Each batch of CBPP vaccine must be shown to be innocuous when inoculated into laboratory animals. Consequently, all batches of the vaccine should be tested for safety/innocuity in laboratory animals before it is released for field use.
I. Materials
II. Procedure
Reconstitute a vial of CBPP vaccine in volume of cold, sterile diluent (e.g. sterile distilled water, PBS or 1M magnesium sulphate solution) such that 1 ml of the vaccine contains 10 field doses. For example a 100 dose vial is reconstituted with 10 ml of reconstitution fluid while a 50 dose vial shall be reconstituted using 5 ml of the diluent.
Inoculate each of 2 guinea pigs with 0.5 ml containing 5 doses of reconstituted vaccine by the intramuscular route into one hind leg.
Inoculate each of 2 guinea pigs with 0.5 ml (5 doses) and each of 6 mice with 0.1 ml (one dose) of the vaccine via the intra-peritoneal route.
Leave 2 guinea pigs and 4 mice as uninoculated controls.
The animals shall be observed for 30 days. If any of the guinea pigs or mice of the control or inoculated groups die from non-specific causes, then the test is not valid and must be repeated.
During the period of observation (i.e. test period), record any signs (including death) that can be attributed to a living organism in the vaccine.
At the end of the observation period sacrifice all animals and carry out complete post mortem. Record the findings.
NB: Any animals that fall sick or show any abnormality shall be examined and investigated to establish the cause of illness.
The vaccine passes the test for innocuity if all the inoculated and control animals in each group remain healthy and survive the observation period and at autopsy, none of the animals shows pathological evidence of the presence in the vaccine of any adventitious agent.
