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Use of ethanol for euthanasia of mice

Article  in  Australian Veterinary Journal · September 1989

DOI: 10.1111/j.1751-0813.1989.tb13590.x · Source: PubMed

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Roger Lord
Australian Catholic University (Brisbane Campus)
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McSweeney CS and Kennedy PM (1987)-ln Herbivore Nutrition
Research, edited by M Rose Australian Society of Animal Produc-
tion, Brisbane, p169
Piercy DWT (1973)-Aust Vet J 49: 444
Rowe J B (1979)-Trop Anim Prod 4: 127
(Accepted for publication 3 February 1989)
Use of ethanol for euthanasia of mice
Queensland Institute of Medical Research,
Bramston Terrace, Herston, Queensland, 4006
R LORD
An alternative and effective method of euthanasia not cur-
rently in practise is the intraperitoneal injection of ethanol.
The effect of ethanol on metabolism has been studied and
reviewed by many researches (Hawkins and Kalant 1972;
Badawy 1984). Ethanol is metabolised mainly in the liver and
the principal enzyme involved in this process is the NAD+-
dependent liver-cytosolic protein alcohol dehydrogenase,
which catalyses the conversion of ethanol into acetaldehyde.
The acetldehyde produced from ethanol oxidation is then con-
verted into acetate by the enzyme NAD' dependent mitochon-
drial aldehyde dehydrogenase and then broken down further
via the citric acid cycle to carbon dioxide and water.
Acute alcohol intoxication occurs when the ethanol concen-
tration exceeds 80 pg/dl causing depression of the central ner-
vous system (Badawy 1984). Mice used in research applica-
tions such as the production of murine ascites can be killed
cheaply with a minimum of discomfort to the animal with this
method.
As a preliminary experiment 8 Balb/c female mice (6 to 7
weeks) were injected intraperitoneally with 500 pl of ethanol*
at various concentrations (30'70, 50%, 70%, 100%). Heart-
beat per minute was taken prior to injection and monitored
until the mouse recovered or died. The behaviour of each
group of mice was also observed to ascertain if any discomfort
was being caused by the ethanol. Only 100% ethanol appeared
to cause some discomfort but this was eliminated by drying the
needle before injection. Mice injected with the 30% solution
recovered while concentrations of 50% to 100% caused death
(Figure 1).
In a study of 10 Balb/c mice, 70% ethanol caused death in
(2 min 41 sec k 52 sec). No discomfort was observable in these
mice. The behavioural changes following injection were
gradual. Each mouse showed a gross loss of muscle control,
with death following coma as a result of respiratory collapse.
Autopsies performed on this group confirmed that a faint
heartbeat did not exist 10 min after pulse cessation.
'Ethanol - analytical reagent grade, May and Baker, West
Footscray, Victoria
268
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160- 1
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140 _-
120 -.
100 .-
80 --
60 -.
40 -.
\1 ,
- ,
0 5 10 15 20 25
Time elapsed (minutes)
Figure 1.500 pl dosage of ethanol at 30% ( V ) and 70% ( 0 )and
effect on mouse heartbeat (2mice in each group). Loss of mus-
cle control was observed at 60 beatslminute and respiratory
collapse evident at 30 beatslmin.
Ethanol has a number of advantages over many of the
techniques normally used in the euthanasia of mice. Firstly, it
is inexpensive, it costs far less to buy ethanol than barbituates.
Barbituates also have the added disadvantage of being S4
restricted drugs and thus cannot be obtained freely by some
research organisations.
Carbon dioxide gas is also used commonly as a method of
euthanasia but unless dry ice is available for gas generation a
costly carbon dioxide cylinder is needed. Diethyl-ether which
is still a preferred method of euthanasia in many laboratories
has the draw-back that it is highly explosive, gives the
laboratory worker a headache and irritates the respiratory
tract of the animal. Other physical methods such as disloca-
tion of the neck require a degree of expertise on behalf of the
research worker, however, in a training situation ethanol
could be used to intoxicate mice prior to a student learning the
technique.
I would like to thank Mrs J Reilly, Animal Welfare Officer,
University of Queensland and Dr JK Blackshaw for much ap-
preciated discussions.
References
Blackshaw JK and Allan DJ (1985) - Principles of Laboratory
Animal Management, Aust SOCfor the Study of Anim Behaviour,
Univ Qld
Badawy B (1984) - In Clinical Biochemktry of Alcoholism, edited by
SB Rosalki, Churchill Livingstone, London
Green CJ (1979) - Animal Anaesthesia, Spottiswoode Ballantyne
Ltd. London
Hawkins RD. Kalant H (1972) - Pharmacdogical Rev 24: 67
(Acceptedfor publication I7 May 1989)
Australian Veterinary Journal, Vol. 66, No. 8. August, 1989