2.0 RESPONSIBILITY: It is the responsibility of all Researchers and Animals Care Personals.
3.0 HUMANE ENDPOINTS/CRITERIA FOR EUTHANASIA:
3.1 Weight loss:
3.1.1 In adult animals, loss of >20 percent of body weight compared to the pre-study weight or to age matched control.
3.1.2 In growing animals, or in animals whose body weight has not been recorded, or in tumor studies, weight loss should be assessed by body condition.
3.2 Decrease in appetite:
3.2.1 Complete anorexia for 24 hours in small rodents and up to 5 days in large animals.
3.2.2 Partial anorexia (less than 50% of caloric requirement) for 3 days in rodents, 7 days in large animals.
3.3 Weakness/inability to obtain feed or water:
3.3.1 Inability or extreme reluctance to stand which persists for 24 hrs.
3.4 Moribund state:
3.4.1 An animal found to be in a state of dying characterized by severe depression, non-modulatory, complete anorexia and hypothermia with little likelihood of recovery.
3.5 Infection:
3.5.1 Infection (either overt or indicated by abnormal body temperature or WBC parameters) which fails to respond to therapy within an appropriate time.
3.6 Tumor growth:
3.6.1 Solid tumor that exceed 10 percent of normal body weight. Assuming 1 cm3 = 1gm, the formula for the weight of tumor is 4/3Ï€r3 i.e. for a 25 gms mouse, a tumor 1.7 cm in diameter = 2.57 gms = maximum allowable size. For tumor not perfectly round, use the average radius.
Loss of body condition indicating that tumor growth is being supported by the body and/or metabolic reverse.
3.6.2 Tumor growth that impedes animal ability to ingest food, water and other normal bodily function or its ability to move about its cage and remain clean and dry.
3.6.3 Tumor that appear to be causing the animal pain or distress that cannot be relieved with analgesics or other palliative measures.
3.6.4 Evidence of tumor necrosis or ulceration indicating the tumor has outgrown its blood supply.
3.6.5 Evidence of organ dysfunction and/ or failure from either primary or metastasized tumors.
3.7 Unrelieved pain/distress:
3.7.1 Signs of significant pain and / or distress which are unresponsive to analgesics, or as determined by the UCB veterinarian.
3.8 Organ dysfunction / failure:
Signs of severe organ system dysfunction non-responsive to treatment, or with a poor prognosis as determined by a UCB veterinarian.
3.8.1 Respiratory: Labored breathing and cyanosis unresponsive to appropriate medical therapy.
3.8.2 Cardiovascular: Acute blood loss resulting in shock or severe anemia; cardiac failure.
3.8.3 Gastrointestinal: Severe vomiting, diarrrhoea, rectal prolapse or intestinal obstruction.
3.8.4 Urogenital: renal failure characterized by elevated BUN or Creatinine; urinary tract obstruction; ruptured bladder; uroperitoneum, vaginal, uterine or penile prolapse.
3.8.5 Nervous: CNS depression; seizures; paralysis of one or more extremities; neurological condition which impede an animal’s ability to ingest food or water or its ability to move about its cage and remain clean and dry inhibiting eating and drinking.
3.8.6 Musculoskeletal: Muscle damage or bone fracture resulting in inability to use the limb or severe pain.
3.8.7 Integument: Non healing wound or severe burns covering more than 10% of the body; repeated self-trauma.
3.8.8 Cancer and toxicological studies
|
PARAMETERS |
WHAT TO LOOK FOR |
|
General
appearance /dehydration |
Decreased
body weight, missing anatomy, abnormal posture, hypothermia, fractured
appendage, swelling, tissue masses, prolapse, paraphimosis |
|
Skin or
fur discoloration |
Urine
stain, pallor, redness, cyanosis, icterus, wound, sore, abscess, ulcer,
alopecia, ruffled fur |
|
Eye |
Exophthalmos,
microphthalmia, ptosis, reddened eye, lacrimation, discharge opacity |
|
Nose,
mouth & head |
Head
tilted, nasal discharge, malocclusion, salivation, respiration, sneezing,
dyspnea, Tachypnea |
|
Urine
discoloration |
Blood in
urine, polyurea, anuria |
|
Locomotors/hyperactivity
|
Hyperactivity,
coma, ataxia, circling, muscle, tremors |
4.0 PROCEDURE:
4.1 Animals should only be euthanized in a quiet, clean environment and preferably away from other animals. In some cases, release of pheromones occurs during induction of unconsciousness. For that reason, other animals should not be present when euthanasia is performed.
