(Ph. Eur. monograph 1956)
1 DEFINITION
Avian viral tenosynovitis vaccine (live) is a preparation of a suitable strain of avian tenosynovitis virus (avian orthoreovirus). This monograph applies to vaccines intended for administration to chickens for active immunisation.
2 PRODUCTION
2-1 PREPARATION OF THE VACCINE
The vaccine virus is grown in cell cultures.
2-2 SUBSTRATE FOR VIRUS PROPAGATION
2-2-1 Cell cultures
Cell cultures comply with the requirements for cell cultures for the production of vaccines for veterinary use (5.2.4).
2-3 CHOICE OF VACCINE VIRUS
The vaccine virus shall be shown to be satisfactory with respect to safety (5.2.6) and efficacy (5.2.7) for the chickens for which it is intended.
The following tests for safety (section 2-3-1), increase in virulence (section 2-3-2) and immunogenicity (section 2-3-3) may be used during the demonstration of safety and efficacy.
2-3-1 Safety
Carry out the test for each route and method of administration to be recommended for vaccination using in each case chickens not older than the minimum age to be recommended for vaccination and from a flock free from specified pathogens (SPF) (5.2.2). Use vaccine virus at the least attenuated passage level that will be present in a batch of the vaccine.
For each test performed in chickens younger than 3 weeks of age, use not fewer than 10 chickens. For each test performed in chickens older than 3 weeks of age, use not fewer than 8 chickens. Administer to each chicken a quantity of the vaccine virus equivalent to not less than 10 times the maximum virus titre likely to be contained in 1 dose of the vaccine.
Observe the chickens at least daily for at least 21 days. Carry out histological examination of the joints and tendon sheaths of the legs and feet at the end of the observation period (as a basis for comparison in the test for increase in virulence).
The test is not valid if more than 10 per cent of the chickens younger than 3 weeks of age show abnormal signs of disease or die from causes not attributable to the vaccine virus. For chickens older than 3 weeks of age, the test is not valid if non-specific mortality occurs.
The vaccine virus complies with the test if no chicken shows abnormal signs of disease or dies from causes attributable to the vaccine.
2-3-2 Increase in virulence
Carry out the test according to general chapter 5.2.6 using 1-day-old chickens from an SPF flock (5.2.2). If the properties of the vaccine virus allow sequential passage through 5 groups via natural spreading, this method may be used, otherwise passage as described below is carried out.
Administer to each chicken of the 1st group by a suitable route a quantity of the vaccine virus that will allow recovery of virus for the passages described below. Euthanise the chickens at the moment when the virus concentration in the most suitable material (for example, tendons, tendon sheaths and liquid exudates from the hock joints, spleen) is sufficient.
Prepare a suspension from this material from each chicken and pool these samples. Administer 0.1 mL of the pooled samples by the route of administration most likely to lead to increase in virulence to each chicken of the next group. Carry out this passage operation not fewer than 4 times; verify the presence of the virus at each passage. If the virus is not found at a passage level, repeat the passage by administration to a group of 10 chickens.
If the 5th group of chickens shows no evidence of an increase in virulence indicative of reversion during the observation period, further testing is not required. Otherwise, carry out an additional safety test and compare the clinical signs and any relevant parameters in a group of at least 10 chickens receiving the material used for the 1st passage and another similar group receiving the virus at the final passage level.
The vaccine virus complies with the test if no indication of an increase in virulence of the virus at the final passage level compared with the material used for the 1st passage is observed. If the virus is not recovered after an initial passage in 5 chickens and a subsequent repeat passage in 10 chickens, the vaccine virus also complies with the test.
2-3-3 Immunogenicity
A test is carried out for each route and method of administration to be recommended for vaccination using in each case chickens not older than the minimum age to be recommended for vaccination. The quantity of the vaccine virus administered to each chicken is not greater than the minimum virus titre to be stated on the label and the virus is at the most attenuated passage level that will be present in a batch of the vaccine. Use not fewer than 30 chickens of the same origin and from an SPF flock (5.2.2). Administer the vaccine by a route to be recommended to not fewer than 20 chickens.
Maintain not fewer than 10 chickens as controls. Challenge each chicken after 21 days by a suitable route with a sufficient quantity of virulent avian tenosynovitis virus. Observe the chickens at least daily for 21 days after challenge. Record the deaths and the surviving chickens that show clinical signs of disease. If the challenge is administered by the foot pad, any transient swelling of the foot pad during the first 5 days after challenge may be considered non-specific. At the end of the observation period, euthanise all the surviving chickens and carry out macroscopic and/or microscopic examination for lesions of the joints and tendon sheaths of the legs and feet, e.g. exudate and swelling.
The test is not valid if:
— during the observation period after challenge fewer than 80 per cent of the control chickens die or show severe clinical signs of avian viral tenosynovitis or show macroscopical and/or microscopical lesions in the joints and tendon sheaths of the legs and feet;
— or if during the period between vaccination and challenge more than 10 per cent of the control or vaccinated chickens show abnormal clinical signs or die from causes not attributable to the vaccine.
The vaccine virus complies with the test if during the observation period after challenge not fewer than 90 per cent of the vaccinated chickens survive and show no notable clinical signs of disease or show macroscopical and/or microscopical lesions in the joints and tendon sheaths of the legs and feet.
3 BATCH TESTS
3-1 Identification
The vaccine virus is identified using a suitable method, for example, an immunostaining test in susceptible cell cultures using monoclonal antibodies, to demonstrate the presence of the vaccine virus stated on the label.
3-2 Bacteria and fungi
Vaccines intended for administration by injection comply with the test for sterility prescribed in the general monograph
Vaccines for veterinary use (0062).
Any diluent supplied for reconstitution of the vaccine complies with the test for sterility prescribed in the general monograph Vaccines for veterinary use (0062).
3-3 Mycoplasmas (2.6.7)
The vaccine complies with the test for mycoplasmas.
3-4 Extraneous agents (5.2.5)
The vaccine is free from extraneous agents.
3-5 Virus titre
Titrate the vaccine virus by inoculation into suitable cell cultures (5.2.4). The vaccine complies with the test if 1 dose contains not less than the minimum virus titre stated on the label.
3-6 Potency
The vaccine complies with the requirements of the test prescribed under Immunogenicity (section 2-3-3) 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 using a vaccinating dose containing not more than the minimum virus titre stated on the label.
