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~ Introctuction
A. GENERAL COMMENTS
Few animals are as valuable for experimental investigations as amphibians
(Kawamura and Nishioka, 1973~: They are ubiquitous in nature, transi-
tory between aquatic and terrestrial life, reasonable in size, ectothermic
and the only naked vertebrates, accessible to direct examination through
all developmental periods, and possess genetic mechanisms amenable to
study by biological and mechanical manipulation.
Traditionally, investigators collected these animals in the wild and com-
pleted their experiments within days. In spite of their ancient history as
experimental animals, care and management of these animals is rudimen-
tary, and few established and tested guidelines for their husbandry are
available (Boterenbrood, 1966; Frazer, 1966; Bardach et al., 1972~. With
few exceptions (e.g., mink, monkeys, hamsters, gerbils), laboratory verte-
brates have been chosen from among those animals previously domesti-
cated for agricultural purposes (e.g., chickens, goats, rabbits, swine), for
pets (e.g., cats, dogs), or have been symbionts of man (e.g., mice and rats).
This is not true of amphibians. While enthusiasts have maintained am-
phibians in limited numbers, no amphibian, except the axolotl, Ambystoma
mexicanum, has a history of mass culture for repeated generations.
B. THE NEED
1. The Demand
Factors that stimulate a demand for amphibians are their utility for current
research problems, the increased cost of avian and mammalian research ani-
mals, and the increased use of lining material in high school and college
instructional laboratories.
Best estimates indicate that 20 million preserved and living amphibians
1
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3
were used for educational purposes alone in the United States in 1971-
1972. Recent studies by the Institute of Laboratory Animal Resources
(ALAR) report the use of 2 million amphibians per year in research labora-
tories (ILAR News, 1972~. Only rodents exceed this figure. Birds exceed
it only if the number of incubated eggs are included in the tally. This
heavy use of amphibians escapes the casual observer, because these animals
are either used within days of their arrival in a research laboratory or lost
prior to the completion of experiments due to inadequate maintenance.
2. The Supply
a. The Natural Resource
Although interest in amphibian resources is long standing (Wright, 1920),
accurate figures on the status of amphibians in nature are unavailable for
most American populations. This question has recently assumed great in-
terest because of the apparent short supply and diseased state of amphib"
fans collected from nature (Maugh, 1972; anonymous, 1973~. American
amphibian dealers find it increasingly difficult to identify high-density pop-
ulations that permit economical collection of animals. Informal surveys
among herpetologists and others interested in amphibians suggest that in
recent times American amphibian populations have decreased significantly.
The best current data dealing with population reduction have been
collected for British anurans (Cooke, 1972~. This survey, based on ques-
tionnaires distributed to educational authorities (schools survey) and pro-
fessional biologists (breeding sites survey), demonstrated that the changes
in these populations were most significantly affected by habitat destruc-
tion and were directly proportional to the density of human populations
and to increasing industrialization (Tables 1, 2, and 3~.
Amphibian populations fluctuate drastically in response to cataclysmic
environmental factors such as food shortage, drought, and early winter. In
addition, the problem of short supply is aggravated by large losses that oc-
cur during shipment and holding periods (Gibbs e' al., 1971~. However,
no clear evidence exists that the survival of any of the amphibian species
commonly used for investigative purposes is endangered by collection
practices. The high reproductive potential of these amphibians assures
that, given optimal environmental and climatic conditions, the popula-
tions rapidly regenerate.
b. Artificial Culture
The demand described above and the increasing difficulty of satisfying
these demands from natural sources has led to a renewed interest in the
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Representative terms from entire chapter:
laboratory lines
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7
artificial culture of amphibians. Serious efforts are currently being made
toward this end. However, costs remain high and the quantity of avail-
able cultured animals remains inadequate to meet the demand. As the
cost of collecting the natural supply increases, artificial culture will cer-
tainly become economically feasible, or even necessary, to maintain the
supply of healthy amphibians for at least scientific users.