4.2 When the animals are euthanatized, an eye is kept on them and one should remain in the room till the last animal is euthanatized and removed from the euthanasia chamber into a biohazard disposal bag.
4.3 Two different species must not be brought into the procedure room at the same time.
4.4 Euthanasia by CO2 (Carbon Dioxide) should be recommended for all rodents and other laboratory (rabbits & Guinea pig) except neonates and rodent used for inhalation toxicity study. In both the cases barbiturate (pentobarbitone) should be used.
4.5 Animals should be euthanized in necropsy room.
4.6 A clear Perspex chamber with lid is used as the euthanasia chamber. Ensure that the lid is left open before starting. If not, then the lid is opened and the air circulation is done for at least 10 minutes.
4.7 The rubber gasket mounted into the perimeter of the chamber lid is checked to ensure that it is in place and not cracked. This rubber gasket reduces the leakage of CO2 from the euthanasia chamber into the procedure room.
4.8 The euthanasia chamber is checked to ensure that it is clean and free of excreta and debris.
4.9 The tubing from CO2 supply to euthanasia chamber should be checked to ensure that it is attached and secured.
4.10 The animals are placed into the euthanasia chamber and the lid is closed gently.
4.11 The instructions given in the CO2 control module should be followed for the release of CO2 into the euthanasia chamber.
4.12 The animal should be observed. Once they appear to be relaxed and are not moving around, CO2 is continued to run until the animals no longer show any respiration or movements. Gas flows should be maintained for at least 1 minute, after the animal no longer shows any respiratory and other purposeful movements.
4.13 The CO2 is turned off by following instructions on the CO2 control module. Depending on the number of animals in the chamber, it may take 5 minutes (for mice) and 10 minutes (for rat) before the animals are sufficiently immobilized to turn off the CO2.
4.14 The lid of euthanasia chamber is opened. The animal is removed and its chest is palpitated to ensure there is no heart beat to confirm death. The webbing is pinched between its toes to ensure there is no withdrawal reflex. If any sign of respiratory or other movements occur, the animal is again put back into the chamber and the above steps are repeated.
4.15 At this point, the animal can be cervically dislocated, or a bilateral thoracotomy should be performed to ensure death.
4.16 Before leaving the procedure room, it is ensured that the lid of the euthanasia chamber is kept open, the chamber should be dry and clean ready for re-use by the next person.
5.0 PRECAUTIONS:
5.1 The animals should be checked carefully for its health status
5.2 The animals should be handled carefully.
5.3 All the checkups of the animals should be done in the presence of Veterinary Personal.
5.4 The exposure of carbon dioxide gas should not exceed 1 minute 30 seconds.
6.0 REFERENCES:
6.1 Anderson LC. Institutional and IACUC responsibilities for animal care and use education and training programs. ILAR J. 2007;48:90–95. [PubMed]
6.2 ACLAM [American College of Laboratory Animal Medicine] Position Statement on Rodent Surgery. 2001. [accessed January 7, 2010]. Available at www.aclam.org/education/guidelines/position_rodentsurgery.html.
6.3 AORN [Association of Operating Room Nurses] Recommended practices for traffic patterns in the perioperative practice setting. AORN J. 2006;83:681–686. [PubMed]
6.4 Arluke A. Uneasiness among laboratory technicians. Lab Anim. 1990;19:20–39. Arndt SS, Lohavech D, van’t Klooster J, Ohl F. Co-species housing in mice and rats: Effects on physiological and behavioural stress responsivity. Horm Behav. 2010;57:342–351. [PubMed]
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