarily with these three animals.
Caging/Lab Conditions -
A cage 30 x 24 x 12 inches is ample space for a breeding
pair of guinea pigs and their unweaned offspring. Smaller
enclosures (18 x 12 x 12 inches) are suitable for two
cagemates of any age. Suggested bedding for guinea pigs is
coarse wood shavings or shredded beet pulp. Recommended
temperatures and relative humidity for the animal enclosures
are 68° to 72°F and 45 to 55 percent (289).
Hamster and gerbil breeding pairs and young do well in 20 x
10x6 inch cages made of almost any material. A 24 x 24
x 8 inch cage is adequate space for eight to ten adults.
Bedding of white pine shavings or some commercially avail-
able substance is suggested and should be changed frequently
Other possible bedding material includes corn cobs, beet
pulp, and peat moss about 65° to 75°F and 55 to 65 percent
humidity are very suitable conditions for the animal
enclosure (289, 667). A good rule of thumb for housing
these animals is that 200 square inches of floor space
216
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should be provided for a breeding pair and their litter.
In a communal situation, 15 square inches per individual
is adequate (668).
Nutrition -
A daily source of ascorbic acid (Vitamin C) is required by
guinea pigs, and it is reported that this species has a
high requirement for folic acid, thiamine, arginine and
potassium (669). Commercial chow (35 gms/day) is an
adequate diet for guinea pigs if the above additions are
made (289) .
Adult hamsters need a daily ration of eight to ten gms of
food (which is commercially available), but females should
receive an extra two to four gms during pregnancy and lac-
tation (289). Gerbil requirements are similar. They have
been maintained on a diet of rolled oats, sunflower seeds,
and guinea pig chow supplemented with fresh vegetables (667)
Both hamsters and guinea pigs need a constant supply of
water, but gerbils use very little (668).
Breeding/Rearing -
The gestation period of guinea pigs is 59 to 67 days. Three
to four litters per year, averaging three offspring per
litter, are possible (289). The young can be weaned at 10
days. Sexual maturity occurs at about five months; the
breeding life of guinea pigs lasts for up to five years.
The hamster's gestation period lasts 15 to 18 days, and
the average litter size is about seven young. Females are
breedable immediately following parturition (289). In
eight to ten weeks they are sexually mature and can produce
offspring for about one year.
For gerbils, the gestation period is 24 to 26 days, and the
mean litter size is 4.5. Sexual maturity is reached at
about 85 days (667). Best reproductive results occur when
pairs of adults are kept separately (668).
Special Needs -
Guinea pigs require Vitamin C daily, but this is included
in special guinea pig chow available commercially (669).
217
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Ecological Role -
Guinea pigs, hamsters and gerbils are all herbivores. Guinea
pigs are basically grass eaters; hamsters and gerbils are
seed eaters. All are possible prey species for predators
of many kinds.
Longevity -
Guinea pigs live to about five years. Gerbils and hamsters
live from two to three years in captivity.
Toxicity Testing -
Substances have been tested on all of these species through
food additives, vapor inhalation and topical application
(449, 621, 670, 671, 672, 673, 674, 675, 676). Substances
tested include pesticides (670, 672, 677, 678, 679, 680,
681), heavy metals (673, 674), carcinogens (682, 683) and
gases of various types (626, 684, 685).
General Suitability -
Guinea pigs, hamsters and gerbils are routinely maintained
in laboratories and are very suitable as experimental ani-
mals. A number of qualities make some of them suitable for
toxicity testing. For instance, inbred guinea pigs exhibit
very homogeneous sexual behavior, while heterogeneous ani-
mals show great individual variations (686) making possible
quantitative studies of behavioral responses to toxic sub-
stances. It should be noted that the homogeneity of the
genetic background of these individuals could bias such
testing. Gerbils are 10 .times more sensitive, in terms of
avoidance responses, than lab rats (668) and behavioral
changes may be more discernible in gerbils than in other
lab animals. None of these species, however, is native to
North America; they have been selectively bred to various
extents for use as lab animals. Therefore, they are prob-
ably less suitable than native species in studies which hope
to extrapolate results to wild populations.
Ecological Alternatives -
Voles, mice.
218
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SUGGESTED SPECIES
Synopsis sheets of the suggested species from each animal
subgroup have been prepared for use as quick reference
sources. These sheets can be found in Appendix D. The
information was largely taken from references used in the
text or from the references enumerated in the synopses
themselves. Much general breeding and life history data
was acquired from texts concerned with specific animal
groups in the wild when there was no information available
from laboratory colonies (57, 133, 144, 196, 221, 310, 321,
330, 687, 688, 689, 690). A summary list of suggested
species from terrestrial animal groups is found on Table 23
Final List
A further distillation has been made from the suggested
species for each group in order to identify terrestrial
animals that are best suited for use in laboratory
experiments. These are recommended for use in cases where
ecological role, phylogenetic relationships and habitat
preferences are not of significant importance for certain
types of testing. They represent good, all-around test
animals and are the most likely selections for preliminary
studies. Most are frequently used species in many labs
at the present time. The list of suggested species is
shown below in Table 22.
Table 22. FINAL LIST OF SUGGESTED SPECIES
Gryllus sp.
Drosophila sp.
Musca domestica
Tenebrio sp.
Rana pipiens
Anas platyrhynchos
Columba livia
Phasian'us colchicus
Colinus vifginianus
CoturnTx coturnix
Gallus~gallus
Mus Musculus
Rattus norvegicus
Canis familiaris
Microtus pennsylvanicus
Peromyscus maniculatus.
field cricket
fruit fly
house fly
mealworm
leopard frog
mallard duck
pigeon
ringnecked pheasant
bobwhite quail
Japanese quail
domestic chicken
house mouse - lab
variety
norway rat - lab
variety
domestic dog
meadow vole
deer mouse
219
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TABLE 23. LIST OF SUGGESTED SPECIES FROM SELECTED TERRESTRIAL ANIMAL GROUPS
INVERTEBRATES
Lumbricua terreatr-ia
Helix aeperea
Milax sf.
Araneua diadematua
Cry Hue penneylvanicue
Praying mantids
Blieeua l&uaoptefus
Leafhoppers
Triboliwn sp.
Tenebrio sp.
Carpocapea pomanella
Agrotia ypeilon
Husaa domestiaa
Drosophila me1a.noga.atev
Apis melifera
Isopods
Centipedes
XERPETOFAUNA
Sana pipiens
Ambyetoma mexiaanum
Terrapene sp.
Chelydra eerpsntina
Anolie aarolinenaie
Phrynoeoma aornutum
Thamnophie eirtalie
Elaphe sp.
AVES
Phalaerocorax auritus
Podilymbus podiaepe
Anat platyrhynahoa
Branta canadensis
Puffinus grieeus
Caemerodiue albus
Fuliaa ameriaana
Philohela minor
Larua argentatue
Melanerpee erythroaephalua
Faleo aparverius
Tyto alba
Columba livia
Colinue virginianua
Phaeianua oolohicua
Dendragupus obecurue
Heloepiza melodia
Riohmondena eardinalia
Turdua migratoriua
Sturnue vulgaria
Paaaer domeatioua
CalT.ua gallua
Coturnix coturnix
Helopaittaoua undulatua
MAM'HALS
brevicauda
EpteeiouB fueaue
Sylvilague floridanua
Dip'odomys ordi
Seotoma lepida
Orysomye paluetrie
Spermophilu3 beeoheyi
Eutanias minimue
Tamiae etriatue
Hiorotus pennaylvanicus
Peromyacua maniculatua
Hue m-uaculua
Rattuo norvegicue
Myocaeter coypua
Vulpee vulpea . •
Mustela vieon
Muatela putoriua
Cania familiaria
Miniature swine (Sue)
Ovie sp.
Odoooileus virginianua
Odoooileue hemionue
Saimiri eaiureue
Didelphis mareupialie
Daeypue novemeinctua
220
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Selection of Animal Species for Testing
In toxicity testing, habitat and ecological role are often
considered important in the selection of species. The
Trophic Triangle shown in Figure 22 represents the basic
food chain/ an important factor in determining the eco-
logical role of a potential subject species. Decomposers
are associated with the substrate, which is an accumulation
of organic detritus. At this level, waste products, and
dead plant and animal material are decomposed and their
nutrients made available to the primary producers, the
vegetation. From the plant level, herbivores, omnivores,
secondary and top carnivores, in turn, utilize foods from
their own level or the ones below. This triangle shows the
trophic level and the general habitats of the suggested
species on that level.
Suggested species, their place in the Trophic Triangle, and
preferred habitat are shown in Table 24. This table can be
of particular importance in the selection of species for
use in multispecies test systems. For instance, if it is
desired to assess the impact of a chemical which has con-
taminated a forest, forest species can be selected from
each food chain and animal group (if suitable species from
each are identified).
The following is a hypothetical system constructed in this
fashion.
Top Carnivore
A Possible Multispecies System
Vulpes vulpes
Falco sparverius
Secondary Carnivore Mustela vison
Omnivore
Herbivore
Decomposer
Melanerpes erythrocephalus
praying mantids
Rana pipiens
Didelphis virgenianus
Sturnus vulgaris
Oryzomys palustris
Colinus virginianus
Terrapene
Helix aspersa
Lumbricus~ terrestris
Isopods
Of course, in order to achieve this level of testing much
221
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preliminary work on lab maintenance must be done on many
species. For example, in Table 24, few carnivores are
listed. They are generally large animals and difficult to
feed in captivity, and very few attempts have been made to
use them in any type of testing. Traditionally, omnivores
and herbivores are most often used as test subjects, and
their numbers are greater both in the laboratory and in
nature. See Appendix C for suppliers of some suggested
species. (Considerable changes may have occured in this
list as it dates to 1971.)
It is suggested that testing in the near future utilize the
List of Species, shown on Table 24, but that serious re-
search be initiated to develop other potentially valuable
species, in order that an efficient, ecologically meaning-
ful, multispecies test program can be initiated. The state
of toxicity testing cannot develop as it should without
research of this kind.
224
-------
SECTION V
METHODOLOGIES
INTRODUCTION
Webster defines scientific method as "a systematic procedure,
technique, or mode of inquiry" employed by the scientific
community. It goes on to add that it is a "way, technique,
or process of or for doing something". A standard procedure
and methodology are tools which help the experimenter achieve
reproducible results. In this section, the methodologies
available for use in toxicological testing will be reviewed
and an assessment will be made of the suitability of each
for use in toxicology studies. Previous use and results
obtained, applicability to wild North American animal
species, experts opinions, and expense are the fundamental
criteria used for determining suitability. Suggestions for
further use are often included in the text.
General Scientific Procedure
Toxicological experimental procedures consist of two basic
steps: exposure of the test animal to the test substance
and analysis of the results. Exposure techniques, though
they may seem straightforward to the layman, have an
important effect on results. Some standard, specific
routes of exposure for laboratory research include:
a) injection
1) intravenous (i.v.)
2) intra peritoneal (i.p.)
3) intra muscular (i.m.)
4) into egg yolk (birds)
b) oral exposure
1) blended in food
2) in loaded food items
3) in water
4) intubation (force feed)
c) topical application
1) dermal
2) ocular
3) in bedding
225
-------
d) in air (vaporized)
The route of exposure should be chosen on the basis of the
test animal's size and anatomy, and in accordance with the
manner in which the animal will possibly encounter the
substance under natural conditions. (Dosages are also
discussed under Section III, Special Problems).
The second major step in testing is analysis of the results.
Analytical procedures available to the scientist are
discussed under two catagories: Physical/Chemical and
Behavioral Methodologies.
Selection of Categories
Selection of these two categories was a difficult task.
As the reader will realize, there is considerable over-
lapping between categories; however, the divisions allow
for a more thorough treatment of all possible methods and
also facilitate use of tables which describes the metho-
dologies available (see Table 25). See Figure 23 for
interrelationships of Physical/Chemical methods.
Another problem in organization was that certain metho-
dologies are specific technical procedures (e.g.Chromato-
graphy, Histology) while others are general methods of
scientific experimentation (e.g. Physiology, Population
Dynamics) in which the specific techniques can be used.
Further distinctions on this point can be found in the
text for each category.
Physical/Chemical Tables
There is a natural division between physical and behavioral
test methods. For this reason, and because of the volume of
information, it seemed advisable to separate the literature
into physical and behavioral methodologies. In the case of
stress, an entry has been made in both tables because it
affects both physical and behavioral aspects of animals.
(See Behavioral Methods for further discussion.)
The tabular presentation of methodologies shown in Table 25
has been developed for a number of reasons. First, it
simplifies the presentation of a great number of methodo-
logies, and pinpoints areas of concentrated study by showin?
the amount of information available for each area. It also
suggests which methods are not frequently used for a given
group. References were gathered with toxicological testing
as the primary objective, and the methodologies selected
reflect this.
226
-------
However, the selection was not limited to toxicological
testing, because certain "unrelated" methodologies may
prove quite suitable for such work.
227
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
INVERTEBRATES
Worms
Snails and
Allies
Spiders and
Allies
Insects
H
a
u a
a c
•H o
,-t H
•H H
U
ft
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N
B
a
905
112
63,
64
Sg
s
27,30
34,35,
39,54,
130,336,
691,692,
693
39
695
322,700
General
Arthropods
HERPS
Snakes
Lizards
Alligators
Turtles
Amphibians
BIRDS
Pelican and
Cormorant
Loons and
Grebes
Waterfowl
Shearwaters
and Petrets
Herons and
Ibis
Cranes, Rails,
and Coots
Woodcock
Gulls and
Terns
Woodpeckers
717,
718
176,
177,
711,
712,
713
141 ,
712
141,
744
714,
715
148
148
139,148,
719,720,
721,722
195
199,200,
203,209
211,283,
723
235,236
O to
~H O
•U-l
a a
II
32
50
54,56,696,
697,698,699
35,39,50,72,7:
76,78,84,85,
96,108,115,
123,124,329,
336,701,702,
703,704,705,
706
50,130,707,
708,710
174
174,716
185
193,197,201,
276,233,724,
725,726,727
185,222
38,229,230
185,231
238
228
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
8,
M 0)
e c
•H «
r-l 14
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n
0
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4J
cn
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91
C.
01
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s§
§s
a E
rl O
3 C
O Q
Q.
Raptors
Doves and
Pigeons
Bobwhite Quail
Pheasant
Miscellaneous
Galliformes
Perching
Granivores
Perching
Insectivores
Perching
Omnivores
Domestic
Gallifonnes
Japanese
Quail
Exotics
728
729,
730
302
732,
733
311,
315,
643
358,
359
256
374,
742,
743
394
389
283
283,293
314
199,314
37
199,347,
738,739,
740
283,744,
745
283,293
276,282,283,
286,368,731
282,283,267,
290,291,301,
303,734,735,
736
308,312
38,238,333,
337,737
225,238,339,
741
201,283,368,
369,370,371,
746,747,748,
749,750,751
283
262
MAMMALS
Moles and
Shrews
Bats
Rabbits and
Hares
Desert Rodents
Wild Rats
Ground Squirrels
Chipmunks, Tree
Squirrels
Voles and
Allies
Deermice and
Allies
Lab Mice
and Rats
754
755,
756
757
438
447,448,
452,580,
677,758
331,'
985
348
256,
532,
766,
767,
768,
769,
770
682,
771,
772,
773,
774,
775
64,
393,
776,
777,
778,
779,
780,
781,
782
783
460
447,448
536,540,
545,677,
681 ,758,
760,784,
785,786,
787,788,
789,790,
791,792,
793
88,423,752,753
430,435
445,759,760
493,498,517,
761,762
332,517,763
5,19,445,543,
761,774,794,
800,801,802,
303,804,805,
806,807,808,
809,810,811,
812
229
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
4J -H * On
H 41 -H >i V.U -H O
a a n n o o 4i-H
n no -u >4J a e
M a n -r> o .e g U-H *j 5
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•HH 4JDS Cg
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
Z
g a
g. » "2
5 e c -•
g1
a -H wo o
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O H-HO O -H e O
«J30 O+J
•
a x) u u
a « c -H
INVERTEBRATES O(2o S •» 3 XWinw
Worms 77 29, 49,
77 834,
835
Snails 834 836 46,47,48,
837,838
Spiders and
Allies
Insects 77, 80, 71,83, 67,69, 12, 65, 113 70,862
234, 125, 111, 73,74, 116, 127,
839 126 834, 82,94, 117, 841,
840, 95, 119, 858,
841, 108, 128, 859,
842 131, 849, 860,
741, 850/ 861
843, 851,
844, 852,
845, 853,
846, 854,
847, 855,
848 856,
857
General 54,
Arthropods 147
HERPS
Snakes 863
Lizards 864 863
Alligators
Turtles 863
Amphibians 147, 150, 12, 870, 873,872,
865 155, 154 871 873,874,
866, 875,876
867,
868,
869
BIRDS
Pelican and 185,
Cororaoran t 249,
877
Loons and 185
Grebes
Waterfowl 234, 276 207, 198, 113
276 212, 879
877,
878
Shearwaters 216, 218 217
and Petrels 877
Herons and 185
Ibis
231
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
0)
a
a
n a
>t o o>
X! 4J -H >,
a* a o en
a u 9 o >
n e c 41 -i b>
tji aao-Hcao o
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4J fl) RJ r-4 -HO O*H gO
10 3 01 O -UXI •HhO-4
S 41 c. ti 15 Ł
54) -H O O C OB
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
" a u
g- n >o &
tt B , c 3 ° &!
u* tutau-Hino o
O O >, .H Ort C >i Ł
B 3 § "n "Jj° -S "^ °
fi *O TO .U fl> U ** U 4J n
0-j« •) C-4 «OO>i
^t«O -P O4J CUaJCjC
Rabbits and 452 626, 640 911, 152, 754, 608,917
Hares 909, 912, 914 915, 918,919,
910 913 916 920,921
Desert Rodents
Wild Rats 475 466 477,
479
Ground Squirrels 486 922
Chipmunks, Tree 488,907
Squirrels
Voles and 466 6, 498, 502
Allies 507 762,
923,
924
Deermice and 907
Allies
Lab Mice 619 925 386, 96, 9,116 858, 113,387, 477, 754, 831,848,
and Rats 522, 256, 519, 969, 401,557, 479, 979, 919,920,
541, 393, 538, 961 784,792, 793, 980, 935,946,
926, 617, 805, 914,948, 975, 983, 985,986,
927, 640, 912, 951,952, 976, 982, 988,989,
928, 679, 952, 961,962, 977, 983, 990,99],
929 767, 953, 963,964, 978 984 992,993,
930, 954, 966,967, 995,996,
931, 955, 968,969, 997,998
932, 956, 970,971,
933, 957, 972,973,
934, 958, 974
935, 959
936,
937,
938,
940,
941,
942,
943,
944,
945,
946,
947,
948,
949,
950,
951
Large
Herbivores
Large 625 560,
Carnivores 561
Small 582 999 3000
Carnivores
233
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
I
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
,
81
*-\
B
INVERTEBRATES
Worms
Snails
and Allies
Spiders
and Allies
Insects
General
Arthropods
HERPS
Snakes
Lizards
Alligators
Turtles
Amphibians
BIRDS
Pelicans
and
Cormorants
Loons
and Grebes
Waterfowl
Shearwaters
and Petrels
Herons and
Ibis
Cranes, Rails
and Coots
Woodcock
Gulls and
Terns
Woodpeckers
Raptors
I
a
cu
tn
O
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1032
&
o
o
rH
8
66
172
1035
169,1035
1035
151,873
1038
165
872
186
192,205
205,213
202
188,232,
237
240,247, 202
1045
187,188,
232,400,
1042,1043
217,219
1044
223
227
188,236,
237,243,
244,388,
1046,
]047,
1048,
1049
134,
844,
]Q33
36
36,89.
F61.1034
178,179,1036
1036,1037
1039
]036
140,
1040,
1041
226
235
-------
TABLE 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
Cn
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o
a
n
3
o
Doves and
Pigeons
Bobwhite
Quail
Pheasant
Miscellaneous
Galliformes
251,258,
259,260
323
295
300
257,260,
1050
294,299
1051
1051,1052
1053,1054
Perching
Granivores
Perching
Insectivores
Perching
Oranivores
DomestJr-
Galliformes
Japanese
Quail
328
372
188
1055
351
399
399
355,356,
371,399,
405,1056,
1057,1058,
1059,1060,
1061,1062,
1063,1064
382,389,
392,396,
400,736,
1066,
1067
363,365,
366,1065
Exotics
MAMMALS
Moles and
Shrews
1068,1069
Bats
Rabbits and
Hares
Desert
Rodents
Wildrats
Ground
Squirrels
Chipmunks,
Tree Squirrels
446,449,
450,451,
671,
1070,1071
918,919,
1072
1073
442,831,
1051,1074
3075,1076
236
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TABLE. 25. METHODOLOGY: PHYSIOLOGICAL
(CONTINUED)
o
a
«J
04
&
o
i-4
O
o
0
A
0.
