1.0 OBJECTIVE
1.1 To lay down a standard procedure to be followed for giving anesthesia to laboratory animals in Animal House Facility.
2.0 SCOPE
2.1 This SOP shall be applicable to the procedure followed for giving anesthesia to laboratory animals in Animal House Facility.
3.0 RESPONSIBILITY
3.1 Study Director
3.2 All personnel involved in giving anesthesia to laboratory animals
4.0 DEFINITIONS
4.1 Nil
5.0 PROCEDURE
5.1 Safety Precautions
5.2 Always wear appropriate personal protective equipment (like an apron, gloves, mask, head cap and shoes cover) before performing the experimental procedures.
5.3 Equipment and chemicals
5.4 Ketamine is classified as an NMDA receptor antagonist. It induces a state referred to as dissociative anesthesia. Ketamine is primarily used for the induction and maintenance of general anesthesia, usually in combination with some sedative drug.
5.5 Xylazine is a drug that is used for sedation, anesthesia, muscle relaxation, and analgesia in animals. It is an agonist at the alpha 2 class of adrenergic receptor.
5.6 Anesthesia Procedure
5.6.1
Ketamine/Xylazine Anesthesia
5.6.1.1
Required Material
5.6.1.1.1
Ketamine,
Xylazine and Sterile Normal Saline
5.6.1.2
Cocktail mixture ratio for different species
|
Species |
Ratio (Ketamine: Xylazine) |
|
Mice |
10:1 |
|
Rat |
15:2 |
|
Rabbit /
Guinea Pigs |
20:3 |
5.6.1.3
Preparation of cocktail for Mice
5.6.1.3.1
Take a clean
sterile vial.
5.6.1.3.2
Take 1ml of
Ketamine (100mg/ml) with the help of a sterile syringe.
5.6.1.3.3
Add 0.1 ml
Xylazine (100mg/ml).
5.6.1.3.4
Make up the
volume up to 10ml with Sterile Normal Saline.
5.6.1.3.5
Seal the
container and store in a sterile place.
5.6.1.3.6
Preferably
prepare fresh cocktail every-time as per need.
5.6.1.4
Preparation of cocktail for Rat
5.6.1.4.1
Take a clean
sterile vial.
5.6.1.4.2
Take 3.75ml of
Ketamine (100mg/ml) with the help of a sterile syringe.
5.6.1.4.3
Add 0.5 ml
Xylazine (100mg/ml).
5.6.1.4.4
Make up the
volume upto10ml with sterile Normal Saline.
5.6.1.4.5
Seal the
container and store in a sterile place.
5.6.1.4.6
Preferably
prepare fresh cocktail every-time as per need.
5.6.1.5
Preparation of cocktail for Rabbit / Guinea
pigs
5.6.1.5.1
Take a clean
sterile vial.
5.6.1.5.2
Take 6.25ml of
Ketamine (100mg/ml) with the help of a sterile syringe.
5.6.1.5.3
Add 0.94 ml
Xylazine (100mg/ml).
5.6.1.5.4
Make up the
volume upto10ml with sterile Normal Saline.
5.6.1.5.5
Seal the
container and store in a sterile place.
5.6.1.5.6
Preferably
prepare fresh cocktail every-time as per need.
5.6.1.6
Note:
Mix properly using vortex
Label the storage container
Store the solution away from light and in a
cool place
5.6.2
Injection of Cocktail into animals
5.6.2.1
Fresh cocktail
should be prepared prior to anesthesia.
5.6.2.2
Inject
0.1mL/10 g intraperitoneally to mice or as per SP.
5.6.2.3
If the animal
is not adequately anesthetized, repeat the injection at a dose of 0.05 ml/10g (approximately
every 30 minutes).
5.6.2.4
Inject
0.2mL/100 g i.p to rats or as per SP.
5.6.2.5
Repeat when
necessary, half a dose at a time after approximately 30 min.
5.6.2.6
Inject 5mL/kg
to rabbits and guinea pigs or as per SP.
5.6.2.7
The leftover
cocktail shall be discarded.
5.6.2.8
Note – The dose-volume for cocktail needs to be standardized depending upon
the stock concentration of ketamine and Xylazine and duration of anesthesia
required.
5.6.3
Inhalant Anesthetics for Rat and Mice
5.6.3.1
Identify the
cage number and animal number to be anesthetized.
5.6.3.2 Take the
animal gently out of the cage, open the lid of the desiccator, and put the animal inside.
5.6.3.3
Keep the
desiccator tightly closed until the animal loses its reflexes.
5.6.3.4
Monitor each
animal closely for signs of anesthesia after placing inside the desiccator.
5.6.3.5
Drowsiness in
the animal and loss of the righting reflex will be indicative of a state of
deep anesthesia.
5.6.3.6
Remove the animal immediately from the desiccator as soon as these signs are observed.
5.6.3.7
Post
anesthesia procedure has to be performed immediately after removing the animal
from the desiccator.
5.6.4 Note: Don’t give over anesthesia as animals may die.
6.0 ENCLOSURES
6.1 Nil
7.0 ABBREVIATIONS
7.1
SOP : Standard Operating Procedure
7.2
QAD : Quality Assurance Department
7.3
AHF : Animal House Facility
7.4 SP : Study plan
8.0 REFERENCES
8.1 Care 101.01 rodent anesthesia (Cornell University)
8.2 Committee
for the purpose of control and supervision of experiments on animals (CPCSEA)
guidelines for laboratory animal facility-2005, CPCSEA, Standard Operating Procedures
for Institutional Animal Ethics Committee (IAEC), Animal Welfare Division,
Ministry of Environment and Forests, January 2010.
9.0
REVISION HISTORY
|
Sr.
No. |
Change
Control No. |
Reason
for change |
|
|
|
|
END OF
DOCUMENT
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