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Euthanasia in
Experimental
Animals
Presenter-
Dr Arun Singh
Senior Resident
Department of Pharmacology
SMS Medical College, Jaipur
Overview
Introduction
Definition
Advantage of Euthanasia
Classification
Chemical Method
Mechanical Method
Conclusion
Introduction
• Animals are used in experimental pharmacological studies and
assessing the drug activity, it requires analyzing the organ (brain,
kidney, lung, etc.), blood (plasma or serum), and body fluid (CSF,
urine, etc.) to conclude the result.
• Hence for this purpose, animals have to be sacrificed to obtain
the said part of the body.
• Therefore, according to the animal care guidelines they should
be euthanized.
Definition
• “Euthanasia means the humane killing (sacrifice) of an animal which
produces rapid unconsciousness and subsequent death without or minimal
pain or distress to the animal.”
• The choice of a method depends on species, age, availability of restraint
and skill of the individual's performing euthanasia.
• It is very important that, in an experimental setting, the method of
euthanasia must be primarily consistent with the experimental goals.
Advantages of Euthanasia
• Reliability and irreversibility
• Minimum pain, distress, anxiety or
apprehension
• Minimum delay until unconsciousness
• Safety and emotional effect on personnel
• Compatibility with requirement and
purpose, including subsequent use of tissue
a
• Compatibility with species, age and health
status
Classification
Euthanasia methods are broadly
classified as:
(I) Chemical method (Inhalants/Non-
inhalants)
(II) Physical methods
(I) Chemical Method
Ex. Halothane, enflurane, sevoflurane, methoxyflurane, isoflurane and desflurane
(a) Inhalant Anesthetics
(i) Carbon dioxide (CO2)
It is an effective and widely used agent to euthanize rodents due
to rapid hypoxia which further leads to depression of vital
centers. (Maintain 20% -70% of the chamber volume per minute)
This method is preferred in several animals but it is dangerous to
use, so precaution should be taken while using it.
(ii) Nitrous oxide (N2O)
It is not preferred due to lack in fast onset of anesthesia but
may produce hypoxemia and cardiac or respiratory arrest.
However, it may be used in combination with other
anaesthetics to speed anesthesia onset.
(iii) Ether
It was formerly used extensively but is now only acceptable conditionally.
The reason is being irritant to mucous membranes and risk of fire and
explosion.
The use of ether is prohibited in many countries.
(b) Noninhalents Anesthetics
(i) Barbiturates
Sodium pentobarbital is the most rapid and reliable
method of euthanasia for most experimental animals.
In non-rodent species, barbiturates are given
intravenously to be most effective.
Intraperitoneal injection of barbiturates is acceptable
for euthanasia in small mammals.
(ii) Potassium chloride (KCl)
• KCl induces immediate cardiac arrest without any significant
depression of the central nervous system.
• Hence, it must only be used after the animal is deeply
anesthetized.
(iii) Magnesium sulphate (MgSO4)
• MgSO4 produces its action through cardiac arrhythmia,
neuromuscular blockade and deep anesthesia, hence ultimately
animal gets euthanized due to cardiac arrest and neuromuscular
blockade.
(iii) Neuromuscular Blocking Agents
Ex. Succinylcholine, Curare, etc.
• These agents induce muscular paralysis
and death because of suffocation.
Distress onset is more, hence less
preferred for euthanasia.
(II)Physical
Methods
Physical methods are performed by skilled and experienced
personnel with appropriate, well-maintained equipment.
(a) Cervical dislocation:
Humane technique for euthanasia which is frequently used for
mice, rats, guinea pig, rabbits (weighing less than 1 kg) and other
rodents.
(b) Decapitation:
Decapitation may be used to euthanize rodents and small
rabbits. Except in neonatal animals, a guillotine is generally used.
(c) Microwave irradiation
• Special instruments designed (appropriate power and
microwave distribution) for this purpose, which is used when an
experiment requires fixation of mouse or rat brain metabolites in
vivo without losing anatomic integrity of the brain.
(d) Penetrating captive bolt
• This method is conditionally acceptable and made for ruminants,
horses, and swine when chemical agents are scientifically
contraindicated.
• This method is not employed in the laboratory animals.
Euthanasia of Poikilothermic (Cold-blooded) Animals
The euthanasia of poikilothermic animals is different due to differences
in the pharmacokinetics, respiration and tolerance to cerebral hypoxia
between these species and homeothermic animals.
Chemical agents: Pentobarbital, Tricainemethane sulfonate or
benzocaine HCl.
Intraperitoneal administration of pentobarbital is an effective method
of euthanasia in amphibians.
Tricaine methane
sulfonate or benzocaine
hydrochloride may be
placed in the water of
amphibians and fish to
produce anesthesia and
prolonged contact may
produce death.
Inhalant anesthetics may
be used for amphibians
and reptiles but due to
the low oxygen
requirements for
amphibian, the onset of
unconsciousness and
death will be significantly
lengthened.
• Physical methods:
Poikilotherms may be euthanized by stunning followed by
decapitation or pithing to ensure death.
In frogs and toads, pithing the brain (single pithing) and spinal cord
(double pithing) are effective and acceptable methods.
Conclusion
• Euthanasia methods are under constant study, and guidelines often change as new
information is gained.
• As new knowledge expands understanding of practices best suited for humane euthanasia,
revised and new guidelines can be expected.
• Veterinarians trained in laboratory animal medicine should use professional judgment to
assess whether a proposed euthanasia method is aligned with the goals of the research and
will provide valid data.
• When necessary, a veterinarian should participate in the assessment and validation of
euthanasia methods on a case-by-case basis.
Euthanasia in Experimental Animals.pptx