&
O
o
as
Voles and
Allies
Deermice
and Allies
Lab Mice
and Rats
Large
Herbivores
Large
Carnivores
Small
Carnivores
Domestic
C arnivores
Horse and
Allies
Swine and
Allies
Sheep and
Allies
Deer and
Allies
Primates
Opossum
Armadillo
Exotics
387,522,
775,805,
925,
1066,
1078,
1079,
1080
110,369,
780,805
601,621
631,1105
654
449,671,
1115
544, 869,956
775,
831,
1006,
1081,
1082,
1083,
1084,
1085,
1086,
1087
586,1006
634 1110
652,1111
1055,
1136
1077
539,805,
919,1088,
1089,
1090,
1091,
1092,
1100
805,819,
919,1083
608
608,611,
614,615,
1103
608
651,1028,
1112,1113
663,664,
665,666
608,1072,
1113,1117
538,1093
This includes shell thinning,
hatchability, growth rate,
mortality.
See also Oology.
570
1104
641
674
JZ
.U
O
626,685,
1094,1095,
1096
1097
33,1098,1099
33
33,594
604
1106,3107,
1108,1109
635,773,831
655,656,1134
677,831,3135,
3338
8
This includes handling, immobilizing,
application techniques, methods of capture,
techniques for assessment, c3ean-up procedure
and instrumentation.
237
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233
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PHYSICAL/CHEMICAL METHODS
Ciliary Transport
The structure and function of cilia are generally consistent.
They are usually engaged in the transport of minute part-
icles or secretions. For instance, in the respiratory tract
of higher animal groups, the cilia transport mucous which
serves as a lubricant and a natural defense mechanism.
Studies of cilia involve a determination of the rate of
flow in "normal" tissue and of the changes that may occur
under the influence of an irritant, e.g. gas. Both in vivo
and in vitro experiments have been devised (359, 765)and
model systems have been developed for mucous flow and
ciliary activity (764). The procedures used are either
surgical, to view in vivo tissue, or extration of epithelial
tissue for an in_ vitro analysis. These studies yield
reliable measures of the effects of gaseous toxicants or
"caustic" liquid.
In previous work (358, 765, 818) chickens and domestic lab
mammals have been used, but because cilia are found in
various systems in many animals, this method could con-
ceivably be utilized on other forms.
Evaluation - Changes in ciliar transport mechanisms can
directly affect necessary functions of related systems
(immunology, food transport) and, if they can be correlated
with reduced vitality, increased illness, reduction in
growth rates, etc. they can be considered significant.
Stress
In a sense, every.environmental or physiological alteration
is a stress. Physiological systems (e.g. adrenocortico -
thyroid system in mammals) have been developed by animals
to cope with stress, modifying physiological functions by
feedback in order to cope with the change (770). They can
be affected by many factors (including toxic chemicals) and
the result is often a "less fit" organism. It is this
reasoning that has led some researchers to test the effects
of stress on subjects previously exposed to a toxic substance.
The effects of stress on exposed animals when compared to
controls is a valid measure of toxicity.
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An advantage of stress testing is that it gives results
which can be extrapolated to a population in nature exposed
to changes in temperature, food availability, day length,
disturbances, social stresses, molting, migration patterns,
etc. (256). Stress testing does not require expensive
equipment nor advanced technology.
Stress testing has used starvation on exposed lab rats (768),
(Peromyscus maniculatus) (348),(Coturnix) (394) , chickens
(374, 742, 743), and pheasants (302). Temperature stress
has also been used (694, 767, 769). These two methods are
the most frequently employed; however, isolation stress
(532), revolving drum stress (766) , and the introduction of
pathogens to exposed subjects (195) have also been used.
The possibilities are innumerable.
Evaluation - Stress testing offers a look at the effects of
a toxic substance on animals under conditions that might be
met in nature. It is applicable to all animals that can be
maintained in the lab. (See Behavioral Methods - Stress).
(For a review of stress - physiology see reference 770).
Respiration
In respiration experiments where study of ciliar transport
is not the objective, the study generally focuses on the
toxicity of gaseous substances measured by physiological
and anatomical parameters. Vapors are usually introduced
into a closed chamber containing the test subject by passing
a controlled amount of air over the emitting liquid.
Periodic sampling/monitoring of the inner atmosphere veri-
fies the gas concentration. Apparatuses are all of the same
basic plan (626, 773) , but highly complex equipment has been
designed for specific uses (such as the study of turtle
respiration - described in reference 718). Long-term,
repeated, or continuous exposure is used with less toxic
substances or with those usually encountered in low con-
centrations (771, 772, 773).
Postexposure analysis can include examination of the gross
anatomy of lungs and other body organs (682, 773), histo-
pathologic scrutiny of the respiratory tract (773), serum
and liver enzyme analyses (773), and observation of
behavioral changes (626, 682).
Previous subjects in respiration experimentation have been
typical lab species—mice, rats, guinea pigs, rabbits, dogs/
goats, pigs, monkeys—but the technique should prove easily
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adaptable to all forms of animal life, even those organisms
which respire largely through their outer integument (i.e.
Helminths).
The methods described here are limited to the testing of
gaseous or airborn substances. In the case of particulate
matter, mists or aerosols can be used. Determination of
the concentration of the test substance in the air is made
difficult when particles of varying sizes occur in the
test atmosphere. Particles of unit density are considered
in these instances. (A caution concerned with interpre-
tation of results - gases and particulate matter can be
inhaled and swallowed as well.) (803).
Evaluation - Respiration experiment techniques are an
excellent route of exposure and can be used to dose animals
for further physical and behavioral tests.
Enzymes
Enzymes are organic catalysts which, like their inorganic
counterparts, function in "the lowering of the activation
energy of a reaction (resulting in) a faster attainment of
the equilibrium point. Presumably all reactions of any
degree of complexity occurring in living cells are catalyzed
by a specific enzyme. Thus the integrity of the enzyme
population of a cell or organism is paramount to its well
being" (1119). Enzymes occur in most living tissue, but
those of the liver have received special study because most
metabolism of ingested materials takes place in this
organ (393, 778).
Almost any enzyme level can be determined in any tissue if
its activity can be measured under controlled conditions.
Review articles on liver microsomal enzymes are available
(777, 780) and entire books have been written on enzyme
methodology. However, biochemical procedures are often
exacting and require an experienced technician. Likewise,
the necessary equipment is expensive. While the results
of enzyme testing give an exact measure of the direct
effect of the test substance, interpreting the indirect
effects may be more difficult. For instance, the stimulation
of an enzyme may cause biochemical side effects that are not
directly related to the test substance. Enzyme inhibition
can do the same. In order to evaluate the activity of an
enzyme and determine the effect of any change in concen-
tration, it is necessary to know the activity of its
products and their subsequent reactions.
241
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Evaluation - Enzyme studies can show variations in levels
and activity, but their real value can only be realized
when it is possible to correlate enzyme changes with changes
in vital metabolic processes.
Anesthesia
The administration of anesthesia to animals can be risky,
especially when little is known about the effects of an
anesthetic on a relatively new species of animal in the lab.
The effects of an anesthesia might change as a result of the
subjects exposure to a toxic substance.
Evaluation - Anesthesia/narcosis can be a good test
method in itself when used as a stress but unless the
compound used is potentially an environmental contaminant
the results would not be directly applicable to natural
stresses. However, reduced tolerance induced by a toxic
substance to any anesthetic may be an indication of reduced
tolerance to other man-made chemicals possibly found in
nature.
Telemetry
The monitoring of radio tagged animals has been used for
some years by wildlife researchers to determine the movements
and body temperatures of free ranging individuals. Grouse
(311, 315), pheasants (732, 733), ducks (825), and large
mammals (824, 826) have been studied in this way.
Evaluation - Radio work can be done with many species, but
its applicability to toxicity testing is unknown because
of the lack of control in open fields. The success at
monitoring body temperature (813, 824) suggests that heart
rate and respiration could also be monitored with radio
equipment. Equipment for large scale programs will be
rather costly; single transmitters can cost $500 to $1000
and receivers, depending on the size and capabilities,
cost upwards of $500.
Observe/Monitor
This general category includes the simple observation of
exposed animals, for a dosage - mortality correlation,
"control" studies in which a toxic substance was tested for
its destructive ability on a given species or larger group I
of animals, residue analyses (seperately listed in the table5
for quick reference), and studies which report general
changes in an animals appearance or activity after
242
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exposure to drugs or other substances. (For studies on
specific behavioral aspects see Behavioral Methods).
General population counts on invertebrates (336), and soil
invertebrates in particular(1047,130,54), have been conducted
Earthworm populations have been monitored in many studies of
toxic substances (30, 34, 35, 37, 692). Other terrestrial
species that have received attention are frogs (721), snakes
(148), pheasants and Japanese quail (293).,. and monkeys
(831). Most of these investigations have been performed
under lab conditions.
Reports of the general appearance of exposed creatures are
limited in number, but external lesions and other
morphological changes have been reported in frogs (139,
1120).
Of special note are the experiments in which brain wave
patterns have been routinely monitored by Iowa State
researchers by using surgically implanted electrodes (628).
This procedure is used with primates as well, and could be
applied to many larger animals. It may be more widely
used than we have been able to detect.
Evaluation - Observation of mortality rates and general
activity of the subject, as well as the palatability of test
substances, is an excellent method for screening potential
compounds. Survival studies are essential for determining
the gross effects of exposure on animals. However, more
detailed biochemical tests, such as enzyme analysis, and
histopathological examination of subjects are required to
elucidate mechanisms of action and the direct causes of
mortality. Behavioral testing is recommended for sublethal
poisoning.
Population Dynamics
Population dynamics is the study of numbers of organisms,
their reproductivity, growth, and survival. It is concerned
with the factors that control the abundance and the
distribution of a species. The effects of toxic chemicals
on the reproductivity of animal population is thought by
many researchers to be a key point of concern in population
studies (20). There has been a large amount of work on
population dynamics (241). Such research can conceivably
be performed with any species desired. Aspects of such
research include fertilization, embryo viability, egg
hatchability, growth rates, and overall survival. Experiments
are designed around the life history and reproductive modes
of the test animal. For instance, birds are particularly
suited for embryological studies in which the egg yolk is
injected with the test substance, a route of exposure
difficult to employ on mammals.
243
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A basic protocol consists of dosing the test animal and then
scrutinizing parameters that may affect reproduction or
reproductivety itself. There are acute studies in which
immediate mortality is a sure measure, and chronic studies
which are concerned with more subtle, long-term effects
due to single or repeated exposures.
While Population Dynamics is closely related to many other
methods (Observe/Monitor, Embryology, Oology) and uses many
procedures from other fields,it is distinguished here to
emphasize its importance. Viable populations are ecological];
important for genetic, social and often economic reasons,
and though the test animals are individuals, it is the
integrity and viability of the entire gene pool that
determines the fate of a species.
Evaluation - Population studies can be performed on entire
populations, or on a few individuals from whom extrapolations
can be made. The methods deal with effects on whole
organisms or populations and are not directly concerned with
individual effects or mechanisms. Population studies are
recommended as screening methods to assess overall effects,
but must be followed by more exact tests if the goal is to
determine mechanisms and causes.
Chromatography
Chromatography is the set of methods used by chemists to
identify organic substances by color and by their positions
relative to each other on chromatographs. Chromatography
is based on the theory that solutes, such as amino acids,
will tend to be distributed between two immiscible liquids
according to their relative solubility in each. There
are many ways of creating this separation. For example,
gas Chromatography vaporizes the mixture by heating it
and blows it over a column of material with a high melting
point. The molecules of the mixture will partition
themselves between the gas and liquid according to their
relative solubilities in the two phases.
Chromatography can determine the presence of chemicals
tested for and a quantitative result can be obtained from
some types of procedures. This method is very useful in
metabolism, residue, and degradation studies where data on
chemical presence and integrity are desired. It is a
standard procedure in many biochemical laboratores, and
exact methods are available in most tests on lab methods.
244
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Evaluation - By itself, chromatography is not a very useful
technique for toxicological testing but used as part of
larger scale testing, such as metabolism studies
(b91, 619) and residue analyses (~J11 220, 276) ^ is a
most useful tool.
Residues
Residue analysis is simply the determination of the concen-
tration of toxic substances in a given place at a given time.
It can apply to soils, plants, animals, or atmosphere - any
part of the environment. However, the analysis of animal
tissues is of specific concern here.
Animal tissues are analyzed for residues through the use
of basic laboratory techniques. Extraction - clean up -
gas liquid chromatography is an oft-used flow in such studies
(29). The results can give a quantitative measure of the
test substance if conditions are right. The significance
of the residue or its effects on the animal is another
problem and not as easily determined. Bioassay, as used
in this report, is similar to residue analysis, but is
concerned with the relative strength of a substance and
its gross effect on the test organism. Residue Analysis,
Observe/Monitor (LD5Q) and Assay methods are interrelated.
Evaluation - Residues in themselves do not show any adverse
effects on the animal. They must be correlated with their
effects on the subject species.
Bioassay
An assay is a chemical analysis to determine the presence,
absence, or quantity of one or more components of a substance.
In the sense it is used here it is practically synonymous
with residues, but is concerned more with the relative
strength of a substance in specific body tissues and the
effect of that substance on the host tissue. This is
usually achieved by measuring the activity of the substance
or related enzyme. (910, 927). Assay is used routinely
in drug research and methods applied in that field are
frequently suitable for toxicological testing.
Evaluation - As with the presence of residues, presence
of a toxic chemical does not give any indication of its
effects on the host animal and therefore is of use only if
levels can be correlated with decreased survival, reproductive
success, behavioral changes, etc.
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Metabolism
Metabolism research is concerned with the study of the way
living systems handle substances^ It is a very broad
biochemical category that analyzes the actual breakdown,
storage, re-use, and clearance of test substances and their
effects on metobolic pathways, including enzyme inhibition
and altered drug metabolism (941, 947).
Radio-isotope labelling is used in many metobolic studies,
particularly those in which the toxic substance is an
organic compound and specific pathways are of interest.
In this test the products resulting from the chemical
breakdown in the system can be located and analyzed.
Metabolism studies are closely related to many categories
in this section. (See Figure 23, The Interrelationship
of Methodologies.) In these investigations, the chemical
fate of toxic substances and their effects on the life
processes of test organisms are the primary objectives and
they often explain the gross effects readily visible by
general observation. Metabolism studies are at the bio-
chemical level and can be performed on all animals.
Evaluation - Metabolic studies are important in toxicity
testing only if alterations at the biochemical level can
be correlated with a reduction in viability of the test
animals. For instance, interference with certain metabolic
pathways can reduce the ability of animals to utilize food
efficiently, defend itself against disease, etc.
Genetics
Genetics is a broad subject which deals with, among other
things, the transfer of information from one generation
to the next. Therefore, embryology, reproductivity,
mutagenicity, and resistance are related to heredity. In
fact, because all characteristics of living systems are
more or less expressions of genetic material, this category
should be the largest. However, we have generally limited
it to those studies mentioned above.
The references under "Genetics" are primarily concerned
with long-term reproductivity (822, 958) carcinogenic studies
(1031, 1081) and resistance (see below). Probably because
so little is known concerning specific genes in most
animals, much of the work of this nature is restricted to
Drosophila (117, 119, 849).
24fi
-------
Resistance to an environmentally selective factor like
a pesticide has been reported in insects (12, 128, 854,
855, 856, 857),frogs (12, 154), and mice (6, 9, 507,
519), and presents a particular problem in the testing
of toxic chemicals (see Special Problems).
The reproductivity of species has received a rather large
amount of interest because many environmental toxicologists
believe it to be one of the best ways to study the overall
effect of toxic substances on animal life (see Population
Dynamics). Some of the specific methods involved in
these studies are included above.
Evaluation - Mutagenic effects on animals as a result of
exposure are considered undesirable, often affecting its
survival. Genetic studies are limited to a few species
at present (e.g. Drosophila, mice, frogs) and may be
difficult to perform on other species less genetically
known.
Anticholinesterases
Acetylcholine is a basic impulse transmitter between
nerve cells outside the central nervous system. The
enzyme which destroys the transmitter is a cholinesterase.
The action of many organic insecticides is in effect to
create anticholinesterases which restrict the enzyme and
may cause muscular fasciculations in the affected animal.
Cholinesterase is also present in red blood cells, which
are more easily monitored than nerve endings.
Purification of the insect cholinesterase for in vitro
experiments is available (860). Anticholinesterases is
separated from enzymes because of the work with neurotoxic
insecticides.
Evaluation - Anticholinesterases can be routinely tested
on all mammals and birds. A cross section of animals should
be used because of variations among species. Though a
very specific test, it is valuable and can show direct
effects on animal survival.
Hepatic Studies
The vertebrate liver has received much attention in
toxicological testing because it serves as a "detoxification
center" where a variety of injurious chemical compounds
are converted by liver enzymes or where vital enzyme
systems such as carbohydrate metabolism can be adversely
affected by toxic substances.
247
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Evaluation - The liver is an excellent organ for the study
of enzyme action and histopathology.
Endocrinology
Traditionally, endocrinology has been the study of the
ductless glands and the physiological adjustments that
their special products facilitate. Because the glands
themselves, and their products can be affected by the
introduction of foreign substances into the system, they
have received some attention- by pharmacologists who have
been involved in studying the effects of drugs (978). Contro
programs of many types have used synthetic estrogens
(e.cr. diethylstilbestrol) as possible control agents (477
479, 1000), and toxicologists have monitored endocrine
levels and gland size to determine the effects of toxic
substances on rats (977).
Lab methods are based on blood chemistry in which the
amount of a specific endocrine is determined in order to
compare the measurement to a standard. Organ weights and
size can also be compared.
The endocrine system is regulated by many factors, among
which the autonomic nervous system has the primary role.
All types of stresses-(drugs, tension, sickness) can affect
the production of endocrines. Certain endocrine levels
are also affected by circadian rhythms, such as breeding
cycles, hibernation, etc. These natural and "unnatural"
variations must be considered in endocrinology work.
Background endocrine information is available for many
mammalian species (762, 923, 924).
Evaluation - The endocrine system is very important in
all animals. Breeding, molting, hibernation, activity,
are a few functions mediated by endocrines. Any test which
shows alterations in levels of endocrines, especially those
related to breeding (e.g. FSH, estrogen,) due to exposure
to toxic chemicals is of value.
Anatomy
This category includes the general anatomical and morphologic'
work in which gross (e.g. hyperplasia-hypoplasia) or
cellular changes have been reported after exposure to a
test substance. The studies included here often overlap
other areas. To avoid redundancy, anatomy is restricted
here to include only those papers dealing mainly with
anatomical features and general subjects that can not be
248
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included under a different methodology. Rodent muscular
efficiency (982), necropsy (585, 863, 984) uterotrophy
(983), heart cells (980), bone marrow (979) , and pulmonary
changes (754) have all been studied in conjunction with
exposure to a wide range of toxic substances. General
nervous system changes (981), and kidney lesions (915)
have been reported after exposure to toxic chemicals in
rats and rabbits.
Taxonomists have developed comparative methods of analysis
(e.g. morphometric, 118) which could prove applicable
to the comparisons of exposed vs. control subjects in
toxicological testing.
Evaluation - General anatomical studies can show changes
due to exposure which are unimportant to a species: for
example, hyperplasia-hypoplasia, or microscopic alterations
in tissue organization (see Histology). However, if survival
or longtime reproductivity are affected by the changes,
then anatomical studies can be valuable in determining
the effects of toxic chemicals.. General anatomical analysis
is part of post mortem work and should be included in that
phase of testing.
Physiology
Physiology is the study of function, whether it be on the
cellular or organ level, in any part of an animal's body.
This very broad category includes the toxicology of the
nervous system (48,872,991,992,1011), circulatory system
(920, 921, 1018), respiratory system (685), muscles (629),
auditory system (1008, 1012), renal system (587, 1084),
general studies (70, 596, 1006), and many more.
The methods used are as varied as the subjects included
under Physiology and there are literally shelves of texts
and lab manuals available which deal with all aspects of the
science. (The major reference is Handbook of Physiology
American Physiological Society 1959, 8 Vols.)
Evaluation - See specific methods for further information
and recommendations.
Histology
The study of tissues, their structure and organization,
is called histology. It overlaps anatomy, physiology,
morphology and other fields, but is strictly defined as
the microscopic studies of tissue arrangements and form. In
this report, histology includes most tissues studies,
249
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including carcinogenic work. Methods employed are microscopy
as well as gross inspection of tissue reactions. (Technical
help in tissue preparation may be necessary for microscopy).
Dermal reactions to toxic substances can be performed on
many animals(449,450,1070,1071,1102). The method is a simple
topical application and succeeding analysis of the exposed
tissue. (A review of the method is given in reference
671) .
Almost all animal tissues can be analyzed for the effects
of toxic substances if a route of exposure is available.
Inhalation studies of the respiratory tract (775, 1080)
have been discussed under respiration. I.V. and I.P.
injections and oral dosing can indirectly expose inner
organs (kidney, 446; muscle,1101;; nervous tissue, 66;
liver, 387, . see Hepatic Studies).
