Recommended Methods of Euthanasia: Mouse / Rat
August 30, 2013
Species
Method
Mouse / Rat / Hamster / Guinea Pig
Asphyxiation: CO2 Dispensed from a CO2 gas flow will be approximately 20% ‐ 30 % of the euthanasia chamber volume per minute, to induce narcosis. regulated cylinder followed by bilateral CO2 flow will be maintained for at least one minute following respiratory arrest. A bilateral thoracotomy will be thoracotomy. performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. Description
NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
ALTERNATIVE: Narcosis is induced by using the gradual fill method for CO2 gas administration by using a commercially available cylinder or tank, pressure reducing regulator, and flow meter, or equivalent equipment. CO2 flow will be maintained for at least one minute following respiratory arrest. A bilateral thoracotomy will be performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections). Asphyxiation: CO2 Dispensed from a CO2 gas flow will be approximately 20% ‐ 30 % of the euthanasia chamber volume per minute, to induce narcosis. regulated cylinder followed by cervical CO2 flow will be maintained for at least one minute following respiratory arrest. Cervical dislocation will be dislocation. performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
ALTERNATIVE: Narcosis is induced by using the gradual fill method for CO2 gas administration by using a commercially available cylinder or tank, pressure reducing regulator, and flow meter, or equivalent equipment. CO2 flow will be maintained for at least one minute following respiratory arrest. Cervical dislocation will be performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia
Recommended Methods of Euthanasia: Mouse / Rat Species
August 30, 2013
Method
Description
Asphyxiation: CO2 Dispensed from a regulated cylinder followed by decapitation.
CO2 gas flow will be approximately 20% ‐ 30 % of the euthanasia chamber volume per minute, to induce narcosis. CO2 flow will be maintained for at least one minute following respiratory arrest. Decapitation (indicate equipment to be used) will be performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections). ALTERNATIVE: Narcosis is induced by using the gradual fill method for CO2 gas administration by using a commercially available cylinder or tank, pressure reducing regulator, and flow meter, or equivalent equipment. CO2 flow will be maintained for at least one minute following respiratory arrest. Decapitation (indicate equipment to be used ) will be performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
Asphyxiation: CO2 Dispensed from a regulated cylinder followed by exsanguination. CO2 gas flow will be approximately 20% ‐ 30 % of the euthanasia chamber volume per minute, to induce narcosis. CO2 flow will be maintained for at least one minute following respiratory arrest. Exsanguination will be performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
ALTERNATIVE: Narcosis is induced by using the gradual fill method for CO2 gas administration by using a commercially available cylinder or tank, pressure reducing regulator, and flow meter, or equivalent equipment. CO2 flow will be maintained for at least one minute following respiratory arrest. Exsanguination will be performed secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia
Recommended Methods of Euthanasia: Mouse / Rat Species
August 30, 2013
Method
Description
Asphyxiation: CO2 Dispensed from a regulated cylinder followed by vital organ harvest. CO2 gas flow will be approximately 20% ‐ 30 % of the euthanasia chamber volume per minute, to induce narcosis. CO2 flow will be maintained for at least one minute following respiratory arrest. (Indicate specific organs to be harvested) will be harvested secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
ALTERNATIVE: Narcosis is induced by using the gradual fill method for CO2 gas administration by using a commercially available cylinder or tank, pressure reducing regulator, and flow meter, or equivalent equipment. CO2 flow will be maintained for at least one minute following respiratory arrest. (Indicate specific organs to be harvested) will be harvested secondarily to assure euthanasia. Euthanasia chamber will not be precharged with CO2. NOTE: CO2 for neonates < 7 days old may require very long exposure times (≥30 minutes), and will still require a secondary method, (consider sharp scissors to use for decapitation, when < 7 days of age, instead of CO2. For guinea pig neonates, see Decapitation with Guillotine sections).
Injectable anesthesia and viable tissue Post induction of anesthesia using (insert anesthetic agent, dose, route, needle size) and verifying the animal has harvest
reached a surgical plane of anesthesia by lack of response to a stimulus (indicate method) , the (indicate organs/ tissues) will be harvested and the animal exsanguinated to assure euthanasia. Inhalant anesthesia and viable tissue harvest
Using a precision vaporizer with induction chamber and scavenger, the gas anesthetic will be administered slowly up to 4.5 % in 100% oxygen until the animal is recumbant respiration has decreased. The chamber is then vented with 100 % O2 and the animal is removed. Anesthesia is continued by face mask using >2% inhaled anesthetic. The (indicate tissues) will be harvested followed by exsaguination to assure euthanasia.
Inhalant anesthetic overdose followed Using a precision vaporizer with induction chamber and waste gas scavenger, the gas anesthetic will be by bilateral thoracotomy.
administered slowly up to > 4.5 % in 100% oxygen and continued until lack of respiration for >1 minute is observed. A bilateral thoracotomy will be performed secondarily to assure euthanasia.
