Euthanasia of Rodent Fetuses and Neonates

Case Western Reserve University
Institutional Animal Care and Use Committee
Based on the intramural NIH guidelines
March 2000

The Report of the AVMA Panel on Euthanasia does not provide specific recommendations for the euthanasia of prenatal or neonatal animals. The following guidelines are suggested to assist individual Animal Care and Use Committees at the NIH in reviewing proposals which involve the use of rodent fetuses or neonates.

Fetuses:

• Fetuses up to 14 days in gestation: Neural development at this stage is minimal and pain perception is considered unlikely. Euthanasia of the mother or removal of the fetus should ensure rapid death of the fetus due to loss of blood supply and non-viability of fetuses at this stage of development.
• Fetuses 15 days in gestation to birth : The literature on the development of pain pathways suggests the possibility of pain perception at this time. Whereas fetuses at this age are not sensitive to inhalant anesthetics, euthanasia may be induced by the skillful injection of chemical anesthetics. Decapitation with surgical scissors, cervical dislocation, or rapid freezing (immersion in liquid nitrogen) are acceptable physical methods of euthanasia. When chemical fixation of the whole fetus is required, fetuses should be anesthetized prior to immersion in or perfusion with fixative solutions. Anesthesia may be induced by hypothermia of the fetus, by injection of the fetus with a chemical anesthetic, or by deep anesthesia of the mother with a chemical agent that crosses the placenta, e.g., pentobarbital. The institute veterinarian should be consulted for considerations of fetal sensitivity to specific anesthetic agents. When fetuses are not required for study, the method chosen for euthanasia of a pregnant mother must ensure rapid death of the fetus.

Neonates:

• Up to 14 days of age: Acceptable methods for the euthanasia of neonatal mice and rats include: injection of chemical anesthetics (e.g., pentobarbital), decapitation, or cervical dislocation. Additionally, inhalant anesthetics (e.g., methoxyflurane used with appropriate safety considerations), may be used. However, neonates have a high tolerance for hypoxia, so exposure must be prolonged (>20 minutes) and death confirmed. Immersion in liquid nitrogen may be used only for newborns; pups older than one day should be anesthetized prior to freezing with liquid nitrogen. Similarly, anesthesia should precede immersion or perfusion with chemical fixatives. Anesthesia may be induced by inhalant or injectable anesthetics; the institute veterinarian should be consulted for appropriate agents and dosages. Alternatively, when adequately justified, hypothermia (l) may be used to induce anesthesia in pups younger than six days.
• Older than 14 days: Follow guidelines for adults.

In all cases, the person performing the euthanasia must be fully trained in the appropriate procedures.

References:

Phifer CB, Terry LM. 1986. Use of hypothermia for general anesthesia in preweanling rodents. Physiol & Behav 38:887-890.