Evaluation - Histology can be effectively used in the
analysis of effects of exposure to toxic chemicals on all
animal tissues. Comparisons of dosed animals to controls
or standards will give some indication of direct effects
on the anatomy of test animals. An evaluation of what
these alterations mean in terms of function, survival
etc. should then be made. (Major reference: Bloom, W.
and Fawcett, D.W. 1968 A Testbook of Histology, 9th ed.
Saunders)
Pathology
Pathology is the study of disease and the structural and
functional changes caused by disease. Study" of "this sort
can utilize methods such as histology and physiology.
Disease can be used as a stress to test the effects of a
substance on affected individuals. Exposed animals can
be treated with a pathogen (195) . Standard microbiological
methods can be obtained from a great number of lab tests,
though their applicability to toxicity testing is limited.
Most of the references under Pathology do not deal directly
with toxicological testing, but the information on diseases
has potential use in dealing with maintenance problems or
in selecting stressors.
Evaluation - The use of disease as a stress is quite suitable
for toxicity testing. It is a natural stress, one that must
frequently be confronted by a species in natural environments-
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Pharmacology
Pharmacology is the science of drug action. Many methods
used in this field are equally applicable to toxicology
because both deal with foreign substances that animals
must cope with. Drug testing performed by the drug
manufacturer is expected to result in knowledge of the drugs
action as well as its safety for animal use. Toxicological
testing must do the same. Drugs can also be used as stressors
in toxicity testing or in studies of their synergistic
effects if there is a chance the two could be used together.
A review of methods for the use of primates in pharmacological
and toxicological tests is available (1110).
Evaluation - The use of drugs as stressors is recommended
only in cases where the animals would be exposed to the
drug, such as domestic species that may receive veterinary
care.
Hematology
Blood is the aqueous suspension of cells that is the
circulating fluid in most animals. It is a very remarkable
body tissue that functions as a medium for the transport
of nutrients, as a regulator of metabolism, as nrotective
tissue, etc. Its elaborate composition is too complex
to describe. It differs from species to species,
often remarkably.
Blood is easily obtained from most animals without requiring
their death and techniques for bleeding mice (1091) , swine
(615) , mink (1100)„• and other animals (1072) are published.
Basic blood chemistry is known for most mammals and some
other vertebrates (611, 651, 663, 664, 1038,. 1088).
Total serum protein, urea nitrogen levels, glucose levels
and cholesterol levels have been a'ffected by food additives
in mice (539). Anticholinesterase changes are determined
from blood levels (608) . Other parameters that can be
measured routinely are hematocrit, leucocyte levels, enzyme
levels, and hemoglobin activity.
Evaluation - Blood studies are efficient and valuable
because of the many meaningful measurements that can be
taken without sacrificing the test animal. Though larger
species (e.g. nutria, sheep, galliformes, turtles, ducks)
are recommended, tests on smaller species are possible,
and insect hematology is fast accumulating background
information that can prove useful for comparison.
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Oology
There are two terrestrial animal groups that produce large
eggs which lend themselves to experimentation: birds
and reptiles. Their eggs are a unique vehicle for
developmental and embryonic study. In this report Oology
has been separated from Embryology to accentuate and more
suitably treat the toxicity studies, all avian, which are
concerned with eaa production (399, 405) , egg shell
characteristics (392, 1063), egg viability and hatchability.
(194,400,1056,1067) , and ovulation lag (736). Actual
development of the embryo and unique responses to chemical
exposures are treated in Embryology.
There are basically two approaches that can be taken in
Oology - Embryology research. The adult organism
can be treated and reproductive/developmental success
monitored, or the embryo itself can be dosed. The second
approach is simple in oviparous groups (See Embryology).
Parental exposure can be accomplished by a variety of
routes (see introduction to Methodologies). Correlations
between brain residue of female parents and egg production
have been reported (396) and ovulation lag in Japanese
quail is reviewed in reference (1107).
Eggshell thinning in larger birds, has been reported since
the early 1960's and been given much attention. A good
review of that phenomenon can be found in reference 1D64.
Evaluation - Oological studies of toxic chemicals are very
important because of the direct effect on reproductivity
that can be estimated from the results. It is limited to
oviparous species at present.
Embryology
The study of developmental biology has utilized frogs'
aggs and birds' eggs in a great many ingenious fashions.
Researchers have studied pesticide distribution in exposed
embryos (1057) , teratogenic effects (226, 363, 641, 1073,1093),
fine structural changes (365), and tissue residues (1062)
Plancental transfer of Cl4 labeled Parathion in sheep
(1104)and eggshell thickness related to embryological
development (1063) are other previous investigations.
Exposure of the embryo is accomplished in bird studies by
injecting the yolk sac via the air sac (355, 356). The
direct exposure, quantification of dose, and quality
control make this method scientifically desirable.
252
-------
Some invertebrate eggs may be used in much the same way as
bird eggs to perform statistically valid experiments.
However, the route of exposure must be different from that
used with bird eggs due to the small size of the eggs.
Topical application (844, 1033),. and exposure of adults
(114), are possible routes.
Evaluation - Embryos and their development are directly
related to the survival and reproductive potential of
a species. These studies are recommended for all potentially
exposed animal species.
Other
Under this heading are included techniques for handling and
testing animals, such as restraints (442, 604), drug
immobilization (33, 655, 1098, 1106, 1107, 1108, 1109), and
hypnosis (1074). Special apparatuses for use in metabolic
cages (1096), for dusting insects (1034), for feeding (656,
831), and for measuring activity (1094, 1097) have been used
for experimental purposes. More sophisticated devices
available include electroantennegrams for insects (89),
body capacitor - olfactometers for small rodents (1075,
1076), and a respiratory apparatus (773) (see Respiration).
Clean-up procedures are necessary for laboratory work
such as chromatography and the literature has much information
available for specific analytical methods (e.g. 1095).
This listing is a general one, because detailed
techniques and methods are quite varied, are often restricted
to specific types of analytical procedures, and may be valid
only for a given species.
253
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BEHAVIORAL METHODS
Introduction
The central premise upon which behavioral toxicology rests
is that an animal's reactions to enviornmental stimuli
(activity, habits, instinct, learning) can be affected by
alterations in its internal environment and that these
changes can be measured, thus directly indicating levels
of toxicity. This branch of toxicology is important
because changes in behavioral patterns affect an animal's
interrelationships with its environment, thereby affecting
its ability .to perform tasks necessary for survival. For
instance, a reduced reaction rate in avoidance responses
indicates a loss of awareness which would make the indivi-
dual less capable of escape from a predator. Reduced
awareness and chance of survival go hand in hand in a
natural environment. Behavioral testing is one method of
measuring subacute toxicity which can be directly related
to an animal's overall fitness. A summary of references
available on various behavioral methodologies is found in
Table 26.
Repellents
In this discussion, a repellent means any chemical compound
that protects a potential food source; it can cause side
effects such as fright, convulsions, decreased productivity,
sterility and death. Known concentrations of repellents
can be field sprayed in solid or liquid form to protect
plants from predators such as slugs (1121) and deer (1182).
Sprayed plots were noticeably more productive than control
plots. Grain baits were used on rodents to measure
repellent effectiveness (508, 509, 1147) or prevent preg-
nancy (478). Treated seed, either fed directly or grown
and then fed, has been used with numerous birds to determine
its potential for reducing pest reproductivity (266).
Encapsulated rodent baits were fed to rats as sterilants
(1146) and to domestic farm animals to measure the dangers
of oral exposure (1177). A toxic, fright-producing chemical
was sprayed on plots in fields and kept blackbirds away
from the whole field (349). With the exception of 349,
these repellents are not species-specific and could
probably be used on invertebrates and vertebrates alike.
Repellents can be used as a behavioral test, comparing the
reactions to those of dosed and control animals.
254
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Evaluation - The use of repellents to measure behavioral
reactions altered by a second chemical is quite feasible for
all species. They can also be used to test avoidance re-
action to evaluate the sensitivity of a test species to a
hazard.
Discrimination
Discrimination is the ability to distinguish one thing from
another by means of one or more of the five senses. Most
laboratory studies have dealt with visual and olfactory
cues.
Olfactory tests done with rodents have demonstrated the
importance of odor in food detection and palatability
(1150, 1151).
Newts can discriminate visually between dosed tadpoles and
controls, attacking the dosed animals far more frequently
(1133). This type of test could be done with any predatory
animal that hunts by sight.
i
Bobwhite quail have been trained in a Skinner box, fed a
toxic substance for a few weeks, and then retested. Their
performance was much lower in the retests (280). This type
of testing could be done on all .animals.
Pigeons (270) and opossums (1186)'have been tested for
visual acuity but this type of test could be done on any
vertebrate species and could readily be adapted to toxicity
testing. For a diagram of the apparatus used see Figure 24.
Behavioral tests of depth perception have been done using
a visual cliff, with young cornish hen chicks (1139) and
first generation offspring of pheasants fed a toxic sub-
stance (734). Movements of the young chicks when placed
in the middle of the visual cliff were recorded. In both
cases, movements were considered partially a produce of
the chick's previous environmentt but the offspring of the
dosed parents differed significantly in performances from
the controls. For a diagram of the apparatus used in these
tests, see Figure 25.
Primates have been tested for visual discrimination and
were rewarded when they made a correct choice (630). For
an idea of the chamber used, see Figures 26 and 27..
A general discussion of chemoreception in gastropods may
also be classified under the category of discrimination
(1122).
262
-------
KEV
PHOTOCELL
BEAMS
Figure 24. Diagram of visual acuity chamber
A partition divides the chamber into two alleys, entrances
to which are marked by hurdles mounted two inches above the
floor. Photocells, illuminated by a white 28V. 0.1-ampere
lamp, record entrances into the alleys. The plastic observ-
ing key and the glass stimulus keys are one inch (2.5 cm)
in diameter and mounted approximately 10 inches (25 cm)
above the floor.
Source: (270) Copyright 1971 by the Society for the
Experimental Analysis of Behavior, Inc.
263
-------
-It-
I-..-—I
Fiqrure 25. Cutaway of a visual cliff
used as a test apparatus.
Source: (1139) Copyright 1964 by the American Psychological
Association. Reprinted by permission.
264
-------
-
Figure 26. Discriminatory device used in primate
testing
Figure 27. Inside of visual discrimination cham-
ber for primate testing
Source: (630)
265
-------
Evaluation - Discrimination tests, comparing control and
dosed organisms, may be directly extrapolated to species
in natural environments. The "less aware" individual, for
example, is more susceptible to predation and other envir-
onmental hazards.
Stress
Included under the general category of stress are such
things as infections, intoxicants, trauma, nervous strain,
heat, cold, muscular fatigue and irradiations. In most
stress studies, a toxic substance is fed, the animals are
subjected to an additional stress, and then survial rate
(348, 394, 742, 743, 1178), reproductive rate (112, 348)
and tissue levels of the substance (374, 743, 768) were
studied.. Painful or emotional stimuli have been used to
study renal response in large mammals (823). Isolation
stress has been studied in lab mice (770). Temperature
gradients have been used to study growth in snails (694).
Mallards have been fed toxic substances and then exposed to
a virus (195). They were less resistant than the control
birds. Rats have been drugged and exposed to cold and
other stresses. Drug action and metabolism were affected
(767) and drug tolerance levels were lowered (766).
Hibernation has been tested as a .stress factor. General
field studies have been done on skunks (1140, 1175), and
lab observations on using the bat and hedgehog (1141). It
has been found that hibernation effects oxygen consumption
and temperature regulation in ground squirrels (1149) and
hamsters (1187). The transition periods at the beginning
and end of hibernation have been studied in pocket mice
(457), ground squirrels and hamsters (482).
^valuation - Animals are stressed in their natural environ-
ments by many factors, for example, disease, temperature,
food shortages, etc. (See Physical/Chemical Methods -
Stress) and their adaptations for coping with seasonal and
daily stresses are of physiological importance. A test
which can compare the reactions of control and dosed organ-
isms to stress situations might be extrapolated to quantify
the effect of toxic substances on the reactions of wild
population. For instance, hibernation occurs in many
mammals (e.g. daily torpor in shrews) and, a few birds, and
diapause occurs in insects. These natural occurrahces are
necessary for the survival of many species and alternations
in this behavior will affect the viability of the organism
during a period of stress. Isolation, disease, and fatigue
are stresses directly applicable to natural situations and
266
-------
are suggested parameters for testing a species reaction to
stress after exposure to toxic chemicals.
Pheromones
Pheromones are chemical compounds produced by animals to
attract mates and communicate sexual receptiveness. They
are reported to occur in many groups of animals, but are
best known in insects. Since their discovery, they have
been used as a means of controlling and eliminating insect
pest species. Methods of extraction, purification, and use
as masks for attractants have been studied in the cockroach
(1125) and many Lepidopterans (91, 1130). A general
discussion of the physical structure, commercial use, and
technology of pheromones is given in reference 1129. The
chemical basis of insect sociality (1128) and the use of
moths (1126) have been studied. The Red-Banded leaf roller
has been used to determine the structural features necessary
for a compound to elicit synergistic or inhibitory activity
(1127). The purification and evaluation of synthetic
attractants for insects has also been studied (1131). Snail
trails as cueing devices have also been studied (1123).
Pheromones might be used in mammalian studies, as mammals
are known to use olfactory cueing.
Evaluation - Pheromones are effective means of insect
control, can be combined with toxic chemicals (1129) and
could be used as a method of collection of insects for lab
use. It is possible to use pheromones in behavioral testing
in which the reactions of dosed animals and control animals
to pheromones is compared.
Neurophysiology
Neurophysiological studies encompass taste reception and
auditory studies, drugs, induced convulsions, EEG's and
EMG's.
Taste reception studies range from analysis of the potentially
important consequences of taste reception (599) to mammalian
species differences in taste sensitivity (1143, 1144), the
electroneural response spectrums of taste fibers in the
domestic cat (1011), electrophysiological studies in the
ungulate (589, 1178), dog (1176), bat and opossum (437), and
electrophysiological studies of synergistic effects of taste
reception in the dog (1009).
267
-------
The auditory responses of sheep have been measured before
and after dosing with a toxic substance (627). Cats have
been given toxic substances in an effort to inhibit cochlea
potential response (1012) and dogs have been used in
similar studies (1007).
Mice have been raised on different types of bedding and
then administered a convulsion producing drug. • It was
found that drug thresholds correlated with the type of
bedding used (527).
Sheep have been given toxic substances orally followed by
EEC's. Gross behavioral observations were also made (628).
Although mammals have been the principal subjects used in
neurophysiological studies, birds, herptiles, and some of
the larger invertebrates could be used. The limiting
factor for invertebrates is the lack of appropriate
electrophysiological equipment.
Evaluation - EEC and other electrophysica.1 methods are
excellent techniques for recognizing changes due to
exposure to toxic substances. However, data on mortality,
impaired functioning, and altered reproduction potentials
or behavior are necessary before altered electrical
responses can be correlated with harm or potential
detriment to the test species.
Other
This category includes general behavior, sleep, circadian
rhythms, and photoperiodism as parameters that can be
important for toxicity testing.
Mites have been exposed to a toxic substance to observe its
effects on their dispersal and oviposition rates (55). The
aggregating behavior of Natrix (water snake) (1132) might
be used in toxicity testing.Metabolism, food capacity
and feeding differences between species have been studied
in shrews (904). Social (511, 1152) and sexual (1153)
behavior have been studied in the genus Peromyscus. A
method of recording burrowing activity (see Figure28 ) has
been devised and used on the mountain beaver (1097).
A means has been worked out for quantifying general ac-
tivity in mice (1094).
268
-------
: 5 Digit Counter
:24V
Mountain Beaver Tunnel
"250/50 V
27K
-vww-
30V
Brass Tuba
6/oss Capillary Tubs Insulator
Flexible Wire
Brass Probe serves
as contact switch. The
flexible feelsr win is bent
by passing animal to
compete the circuit.
Figure 28.
Wiring diaaram of traffic counter.
Mountain beaver burrow with picto'rial
drawing of burrow probe in place.
(1097)
269
-------
Various methods of dosing Coturnix with toxic substances
have been used to determine (a) the effects on sleeping
time and (b) if the route of exposure made any difference
in sleeping time (384, 398, 403) .
Mallard and black ducks, though closely related species,
have been shown to have striking differences in their
responses to different photoperiods (194). Differences due
to circadian rhythms have been demonstrated in drug-induced
mortality (1159) and heart rate (1158) in mice and rats.
The effects of photoperiodism on reproductive cycles have
been studied in birds (254).
Most of these methods did not deal directly with toxicity
testing. However, they point out various aspects of
animal behavior that should be taken into account when an
animal is used in such testing. Even though these .methods
were used on a limited number of species, there seems to be
no reason why they could not be applied to all vertebrate
species and some invertebrates.
Evaluation - The general behavior of most species can be
monitored and specific alterations caused by toxic substances
can be measured. The results may be extrapolated to wild
populations to evaluate the effect of the toxic substances
on their viability.
Instinct
Instinct is defined as behavior that is innate, not learned.
Because of its universality and its uniformity within a
species, instinct can be a useful parameter in measuring the
effects of toxic substances.
The reaction of Gallinaceous chicks to frightening stimuli
has been measured in terms of the distance and duration of
the chicks' movement away from the stimuli (1135). For
pictures of the apparatus used, see Figure 29. Another
test of Gallinaceous chicks exposed the chicks to a tape
recording of a hen's call and measuring their speed of
response (1135). For pictures of this apparatus, see
Figure 30.
Spiders have also been used in studies of instinct. Their
webs were photographed, measured and compared by computer
to controls. /For pictures of the examination table used,
see Figure 31. For pictures of the photographic setup
used, see Figure 32. A general discussion of methods of
preparing webs for photography and of calculating the
270
-------
igure 29 (a) . Runways
ith frightening device
t one end.
Figure 29(b). Runway mech-
anism for operating frighten-
ing device.
-gure 29(c). Runways,
:ightening device, and
<*mera used to record
Uck's response.
(1135)
271
-------
Figure 30(a). Enclosure for 10 runways.
Figure 30(b). Runway with speaker at far end,
Source: (1135) 272
-------
Figure 30 (c). Holding area - point of release
Source: (1135)
273
-------
Figure 31(a). Examination table prior to
photographing web.
Figure 31(b). Examination table prior to
photographing web.
Source: (53)
274
-------
•
Figure 32. Apparatus for photographing web,
Source: (53)
275
-------
changes in structure of the webs is found in 1124. A
discussion of the variables that affect web building and
web building's applicability to pharmacological testing is
also included. Reference 53 describes a study of spiders
exposed to drugs and allowed to spin webs.
Evaluation - Innate behavior is uniform in a species and
universal in the animal kingdom. It can be used to test an
animal's viability when exposed to toxic substances. For
example, behavior related directly to mating (e.g., court-
ship displays in grouse, ducks) could be monitored (perhaps
in cage field enclosures). Instinctual behavior related to
nesting (all birds), migration (ducks), territoriality
(Dipodomys), maternal care of young (birds and mammals), and
breeding (all species) could also serve as parameters.
Learning
In the laboratory, learning is studied by teaching tasks or
skills to a subject, then testing its ability to accomplish
them. Data are available on learning for many species and
experiments concerned with the effects of toxic substances
on learning are easily carried out.
The effects of various drugs on mice have been studied via
the learned skill of walking a revolving rod (1173). Rats
have been tested in a Skinner box, drugged, then retested
(547, 1165, 1171).
Rats and mice have been trained and then tested to determine
their reaction to increasing amounts of toxic vapor (1168).
Rats have also been drugged, taught and then retaught with-
out drugs and their performances compared (1164). Rats have
been injected with an endotoxin and a dose-dependent
decrease in activity noted (1163). Pavlovian procedures
have been used to produce aggression in rats as a response
to a tone stimulus (1162). Gerbils have been taught dis-
criminated avoidance response to a Skinner box (1166).
Opossums have been taught to solve the Guthrie-Horton puzzle
box (1189). They were also taught to give an avoidance
response (1190). For an automated method for studying
aggression in primates (1185) see Figure ."33. Primates were
also trained, given toxic substances, and impaired operant
behavior was noted in all cases (104, 547, 1184). Sheep
were trained, then electrodes were surgically implanted in
their skulls and they were orally dosed with a capsule of
toxic material daily. Their general behavior and results
of post-operation tests were evaluated (1179). Coturnix
chicks were fed toxic substances and changes in their
276
-------
Fiqure 33. An automated method for studying aggression
in primates.
r
_ MOH
ium»t
to* view
b. Bite Sensor Assembly
a. Bite Sensor and Hinged Neck Yoke
MONt VltW
The biting sensor is
a piece of pure gum rubber,
general purpose pressure-vacuum
laboratory hose (E. H. Sargent
and Co., Catalogue No. S-73515,
size F). Its outer diameter is
5/8 in. and the wall thickness
is 1/8 in. The ends of the hose
were forced over 1/2 in. diameter
brass nipples, which were soldered
at right angles to the 1/4 in.
brass hose supports. These hose
supports were long enough to allow
the hose assembly to be moved
toward or away from the monkey's
face. One of the support arms
was drilled out to permit trans-
mission of pressure changes to
the biting transducer.
c. Biting Transducer Assembly
Source: 1185
277
Copyright 1966 bv the Society for the Experimental
Analysis of Behavior, Inc.
