feel comfortable. Laboratory benches should be avoided as the rabbit will constrict its muscles on any smooth surface, however wrapping the rabbit in a towel is often sufficient and this may serve as a restricting device. Use of the rabbit restrainer may be necessary. In general, routine sedation is not necessary for antigen injections but is recommended for blood collection (see below).
In general, boosters are recommended to be given at 2- 4 week intervals. Blood collections should be performed 7-10 days following a booster injection for maximum results, depending upon the titer response curve generated per antigen or animal.
Day 0 Day 14 - 21 Day 28 - 35 Day 35 – 42 Day 42 – 56 Day 49 – 66
Pre-immunization bleed + initial antigen injection First antigen booster Second antigen booster Test bleed Third antigen booster Blood collection
of antigen injections and blood collections may be done at additional times as
needed for antibody production with the exception that if an animal fails to produce a satisfactory titer after the fourth booster it should be considered unsuitable for the antigen and taken out of the study. Variations differing from the above recommendations must be described in the individual investigator’s protocol.
Observe animals for a minimum of 15 minutes after the final injection or blood collection that day. Contact LAR staff immediately if an abnormal reaction is noticed.
Animals must be checked at least three times per week for 4 weeks after immunization.
If complications are noted, animals must be checked daily.
Investigators, veterinary staff and animal technicians should monitor rabbits for signs of pain or distress as well as swelling, abscesses or ulceration at the injection sites.
Staff (investigator or animal care) should contact LAR veterinarians if any of the above signs are noticed in an animal.
Intradermal injections for rabbits
Restrain the rabbit in an appropriate manner.
Intradermal injections for rabbits should be 0.05 ml or less per injection site; maximum of 20 sites for a total per injection session not to exceed 1 ml.
Do not place injections in sites that are used for grasping or physical restraint. Sites should be far enough apart to prevent coalescence of the local inflammatory reaction.
Injections sites must be clipped and disinfected with 70% alcohol before injections are made.
To give an ID injection a 25-gauge needle attached to a tuberculin syringe is inserted at a 10-15° angle, bevel up, just under the epidermis.
Verify that the needle is not in a blood vessel by pulling back on the plunger. If no blood appears in the needle hub, then the needle is not in a blood vessel.
Depress the plunger to inject the desired amount. An intradermal injection will cause an obvious small wheal or bump to form if the needle has been placed correctly. Following a few seconds pause, the needle is withdrawn. Do not continue to apply pressure to nor draw back on the plunger to avoid contamination of the needle track with the antigen/adjuvant mixture.
Repeat this procedure until the total volume has been delivered.