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OCR for page 108
V! ~ ~ Records ant'
Information Control
A. IDENTIFICATION OF INDIVIDUALS
1. Numbering
The identification of individual animals-often necessary for the manage-
ment of experimental animals-is mandatory in breeding and genetic pro-
cedures. Several techniques can be used to identify individual amphibians.
Selection depends on the species and the developmental stage. Ideally, the
system of choice would allow identification of individual animals at all
stages of development from the time of fertilization through larval and
postmetamorphic development. However, aside from isolation in indivi-
dual enclosures, no system adequate to accomplish this objective is avail-
able. In practice, a clutch of eggs or a shipment of animals may be given
a serial identification number such as 35,000 with subsequent numbers in
the series reserved in proportion to the number of clutch or shipment
members to which individual numbers will be assigned. As animals are
assigned for specific experimental purposes, the experimental group or
individual may be given numbers from among those reserved. Thus,
35,XXX identifies the clutch or shipment; the hundred's digit may be used
to identify groups and the ten's and unit's digits to identify individuals.
Serial numbers that do not include letters are recommended to facilitate
adaptation to computer techniques, should these be found useful (see Sec-
tion B below). Identification numbers may be written in waterproof ink on
waterproof labels that, when appropriate, may be easily transferred from
one enclosure to another. In the Amphibian Facility of the University of
Michigan, it has been found useful to assign unique group numbers to
members of a clutch or shipment that share an enclosure. This permits
108
OCR for page 109
109
tracing enclosure mates and identifying individuals with common environ-
mental experiences.
2. Tattooing
Tatooing by ink injection techniques has proved successful. It is particu-
larly useful for larvae with well-developed tail fins, although its use is not
limited to them. in this technique the animal is lightly anesthetized (see
Chapter IX, Section F) and a fine needle attached to a hypodermic syringe
containing tattoo ink is inserted into the tail fin or into the skin. The ink
is injected as the needle is withdrawn, leaving a "bar" of ink. By using inks
of several colors and forming a series of parallel bars, unique identification
codes may be constructed. Recently, several useful variations on this prin-
ciple have been published (Woolley, 1973~.
Tattooing by the use of vibrating needles, as used in human tattooing,
cannot be applied to larvae or small postmetamorphic animals; in fact,
this method can only be used for larger postmetamorphic animals and
even then must be renewed frequently. R. pipiens and bullfrogs retain
such a tattoo for only 3-4 months. Because these and other amphibians
shed their skin, it is difficult to assure that the needle-driven tattoo pene-
trates into skin layers that are not lost.
3. Branding
Wolf and Hedrick (1971) describe "chemical branding" as a procedure for
marking Xenopus; these are permanent for at least a year. Using a cotton
swab dipped in a solution of 0.5 percent amido Schwartz in 7 percent
acetic acid, label figures are formed on the back of an animal after mucous
secretions are removed by repeated wiping with paper tissues. After 1 min
of contact the animals are returned to an enclosure. The dye provides a
temporary identification until scar tissue is formed a few days later in
response to the acid treatment.
The cold branding technique (Farrell and Johnson, 1973) has proved
satisfactory for adult R. pipiens and R. catesbeiana and may be applied
to the dorsal surface where the label can be easily read. The advantage of
this technique over tattooing is the longer period of label retention. How-
ever, cold branding also disappears with time and cannot be applied to
juvenile animals. Tattooing and cold branding are preferable to heat brand-
ing because they result in less trauma to the subject.
Urodeles can be tattooed or branded at relatively early stages if it is
done under light anesthesia (see Chapter IX, Section F). Such markings
are lost, however, as the skin is shed and must be renewed periodically.
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110
4. Toe Clipping
Toe clipping is an effective technique for amphibian identification and is
appropriate for most anurans that do not readily regenerate lost digits.
However, it is inadequate for urodeles and Xenopus in which the regen-
eration process replaces the removed digits, unless the regeneration is in-
hibited by treatment with berilium nitrate (Heatwole, 1961~.
Ranidae may be toe clipped shortly after metamorphosis using light
anesthesia and cuticle scissors. Although older R. pipiens may be toe
clipped with scissors, mature R. catesbeiana, because of their size, may
bleed excessively and danger of local infection occurs when toes are
clipped in this manner. However, toe clipping large anurans may be ac-
complished by using an appropriate cautery knife or loop to seal the
wound at the time of amputation. A coding system for toe clipping based
on the system for punching edge coded cards and that allows 9,999 in-
dividuals to be distinguished with only two amputations per foot is
illustrated in Figure 24.
5. Other Marking Systems
Various systems for labeling amphibians by the insertion of plastic rings
and other devices have been attempted but without notable success.
4
7~'
1, 000's
Frog's Left
ADPLaNTAR VIEW
(Dorsal )
FORE 4
CLIP TO ~J
DOTTED LINE ~/
Frog's Right
IO's
/\20,000 10,000l?(! ~
Tr9 ~'/1 A \
HIND
PLANTaR VIEW
(Ventral)
units
30,000 · BOTH 20,000 AND 10,000 CLIPPED
ABOVE 39,999, TREAT AS FROM I TO 39,999
FIGURE 24 Toe clip conventions
(Nace et a/., 1973; reproduced with
permission from the American
Zoo/ogist).
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111
6. Drawings and Photographs
The most foolproof method of identifying individual amphibians is the
use of photographs that record the details of natural disruptive patterning.