-------
avoidance responses were noticed (380) . The effects of
drugs on mixed reinforcement tests were studied in pigeons
and it was found that endogenous stimuli were affected (264).
The effects of an amnesia producing drug on repeated
reversals of discriminatory testing were also studied in the
pigeon (1134). A general study of conditioned and uncondi-
tioned aggression without the use of toxic substances was
also undertaken with the pigeon (269). A general study of
factors that influence learning has been done with land
snails (42).
Evaluation - Learning involves the acquisition of a task or
skill or other capability on the part of the subject. The
ability to respond or to perform the task can be hindered by
exposure to toxic chemicals and the degree of confusion or
change. Also, the ability to learn new tasks can be tested
on dosed subjects and controls and the effects of exposure
on the learning process can be evaluated.
This type of testing can reveal an animal's ability to cope
in its natural environment and can conceivably be performed
on any animal that can learn.
278
-------
SUGGESTED METHODOLOGIES
Introduction
Scientific experimentation is designed to produce results
that prove or disprove a hypothesis. The general hypothesis
in toxicity testing is very often that the test chemical is
adversely affecting an animal or animal species. Conse-
quently, the methods used must distinguish toxic effects.
After years of study, many environmental toxicologists have
concluded that any change caused by a chemical that alters,
injurs, reduces, or otherwise affects the immediate or long
term reproductivity of a species is a significant toxic
effect; reproductive fitness is the major factor determining
the success of a species (20, 822). The parameters used to
determine the effect of toxic substances on reproductivity
can include decreased reproductive life span, sterility,
reduced litter/clutch size, early mortality, behavioral
changes affecting intra and interspecific communication and
so on.
A distinction must be made between toxic effects which can
be correlated with decreased reproductivity and those which
are not or cannot. For example, residue analyses and bio-
assays are useless .unless they can be correlated with acute
toxicity, chronic toxicity or decreased reproductivity.
LDso's often do not reflect environmental conditions because
concentrations of the test chemical do not match those
found in nature nor do the routes of exposure used fit the
exposure routes found in natural environments.
Suggested Approaches to Toxicity Testing
The methods selected in this report yield effects that are
directly or indirectly related to the reproductive fitness
of a species. The list does not include all possible test
methods, but was assembled using the information found in
the literature and the recommendations of environmental
toxicologists.
A list of selected toxicity testing methods is presented in
Table 27. (See evaluations of each method in previous
section for further discussion.) To obtain comprehensive
results, most methods should be used with both adults and
juveniles. Some methods, however, are suited for embryos;
this is specified under the heading "Routes of Exposure."
Routes of exposure were selected to best reflect those that
279
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o
55
H
EH
CO
W
EH
EH
U
H
O
EH
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ffi
-------
A / Mortality
B
Morbidity
C / Functional Change
Biochemical Change
Threshold Level
(residue)
Organis
Tissue
Molecule
Structur(
FIGURE 14
Succession of Toxic Response
282
-------
may occur naturally. Figure 34. Succession of Toxic
Responses, is keyed to Table 27 to show which methods are
best suited for investigating toxic effects at each level
of response.
General Studies
Long term generation studies can use aspects of all selected
methods, and can be conducted on any species that is readily
bred under laboratory conditions (See final list in Sugges-
ted Species). When short generation times are desired, in-
sects (Drosophila, Musca), mice (Mus, Peromyscus), voles
(Microtus), rice rats (Oryzomys), and Japanese quail are
recommended. Generation studies are important for toxicity
testing because their results are directly applicable to
the long term reproductivity of animals. The costs and time
involved are considerable, but the results are well worth
the effort. Previous experimental work can be used as a
guide to further study (see reference 822).
The Selection of Methods for Toxicity Testing
There are two important decisions to make in testing of
toxic chemicals: (a) choosing the test that will tell the
most about the specific chemical substance; and (b) choosing
the animals that will be used for subjects. These decisions
are frequently interrelated.
Selection of test method should be made in light of previous
work with chemicals of the same class as the test substance.
The test selected should give preliminary information. For
example, chemical A is a member of a class of compounds
which are known or suspected of being neurotoxic, but the
biochemical mechanism is unknown. These chemicals are also
contaminants of forested lands and are taken into plants in
large quantities. Initial testing should then utilize
herbivores of forest inhabitats and, because chemical A may
be neurotoxic, behavioral tests applicable to these species
should be used in conjunction with neurophysiological-
biochemical lab work to determine the mechanism of action.
Certain approaches to toxicity testing require basic know-
ledge about the test animal and its special characteristics
(see Table 27). For example, mutagenicity testing can most
easily be done on genetically well-known animals (e.g.
Drosophila) while dosing embryos in teratogenic experiments
is best accomplished with easily accessible embryos (e.g.
those of oviparous species). On the other hand, if a route
of exposure is available, most tests can be performed on any
species.
283
-------
A description of the levels of toxicity testing is provided
in Figure 34. The organism as a whole and survival are of
primary importance while morbidity research, functional
studies, biochemical analyses and residue determinations
are sublevels of interest. The survival of each individual
is dependent on what occurs at each lower level and testing
at each one is a way of obtaining information about the
whole organism. All information which can be related to
the survival of individuals can be a means of assessing what
will happen at the population level.
The level selected for testing should be chosen on the basis
of known information about the chemical and the suitable
species.
State-of-the-Art
Feelings are mixed throughout the scientific world about the
state-of-the-art of toxicity testing. Some experts in en-
vironmental toxicology feel that the field is on a par,
scientifically and technically, with most of the biologically
related disciplines. On the other side, there are scientists
who feel that present efforts fall short of sufficiency and
that more areas of testing should be introduced to the field.
They cite the lack of background residue data with which
most testing is done as one example of inadequate testing.
They advocate the routine inclusion in preliminary work of
total in - total out, total residue and clearance work and
100 percent of the dose accounted for after exposure.
Other suggestions and criticisms follow. They summarize
the general feelings we have encountered in the literature
and from contact with scientists.
1. testing juveniles as well as adults (all age classes)
2. more tests required on species actually affected by the
test chemical in natural situations, not just lab ani-
mals
3. increase federal funds for environmental toxicology re-
search and education programs
4. more long term generation, chronic, and subacute,
studies for wild species
5. poor knowledge of test animals' background (i.e. pos-
sible previous exposure) before testing
6. most tests performed in the past were not sensitive to
284
-------
the effects of test chemicals on the animal (e.g. LD5Q/
residue) and prove little more than exposure
7. failure of test methods, especially routes of exposure,
to coincide with actual concentrations and rates en-
countered in nature
The Future
At the present time there is research taking place in an
attempt to develop a computerized methods of quantitatively
analyzing the biochemical structure of drugs and predicting
from that structure, the mechanism of their action, without
microscopic knowledge of complex processes occurring in vivo.
This is theoretically possible because all compounds have a
chemical structure which determines their physical activity
or reactions with other compounds. If The Hansch Approach,
as this is called, ever becomes a usable system, it could
concievably be adapted for use with whatever class of chem-
icals one desires, including toxic chemicals. However,
the difficulties in the development and operation of such a
system are immense and much further research is necessary
before the Hansch approach can be applied.
Another approach to testing which appears to have greater
feasibility is the use of model ecosystems for toxicity
testing. The EPA labs in Corvallis, Oregon, are putting
forth a major effort to build systems that can be readily
set up for widespread use. These systems are being de-
signed for specific problems or for areas where certain
chemicals are applied (e.g. agricultural systems and pesti-
cides) . The problems associated with model ecosystems and
toxicity testing are presented in the Special Problems Sec-
tion of this report.
Many environmental toxicologists feel that the future of
toxicity testing should include a greater standardization
of testing methods/procedures and a greater emphasis on the
effects of chemicals on natural animal populations. LE^Q
and residue studies are of preliminary importance, but the
subacute physical, chemical and behavioral effects of real-
istic doses administered under conditions found in the en-
vironment on animals that are actually affected are now in
order and have been initiated in various institutions a-
round the country and the world. We must learn the sub-
lethal effects of chemicals in order to determine the over-
all hazard they present and to quantitatively and realis-
tically predict the level of toxicity of any chemical that
is applied to our environment.
285
-------
APPENDIX A
CONTACTS AND CONSULTANTS
-------
CONTACTS AND CONSULTANTS
James Akerman
Richard Aulerich
Mr. Woodrow Benson
Joel Bitman
Woodrow Benson
William Buck
Ron Borchard
EXPERTISE
Wildlife
Biologist
Mustelid
Toxicology
Pesticide
Monitoring
Avian
Toxicology
Pharmacology
Behavioral
Toxicology
Psychologist-
Herpetologist
ADDRESS
KPA
Ecological Effects
Branch
Washington, D.C.
Department of Poultry
Science
Michigan State Uni-
versity
East Lansing, MI
Director
Idaho Community
Pesticide Project
State House
Boise, Idaho 83720
Agricultural Research
Station
USDA
Beltsville, MD
Professor
Department of Physi-
ology
College of Veterinary
Medicine
Washington State Uni-
versity
Pullman, Washington
Director
Behavioral Toxicology
Laboratory
Veterinary Medicine
School
Iowa State University
Ames, Iowa 50010
University of
Tennessee
Knoxville, TN
-------
CONTACTS AND CONSULTANTS
(continued)
James Cannelli
Charles Cargille
Helene C. Cecil
Richard Crawford
Cirpriano Cueto
Armondo de la Cruz
William Diechman
Richard Dorn
EXPERTISE
Avian
Toxicology
Model
Ecosystems
Avian
Toxicology
Pathology/
Epidemiology
Toxicologist
Effect of
Pesticides
on isopods &
grasshoppers
Beagle
Toxicology
Veterinarian
Toxicology/
Epidemiology
ADDRESS
Massachusetts Coop
Wildlife Research
Unit
Amherst, MA
NICH
NIH
Bethesda, MD
Agricultural Research
Station
USDA
Beltsville, MD
Professor
Diagnostic Pathology
College of Veterinary
Medicine
Washington State Uni-
versity
Pullman, WA
National Center of
Toxicology Research
Pine Bluff, AR
Professor
Department of Zoology
Mississippi State
University
State College, MS
39762
University of Miami
Medical School
Coral Gables, FL
Professor
Department of Veteri-
nary Micro
Veterinary Medicine
School
University of
Missouri
Columbia, MO
-------
CONTACTS AND CONSULTANTS
(continued)
Virqil Freed
Milton Frifind
A. R. Gaufin
Jerry B. Graves
D. Gerberq
Yvonne Greichus
Max A. Haegele
R. L. Harris
EXPERTISE
Environmental
Toxicologist
Avian
Toxicology
Limnology
Avian
Toxicology
Insect
Toxicology
Biochemistry/
Wildlife
Toxicology
Avian
Toxicology
Veterinary
Toxicology/
Pesticides
ADDRESS
Environmental Health
Science Center
Oregon State Univer-
sity
Corvalis, OR 97331
Denver Wildlife
Research Center
Federal Center
Denver, CO
Professor
Department of Zoology
University of Utah
Salt Lake City, UT
Louisiana State Uni-
versity
Baton Rouge, LA
Insect Control and
Research
Baltimore, MD
Experimental Station
South Dakota Univer-
sity
Brookings, SD 57006
Denver Wildlife
Research Center
Federal Center
Denver, CO
Research Leader
Veterinary Toxicology
and Entomology
Laboratory
USDA
P. 0. Box GE
College Station, TX
77840
-------
CONTACTS AND CONSULTANTS
(continued)
Dale Hattis
Robert Heath
Glenn A. Hood
Robert Horton
Howard L. Hunt
David Hutchenson
Harold Kaplan
Philip Kearny
Irving Klaus
EXPERTISE
Genetic
Toxicology
Avian
Toxicology
ADDRESS
Case Western Reserve
University
Cleveland, OH
Patuxent Wildlife
Research Center
Laurel, MD
Rodenticide, Denver Wildlife
Repellent and Research Center
Chemosterilant Federal Center
Development Denver, CO
Senior
Research
Health
Advisor
Insect
Toxicology
Nutrition/
Toxicology
Toxicology/
Physiology
Avian
Toxicology
Avian
Toxicology
EPA
Research Triangle
Park, NC
EPA
ARC-East
Beltsville, MD
Associate Professor
Veterinary Medical
School
Sinclair Research
Farm
University of
Missouri
Columbia, MO
Southern Illinois
University
Carbondale, IL
Agricultural Research
Station
USDA
Beltsville, MD
Patuxant Wildlife
Research Center
U.S. Fish & Wildlife
Laurel, MD
-------
CONTACTS AND CONSULTANTS
(continued)
Seorge Levinskas
Mark Lewis
Ray Linder
Keith R. Long
William Lower
John Luteman
John McCanne
R. L. Metcalf
EXPERTISE
Avian
Toxicology
Behavioral
Toxicologist
Effects of
Pesticide
Residues on
Pheasant
Population
Environmental
Toxicology
Genetics
Supervisor or
Director -
Beltsville
Laboratories
Rodenticide
Research
Pesticide &
Aquatic
Model
Ecosystems
ADDRESS
Monsanto Corporation
St. Louis, MO
FAA Civil
Aeromedical Insti-
tute
AAC - 1186 6
P.O. Box 25082
Oklahoma City,OK
73125
South Dakota Coop-
erative Wildlife
Unit
South Dakota State
University
Brooking, SD 57006
Director
Institute of Agri-
cultural Medicine
University of Iowa
Iowa City, IA 52241
Environmental Trace
Substance Center
University of
Missouri
Columbia, MO
Building 402
ARA - East
EPA
Beltsville, MD
EPA
Beltsville, MD
Professor
Department of
Entomology
University of Illinois
Urbana, IL 61801
-------
CONTACTS AND CONSULTANTS
(continued)
David Mick
Howard L. Morton
John Mulder
L. D. Newsom
John J. Raisweiler
Jane F. Robens
John Santolucito
Peter Savarie
Ed Schafer
EXPERTISE
ADDRESS
Environmental Department of Envi-
Toxicologist ronmental Quality
State of Iowa
Des Moines, IA
Toxicologist- Agricultural Research
Honeybees
Director,
Animal
Laboratory
Resources
Environmental
Toxicology
Bats -
Breeding &
Maintenance
Toxicologist
Behavioral
Toxicity
Pest Control
Evaluation
Bird Damage
Control and
Chemical
Development
Station
USD A
Tucson, AZ
Michigan State Uni-
versity
East Lansing, MI
Louisiana State
University
Baton Rouge, LA
Department of Medi-
cine
Columbia University
New York, NY
Hoffman-LaRouche
Nutley, NJ
National Environ-
mental Research
Center
EPA
Research Triangle
Park, NC
Denver Wildlife
Research Center
Federal Center
Denver, CO
Denver Wildlife
Research Center
Federal Center
Denver, CO
-------
CONTACTS AND CONSULTANTS
(continued)
Dale Schwindamer
Joan Sovker
William Stebbins
Lucille Stickel
William Stickel
Gary Van Gelder
S. Bradleiqh
Vinson
Ralph C. Wandes
EXPERTISE
USDA Registry
List of
Animal Care
Facilities
Rodent
Toxicology
Oto
Toxicologist
Avian
Toxicology
Avian
Toxicology
Behavioral
Toxicology
Entomologist
Director,
Toxicity
Information
Center
ADDRESS
UCDA
Washington, D.C.
University of
Virginia Medical
School
Charlottesville, VA
Kresge Hearing
Research Institute
University of Michi-
gan College of
Medic Jne
University of Michi-
gan
Ann Arbor, MI 48104
Patuxant Wildlife
Research Center
U.S. Fish & Wildlife
Laurel, MD
Patuxant Wildlife
Research Center
U.S. Fish & Wildlife
Laurel, MD
Chairman
Department of Anatomy
and Physiology
Veterinary Medical
School
University of
Missouri
Columbia, MO
Texas, A & M
College State, TX
National Academy of
Science
-------
CONTACTS AND CONSULTANTS
(continued)
Bernard Weiss
Larry Wing
Peter N. Witt
Robert Wolf
James C. Yarbrough
Robert Yeager
Ray Zimmerman
EXPERTISE
Behavioral
Toxicology
Wildlife
Management/
Pesticide
Residues
Spiders -
Drug Testing
Infectious
Disease
Effect of
Pesticides
on Aquatic
Vertebrates
Executive
Secretary
of Animal
Division
Avian
Research
ADDRESS
University of
Rochester
Rochester, NY
Associate Professor
Department of Zoology
Science Hall II
Room 105
Iowa State University
Ames, IA
Division of Research
North Carolina Depart-
ment of Mental
Health
Raleigh, NC
Delta Primate Re-
search Center
Tulane University
Covington, LA 70435
Professor
Department of Zoology
Mississippi State
University
State College, MS
39762
National Academy of
Science
Washington, D.C.
Ralston Purina Co.
St. Louis, MO
-------
APPENDIX B
LIST OF SUPPLIERS
-------
SCIENTIFIC
COMMON
KEY
INVERTEBRATES
Lumbricus terrestris
Helix sp
Gryllus sp
Apis melliferra
Drosophila sp
Praying mantids
Tenebrio sp
Musca domestica
HEKPETOFAUNA
Rana pipiens
Ambystoraa mexicanum
Chelydra serpentina
Terrapene sp
Anolis carolinensis
Phrynosoma cornutuia
Thamnophis sirtails
BIRDS
Anas platyrhynchos
Branta canadensis
Ardeola sp
Tyto alba
Columba sp.
Colinus virginianus
Phasianus sp
Sturnis vulgaris
earthworm
land snail
cricket
honey bee
fruit fly
mealworm
housefly
Leopard frog
Mexican axolotl
Snapping turtle
box turtle
Anole
horned lizard
garter snake
rat snake
mallard
Canada goose
egret
barn owl
pigeon
bobwhite
pheasant
starling
CON, GEL, HER, LEM,
NBL, STE
STE
LEM, MAZ, NBL, STE
STE
CON, GEL, MAZ, NBL
STE
MAZ, STE
CON, GEL, LEM, MAZ,
NBL, STE, TAR
MAZ, STE
BEI,
LAJ,
NIC,
WES
HAC
LEM,
LEM,
ZOO
CHC,
MAZ,
MAZ
CHC,
STE,
CON, GEL, HOP,
LEM, MAZ, NBL,
SHE, STE, TAR,
MAZ, STE, ZOO
MAZ, STE, TAR,
GEL, HER, LEM,
TAR
MAZ, PET, SAI,
TAR, WHI
PET, WHI, ZOO
ADV, MAZ, TRS
ADV, MAZ, TRS
SAI
CHC, MAZ
AIM, LEM, MAZ, RAI,
RID, STE, TOT, TRS
BRN, MAX, THR, TRS
RED
BRN
Source: 7
-------
(CONTINUED)
SCIENTIFIC
BIRDS(continued)
Passer domesticus
Callus domesticus
MAMMALS
Eptesicus fuscus
Sylvilagus sp.
Dipodomys sp.
Neotoma lepida
Slgmodon hispidus
Spermophilus beecheyi
Tamias striatus
Eutamias sp.
Peromyscus maniculatus
Myocaster coypus
Hustela vison
Ovis sp.
Sus scrofa
Saimiri sciureus
Didelphis virginianus
Daysypus novemcinctus
COMMON
house sparrow
chicken
big brown bat
cottontail
kangaroo rat
pack rat
cotton rat
California ground
squirrel '
Eastern chipmunk
Western chipmunk
deer mouse
nutria
mink
sheep
swine, miniature
squirrel raonkey
opossum
armadillo
KEY
MAZ
BAB, COF, HAB, MAZ,
RED, THE, TOW
MAZ, NIC, ZOO
ANM, MAZ
PET, PFI, RID, WHI
MAZ
PET, RID
LOK, MAZ, RID, TOT
RID, TOT, WHI, ZOO
2OO
MAZ,
ANM,
mink
ADV,
STM
HRL,
VIV
BRP,
SDA,
ZOO
PET
CHC, MAZ
ranches
BAR, BUB, CLA,
LIT, MAZ, TRF,
FDI, PRL, RID,
WAC, WHI, ZOF,
ANM, MAZ, RID, SAI
ANM,
LOK,
SAI,
BRN, CHC, FRE,
MAZ, PFI, RID,
TAR, ZOO
-------
(CONTINUED)
ADV
ANM
BAB
BAR
BEI
BRN
BRP
BUB
CHC
CLA
COF
CON
FDI
FRE
GEL
Adventure Hill
Edith Tramutola
R.F.D.
Clinton, New Jersey 08809
Animals, Inc.
Dr. Jon Kenneth Ferguson
Route 1, Box 395
Wharton, Texas 77488
Babcock Poultry Farm, Inc.
Dr. Jack F. Hill
P.O. Box 280
Ithaca, New York 14850
Bar-Wan Rabbitry & Kennels, Inc.