3. Recommendations
We recommend that
a. A survey of the natural amphibian resources, parallel to that con-
ducted in England (Cooke, 1972), be conducted on the North American
continent to allow adequate evaluation of the supply base.
b. Increasing efforts be made to develop the culture of amphibians,
both for educational and research purposes.
c. Improvements in the care and management of amphibians be insti-
tuted as early as possible to minimize the losses that now occur between
the supply and the ultimate use of amphibians and to maximize the
quality of research animals.
C. USE RS OF AMPH IBIANS
Amphibian users are best described by the type of agency Mat supports
their work (Table 4), the location of the research (Table 5), and the topics
under investigation (Table 6~. These tables were compiled from Science In-
formation Exchange Notices of Research Projects, active as of January
1969, and were originally published in Nace (1970) where they appear
with more detail and discussion.
Federal, state, and foundation agencies funded 277 projects relevant to
this document (Table 4~. In addition, much research on amphibians is con-
ducted on low-budget projects. Although there is no accurate method of
estimating the numbers of such projects, reviews of the index journals,
including Dissertation Abstracts, would reveal these numbers. At the Uni-
versity of Michigan and at the time in question, however, for each listed
project about 15 investigators were conducting research involving the use
of amphibians on unlisted projects. This 1: 15 ratio of listed to unlisted
projects may be conservative when it is recalled that many investigators
at smaller colleges not funded by nationwide granting agencies use am-
phibians. Thus 3,000-4,000 amphibian projects may be active, a figure,
when divided into the ILAR survey on amphibian use, representing 500-
600 amphibians used (received) per project. This figure seems high for
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11
many projects, but may be a reasonable average when known projects
using 10 times that number are considered. Also, pituitaries from 3 to 10
animals are used for the induced ovulation of a single female frog, and a
conservative estimate suggests no more than 50 percent of the amphibians
received for research purposes survive the period between receipt and use.
D. LABORATORY-DEFINED AMPHIBIANS
Defined laboratory amphibians (Gay, 1971; Committee on Animal Nutri-
tion, 1972), though essential for high-quality investigations, remain largely
a hope for the future. To date, the best definition (nomenclature described
in Chapter III) has been attained with the axolotl (Boterenbrood, 1966~.
Some colonies of laboratory amphibians-such as Xenopus in the labora-
tory of Fischberg (Blackler and Fischberg, 1968) and Pleurodeles in the
laboratories of Gallien (Guillet et al., 1971) and Beetschen (1971)-do
exist and meet the criteria defined for laboratory-reared and laboratory-
bred animals; however, these have not attained the status of defined lab-
oratory lines. For Rana pipiens efforts are now being made to establish
laboratory lines (N. ace et al., 1965, 1966; Nace, 1968, 1970; Nace and
Richards, 1969, 1972a,b,c), but animals in these lines are not yet avail-
able in significant numbers.
The importance of nomenclature cannot be overstressed during this
period when defined strains of amphibians are still under initial develop-
ment. Any nomenclature that is adopted should include a designation of
the laboratory where the animals were developed and a key to the criteria
satisfied. The geographic designation should be as precise as possible. It
should specify a geographic region in sufficient detail to suggest the
parent population.
It is hoped that those strains currently under development will soon
become sufficiently well defined to justify publication of a standard no-
menclature and growth tables as exemplified by those listings that have
appeared for mice (Staats, 1972) and other laboratory animals (Poiley,
1972~. Since the laboratory use of amphibians will continue to depend on
animals from wild populations, terminology to designate the history of
such animals must be introduced. These terms (defined in Chapter III,
Section B) include wild, wild-caught nonconditioned, wild-caught condi-
tioned, laboratory-reared, and laboratory-bred. Types of laboratory-
reared and laboratory-bred populations and lines are defined in Chapter
III, Section C.