Figures 25, 26, and 27 illustrate disruptive patterning in R. pipiens and
how these patterns are coded for identification purposes in the Amphibian
Facility of the University of Michigan (Nace et al., 1973~. Similar classifi-
cation of the patterns of other amphibians is possible, but has not as yet
been completed.
In laboratories with small numbers of animals, it is adequate to prepare
duplicated outline drawings of the amphibian and to fill in these outlines
with drawings of the disruptive patterning. This technique, however, is too
time-consuming for a large colony of animals; thus, photographs are rec-
ommended. Using the classification system, it is possible to identify quickly
the class of patterns to which an animal belongs and then, by comparison
with the appropriate specific photographs or drawings, to identify the
individual animal. When the pattern classification and recorded pattern
are used in conjunction with other characteristics-such as sex, last recorded
snout-vent length, and other-unique characteristics-a very large number of
individuals may be identified.
B. INFORMATION CONTROL SYSTEMS
The management of animal colonies or of data collected in even small col-
onies becomes increasingly complex as the numbers of animals or the his"
tory of the colony increases. This problem can be greatly alleviated by the
adoption of computer-based techniques. Recent improvements in available
computer programs and time-sharing techniques, even over long-distance
telephonic connections, permit even those inexperienced in computer use
to adopt these procedures readily. Although they may be adopted at any"
time in the history of a colony, the earlier they are used, the greater the
economy that is realized. Current technology is sufficiently advanced that
standardization of the data base or of the software is not necessary. It is
sufficient to record the available data in machine-readable form; the com-
puter itself can restructure the data in accordance with the requirements
of specific current or future software.
A computer-based system currently in use for the management of am-
phibians is described in some detail in Nace et al. (19731. This system is
in the public domain and accessible to long-distance users. Other systems
that may be adaptable to users of amphibians include the ARE system
(Laboratory Research Enterprise, Inc., Kalamazoo, Mich.) for Beagles
and a system used for microorganisms (Bachmann et al., 1973~.
OCR for page 112
112
NO SPOTS ON ~ IDLI NE
6-10
NO.
OF
SPOTS
HEAD PATTERNS
r ~
!1 ~
i I ,0_ NO ~ ~SPOT ON
if ~\ SPOTS ~SNOUT ONLY I
, i
r
' <2 Eye Spots Unequol byFoctorof Two A _ I
SPOT ON SPOT ON |
I `~\ SNOUT | SNOUT
EYE RIGHT LEFT t3OTH
SPOTS NO SPOT
~5 ^6 -7
`~. ~ON SNOUT
EYE RIGHT LEFT BOTH
SPOTS ~ 8 ~ 9 ^10 SPOT ON
p~t [ ~{~\ SNOUT
OTHER EYE SPOT SNOUT SPOT
_ 15; ~3/4 of spot 3/4 of spot between
Em . ~on eye or posterior to nostrils
_ ^~. ond anterior to eve
BODY PATTERN
SPOTS ON MIDLINE
134
['
.i' - N ~
i \d
REGU LAR
it'
lb.'
·,'
L'' at.
I PREGULAR
BODY
PATTERN
SPOTS
BETWEEN
PLICAE
POSTERIOR
TO EYE
ANTERIOR
TO VENT
15
OTH ER
(NO EXAMPLE)
FIGURE 25 Fiana pipiens head and body pattern code characteristics. The num-
bers appearing by each pattern in Figures 25 and 26 are used to write an identifica-
tion formula. Thus, 7~24 identifies the animal shown with body pattern "6"
Mace eta/., 1973; reproduced with permission from the American Zoologist).
OCR for page 113
113
NO MARKING
tiP
BACKGROUND PaTTERNS
`-' 1
Hi''
ir~.'lii
i,,,'
FEW,COUNTABLE (~100)
,PINPOIN~
[,..'
~ ~ text
MANY (~100)
PINPOINT, DISCRETE
OR FUSED
l
bit
lo.
A
,~ ~
COMPLEX COMPLEX
REGULAR IRREGULAR
SCULPTI NG SCU LPTI NO
(os Kandlyoh i )
OTHER PATTERNS
-_
3
- 1
t~
1 ~
an. *~
~ Ski
~ ~ ~ I ~
NO SPOTS ON ARMS
NONE ON | ON LEGS
LEGS I
L:l
i.
he
F
If
Or
SPOTS ON ARMS
NONE ON | ON LEGS
LEGS ~
FIGURE 26 Background and other patterns (Nace et a/., 1973; reproduced with
permission from the American Zoologist).
OCR for page 114
114
SPOT:
DETERMINED BY
CHARACTER OF
SURROUNDING SPOTS
FUSED SPOT
= ONE SPOT
- IRREGULAR
SPOTONMIDLINE:
WHEN 1/3 OR MORE OF
SPOT OVER MIDLINE
Background y'<
I.~!
REGU LAR: APPROXI MATES
A SOUARE'OVALOR CIRCLE
IRREGULAR:
NOT SYMMETRICaL
I N DENTED OR LONG
LONG SPOTS
3 x (or more) AS LONG
AS AVERAGE WIDTH
i.,
Irregular
One Spot
ill:
I-,
lo; in,
Hi\\
Irregular
my'
Reg ul tar ~ ~ l irte
FIGURE 27 Conventions used to characterize spots (Nace et a/., 1973; reproduced
with permission from the American Zoologist).
Representative terms from entire chapter:
toe clipping