L. M. Barnfield
Route 1, Box 60
Crocker, Missouri 65452
Beauraanor Farms
Thomas' G. Herrick
1712 Sheridan Road
Cleveland, Ohio 44121
Bronson Tropical Birds
Julien Bronson
70 Nagle Avenue
New York, New York 10040
Blue Ribbon Pet Farm, Inc.
John H. Marolf
8772 S.W. 131st Street
Miami, Florida 33158
HAB
Richard Bubolz
Route #2
Rio, Wisconsin
53960
Inc.
Charles P. Chase Company,
Charles P. Chase
7330 N.W. 66th Street
Miarai, Florida 33166
George Clauss
18-19 Saddle River Rd.
Fairlawn, New Jersey 07410
Colonial Poultry Farms, Inc.
M. R. Irwin
Pleasant Hill, Missouri 64080
Connecticut Valley Biological
Supply Co., Inc.
Michael L. Taylor
Valley Road
Southampton, MA 01073
Fauna Distributors, Inc.
Miguel A. Campo
2286 N.W. 36th Street
Miami, Florida 33142
Alton V. Freeman
Spruce Pine, NC 28777
CCM: General Biographical, Inc.
Order Desk
8200 South Hoyne Avenue
Chicago, Illinois 60620
HER
HOF
HRL
LEM
LIT
LOK
MAZ
NBL
NIC
PET
PFI
PRL
Hall Brothers Hatchery, Inc.
Mrs. A. B. Hall
200 Cool; Hill Road
Wallingford, Connecticut 06492
Hacienda Aquatics
John A. Kopec
P.O. Box 218
La Puente, CA 91747
Hermosa Reptile and Wild Animal
Farm, Inc.
Charles McClung
P.O. Box 182
219 Pacific Coast Highway
Hermosa Beach, CA 90254
E. G. Hoffman & Son
Earl A. Hoffman
P.O. Box 815
Oshkosh, Wisconsin 54901
The Hormel Institute, University
of Minnesota
J. Hobart Belknap
801 16th Avenue N.E.
Austin, Minnesota 55912
The Lemberger Company
Sheldon Antall
1222 W. South Park Avenue
Oshkosh, Wisconsin 54901
Norman B. Little
P.O. Box 382
Rocky Hill, Connecticut
06067
Otto Martin Locke
P.O. Drawer 731
New flraunfels, TX
78130
George Mazur Enterprises, Inc.
Ben Sherman
77 Eye Street, S.E.
Washington, D.c. 20003
National Biological Laboratories, Inc.
Frank S. Blasdell
P.O. Box 511
236 Dominion Road
Vienna, Virginia 22180
J. C. Hicholls, Jr.
Biological Collector
Murphy, NC 28906
The Pet Corral
Norman Nye
4146 Oracle Road
Tucson, Arizona 85705
Pet Farm, Inc.
Dr. B. M. Levine
3310 »W South River Drive
Miami, Florida 33142
Primelabs, Inc.
Dr. James H. Vickers
Monmouth County Airport
Farmingdale, NJ 07727
-------
(CONTINUED)
RAI Research Animals, Inc.
Norman or Pete Weissraan
Box 405
Braddock, PA 15104
RED Redwood Game Farms and Laboratories
David G. or Norzna S. Lewis
1955 North Redwood Road
Salt Lake City, UT 84116
RID Rider Animal Company, Inc.
V. D. Rider
R.R. *2, Box 270
Brooksville, FL 33512
SAI Safari Animal Imports, Inc.
Frederick W. Thorpe
7703 N.W. 36th Avenue
Miami, FL 33147
SDA San Diego Animal Supply
James M. P.obinson, Jr.
P.O. Box 544
Lemone Grove, CA 92045
SHE J. R. Schettle Biologicals
J. R. Schettle
P.O. Box 184
Stillwater, MN 55082
STM Stumbo Farms
Donald L. Stumbo
O'Neil Road
Lima, NY 14485
TAR Tarpon Zoo, Inc.
Trudie Jerkins or Fred Penman
P.O. Box 847
Tarpon Springs, FL 33589
THR Three Springs Kennel Company, Inc.
W. J. or Paul R. Haas
R.D. II
Zelienople, PA 16063
TOT Tote Em In Zoo
Mr. Tregembo
Route 2, Box 368
Wilmington, NC 28401
TOW Town Line Poultry Farm
Henry E. Geerlings
4198 96th Avenue
Zeeland, MI 49464
TRF Thompson Research Foundation
Crosby L. Brownson
Box 97, Postal Route II
Monee, IL 60449
TRS Truslow Farms, Inc.
John U. Truslow
Chestertown, MD 21620
VIV Vita Vet Laboratories
Forest E. Conder
1600 West 26th St.
Marion, IN 46952
MAC Woodard Asiatic Corporation
Dr. Geofli«.y Woodard
.12310 Pinecrest Toad
Herndon, VA 22070
WES Weston Research Laboratories, Inc.
Fritz Oyen
Route 1, Box 33
Purcellville, VA 22132
WHI White Animal Farm
Helen D. Perley
Seavey's Landing Road
West Scarboro, ME 04074
ZOF Zoological Fauna
J. Patrick McHale
1526 West Highland Avenue
Chicago, IL 60626
ZOO Zoological Center, International
Ray Pawley
15W506 W. 63rd Street (Burr Ridge)
Hinsdale, IL 60521
-------
APPENDIX C
SUGGESTED SPECIES SYNOPSIS SHEETS
-------
LUMBRICUS TERRESTRIS
Description Segmented, cylindroid invertebrate with body length
from 6 to 12".
AVail3Qility Found in most moist soil areas of the world; readily
available from biological supply houses and bait dealers,
Readily available; breeds easily in lab; easy and cheap
maintenance; ecologically important.
Light - natural photoperiod; temperature - 50°F for
breeding.
Any size container having drainage holes covered with
Cage screen and filled with gravel, dung, soil, and peat
moss is sufficient'for a limited number of worms; 4x8x2'
bin can support 50,000 worms.
FOOd Peat moss, manure, leaf mold, sod, decaying organic
material, chick starter, cornmeal; special needs
include moisture and substrate mixture of pW 7.
Water Moisture necessary at all times.
Parasites & Disease
_ . _ Breeds well in loose substrate of one part
Breeding & Rearing dung, 3 parts soil, and 5 parts peat moss if
space, moisture & temperature is optimal;
eggs deposited in small cocoons throughout
substrate.
Special Costs
Restrictions
Toiicological Studies 2?, 28, 29, 30, 32, 33, 34, 35, 36, 37, 38,
39
Ecological Alternatives snails.
Physiological Parameters
-------
HELIX ASPERSA
Food
Land snail; commonly called the garden snail; life
expectancy-unknown.
Found in temperate areas of world; relatively
abundant in most moist areas.
Mainly nocturnal; can be bred and reared successfully
in the laboratory.
Temperature, 23°C; constant moisture required by
active snails.
Colonies kept in wood boxes, 55x40x28cm; 5cm layer
of damp soil; box covered with aluminum screen; quart
fruit jars with 5cm of damp soil in bottom can be used
for breeding.
Lettuce and fresh vegetables.
Sufficient moisture must be present at all times
Parasites & Disease
Sexual maturity-4 months; avg. number of eggs
& Rearing laid-53 (about 10 eggs survive hatching); eggs
are laid in 5cm of soil in bottom of jar and
are 3mm in diameter; usually laid during wet
season; photoperiod important in egg laying.
Special Costs
Restrictiois
lexicological Studies 42, 46, 4?, 48, 49, 50
Alternatives Earthworms, some herbivorous insects.
Physiological Parameters
-------
Availability
MILAX sp.
Shell-less mollusc, body 1/2 to 2" long,
Some suppliers.
Snail-like, active day and night, produces large
amounts of mucous
Light-natural photoperiod; temperature - should
fluctuate around 20°; relative humidity - over 70%
Cage Material-all glass aquaria, any large waterproof
container; substrate - peat-soil.
Vegetables, such as carrots, potatoes.
Unchlorinated, unpolluted.
Parasites & Disease
& Rearing Reproduction stimulated by fluctuating
6 temperature around 20°C.
Special Costs
Restrictions
lexicological Studies
Ecological Alternatives snails.
Physiological Parameters
-------
ARANEUS DIADEMATUS
Small orb-weaving spiders about 2/5 to 1" long.
Availability Common orb-weaver of the Eastern U.S.; most dependable
HI dlldUllllJ source of spiders is Mr. Leonard Pankhurst, 204
Stroud St., Canastota, NY, 13034 (52) .
Easy maintenance; ecologically important; web-building
is a useful experimental tool.
Light - 13L:11D photoperiod; temperature - temperature
change important (colder at night) .
Cage 50 x 50 x 10cm aluminum enclosures with removable
glass windows suitable to study web patterns (52).
Food Flies, mealworms.
Provided by syringe administration or saturated cotton
ball.
Parasites & Disease
Breeding & Rearing Minimal breeding in lab; cocoons of fall
may be hatched in the lab under correct
conditions.
Special Costs
Restrictions
lexicological Studies 53. 1124
Ecological Alternatives centipedes.
Physiological Parameters
-------
GRYLLUS PENNSVLVANICUS
Dark-colored insect with an adult body lenath of 1/2"
large hind legs adapted for jumping (57) .
Found ^n pastures, meadows, roadsides, fields and homes
all Qver the 0>s> (316) . readily available through com-
ercial suppliers and bait dealers.
r63tlir6S Readily available; easy maintenance and breedability
in lab.
Light - 16L:8D photoperiod; temperature - 24°C; relative
humidity - 60 to 65%
Cage
42 x 18 x 18" stock aluminum cages for 250 adults;
11 x 7 x 8" glass aquarium for 500 nymphs; (Note: moist
sand substrate should be avaiblable for oviposition).
Food
Mixture of biscuit meal, dried milk, Bemax and dried
yeast in ratio 50:5:40:5; greens should be supplied
once a week (62).
Constant supply.
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
Some crickets breed all year while others
breed once a year (58); oviposition should
occur in moist sand; hatching may occur in
petri dishes; incubation - 15 days at 24.0°C;
hatching takes place after 3 month diapause
period at 4.0°c (62).
lexicological Studies 71
lliernotiiior
fUicrndllTCS
Physiological Parameters
Roaches ' grasshoppers, praying mantids, pre-
daceous beetles and other herbivorous insects,
-------
STAGMOMANTIS CAROLINA - praying manticl
Large, elongate, slow-moving insects with modified
front legs (57); body length - about 2".
Availability
Found mainly in the Southern U.S., readily obtainable
from commercial suppliers.
Features
Fairly available, good breedability in lab (61).
Laboratory
Light - natural protoperiod; temperature - 75 to 80°F;
relative humidity - 50 to 70%.
Cage
Glass jar or box closed with small mesh screen and
provided with small sticks and a potted house plant
is sufficient for a pregnant female (61) .
Food
Any living insect of appropriate size, for example,
live house flies, blue-bottle flies, grasshoppers,
grubs.
Should be given to mantid everyday with medicine
r2raSlt6S & UlSCaS6 Host for certain parasitic flies and
some mites.
Breeding & Rearing
Special Costs
Restrictions
Mantids lay a dozen to 400 eggs enclosed
in ootheca; trees are ootheca deposition
sites; eggs over winter and hatch in spring
and early summer; eggs may be hatched in
lab under correct light, temperature, and
nutrition conditions.
Cannabalistic tendencies.
Toxicologies! Studies
Ecological Alternatives
Physiological Parameters
Crickets, roaches, grasshoppers, other
herbivorous insects, predaceous beetles,
-------
BLISSUS LEUCOPTERUS
Pest insect with body length of 3.5mm (57); nonflying.
Mdlldullliy Found in wheat, corn rye and barley fields in U.S. with
some abundance.
Fairly abundant; economically important; probability of
easy maintenance and breedability in lab is good.
Light - natural photoperiod; temperature - "room".
t
Cage No special cage requirement; Blissus may be reared in
small glass tubes (as in done with milkweed bugs) .
Organic plant fluids (57) .
Water with food.
Parasites & Disease
ing & Rearing Eggs laid in May in ground; several hundred
eggs hatch a week to 10 days 'after laying;
nymphs mature at 4 to 6 weeks.
Special Costs
Restrictions
lexicological Studies
Alternatives Milkweed bugs_and other herbivorous
„,.,.,_ invertebrates.
Physiological Parameters
-------
EMPHOASA FABAE, XEROPHLOEA MAJOR - leaf hoppers
Small hopping insects about 1/4 to 1/2" in length (57).
I "I h'l'f Found all over U.S. on all types of plants; easy to cap-
AV3ll3Qllliy ture with insect net. (Note: use only one abundant
species specific to geographical area in testing (121) .
Abundant; high probability of ease of maintenance and
breedability in lab; economically important pest.
Light - natural photoperiod; temperature - "room".
An aquarium of sufficient size with small mesh screen
roof; soil, water, and specific food of selected leaf-
hopper should be available.
Specific food plant depends on leafhopper used (for ex-
ample, Erythroneura prefers apple leaves (121).
Available with food.
P2rasites & Disease
& Resting Most have one generation per year; some
have two or three generations per year.
Special Costs
Restrictions
Toxicologies! Studies
Alternatives Tree hoppers, froghoppers, planthoppers.
Physiologies! Parameters
-------
TRIBOLIUM sp.
Description
Flour beetle; length-to 5mm,
Availability Abundant in flour waste; suppliers.
Features
Cage
Light-unknown; temperature-21 to 27°C; relative
humidity-unknown.
Glass jars.
Flour, dog food, fruits .
Obtained from food
Parasites & Disease
Ł Doorinn 10° adults in 2 liter jar with flour; life
ft Rearing
cycle compieted in 8 weeks.
Special Costs
Restrictions
lexicological Studies so
Alternatives other granivorous- beetles.
Physiological Parameters
-------
TENEBRIO sp.
Mealworms; length 13 to 17 nun; dark color-larvae yellov
Very abundant around food stores, suppliers
Features
Light-natural photoperios; temperature-over 30°C;
relative humidity-70%.
Cage Size-60x50x50cm; material-anything easy to clean
ventilation necessary.
pflnH Grain, dog food, fruits.
Obtained in moist food.
Parasites & Disease
& Rearing Sexual maturity-depends on temp, and r.h.;
life cycle-complete in 4 to 6 months; breed
year-round,
Special Costs
Restrictions
lexicological Studies
Alternatives Other phytophagous beetles.
Physiological Parameters
-------
CARPOCAPSA POMONELLA
Small pest moth (57)
Availability
Found all over U.S., usually on foliage, fruits or
nuts (57) .
Fe3tlireS High potential for easy breeding and maintenance in
lab; economically important pest.
Careful balance of temperature and humidity (87, 88)
Cage
53.3 x 30.5 x 7.6cm stainless steel trays with 10-13mm
layer of burrowing medium on bottom (87).
Food
Wheat germ, soybean meal, rice flour, pinto beans,
alfalfa meal, cottonseed meal, corn meal, carrot
powder (87) .
Should be available in food.
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
Larvae may be reared in lab from eggs in
6 oz. wax paper cups with cardboard lids
(88).
Cannibalistic larvae.
lexicological Studies
Ecological Alternatives
Physiological Parameters
-------
AGROTIS YPSILON
UBS Cl ijJliuli Heavy-bodied moths having a wingspan of 1 to 2 inches
(57).
In the largest family of lepidopterans ; attracted
to lights at night for easy capture.
Have potential for routine breeding and rearing in
lab; economically important.
Light-light: dark photoperiod; temperature - 80+2°F.
Cage 20 adults held in 12 x 12 x 16" glass panel cages.
rOOu Rearing medium includes lima beans, pinto beans,
brewer's yeast, ascorbic acid, agar & water (88);
dental cotton plugs soaked in 10% dextrose solution
is good nutrient (86)-; larvae fed red clover or
Wa»pr tobacco.
ndm Available in food.
Parasites & Disease
& Rearing Gallon jars with moist paper serve as laying
environments; larvae may be placed in 12 x 8
x 2" pyrex trays with 1 inch of moistened soil:
post fifth instar larvae placed in small flint
fitctc jars with 1.5" layer of substrate containing
lOSIS 6% moisture (86).
KcSinCllunS Larval forms are cannibalistic.
lexicological Studies
Ecological Alternatives other noctuid moths
Physiological Parameters
-------
MUSCA DOMESTICA
house fly with body length of 1/2"; longevity
may be one year.
Availability Sometimes considered the most couonon animal known to
man.
Easy maintenance and breedability; veteran of previ-
ous research; economically important.
Light - natural photoperiod; temperature - 27.5+l°C;
relative humidity - 50 to 55%.
45x45x 45cm wooden cage with screen side for easy
cleaning (98). .
rftn|i Adults - shallow dish of sugar; larvae - dog bisguits,
• "00 fish, wheat bran, alfalfa meal, baker's yeast, malt
extract, manure, sugar and water.
Water-soaked cotton balls should be available at all
times.
P2r3SiteS & Disease Serve as vectors for numerous diseases
(b /) •
Adults transferred to gauge-lined plywood
Breeding & R63riflŁ cage; milk-water soaked cotton balls are
placed in cage for oviposition site; egg-
laden cotton balls placed in muslin-covered
jars where hatch larvae pupate; pupae placed
Special Costs in adult cage to emerge (98)"
Restrictions
lexicological Studies so, ioa, 109, in, 113, 114, 115, eis
r . . . ... .. Blowflies, stable flies, other suitable
Ecological Alternatives diPtera.
Physiological Parameters
-------
DROSOPHILA MELANOGASTER
Small flies about 3 to 4mm in body length (57) .
Common throughout U.S.; college campuses use cultures
Availability or <3enetic demonstrations; Contact; Drosophila Infer
AT dlldUllllJ mation Science Department of Biology, University of
Oregon, Eugene, Oregon 97430.
Easy to breed and maintain in lab; veteran of previ-
ous research; good model for population studies.
Light - normal photoperiod; temperature - "room".
Cage
Drosophila can be raised in baby food jars with no
problems.
A banana-agar or banana-meal mixture in the bottom of
a jar is suitable nutrition (102).
Water
Parasites & Disease
ing & Rearing
Place adults in baby food jar with correct
medium and culture becomes self-pertuating
Special Costs
Restrictions
lexicological Studies
116' 117' us, 119,
Shore flies, other small dipterans.
Physiological Parameters
-------
APIS MELLIFERA.
Social insect having a body length of approximately
3/4".
AfdlldDllliy only honeybee to occupy the U.S.; readily obtainable
from beekeepers throughout the U.S. (57).
Easy to keep and work with; good model for study of
the effects of chemicals on social organizaiton, pop-
uation dynamics and behavior; ecologically important.
Light - natural photoperiod; temperature - ambient.
6x6x2" cage for 100 bees (121) in field or lab.
60% sucrose and pollen (121) .
\jjf3tfjf Available in food.
Parasites & Disease
Self-perpetuating in most cases for at least
ina Ł Daarina a season w^en established outdoors; reproduc-
liig « n63nng tive uncertainty arises when colonies are
brought into the lab; nest boxes are necessary
for colony (57) .
IBI COStS Special bee handling equipment.
Restrictions
lexicological Studies us, 122, 123, 124, 126, 128, 129
Ecological Alternatives Bumblebee.
Physiological Parameters
-------
ISOPODA
Gray to black crawling crustaceans; doros-ventrally
flattened body; length, 5 to 15 cm (687).
AV3il3biHtV 400° marine species; only pill bugs or wood lice are
' truly terrestrial; wood lice found in great numbers
in moist areas under boards and in logs.
F63ttireS Some terrestrial isopods have ability to roll
up in bail for protection and prevention of water
loss (687).
Light-natural photoperiod; temperature "room".
Terreriums of any size filled with moist soil and
having abundant detritis-
Algae, moss, bark, decaying vegetable and animal ^
matter; some terrestrial species are parasitic on
ants, and some are carnivorous.
Water Moist areas; organisms reac^; to water loss by rolling
n .. B ijxto ball (687).
Parasites & Disease
0 Rearing Eggs brooded in a marsupium after fertilization
» postlarvae hatching stage with last pair of !
legs incompletely developed (687).
Special Costs
Restrictions
lexicological Studies
131
Ecological Alternatives Millipede, termite.
Physiological Parameters
-------
CHILOPODA - centipedes
Wormlike, or heavy-bodied; length from 1' in tropics
to 1" or 2" in North America (687).
Most are confined to the moist environment beneath
stones and wood and in soil; found the world over (687)
Poison gland in claws; only active on surface of ground
at night; slow but prolific reproducers (687).
laboratory
p Terrariums of any size, filled with moist soil, having
"•§8 plenty of cover (rocks, wood) and small arthropods.
FflOd Small arthropods mainly; will eat toads, small snakes
and small mice (687).
Water
Parasites & Disease
Ł Dgaring Sexes distinguishable; some species lay and
" brood their eggs, others simply lay the eggs;
years are required to reach sexual 'maturity.
Special Costs
ftestrictiois
lexicological Studies 130
Ecological Alternatives spiders.
Physiological Parameters
-------
RANA PIPIENS
leopard frog, head and body 2" to 5"; life expectancy
2 to 3 years.