Inhalant anesthetic overdose followed Using a precision vaporizer with induction chamber and waste gas scavenger, the gas anesthetic will be by cervical dislocation.
administered slowly up to > 4.5 % in 100% oxygen and continued until lack of respiration for >1 minute is observed. Cervical dislocation will be performed secondarily to assure euthanasia.
Inhalant anesthetic overdose followed Using a precision vaporizer with induction chamber and waste gas scavenger, the gas anesthetic will be by decapitation.
administered slowly up to > 4.5 % in 100% oxygen and continued until lack of respiration for >1 minute is observed. Decapitation with (indicate equipment to be used ) will be performed secondarily to assure euthanasia.
Inhalant anesthetic overdose followed Using a precision vaporizer with induction chamber and waste gas scavenger, the gas anesthetic will be by exsanguination.
administered slowly up to > 4.5 % in 100% oxygen and continued until lack of respiration for >1 minute is observed. Exsanguination will be performed secondarily to assure euthanasia.
Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia
Recommended Methods of Euthanasia: Mouse / Rat
Species
Method
August 30, 2013
Description
Inhalant anesthetic overdose followed Using a precision vaporizer with induction chamber and scavenger, the gas anesthetic will be administered slowly by vital organ harvest.
up to > 4.5 % in 100% oxygen and continued until lack of respiration for >1 minute is observed. (Indicate organs) harvest will be performed secondarily to assure euthanasia.
Inhalant anesthetic overdose by using a Using a bell jar in a fume hood, animal is exposed to Isoflurane or Sevoflurane in oxygen until lack of respiration for >1 minute is observed. Animal will be placed on elevated surface to prevent direct contact with liquid bell jar followed by bilateral thoracotomy.
anesthetic. Bilateral thoracotomy will be performed secondarily to assure euthanasia. Inhalant anesthetic overdose using a Using a bell jar in a fume hood, animal is exposed to Isoflurane or Sevoflurane in oxygen until lack of respiration bell jar followed by cervical dislocation. for >1 minute is observed. Animal will be placed on elevated surface to prevent direct contact with liquid anesthetic. Cervical dislocation will be performed secondarily to assure euthanasia. Inhalant anesthetic overdose using a bell jar followed by decapitation.
Using a bell jar in a chemical fume hood, the animal is exposed to Isoflurane or Sevoflurane in oxygen until lack of respiration for >1 minute is observed. The Animal will be placed on a perforated plate to prevent direct contact with liquid anesthetic. Decapitation with (indicate equipement to be used ) will be performed secondarily to assure euthanasia. Inhalant anesthetic overdose using a bell jar followed by exsanguination.
Using a bell jar in a chemical fume hood, the animal is exposed to Isoflurane or Sevoflurane in oxygen until lack of respiration for >1 minute is observed. The animal will be placed on a perforated plate to prevent direct contact with liquid anesthetic. Exsanguination will be performed secondarily to assure euthanasia. Inhalant anesthetic overdose using a Using a bell jar in a chemical fume hood, the animal is exposed to Isoflurane or Sevoflurane in oxygen until lack of bell jar followed by vital organ harvest. respiration for >1 minute is observed. The animal will be placed on a perforated plate to prevent direct contact with liquid Anesthetic. The Vital organs to include (state organs) will be harvested to assure euthanasia. Decapitation by Guillotine (Juvenilles / Post induction of anesthesia or sedation using (indicate drug, dose, route, needle size if applicable) the animal is Adults) with sedation. introduced into the cone with the other hand so that the nose is at the breathing hole at the small end. Holding the cone at the rear, the small end of the cone is inserted into a commercial guillotine to the level of the cervical vertebrae and the guillotine is activated.
Decapitation by Guillotine (Juvenilles / Decapitation with guillotine using a decapicone. While holding the decapicone in one hand, the animal is Adults) without sedation.
introduced into the cone with the other hand so that the nose is at the breathing hole at the small end. Holding the cone at the rear, the small end of the cone is inserted into the decapitator. NOTE: Scientific Justification is REQUIRED for the use of this method without sedation / anesthesia on juvenille or adult animals. Decapitation by sharp scissors of Neonates < 7 days of age: decapitation performed with sharp scissors without sedation or anesthesia. Neonates (only for use with Mouse, Rat or Hamster)
NOTE: Not to be used for precocial neonates (e.g., guinea pig)
Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia
Recommended Methods of Euthanasia: Mouse / Rat Species
August 30, 2013
Method
Description
Cervical Dislocation with anesthesia / sedation. Post induction of anesthesia or sedation using (indicate drug, dose, route, needle size if applicable) , the thumb and index finger are placed on either side of the neck at the base of the skull or, alternatively, a rod is pressed at the base of the skull with the animal laying on a table surface . With the other hand, the base of the tail or hindlimbs are firmly and steadily pulled to cause separation of the cervical vertebrae and spinal cord from the skull.
