Canine Parainfluenza Virus Vaccine, Live
Canine Parainfluenza Virus Vaccine, Live is a freeze-dried preparation containing one or more attenuated strains of canine parainfluenza virus grown in suitable cell cultures.
Production
The virus is propagated in suitable cell culture. The viral suspension is harvested, titrated and may be mixed with a suitable stabilizing agent. The vaccine is then freeze-dried and can be used either with any suitable diluent or after reconstitution with licensed liquid canine vaccine components.
Choice of vaccine strain. A well characterized strain obtained from an authentic source shall be used for the vaccine production. The master seed which has been established as pure, safe and immunogenic for the species for which it is intended shall be used for vaccine production.
Identification
When inoculated into dogs, the vaccine stimulates the production of specific neutralizing antibodies against canine parainfluenza virus determined by suitable serological tests.
Tests
Mycoplasmas (2.7.8). Complies with the test for mycoplasma.
Extraneous agents. Neutralize the vaccine virus with a suitable mono specific antiserum against canine parainfluenza virus and inoculate into cell cultures known for their susceptibility to viruses pathogenic for the dog. Carry out 2 passages with an interval of 6 to 8 days. The vaccine complies with the test if no cytopathic effect develops.
Sterility (2.2.11). Complies with the test for sterility.
Safety. Carry out the test for each route and methods of administration to be recommended for vaccination. Use vaccine virus at least attenuated passage level that will be present between the master seed lot and a batch of the vaccine. Inject 10 times the minimum dose into each of 6 dogs of the minimum recommended age that are shown to be preferably free of canine parainfluenza virus antibodies. Observe the dogs for 21 days. None of the dogs shows abnormal local or systemic reactions or dies of any causes attributable to the vaccine.
If the vaccine is intended for use in pregnant bitches, administer the virus to not fewer than 5 bitches at the recommended stage or stages of pregnancy and according to the recommended schedule. Prolong the observation period until 1 day after whelping. The dogs remain in good health and there is no abnormal local or systemic reaction. No adverse effects on the pregnancy or the offspring are noted.
Increase in virulence. Administer intranasally and by a recommended route to each of 2 puppies, 5 to 7 weeks old and which do not have antibodies against parainfluenza virus of canine origin, a quantity of virus that will allow recovery of virus for the passages described below. Use vaccine virus at the least attenuated passage level that will be present in a batch of the vaccine. Collect nasal swabs from each dog daily from 3 to 10 days after inoculation. Inoculate the suspension from the swabs into suitable cell cultures to verify the presence of virus. Use the suspension from the swabs that contain the maximum amount of virus and administer intranasally 1 ml of the suspension into each of 2 other puppies of the same age and susceptibility. This operation is then repeated at least 5 times. If the virus is not recovered at a given passage level, a second series of passages is carried out. Inoculate virus from the highest recovered passage level to not fewer than 5 puppies, observe for 21 days and compare any reactions that occur with those seen in the test for safety described above. There is no indication of an increase in virulence as compared with the non-passaged virus.
Immunogenicity. Inject each of eight susceptible dogs, between 8 and 14 weeks old that have been previously tested and shown to be preferably free from canine parainfluenza virus antibodies with a dose of the vaccine stated on the label. Use another two dogs of the same age group as unvaccinated controls. Observe the animals for a further 21 days. Challenge all the dogs with sufficient quantity of a suspension of canine parainfluenza virus by intranasal route. Observe the animals for a further 14 days. Collect nasal swabs from day 5 to 10 days after challenge and test the samples for the presence of excreted virus. Use a scoring system for recording the incidence of coughing in each dog. The control dogs show typical signs of coughing or excretion of the virus. The vaccine complies with the test if the scores for coughing or virus excretion in the vaccinated dogs are significantly lower than the controls.
If the potency test has been performed with satisfactory results on a representative batch of the vaccine from the seed lot, it may be omitted as a routine control test during production on other batches of the vaccine prepared from the same seed lot.
Virus titre. Not less than 103 TCID50/CCID50 per dose, determining the titre of the vaccine in a suitable cell culture with suitable medium.
Manufacturer’s tests
Virus titre. Virus titre is determined in final bulk harvest in a suitable cell culture with suitable medium.
Identification. Vaccine complies the requirements of the test mentioned under production.
Extraneous agents. Vaccine complies the requirements of the test mentioned under production.
Mycoplasmas. Complies with the test for freedom from mycoplasmas (2.7.8).
Sterility (2.2.11). Complies with the test for sterility.
Batch tests
Identification
Vaccine complies the requirements of the test mentioned under production. Alternatively, identification on the final lot by molecular techniques are acceptable and can be used in the routine batch release tests after proper validation (2.8.1).
Water (2.3.43). Not more than 3.0 per cent.
Extraneous agents. Vaccine complies the requirements of the test mentioned under production. Alternatively, molecular techniques for detection of pathogenic viruses of dog are acceptable batch release test after proper validation (2.8.1).
Mycoplasmas (2.7.8). Complies with the test for freedom from mycoplasmas. Alternatively, molecular techniques for detection of mycoplasma nucleic acid are acceptable batch release test after proper validation (2.8.1).
Virus titer. Not less than 103 TCID50 of the virus per dose, determining the titre of the vaccine in a suitable cell culture with suitable medium or one dose of vaccine contains not less than quantity of virus equivalent to the minimum virus titre stated on the label.
Sterility (2.2.11). Complies with the test for sterility.
Safety. Inject each of two susceptible dogs, between 8 and 14 weeks old, free from canine parainfluenza virus antibodies with a dose of the vaccine reconstituted with the sterile diluent equivalent to 10 times the dose and by the route stated on the label. Observe the animals for 14 days. None of the dogs shows any systemic or local reactions.
Potency. The vaccine complies with the requirements of test mentioned under immunogenicity when administered by a recommended route and method. It is not necessary to carry out the potency test for each batch of the vaccine if it has been carried out on a representative batch of the vaccine using a vaccinating dose containing not more than the minimum titre stated on the label. The virus titer is considered for a routine batch release provided the traceability of the vaccine strains used is from the same master seed.
Labelling. The label states (a) the minimum dose; (b) the recommended routes of administration; (c) storage temperature (d) virus titre (e) expiry period.