AV3ii3biiity Numerous suppliers; ponds/ quiet waters throughout
U.S.
F63tlir6S Amphibian, insectivorous; reacts to movement.
Light-regular photoperiod; temperature of water,
18 to 20°C.
Large tanks with aquatic and terrestrial areas
(22"xl4"xl4" for 25 adults for short periods).
p g Q (j Cooked lettuce, cooked greens, trout chow, liver
bits, pond plankton, larvae, mosquitoes, flies,
crickets.
Wot or Unchlorinated, unf lour ida ted, unpolluted; moist
conditions necessary.
r3r3SlteS a UIS63S6 Many natural internal and external parasites
New arrivals should be quarantined and
checked.
DT66Qtng & Rearing External insemination in water, embryos
in shallow enamel pans; 100's of eggs from
each female; generation time is 13 to 15
months•
Special tOStS Large operation may need special water
treatment.
ReStriCllOfiS No spinach in diet (may cause kidney stones);
clean water necessary.
lexicological Studies 147, i4a, 149, iso, 151, 152, 153, 154, 155,
758
ECOlOgiC3l Alternatives Turtles, lizards, toads, salamanders.
Physiological Parameters
-------
AMBYSTOMA MEXICANUM
Description
Availability
Features
Partially or completely nebl-onous slamander having a
length of 30cm (142).
Found in nature only in Mexico; may be obtained from
commercial suppliers (142) ; being bred at University
of Michigan Amphibian facility (137) ;; veteran of
scientific research.
Feirly available; breedable in lab (artificial ferti-
lization) ; veteran in lab situation (142) .
Laboratory
Light - natural photoperiod; temperature - water tem-
perature of 14° to 18°C.
Cage
50 x 100 x 35cm asbestos cement container houses 10
adults; no sand, gravel or plants are needed (142).
Food
Beef heart, liver; vitamins (142).
Unchlorinated water necessary (142).
Parasites & Disease rungai infections (142).
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity reached in one year; breeding
season from December to June; eggs laid vary
between 200 to 2,000 per spawning; "hatching"
occurs after 2 weeks (142) ; usually 2 spawn-
ings per year.
lexicological Studies ieo, 152
Ecological Alternatives °ther
Physiological Parameters
-------
TERRAPENE SP.
Dry land-turtles with body lengths of 4 to 6.5"; ages
of 30 to 40 years are common; some live to be a century
old (133).
Availability Found all over most of the U.S. in woodland areas; avail-
able in pet stores.
Available; long lived for chronic testing; hardy, main-
tains extremely well in captivity.
Light - natural photoperiod; temperature - "room".
Dry tanks (130 x 72cm) with water available for drinking
(165) ; terraria with dirt for turtle to dig in, and wate
for occasional immersion.
Gound horsemeat and lettuce (165) ; raw hamburger and
table scraps (133) .
Water M libitum
& DiSeBSe Tuberculosis, Salmonella, assorted
parasites.
9 Dairinn Spring mating is usually the case; 3 to 6
& Rearing leathery eggs are laid in June or July;
eggs hatch in September (1192); eggs may
be collected from natural nest sites for
rearing.
Special Costs
ReStriCtlOflS Breeding in captivity difficult.
Toiicological Studies 167' 18°
Ecological Alternatives species of
Physiological Parameters
-------
CHELYDRA SERPENTINA
Large, short-tempered turtle; 8 to 12" body length;
10 to 35 Ibs. (133).
JtYdil3biHty Southern Canada to Gulf of Mexico; Atlantic Ocean to
Rockies; generally occupies a permanent body of fresh
water (133).
Hardy; good test animal when young; easy to maintain
under lab conditions (175); economically important
(133).
Light - natural photoperiod; temperature - "room".
pa_ Enclosed outdoor area (5 x 8") with shallow stone or
"•6* concrete pond (12 to 15" deep) and shelters in the
form of logs or stumps (133) .
Insects, small fish, earthworms, dog food, vegetation
(175).
Ad libitum
& DJS63S6 Tuberculosis; Salmonella; assorted
parasites (166) .
Rraaiiinn 9 Datrlnn E99S laid in June'" clutch size between 20
DlCcUlllg « Rearing and 30 eggs; eggs hatch in September and
October (1192).
Special Costs
Breeding in lab is questionable; handle
with care.
lexicological Studies 147, 175,
Alternatives Macroclemys temmincki (alligator snapping
«. «. turtle) .
Physiological Parameters
-------
ANOLIS CAROLINENSIS
The false chameleon with a body length of 5 to 7 1/2"
(133).
Availability Common in Southeastern U.S.; may be purchased in pet
stores (133).
Available, easy to maintain in lab.
Light - natural photoperiod; temperature - "room".
Standard aquarium filled with sandy soil, some plants
and small branches (133).
Mealworms, wide variety of insects.
Spray cage with water everyday to supply lizards with
water.
Parasites & Disease
& Rearing 1 to 2 eggs per clutch; 6 to 9 clutches
per season (1193).
Special Costs
Do not breed well in captivity.
lexicological Studies 175, iso, 148
Alternatives Eumeces, toads, snakes.
Physiological Parameters
-------
PHRYNOSOMA CORNUTUM
Spiny lizard; body length - 2.5 to 4" (133).
RY3ll3Ulllty Found in parts of Kansas, Texas, Arizona and northern
Mexico (133) can be bought from pet stores.
F62tlir6S Available; easy to maintain under lab conditions.
Light - natural photoperiod; temperature -"room".
Cage Aquarium filled with sandy soil, some plants, small
branches, rock pile, and water dish (133).
Spiders, sowbugs, insects (133).
Ad. libitum
Parasites & Disease
Breeding & Riarwg 14 to 37 e^gs ?er clutch; i or more
clutches per year (1193) .
Special Costs
Poor breedability in lab.
Toxicological Studies
Alternatives Toads, snakes, Eumeces.
Physiological Parameters
-------
THAMNOPHIS SIRTALIS
striped, thin-bodied snake, 18 to 26 inches long (133);
Eastern half of U.S. (133); found in a wide variety of
habitats in the city and country.
Features
Laboratory
Available; easy to maintain and handle in lab; breeds ]
well in captivity giving birth to many young at one
time (175).
j
Light-natural photoperiod, needed for heating purposes;1]
temperature - 26o to 36°C; relative humidity - 33 to 661!
(171).
Cage
Food
Terrariums with tightly sealed lids; pea gravel or
artificial grass substrate; rock or branches necessary
for ecdysis (133, 171, 173).
Frogs, toads, salamanders, fish tadpoles, earthworms,
chopped raw fish (133).
ad libitum in open bowls.
& DiSeaSe Ectoparasites common.
Breeding & Rearing
i
Sexual maturity reached in 1 to 2 years; *
3 to 85 young per clutch per season (1193). <
Special Costs
Restrictions
lexicological Studies I48'
» 715
Ecological Alternatives
Physiological Parameters
Lizards, turtles
-------
ELAPHE spv
Large, constricting, handsome snakes having body
lengths ranging from 24 to 72 inches (133) . ,
AV8il8l)ility Found throughout most parts of U.S. in trees, cliffs,
fields, etc. (133).
Wide distribution; maintains extremely well in
captivity.
I ah flf at (try Light-natural photoperiod, needed for heating purposes,
" temperature - 26° to 36°C, relative humidity - 33 to 60%
(171).
Terrarium with tightly sealed lids; pea gravel or artificial
grass substrate; rocks or branches necessary for
ecdysis (133, 171, 173).
Mice, rats, small birds, lizards, frogs (133).
ad libitum in open bowls.
& DJSeaSe Ectoparasites common
Breeding & Rearing i to 44 eggs per dutch.
Special Costs
Breedability in lab questionable,
Toiicological Studies 175, iao, 715
Ecological Alternatives Lizards' turtle*.
Physiological Parameters
-------
PHALACROCORAX AURITUS
D6SCript'lOn Large, dark/ water birds with long necks and tails;
30 to 35" body length (221) .
Common on coasts, inland lakes and rivers, generally in
the Eastern half of the U.S. (221). ;
top of their food chain; intelligent; good short-
term test bird; easy to obtain adults, young and eggs;
typical of fish-eating birds (181) .
L3BOr3tOry Light - natural photoperiod; temperature - young birds
kept warm with heating lamps (181) .
6x6' cage made of wood, cement, chicken wire and
indoor-outdoor carpet will house 6 cormorants; cage
should be equipped with tree trunks and large water
tanks for bathing and drinking (181) .
Fish (181, 182) and vitamin B supplement and ferrous j
iron supplement; grit required as a digestive aid and j
mineral supplement (182) . I
Ad libitum (See "cage"). !
Parasites & Disease
& Rearing Clutch size - 2 eggs; one clutch per year;
colonial nesters (182, 183).
Special GOStS Food costs high due to bird's appetite.
KeStMCtlORS Somewhat hard to breed in captivity; takes
a long time to reach sexual maturity; largf
bird with enormous appetite.
lexicological Studies 188, 189
Ecological Alternatives GUHS and terns.
Physiological Parameters
-------
PODILYMBUS PODICEPS
A duck-like diver; body length - 12 to 15" (221)
fiV3ll3ulllty Found throughout the U.S. and Canada, near or in ponds,
creeks and marshes (221) .
Diurnal and migratory; fairly abundant.
Light - natural photoperiod; temperature - ambient.
Cage Mallard specifications.
In nature, they eat vertebrate and invertebrate aquatic
organisms; grit needed as a digestive aid and mineral
supplement.
Ad libitum.
& Disease Worms, ticks, fleas, lice, botulism.
& Rearing 2 eggs per clutch; one clutch per year;
young are precocial and nidifugous (183)
Special Costs
Poor breeders in captivity
lexicological Studies 185
Ecological Alternatives Diving ducks .
Physiological Parameters
-------
ANAS PLATYRHYNCHOS
A familiar surface-feeding duck; body length 20 to 28";
Wei9ht 2 V2 to 3 1/>2 ^s-> life expectancy - 20 years
in captivity, 16 years in wild.
Found in wooded swamps, marshes and ponds throughout me
of the U.S. (221); legislation exists which may limit
wild bird testing.
Cage
Food
Water
Parasites
Good availability; breeds easily in captivity; large
enough for large samples; representative waterfoul; adap
well to small areas; healthy, stable and vigorous; (3811
important in sports and recreation.
Light - 10D:14L photoperiod (207) ; temperature - "room!!
8 x 14 x 7' open wire mesh cage for natural environment i\
testing, 20 to 25 birds per cage (381); 30 x 15 x 6'
recommended pen size for 5 adult birds, must be provided-
with water area of 10" depth (192); 30 x 15 x 6' pen witti:
nest boxes, 250 gallon water troughs and feeders, used ti
breed mallards (194). f
Natural-grain, seeds, roots, water plants, snails, insed
small aquatic organisms (1194) ; lab-commercial game bird|
ration usually supplemented with millet, cracked corn, vb
milo and barley; grit needed as digestive aid and dietarf
supplement (191, 192).
Water available at all times for drinking and recreation*;
& Disease
Flies, true bugs, lice, protozoans, worms,
flukes; botulism.
Breeding & Rearing
Special Costs
Restrictions
Toxicologica! Studies
Sexual maturity - 1 to 2 yrs.; 5.1 to 9.3 eg|
per clutch; renesting probability high; incubj
tion period of 21 to 28 days; 49 to 60 day f|
ing period; newly-hatched young display impr
behavior; easy breeders in captivity (381).
Cages have to be cleaned on a regular basis;]
cial floor covering materials keep cleaning ^
quency and expense minimal (381). j
Territorial during breeding; crowding should
avoided; migration restlessness during certai
periods of year.
184, 192, 195, 197, 198, 199, 200, 201, 203,
206, 208, 209, 210, 213, 1195
Ecological Alternatives
Physiological Parameters
Other waterfowl.
-------
BRANTA CANADENSIS
Largest of wild aeese; length - 35 to 43"; weight -
8 to 13 pounds; (1194); Longevity - 33 years in cap-
tivity, 23 years in wild.
Availability Found in wet lands in most of North America during
some part of the year (221).
Canada geese keep well in captivity (1196).
See mallard (Anas platyrhynchos) synopsis.
Cage See mallard (Anas platyrhynchos) synopsis.
Natural food - grain, grasses, green parts of plants
Food and small bits of animal matter (1194); Lab food -
commercial game bird ration; grit needed for digest-
ive aid and mineral supplement.
UK 3 {Of Should be available at all times for drinking or bathing.
ParaSiteS & DiSeaSfi Flies, true bugs, lice, worms and flukes
(1194); may be susceptable to botulism,
fowl cholera, avian tuberculosis, and malaria.
BrPPfiino Ł Pparino Becomes sexually mature in 2 to 3 years; Nest
Dieeumg & wearing in earl^ spring/ usually mid-Aprii; 4 to ?
eggs laid per nesting; 28 day incubation per-
iod; male and female guard nest; 63 to 86 day
fledging period; young take to water
Special COStS immediately (1194).
iieStriCtiORS Migratory restlessness; territorial during
mating season; protected by legislation.
TGXiCOlOgiCal StUdieS 205,206,210,213, 248 (Note: See mallard
(Anas platyrhynchos) section for similar
references].
ECOlOgiCal Alternatives Other waterfowLof the world.
Physiological Parameters
-------
PUFFINUS GRISEUS
Description
Gull-like sea bird; body length 16 - 18"
Availability
Features
Sea bird found in Northern and Southern hemisphers
along Atlantic, Gulf and Pacific coasts; most abun-
dant in Pacific area (221, 1196).
Migratory and diurnal; nasal salt glands allow bird
convert sea water to fresh water.
Sea birds do not do well in the laboratory.
Cage
Cage provisions have not been fully documented for
sea birds.
Food
Water
In nature, they feed on fish, Crustacea, cephalopods |J
and macrophy ton ; grit needed in diet as a digestive ai«
and mineral supplement.
Salt or fresh water should be provided at all times. [\
Parasites & Disease
Ticks , worms, mites, fleas, lice.
Breeding & Rearing
Special Costs
Restrictions
Colonial nesters; usually one egg laid
year; incubation - 6 to 11 weeks; 13 to
week fledging period; sexual maturity usu-j
ally reached by second year; renesting poŁ'
sibiltiy high (183, 215); young are
altrical and nidicolous.
Not easily maintained or bred in captivity
slow reproducers.
lexicological Studies 218
Ecological Alternatives
Physiological Parameters
Or terns.
-------
CASMERODIUS ALBUS
Wading birds with long necks and legs and pointed bills;
body length of 37-40" (221).
Availability Found in marshes, coastal areas and rivers of the
Eastern half of the U.S. (221).
Fairly easy to maintain in captivity.
Light - natural photoperiod; temperature - "room".
Outdoor duck pens of sizable dimensions may be used for
holding Egrets.
FOOD Feed on both vertebrate and invertebrate aquatic
organisms.
Ad libitum.
& DJSeaSe Ticks, worms, mites, fleas, lice.
& RearillŁ 3 eggs per clutch; 1 clutch per year; young
altricial and nidiculous, young fledge at
5 weeks of age (183, 222) colonial
nesters.
Special Costs
Slow breeder, large size, protected by law.
lexicological Studies
Cranes, rails, coots.
Physiological Parameters
-------
FULICA AMERICANA
Gray duck-like bird (221); body length - 15" (1196)
Availability Found in marsh areas mainly in the Eastern U.S. (221)
Readily available for study.
Light - natural photoperiod; temperature - ambient.
Cage Mallard cage specifications.
F00IJ Carnivorous in wild, need grit for digestive aid and
mineral supplement.
Ad libitum
Parasites & Disease
& Rearing Young are precocial and nidifugous,
Special Costs
Protected by legislation
lexicological Studies
Ecological Alternatives Herons, ibis, ducks.
Physiological Parameters
-------
PHILOHELA MINOR
A large, husky, neckless, long-billed brown bird;
lenght of body - 10 to 12" (221).
Found in swamps, wetlands and thickets throughout most
of the U.S.
Migratory and territorial; available; socially tolerant
of one another; hardy, withstands heat and cold well
(225).
Light - natural photoperiod; temperature - "room".
20 x 40 x 8' field enclosure; 4x4x5' ground cage;
22 x 29 x 30" steel cage for lab (225). All cages are
provided with food and water; they should be covered
with burlap to prevent flushing and injury to the bird.
In the lab they feed on earthworms, mealworms, fly lar-
vae placed in peat moss in food pans at all times; in
nature, they feed on invertebrates near lakes, ponds
and streams.
Ad libitum.
&
Ticks, mites, fleas, lice, worms.
Breeding & Rearing
COStS
Restrictions
lexicological Studies
Cages must be kept clean to prevent disease.
Nervous and therefore prone to injure them-
selves in captivity.
27, 226, 22?, 223, 229, 230
Ecological Alternatives snipe.
Physiological Parameters
-------
LARUS ARGENTATUS
Known as common seagull; body length - 23 to 26" (221)
AV3Jl3bJiJty Ranges in most of the Eastern half of the U.S. near the
coast and along river banks (221).
Pa of I) roc Easy to obtain eggs and adult birds; good subjects for
short term studies and oological studies because of
colonial nesting; migratory and diurnal.
LdDOr3tOry Light - natural photoperiod; temperature - "room".
Cage dimensions used for ducks may be suitable for the
herring gull.
Invertebrate and vertebrate aquatic life should be
supplied at all times; grit should be supplied as a
digestive aid and mineral supplement (182) .
Ad libitum .
Par3Sit6S & DiSeaSe Fleas, mites, ticks, lice.
ing & Rearif]Ł 4 eggs per clutch; 1 clutch per year; high re-
nesting potential; incubation period - 27 days;
young precocial, nidicolous, fledge at 43 days
(183, 184, 231).
Special Costs
ReStriCtIORS Hard to maintain in lab; protected by
legislation.
Toxicological Studies i88, 231, 232, 235, 235, 249
Ecological Alternatives Terns, other guns, Pei
Physiological Parameters
icans.
-------
MELANERPES ERYTHROCEPHALUS
chisel-billed tree-climbers; stiff, spiny tails;
8-8 1/2 body length; head completely red (221) .
Found throughout U.S. in woodlands (221). .
Diurnal and migratory; hole-nesters.
Light - natural photoperiod; temperature -; "room".
Cage
Bobwhite quail cages with modifications.
Food
In nature - seeds and terrestrial insects - grit need-
ed in diet to aid in digestion and supplement minerals,
Water
Parasites & Disease
Hosts of fleas, mites, lice.
Breeding & Rearing
Special Costs
Restrictions
2 to 3 eggs per clutch; 12 day incubation
period; young are nidicolous and altricial;
24 day fledging period.
lexicological Studies 233
Alternatives Brown creeper, nuthatch
Physiological Parameters
-------
FALCO SPARVERIUS
Small sparrow-like falcon; body length - 9 to 12";
long life spans.
AĄ3liability Most abundant hawk found throughout North America (241)
Features
Laboratory
Cage
Food
Ad libitum
Water
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
Available, fairly easy to maintain; good indicators of
pesticide level in the environment, small and fairly
easy to handle in comparison with larger raptors.
Outdoor pen - 8 x 12 x 7' enclosure with roof made of
1x1 1/2" mesh with box 10 x 10 x 15" with 3" diameter
opening; larger dimensions (50 x 20 x 6") can house mul-
tiple pairs of hawks (239); alternate cage - 6 x 6x8'
solid wire cage for a pair of sparrow hawks.
Dead cockerels, ground beef, vitamins, minerals, and tur-
key breeder crumbs (239, 240); a combination of hamsters
and chicken heads and necks may be substituted for ground
beef; roughage in diet is needed for formation of fecal
pellets.
Hosts for external parasites to a limited
extent (182).
Sexual maturity - end of first yr.; clutch
size - 3 to 7, average 4 or 5 eggs; incuba-
tion period - usually 30 days; fledging per-
iod - 30 days; nesting success - 3.88 young
hatched per nest and 3.81 young fledged per
nest (215, 241).
Equipment for handling, fairly expensive diet
Violence during handling; slow reproducer
requiring large pens.
lexicological Studies 134, isa, 243, 249
Alternatives Kites, shrikes, buteos, accipiters and falco-
Physiological Parameters
-------
TYTO ALBA
Long-legged, light-colored owl with while, heart-shaped
face; body length - 15 to 20"; claws and beak modified
to handle prey (221) .
AV3II3DUIIJ Found mainly in the Eastern half of the U.S., particu-
larly near barns and old buildings (221) . .
Fairly good availability; fairly easy to maintain; good
indicator of pesticide levels in environment; nocturnal,
L3BOr 3tOry Light - natural photoperiod; temperature - "room".
6 x 10 x 50' chicken wire and wood frame cage with por-
ches and nest box for one breeding pair (20) .
Carnivorus 'habits in nature and in lab; small whole rod-
ents - especially while brooding (20) ; roughage needed
for fecal pellet formation.