Cervical Dislocation without anesthesia The thumb and index finger are placed on either side of the neck at the base of the skull or, alternatively, a rod is / sedation ( Mouse, Hamster or Rat) pressed at the base of the skull with the animal laying on a table surface . With the other hand, the base of the tail or hindlimbs are firmly and steadily pulled to cause separation of the cervical vertebrae and spinal cord from the skull.
NOTE: Scientific Justification is REQUIRED for use of this method without sedation/ anesthesia. Personnel perfoming this procedure must have documented training. Rats , must be <200 grams .
Induction of surgical anesthesia Using (indicate drug, dose, route, needle size if applicable) to induce General Anesthesia, vital perfusion will be followed by vital perfusion with XXXX performed using (indicate name) perfusate injected into the (insert point of vascular access ). Perfusion will be performed in a chemical fume hood if required by EHS. Perfusate waste will be disposed of by (indicate method of disposal).
p. 38, 49
p.48
Pentobarbital (Barbituate) anesthestic Administration of ≥ 100mg/kg Intraperitoneal (IP) or Intravascular (IV). Monitor animal until lack of respiration is overdose noted for ≥ 1 minute prior to tissue harvest or carcass disposal. Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia
Recommended Methods of Euthanasia: Fish
August 30, 2013
Species
Method
Description
Fish Benzocaine Hydrochloride Fish are bathed in > 250 mg/L buffered solution for a minimum of 10 minutes after cessation of opercular movement (anesthesia to respiratory collapse) followed by decapitiation. Benzocaine Hydrochloride Fish are bathed in > 250 mg/L buffered solution for a minimum of 10 minutes after cessation of opercular movement (anesthesia to respiratory collapse) followed by pithing. MS‐222 (Tricaine Methane Sulfonate)
Fish are bathed in ≥ 250 ‐ 500mg/mL buffered solution for a minimum of 10 minutes after cessation of opercular movement (anesthesia to respiratory collapse) followed by pithing.
MS‐222 (Tricaine Methane Sulfonate)
Fish are bathed in ≥ 250 ‐ 500mg/mL buffered solution for a minimum of 10 minutes after cessation of opercular movement (anesthesia to respiratory collapse) followed by decapitation (indicate equipment used) .
Progressive chilling to 2°‐ 4°C followed by Fish are rapidly transferred (indicate method of transfer) from the home tank to the chilled waterbath. Fish will pithing.
not be in contact with any ice that may be present in the chilled waterbath by (indicate method of prevention of contact with ice). The water temperature will be measured and verified with (indicate equipment used) . Fish will remain in the chilled waterbath for a minimum of 20 minutes after the loss of orientation and opercular movement followed by pithing.
Progressive chilling to 2°‐ 4°C followed by Fish are rapidly transferred (indicate method of transfer) from the home tank to the chilled waterbath. Fish will decapitation.
not be in contact with any ice that may be present in the chilled waterbath by (indicate method of prevention of contact with ice). The water temperature will be measured and verified with (indicate equipment used) . Fish will remain in the chilled waterbath for a minimum of 20 minutes after the loss of orientation and opercular movement followed by decapitation.
Pentobarbital (Barbituate) anesthetic overdose Adminitstration of ≥100mg/kg intra abdominally. Monitor animal until lack of heart beat for a minimum of 2 minutes prior to tissue harvest or carcass disposal. NOTE: It may take 30 minutes or more after barbituate administration until heart stops beating . Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia
Recommended Methods of Euthanasia: Xenopus
August 30, 2013
Species
Method
Description
Xenopus MS222 ( "TMS"or Tricaine Methane Sulfonate)
Animal is placed in a covered pan with 0.1% / 400‐500mg/L buffered solution until a level of surgcial anesthesia, verified by lack of response to a stimulus (toe pinch), is reached followed by decapitation (indicate equipment used) .
MS222 ( "TMS"or Tricaine Methane Sulfonate)
Animal is placed in a covered pan with 0.1% / 400‐500mg/L buffered solution until a level of surgcial anesthesia, verified by lack of response to a stimulus (toe pinch), is reached followed by decapitation (indicate equipment used) .
Pentobarbital (Barbituate) anesthetic overdose Administration of ≥ 100mg/kg intravenously (IV), intraperitonealy (IP), intracoelomically (IC), subcutaneous lymph space, or dorsal lymph sac. Monitor animal until lack of repsonse to a stimulus (toe pinch) for at least 2 minutes prior to tissue harvest or carcass disposal. Revision #: 1
Last Update: 8/30/2013
Revised by: Annabelle Crusan
Courtesy of Dr. Annabelle Crusan, University of Maryland School of Medicine Animal Care and Use Program, [email protected]
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Recommendations on Euthanasia