Ad libitum
Parasites & Disease
Breeding & Rearing 5 to 1]- e^s ?er clutch
Special Costs
Hard to tame and breed in captivity; capable
of causing injury to handler.
lexicological Studies 245, 243
Alternatives Kites, shrikes, buteos, accipiters and falcons,
Physiological Parameters
-------
COLUMBA LIVIA
Cage
Domesticated bird; body length about 13", weight - about
450g. ; sexual monomorphic (221, 1196) .
Common throughout U.S. in association with man; captured
with traps or mist nets; can be raised in captivity or
obtained commercially.
Large enough for good samples; can be breed easily in
captivity; easily housed and caged; good temper ment,
can be conditioned and trained (182, 267, 272).
Light - 14L:10D photoperiod; temperature - 20°+1°C (251)i.
20 x 29 x 18cm steel rat cage houses one bird (250)
40 x 64 x 40cm cage houses one bird (251) ; 45.7cm cage;
made of 7.3 x 2.5cm gage welded wire with perch, food caa
automatic watering device and nest box should house a pai
(252) 115 x 134 x 229cm cage houses a pair of birds (253
rfi j Commercial pellet diet or a mixture of wheat, maize, and
'000 tick beans (251); grit should'be available to birds at
all times (182); eats seed and grain in the wild. ;
Ad libitum . |
Parasites & Disease
Breeding season - all year; clutch size -
Breeding & Rearing 2 e^s> laying interval between eggs - 1.5 tc
6 6 2 days; incubation period - 15 days; fledgin?
period - 17 days; fertility - 83.3 to 92.9%
eggs; (215, 252).
Special Costs
ReStriCtiORS Protected by legislation in some areas.
T - , • , Pl .. 152, 199, 252. 255, 259, 260, 261, 262, 263,
lexicological studies 264, 266, 267; 268, 259, 270, 271, 272, 273,
885, 1134
Ecological Alternatives Rin9 dove' mourning dove.
Physiological Parameters
-------
COLINUS VIRGINIANUS
Chicken-like bird; body length 8.5 - 10.5".
Availability
Eastern Farmlands from Gulf of Mexico to Ontario
Breeds well in captivity; requires minimal housing space;
readily available and easy to maintain.
Light - natural photoperiod; temperature - "room".
i 000
Water
6 x 10 x 8" cage constructed of 1" wire mesh;
4 x 6 x 1.5" pen designed for a large number of birds;
3x6" outdoor pens made of wire net (276).
F & M game bird chow, turkey starter mash, Purina
pigeon chow (274, 277, 278); grit needed to aid di-
gestion and to supply dietary minerals (182) .
Ad libitum.
& Disease
Fleas, mites, ticks and lice.
25 day incubation period; young are
precocial and nidfugous.
Special Costs
lexicological Studies
Cannibalism in crowded quarters; protected
by legislation.
202, 274, 275, 275, 273, 279, 28.0, 231, 28.2,
283, 284, 285, 286
Alternatives Cotumix, pheasant, grouse.
Physiological Parameters
-------
PHASIANUS COLCHICUS
Large gallinaceous bird with long pointed tail; body
length - male, 33-36"; female - 20 1/2" (221).
Availability Found in farming country north of Mason-Dixon line.
Breeds easily in captivity; easily maintained; migra-
tory and diurnal.
Light - normal photoperiod; temperature - "room".
Pen complex 22 x 6.1 x 1.7m. can be partitioned into
27 individual cages with dirt runways and plywood
shelters (287).
p00d Diet recommended in Poultry Formula guide (288) ; grit
needed (182) .
Ad libitum
Parasites & Disease Ticks, iice, mites,
& Rearing Precocial and nidifugous young.
Special Costs
Cannabalistic in crowded quarters; protect-
ed by law.
lexicological Studies 2i°/ 248' 281, 232, 290, 291, 293, 294, 295
296, 297, 298, 299, 300, 301, 302, 303, 304
305, 306, 734, 735, 1197, 1198
Ecological Alternatives Grouse, turkey.
Physiological Parameters
-------
DENDRAGAPUS OBSCURUS
Plump-breasted, short-winged birds approximately 17-21"
long (1196).
nV3!l3ulllty Lives in coniferous forests of humid Pacific coast and
mountains (1196) .
Easy to maintain and breed in captivity; useful in field
and lab telemetric studies due to large size.
Light - natural photoperiod; temperature -"room1 .
Cage
Food
24.5 x 2.5 x 3.25' concrete tank with wire mesh top;
20 x 10 x 6' pen made of fish netting and steel posts;
8.1 x 16.2' pen with turf floors and mesh roof;
1x1x2' wire cage used for the field.
Game bird chow in pellet form (308); grit needed as a
digestive aid and mineral supplement (182).
Water
Ad libitum.
&
Ticks, mites, fleas, worms.
Breeding & Rearing
Sexual maturity reached in 112 days (309)
young are nidifugous and precocial (183).
Special Costs
Restrictions
Cannabalistic in crowded quarters; protected
by legislation.
lexicological Studies 312, 313
Alternatives Coturnix, bobwhite quail, pheasant.
Physiological Parameters
-------
MELOSPIZA MELODIA
Sparrow; length - 5 to 6 3/4"; short, stout bill used for;
seed cracking; longevity of approximately 2 years (221). "
AV3ll3Dllliy Found throughout most of the U.S. in woods and non-
aquatic areas (221) .
F62tlireS Readily available in the wild.
L3BOr3tOry Light - natural photoperiod; temperature -"room".
fnnij
120 x 60 x 60cm cage made of wood and 12mm mesh; single j
bird cage of 41 x 61 x 38cm; 3x3x2' cages and
6x5x2' cages used .
Lab food - pigeon chow, canary or millet seed, or a mix-
ture of biscuit meal, dried milk, soya bean meal and maw
seed; Natural food - seeds; cuttle bone necessary for
beak trimming purposes, and grit should be provided as a
digestive aid and mineral supplement.
Should be available at all times.
& DiSeaSe Lice, flies, mites and ticks.
RrPPfiino Ł DP or in a
Dl CCUlllg ft ncdllilg
Special Costs
lexicological Studies
Usually four nestings a season; 2 to 5 egg'
Per nesting; incubation period of 12 to 13
dayg. young leave nest after 9 to 11 days;
young fledged after 45 days; difficult to
breed in captivity.
Highly territorial and migratory; hard to
breed in captivity; subject to parasitism.
2?s
Ecological Alternatives
Physiological Parameters
Blackbirds.
-------
RICHMONDENA CARDINALIS
Solid red-crested bird about 8 to 9 inches long (221).
life span of 3 to 6 yrs. in nature (321).
AV3Jl3bilitY Found in U.S. east of plains near towns, farms, roads,
* swamps, etc. (221).
Features
Readily available; one of the best "model" granivores.
Light-natural photoperiod; temperature-room,
Cage
Food
Water
Parasites & Disease
120x60x60cm wood and 12 mm mesh cage (317); 3x3x2" to
6x5x2' cages have been used with enclosed nest box (318);
61x41x38cm cage has been used for single bird (319) .
Pigeon chow (275) canary or millet seed (319);
bisquit meal, dried milk, soya bean meal, maw seed
(319); cuttle bone needed for beak trimming and grit
needed also (182, 319).
Ad libitum .
Lice, fleas, mites, and ticks (321).
Breeding & Rearing
Special Costs
Restrictions
Toxicological Studies
Cardinals become sexually mature at one year;
2 to 5 eggs per clutch; 4 to 5 clutches per
year (241); 12 to 13 day incubation; fledging
period is 45 days (288).
Highly territorial and protected by legis»
lation; does not adapt well to lab.
Ecological Alternatives Blackbirds,
Physiological Parameters
-------
TURDUS MIGRATORIUS
Long-legged, short-billed bird; body length - 8.5 - 10.5"
(221) ; longevity in wild - 1 to 3 years; longevity in I
captivity - 12 years.
Availability Found throughout United States (221).
P63tlir6S Song bird; monogamous mating; migratory and diurnal.
Light - 15L:9D photoperiod; temerature - 70° to 80°F.
jjapg 9 x 12 x 7' communal cage for 20-70 birds;
8 20 x 15 x 22" cage for 10 birds;
9.75 x 7.0 x 8.0" metabolism cage for one bird.
Food Beetles, true bugs, spiders, caterpillars, ants, small
hymenoptera are major food items; grasshoppers, crickets,
craneflies, moths, millipedes and snails are minor food!
Water Should be available at all times.
& UlSeaSe Hosts for mites and ticks.
Rroeriino Ł Dearino 3 to 4 eggs per clutch; usually 3 clutches pf
DICeUlllg ft nCdllllfc year (241); !3 to 16 day incubation period;
young are altricial and nidicolus; fledging
takes 13 days on the average.
Special Costs
ReStriCtlOIIS Territorial, hard to breed and maintain in
lab; protected by legislation.
Toxicologicai Studies
37, 326, 333, 334, 335, 336, 337, 338
Alternatives Vireos, warblers, goatsuckers.
Physiological Parameters
-------
STURNUS VULGARIS
Similar in shape to a Meadowlark; 7 1/2-8 1/2" body
length (221).
Common resident species throughout continental U.S.;
readily captured in Japanese mist nets.
Features
Laboratory
Mean annual adult mortality in wild, 51-58% (344) ;
largely insectivorous in the wild; not protected
by law.
Temperature -"room", 70°-80°F (329); light-natural
photoperiod-
Cage
Food
Outdoor individual holding cages 2x1x1"; (342)
Metabolism cages 9 3/4x7x7" (329).
Chick mash, ad libitum (329)
i -i i. Pr,Qtfiin Fiber Fat NFE Ash
pellets: 23.9? 5.6% 3.4 57.2 10%
mealworms:50.4% 4.9 34.9 6.7 3.2%
Ad libitum (329).
% highly
ib, consumption digestable
(g)/day) 3.4%
6gm
85%
Hater
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
Young altricial (344) ;first clutch 5.5*0.9,
second clutch 4.1-0.96 (344); Incubation -
12 days (344) ;Fledging - 21st day after
hatching (344).
Highly territorial; require separate cages
for each breeding pair (182).
^tnriioc Used in nationwide monitoring of levels of
31UUIB5 certain pesticides and pollutants (342, 347,
739, 740).
Alternatives Red~win
-------
PASSER DOMESTICUS
5 to 6 1/4" body length (221); life expectancy - 3
years with exceptions up to 12 years (331) .
Availability Found in great numbers around cities, towns, and
farms in the U.S. and Canada (221).
Familiar bird, distributed where man lives.
Light - natural photoperiod; temperature - "room".
n
w«gB
rOOu
7x7x7' cage - 50 to 100 birds for 4 to 6 weeks;
2x2x2' cage - 7 to 10 birds for 3 days; 6x9x6'
cage - individual test cage; all cages made of 1" mesh
poultry netting.
Pigeon chow checkers and chick starter mash; grit needs
as a digestive aid and mineral supplement.
Ad libitum
& Disease
Lice, flies, mites and ticks; serves as
a carrier of encephalitis.
ing & Rearing
Special Costs
3 to 6 eggs per clutch; ll day incubatiott
period, 14 day fledging period; 2 to 4
clutches per breeding season.
Birds are highly territorial and require
separate cages for each breeding pair.
lexicological Studies
210, 275, 335, 339, 3*2
Ecological Alternatives
Physiological Parameters
-------
CALLUS CALLUS
Tlie common domestic chicken.
nY3ll3Dlllty Can be obtained in large numbers from commercial breeders
in any part of the country.
Large yet easily handled; easily obtainable; continuous
nesters; can be housed in limited space and cheaply.
Light - 14L:10D photoperiod; temperature - 23.9°C;
Humidity - 45 to 78% depending on age of bird.
Cage
Food
30 x 20x 8" cage for 10 to 20 chicks; 15 x 10 x 8"
experimental cage for 10 to 50 chicks; individual and
brooder units with shavings, peat moss or commercial
litter are available commercially.
Commercial chow - ad libitum; grit needed for mineral
requirements.
Ad libitum.
Water
Parasites & Disease
Newcastle disease, bronchitus, fowl pox,
coccidiosis, other parasites common.
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity - 20 to 22 weeks; continuous
laying throughout most of year; incubation
period - 21 to 23 days.
Cannabalistic tendencies.
lexicological Studies
350, 355, 356, 358, 359, 360, 361, 362, 363, 365,
366, 367, 368, 369, 370, 371, 372, 373, 374, 375,
376, 377, 466, 619, 1199
Ecological Alternatives Jungle fowl, wild turkey.
Parameters Spontaneous motility in chick embryos (1200);
heart rate of chick embryo (357); monitoring
system for cardiac and gross motor activity
(378).
-------
COTURNIX COTURNIX
Small quail; weight - male, llOg. (90-120g.)/ female
130g. (110-150g.) (383).
Availability Exotic species; easy to breed in laboratories.
6 week reproductive cycle; cheap to breed and raise in
laboratory; small for easy handling in lab yet large
enough for good sample (406); representative of upland
game birds (199, 406).
LabOr 3tOry Light - 14L:10D photoperiod; relative humidity - 60 to
70%.
Cage
Food
3 x 6 x 1.5' with 1/2" mesh breeder cage for 5 males and
15 females (379); 12 x 12 x 7" cage by GOF Mfg. Co., P.Oi
8152, Savannah, GA 31400, recommended for breeding.
Seeds, commercial game bird chow.
WatBf Ad libitum.
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
Toxicological Studies
Sexual maturity - 6 weeks; recommended sexual
ratio- 1 cock:3 hens, 2 cocks:7 hens; laying
schedule - continuous all year approximately
200 to 300 laid per year with artificial H$
source; incubation time and temperature - 16
days±8 hours at about 100°F; Fertility - 90%
after 50 days (379); up to 4 generations per
year.
Migrating restlessness observed in April and
September under natural conditions.
L99, 249, 255, 293, 326, 371, 380, 382, 384,
385, 386, 389, 390, 391, 392, 393, 394, 395,
396, 397, 398, 399, 400, 401, 402, 403, 404,
406
Ecological Alternatives eobwhite
Physiological Parameters
-------
MELOPSITTACUS UNDULATUS
DeSCriptiOl) Small, slender bird; about 4 - 6"long,
Availability Native to tropical Australia; coiruucrcially available at
pet stores and distributors.
Easy to breed and maintain in the laboratory.
Light - natural photoperiod; temperature - "room".
Gage Standard breeding cages, nest boxes and bowls are avail-
able commercially.
lOOu Commerical chows; sunflower seeds supplement diet; iodine
necessary in diet; tree branches necessary for bill trim-
ming purposes.
KKater M libitum.
& DJSeaSE Fleas, ticks, lice and mites.
& Rp/milO Sexually mature at 6 months; will breed all
« neamig year round. 3 to 4 eggs per clutch, can have
as many as 6 eggs per clutch; 18 to 23 day
incubation period; fledged at 6 to 9 weeks.
Special Costs
Very noisy, sanitation problems common.
lexicological Studies
Ecological Alternatives parrot.
Physiological Parameters
-------
BLARINA BREVICAUDA
large shrew, head and body 3-4 inches; adult females
16.5 g, males 18. g (416) life expectancy: 20 months
(414).
Availability common in eastern U.S.A.; pit traps; some dealers.
Features
Laboratory:
Cage
Food
Water
Parasites &
Food and water must be constantly available. Active
day and night. Individual variation to temperment.
Odor may be problem. Possess poison glands (saliva).
Light - ?; Temperature - "room1; Realtive humidity -
? +70%; Air quality - watch amnonia and musk levels;
Sound - may be sensitive.
Density - 1, more with care; Size - variable; don't
jump, do dig; Material - variable, will climb screen;
Nest - provide container and lining or_ allow soil cham-
ber and provide lining; Bedding - sphagnum or soil,
3-6 inches deep; Sanitation - ?; Furniture - ?.
Ground horse or beef; canned dog food; earthworms; mice;
mealworms. CAUTION: Very sensitive to food shortages.
Feed ad libitum.
CAUTION: Sensitive to water shortages. Water ad libitui
UISedSe Sensitive to exposure, especially getting fur
wet. Mites. i
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity at 1-2 months. Gestation about l
21 days. Pcst-partum estrous - (?); Litter size
5-7; Litter number 2, possible 3 per year. Sea-
son - peak in April. Weaned - about 22 days.
Initial trapping program and transport, or
Consume less than 1/2 body weight in food/day.
Difficult to maintain in lab.
lexicological Studies
Ecological Alternatives
Physiological Parameters
Cryptotis
906
Sorex cinereus
-------
EPTESICUS FUSCUS
Large bat, head and body 3" to 4" forearm 1" to 2";
weight 10 to 16 g; life expectancy unknown.
AVail3ullity Available locally throughout U.S.; collected with
mist nets •
Nocturnal; usually solitary, sometimes colonial.
laboratory Light - 13L:11D photoperiod; temperature, 21 to 28°C;
relative humidity, 55-92%.
tage Density - 5 to 10 per cage; size is variable,
20 x 20 x 25 cm to 80 x 92 x 138 cm with roosting
box 80 x 92 x 57 cm; material - wood, hardware cloth
(bats hang from hardware cloth ceiling).
FOOd Young birds, mealworms and other insects, high
vitamin-protein mixtures-
Water Ad_ libitum.
Parasites & Disease Mites,
Breeding & Rearing Sexual maturity- 1 year; gestation - unknown
(delayed implantation); post-partum estrous -
no; litter size-usually two; litter number-
one per year; season - May through June
Special COStS Initial supply .
Restrictions Breedablllty in captivity unknown.
lexicological Studies 438
Alternatives Other insectivorous bats.
Physiological Parameters
-------
SYLVILAGUS FLORIDANUS
(S. AUDUBONI)
Description
Availability
Features
Cottontail rabbit; head and body 12 to 17"; tail
1 to 2"; weight, 1.5 to 4 Ibs; life expectancy -
over 2 yrs.
Abundant throughout brush, grasslands in Eastern
and Southern U. S. (deserts and foothills in South-
western U. S. ).
Active day and night, game animal.
Light-"natural photoperiod"; temperature-"room";
relative humidity - over 50%.
Cage
Food
Density - over 4; Size - variable (minimum 1 sq.
ft. per animal); Material - wood, plastic, metal,
fiberglass.
Green vegatation, commercial rabbit diet.
Water
Parasites & Disease
Ad libitum.
Tularemia.
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity - about 1 yr.; Gestation
26 to 30 days; Postpartumestrous - ?;
Litter size - 2 to 7; Litter number - 2
to 4 per year; Season - spring to fall.
Initial supply.
lexicological Studies
Ecological Alternatives Hares, voies.
Physiological Parameters
-------
DIPODOMYS ORDI
Large kangaroo rat, head and body 4" to 5"; weight,
43 to 70g; light-colored desert rodent with large
head and long tail, 5" to 6"; front legs weak, sal-
tatorial; fur-lined cheekpouches (412).
nVdlldDllliy Common in sandy soils throughout western plains and
deserts of U.S.; some dealers.
Features
Laboratory
Cage
Food
Water
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
lexicological Studies
Nocturnal, active throughout year, stores seeds,
burrows, temperament varies individually, often
docile and easy to handle.
Light- 12L-.12D; temperature- 24 to 28°C; relative
humidity-- very low (as in natural environment).
Density- 1, but depends on cage size; size- variable,
10 xll x!6" to 2 x2 x3'; breeding cage- 5'x7'x2f;
must be covered; material - variable, glass or metal
(will gnaw wood); nest- mailing tube, small boxes;
bedding, dried leaves, grass; substrate- dried leaves;
sand important for cleaning pelage.
Cereals, ground meat meal, various seeds; greens may
be important nutritionally and for proper breeding.
Water requirements negligible though may increase
if kangaroo rats become accustomed to drinking.
Sexual maturity 90 days; gestation- 29 to
32 days; post-partum estrous, unknown; lit-
ter size- 2 to 5; litter number- 1 to 2 per
year; season, May through June and late
summer - weaned 20 to 25 days.
Initial trapping program and transport;
supply.
Heteromyid rodents are very territorial;
colonial breeding may be a problem.
Ecological Alternatives
Physiological Parameters
s sp.; Perognathus hispidus;
us flavus .
454, 45?
-------
NEOTOMA LEPIDA
Desert wood rat, head and body 6" to 7", tail, hairy,
4.5" to 6.5"; weight, 100-17 Og. ; life expectancy - 5 yea
Locally common on southwestern deserts and rocky
slopes, some dealers-
Generally nocturnal
Light - normal photoperiod; temperature - 23°C+; relati\
humidity - 60 to 70%.
Density - depends on cage size; size - variable; materia
metal, glass, wire mesh; nest - cotton, grass, etc.
Seeds, fruits, acorns, cactus, food scraps
Ad libitum.
Parasites & Disease
Sexual maturity - 60 days; gestation - 30 to 36
days; post-partum estrous - unknown; litter }
size - 1 to 5 (avg. 3); litter frequency - ovei
4 per year; season' - warm months; weaned - 15 di
lOStS Initial acquisition of breeding stock. ;
Restrictions
lexicological Studies
Ecological Alternatives other
Physiological Parameters
-------
ORYZOMYS PALUSTRIS
small rat; head and body 4.5" to 5.5", tail 4.5" to 7";
weight 40 to 75g.; life expectancy - unknown.
Common in marshlands in Southeastern U. S.
fCatUreS Nocturnal, semiaquatic.
Light- natural photoperiod; temperature- "room";
relative humidity- unknown.
Cage Density- depends on cage size; size variable-
27x17x14" to 3'x8'xl5" cage with smaller compartments;
material- wire mesh.
Food Green vegetation, seeds, dog food.
Water Ad libitum.
Parasites & Disease
Ł Peering Sexual maturity - 50 days; gestation - 25 days;
» post-partum estrous - unknown; litter size -
1 to 7 (avg. 3.5); litter frequency - 3 to 5 per
year; season - warm months; weaned at 13 days.
Special COStS initial trapping program.
Restrictions
lexicological Studies
Alternatives Any small herbivore
Physiological Parameters
-------
SPERMOPHILUS BEECHEYI
DeSCriptiO(l Large ground squirrel, head and body 9" to 11";
tail 1.25" to 2.5"; weight 45-70 g.; life expectancy-
unknown-
Availability Locally abundant along Pacific coast.
Diurnal burrowing rodent, openland inhabitant.
Laboratory
Cage
Food
Light-normal photoperiod; temperature-"room";
relative humidity-over 50%.
Density-depends on cage size; size-variable, 12 ft. ;
packing crates with wire mesh, 4 to 10 ft metal sus-
pension cages; material-metal, wire mesh (wood not '
suitable); nest-boxes; bedding-cotton; substrate-
burlap sacks, wood shavings; furniture-exercise wheels;
sanitation-daily; breeding cage-4x8x2' of 1/2" wire mesh
nest box, 12x12x12".
Seeds, greens, fresh vegetables, dog food, raw meat,
lab chows-
Ad libitum.
Water
Parasites & Disease Mites.
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity-1 year; gestation-25 to
30 days; post-partum estrous-unknown;
litter size-4 to 15 (avg. 7); litter
frequency-1 year; season-usually March to
April; weaned-35 to 40 days.
Initial trapping program.
Possible social problems in colonies •
lexicological Studies
Alternatives Other ground squirrels, Eutamias
Physiological Parameters 1149, 1149
-------
EUTAMIAS MINIMUS
Small chipmunk, head and body 3.5" to 4.5"; tail, bushy,
3" to 4.5"; weight, 30-60g.; life expectancy-5 to 8 years.
Common throughout various habitats of west central
lowlands and upland U.S. and across most of Canada.
Diurnal; very active, alert, agile; climbs readily;
lives in burrows; stores seeds.
lahnrotnrw Light-normal photoperiod; temperature-"room"; relative
LdBQIdlUry humidity-over 25%.
Density-pair or more; size-variable, 2'x2'x2' must be
covered; material-glass, metal, wood .may be unsatisfactory;
Cage nest-small box, mailing tube; bedding-dried leaves,
c^iall twigs, grass; substrate-soil paper, wire mesh;
furniture-exercise wheel.
Cftfld Fruits, nuts, seeds insects, greens, chopped raw
uu meats, cod liver oil.
Ad libitum.
Parasites & Disease
Breeding & Rearing Sexual maturity-1 year; gestation-3t) days;
post-partum estrous-no; litter size-2 to 6;
litter number-1 per year; season-late spring;
weaned-25 days.
Special Costs
Restrictions Poor previous success at breeding in captivity.
lexicological Studies
Alternatives Tamias striatus;_Spermophilus sp.; Peromyscus sp.
Physiological Parameters
-------
TAMIAS STRIATUS
Description
Availability
Woodland terrestrial squirrel; head and body 5" to
6"; tail bushy, 3" to 4"; weight, 65 to 130g.; life
expectancy-8 years.
Locally common throughout deciduous forests of
eastern North America.
Features
Diurnal, quick, alert, hibernates in winter.
Laboratory
Cage
Food
Light-500 to 2400 lux; temperature-20° to 25°C;
relative humidity-70% .
Density-1, possible more; size-2x2x2'; material-
glass, metal, wire mesh; nest-box, flower post,
mailing tube; bedding-cotton, grass dried leaves;
furniture-exercise wheel, climbing poles.
Seeds, corns, fruits, insects.
Ad libitum.
Water
Parasites & Disease
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity-6 months to 1 year; gestation-
31 days; post-partum estrous-no; litter
size-2 to 8 (avg. 4.5); litter frequency-
2 per year; season-April and August.
Initial trapping or supply.
Unsociable habits may cause difficulty in
group cages.
lexicological Studies
Ecological Alternatives
Physiological Parameters
squirrels.
-------
MICROTUS PENNSYLVANICUS
Availability
Features
Meadow vole; head and body 3.5" to 5"; tail 1.5" to
2.5"; weight 30 to 70g.; life expectancy-unknown.
Abundant in low moist areas, grasslands of all
types, orchards, etc. across northern U. S.
and Canada.
Active day or night.
Laboratory
Light-natural photoperiod; temperature-"room" ;
relative humidity-over 10%.
Cage
Food
Density-depends on cage size; size-12x8x5" for breeding
pair, 6x25x2' with 48 2x3x6" nest boxes; material-
wire mesh, metal, plastic, fiberglass; nest-box 2x3x6";
bedding-cotton, fine grass; substrate-peat moss, sand
wood shavings, corn husks; furniture-exercise wheel
Grasses, seeds, bark; greens may cause diarrhea.
Water ^ libitum.
Parasites & Disease Mites' fleas-
Breeding & Rearing
Special Costs
Restrictiois
Sexual ma'turity-40 to 50 days; gestation-
21 days; post-partum estrous-yes; litter size-
1 to 9 (avg. 3 to 5); litter frequency-
monthly; weaning-21 days .
Initial supply-
lexicological Studies
Ecological Alternatives other voles*
Physiological Parameters
-------
PEROMYSCUS MANICULATUS
Typical mouse, brown to gray back and sides, white under
parts; head and body 3" to 4"; tail 2" to 5"; weight
19 to 35g.; life expectancy-5 to 8 years.
Availability Common to abundant throughout the U. S. except the
Southwest; numerous dealers.
Primarily nocturnal, some diurnal activity;
limited climbing.
Laboratory
Cafe
Food
Light-unknown; temperature-"room"; relative
humidity-over 10%.
Density-dependent on cage size; size-variable, 9x7x5",
or 15x14x12"; material-wire mesh, wood, metal; nest-
fruit jar on its side; bedding-cotton, dried leaves;
substrate-woodchips, sand; furniture-exercise wheel.
Commercial diets available, lettuce, rolled oats,
meat scraps, seeds, fruit.
Water
&
Ad libitum
Mites, ticks, fleas, nematodes.
Breeding & Rearing
Special Costs
Restriction
Sexual maturity-60 days; -gestation-25 days;
post-partum estrous-yes; litter size-3 to
6; litter frequency-5 to 9 per year; season-
year round; weaned-25 days.
Initial supply.
lexicological Studies 167> 348' see, si?
Alternatives
Physiological Parameters
sma11 ominovorous rodent,
-------
MUSTEIA VISON
Description
Carnivorous mammal with long slender bodies and short
legs; usually dark brown with white throat patch; total
length 20" to 25" (Male) (1201); 17" to 20" (Female);
weight-1-1/2 to 3 Ibs. (Male); 1-1/4 to 2-2/5 Ibs. (Fe-
male) ; life expectancy-3 years-
Availability Available from pelt industry farming; numerous mink
ranches in U.S. raising 5000 or more aniiaals per year
(1202); have been trapped by professional trappers; range
entire N. American continent except southeastern U.S.
Aggressive; chiefly nocturnal; at home on land or in
water; usually lives in banks of stream or lake or under
tree stumps or logs; usually solitary.
Light-natural photoperiod; temperature-"room" ;
relative humidity-unknown-
Laboratory
Cage
Density-l; observation cage-40x60x40cm.
Food
Cotton rats; raw meat; mice; fish; muskrat; frogs;
special mink food when sold commercially
mixed diet of horsemeat, bonefish, ocean fish, liver
and cereal.
Ad libitum.
Ł Hicaaco
6 UlSCdSe
& Rearing
Special Costs
Restrictions
Aleutian disease (viral disease causing lesions);
Chediak-Higashi syndrome, leucodystrophy,
hemivertebrae, Ehlers-Danlos syndrome,
encephalopathy
Sexual maturity-1 year; gestation-40 to 75
days; litter size-2 to 10 per year; litter
frequency-2 per year; season-March to April.
Initial supply.
Very nervous; can be vicious, will fight
cage-mates•
lexicological Studies 579
Ecological Alternatives
Physiological Paraaeters
Other Mustelidae..
-------
MYOCASTOR COYPUS
Muskrat-like; head and body 22 to 25"; tail 12" to 17";
weight 15 to 20 Ib.; hind feet webbed; life
expectancy - 12 years -
Availability Locally abundant in southern Mississippi River Valley,
* Texas.
Nocturnal, feeder on aquatic plants, burrows in banks.
Light- natural photoperiod; temperature- about 70 F;
relative humidity- over 50%,
CaŁ6 Density - 2; size - variable (3x2x1'); material -
metal, hardware cloth; nest (for breeding)
20x2Ocm; bedding - straw; substrate - sawdust.
Sugarbeets, carrots, cabbage, grass, bread, cereals
Ad libitum.
& Disease Salmonella, tuberculosis, tularaemia, ticks,
fleas (550).
0 ., . Sexual maturity - 5 to 9 months; gestation -
& Rearing avg. 128 days; post-partum estrous - yes;
litter size - 2 to 11; litter number - over
2 per yr . ; season - year round ; weaned -
24 hours.
LOStS initial supply, metal cages
Metal cages only; ponds for swimming may be
necessary.
lexicological Studies
Ecological Alternatives voies,
Physiological Parameters
-------
MUSTELA PUTORIU5
Description
Maitabitity
Carnivore; head and body about 20", tail about 10",
bushy; weight 400-3,500g; life expectancy - 3 to over
5 years.
Suppliers.
Features
Domestic form, long fur, easily handled in lab.
Laboratory
Light - natural photoperiod; temperature- 10° to 30°C
Gage
Food
Density - dependent on cage size; size - variable
(60cm2 for 3 females or 2 males; material - wood
metal, concrete; bedding - wood shavings.
Meat, bread, milk, commercial diets.
&
Ad libitum.
Pneumonia, gastric ulcers, virae, bacteria,
mites, ticks -
Breeding & Rearing
Special Costs
Restrictions
Sexual maturity - 4 to 5 mos.; gestation - 51
to 43 days; post-partum estrous - unknown; litter
size - 3 to 15; litter number - 2 per yr.;
season - spring to July, August.
Initial supply.
lexicological Studies
Ecological Alternatives weasels
Physiological Paraaeters 565
-------
VULPES VULPES
Red fox; head and body 22-25"; tail 14-16", wt. 30-
80 Ib.
Availability Most of U. S., fur ranchers.
Doglike animal, reddish fur; bushy tail, active day
and night.
Light-natural photoperiod; Temperature - "room";
Relative humidity - 50%.
Cage Density-variable; size-large enough for exercise;
Material-wood, metal, wire mesh; Nest-dog house;
Bedding-straw.
rOOd Fish, cooked cereals, berries, dog food.
Water Ad libitum .
Parasites & Disease Ticks, fleas/ rables.
Breeding & Rearing Sexual maturity - 1 yr.; Gestation - about
51 days; Postpartumestrous - No; Litter size
4 to 9: Litter number - 1/yr. Season - March
to April.
Special Costs
Restrictions
Toxicological Studies 560
Ecological Alternatives coyotes.
Physiological Parameters
-------
CANIS FAMILIARIS
Domestic dog; head and body 12 to 45"; tail 5 to 25",
life span - to 15 years.
nTalldUIIll) DOg pounds, breeders, supply houses.
Features
Docile, domestic.
Laboratory Light-"natural photoperiod"; temperature-"room" ,
relative humidity-60-80%.
Cage
Food
Density 1 to 5; size-depends on size (from Ito3 sq. ft.
per individual); material-wood, metal, concrete;
bedding-shavings, rugs; furniture-nest box for mothers.
Commercial dog food.
Water
Ad libitum .
Ticks, fleas, tapeworms, heartworms
Breeding & Rearing
Special Costs
Restrictiois
Sexual maturity 1 yr.; gestation-9 wks.;
post partem estrous-no; litter size-3 to 10;
litter number-1 to 2 per year; weaned-about
28 days •
Purebred animals are expensive.
lexicological Studies 822
Ecological Alternatives Domestic cats.
Physiological Parameters
-------
SUS SCROFA - miniature swine
Small pig; weight - 100 to 200 pounds (1203).
Through Sinclair Comparative Research Farm, Univer-
sity of Missouri, Columbia, MO 65201; $70-$100 per
animal.
Breeds and maintains fairly easily in lab; good
"model" animal; economically important (1203) ;
veteran test animal.
Light - natural photoperiod; temperature - "room"
8 x 12" fenced-in holding pens, 3 to 4 sq. ft. per
CaPB animal; 6 animals per cage(1203); experiment enclo-
sure 2 x 3.5' (1203).
Various mixtures of cornstarch, sugar, soybean meal,
FOOd corn ground shell, meat and bone meal, alfalfa meal,
wheat shorts, dical phosphate, limestone, salt, vita-
min premix once a day (1204);
Should be available at all times
Parasites & Disease
ing & Rearing 114 day gestation period; 3 litters per
year.
Special COStS Cage cleaning.
Restrictions
Toxicological Studies 612
Ecological Alternatives °°9' other Pigs<
Physiological Paraneters
-------
OVIS - domestic sheep
Domestic ungulate wool producer
Availability
Features
Laboratory
Has wide distribution in U.S.: found in almost all
agricultural areas.
Plentiful; adapts to lab easily; not too expensive;
model ruminant; can tolerate surgical procedures;
economically important; easy to maintain and breed;
behaves well; good animals for physiological and
behavioral studies (613).
Light-natural photoperiod; temperature-"room".
Cage
Food
Any type of large outdoor or indoor pen is sufficient
as long as it gives each animal 64 sq. ft. of
territory (613).
Hay supplemented by corn and protein (613).
Ad libitum
Water
Parasites & Disease
Much information available; wide variety
of parasites and diseases.
& Rearing
Special Costs
Restrictions
Usually breed in fall and give birth in
early spring to early summer; 1 to 2
offspring per yr. (613).
ToxiCOlOgical Studies 590, 592, eia, eie, si?, sis, 619, 620,
627, o2o,
Ecological Alternatives other ruminants.
Physiological Parameters
-------
ODOCOILEUS sp.
H_. • _»;-„ Deer; height 3" to3.5'; weight-females 100 to 250
UcSulpllOn Ibs., males, 75-400 Ibs.; carry antlers during part
of year, large tail; life expectancy-15 years.
1 "I IT!'* White-tail deer found through N. America except Calif.,
AVail30lllty Nev. , Utah; mule deer found in N. America west of Great
Plains; both species abundant locally.
Active day and night, seen often in herds; very alert
creatures; generally found in forested regions.
Outdoor enclosures most suitable; cover from sun and
cold essential.
p Any enclosure of suitable areal extent and height
«3ge will suffice; walls-over 8 feet.
Pgnij Twigs, berries, acorns, apples, cereals, dairy chow.
Ad libitum
Water
Parasites & Disease
n .. « Q . Sexual maturity-1 year; gestation-6 to 7
Breeding v Hearing months; post-partum estrous-no; litter size-
1 (sometimes 2); litter number-1 per year;
season-breed in fall; birth in spring.
Soecial GOStS Large enclosures, initial supply.
ReStnCiiORS Large size, long generation time.
lexicological Studies 625' 631
Ecological Alternatives ^bbits,
Physiological Parameters
-------
SAIMIRI SCIUREUS
Sma11 New World monkey; prehensile tail; head and
body 9 to 12"; weight - 2 to 4 Ibs.
Imported from Colombia, S.A.; available through Primate
Imports Corporation, 34 Munson St., Port Washington, L.I.,
N.Y. (613, 1205).
Features
L3iOr3tQr]f Light-natural photoperiod; temperature - 74°F (1205).
15x20x30" individual cage (about the same price as a
dog cage (613); 24x24x28" metal and plastic test cage
(1205) ; 14x19x24" individual maintenance cages (1205).
Commercially prepared chow plus oranges, fresh fruit
and other supplements (613).
Water
&
Worm parasites common; enteric and respiratory
problems common (613) •
Breeding & Rearing
16° to 17° day gestation period.
Special Costs
Dartr jrtifliC
Animals fairly expensive and rare; will bite
lexicological Studies 613' 1205
Ecological Alternatives
Physiological Parameters
Special Reference The Squirrel Monkey, Cooper and Rosenblum
-------
DIDELPHIS MARSUPIALIS
Marsupial; head and body 15" to 20"; tail 9" to 20";
weight, 9 to 13 Ibs.; life expectancy-over 7 years.
Locally abundant throughout most of U. S.
F62tUrtS Nocturnal, slow moving, will act dead if molested,
odorous prehensile tail and opposable first digits.
Light-natural photoperiod; temperature-over 15°C;
relative humidity-over 50%.
Density-2; size-20x22x!5" and larger; material-
Cafe galvanized metal; nest boxes-5x5x3' in large
outdoor cages.
Fruits, vegetables, eggs, carrion, dog pellets, table s<
Ad libitum.
4 Disease Many and varied diseases (641, 642, 647, 649
650, 651, 653).
Sexual maturity-1 year; gestation-13 days,, j
RroeHino Ł fiiarintr followed by 2 months in pouch of mother; posi^
DIBEUIIIg ft RBdMIIg partum estrous-no; litter size-up to 14j'li^
number-2 per year possible; season-warm montM
weaned-2 months; best results in large,
outdoor pens.
Special Costs initial suPPiy.
Opossums are very susceptible to disease.
lexicological Studies
Ecological Alternatives
Physiological Parameters esi, 1023, 1112
-------
DASYPUS NOVEMjCINCTUS
Armadillo; armored body; head and body 16"; tail
15,,; wieght 8 to 17 lbs>; life expectancy 6 years.
Southern U. S., Texas area.
Features
Nocturnal, digger, polyembryonous.
Laboratory
Cage
Light-natural photoperiod; temperature-over 20°C;
relative humidity-unknown.
Density-unknown; size-9x9x4' and 18"x4'x4' compartments
in one cage; material-concrete floor plywood walls;
nest-unknown; substrate-cedar shavings; furniture-pool
of water, logs for scratching.
Food
Berries, fruits, roots, dog food, raw beef, milk,
eggs, liver.
Water
Ad libitum.
infection beneath dermal plates is common
Breeding & Rearing
Special Costs
Restrictiois
lexicological Studies
Sexual maturity-unknown; gestation-(delayed
implantation); post-partum estrous-no; litter
size-1 to 12; litter number-1 per year;
season-breed in fall-
Initial supply; special caging-
Armadillos easily injure themselves on metal,
screen, etc., and armor may restrict certain
methods.
Alternatives Other armadillo species.
Physiological Parameters 662, 663, 664, 665, 666
-------
SECTION VII
BIBLIOGRAPHY
1. Rudd, R. L. 1964. Pesticides arid the Living j
Landscape. The University of Wisconsin Press,
Madison, Wisconsin j
2. Animal Welfare Act, Statutes at Large,
80 (1966) PL 89-543
3. Animal Welfare Act, Statutes at Large,
84 (1970) PL 91-579
4. Reitz, A. W., Jr. 1972. Environmental Law,
North American International, Washington, D. C.
5. Chance, M. R. A. 1947. Factors influencing the
toxicity of syrnpathomimetic amines to solitary mice.
J. Phar. and Exp. Ther. 89:289
6. Webb, R. E., Hartgrove, R. W., Randolph, W. C., et
al. 1973. Toxicity studies in endrin-susceptible
and resistant strains of pine mice. Tox. and App.
Phar. 25:42-47
7. National Academy of Sciences. 1971. Animals
for Research. Institute of Laboratory Animal
Resources, National Research Council, Washington,
D. C.
8. Kendeigh, S. C. 1964. Animal Ecology. Prentice-
Hall, Englewood Cliffs, N. J.
9. Ozburn, G. W., and Morrison, F. 0. 1962. Development
of a DDT-tolerant strain of laboratory mice. Nature
196:(4858):1009
10. Georghiou, G. P. 1972. The evolution of resistance
to pesticides, pp 133-167 in Annual Review of
Ecology and Systematic^, Vol. 3, ed. by R. F.
Johnston.Annual Reviews, Inc. Palo Alto
11. Hart, L. G., et al. 1963. Stimulatory effects of
chlordane on Hepatic microsomal drug metabolism in
the rat. Tox. App. Pharm. 5:371
12. Cory, L., Fjeld, P., and Serat, W. 1971. Environ-
mental DDT and the genetics of natural populations.
Nature 229:128-130
-------
13. Sterburg, J., and Kearns, C. W. 1952. The
presence of toxins other than DDT in the blood
of DDT-poisoned roaches. Science 116:144
14. Sternburg, J., et al. 1950. Absorption and
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