The duration of immunity in cattle following inoculation of rinderpest cell culture vaccine.

The duration of immunity following a single administration of rinderpest cell culture vaccine, of 90 or more monolayer passages, was studied in E. African zebu (Boran) and grade (cross-bred European) cattle. All animals were kept for periods of 6-11 years in rinderpest-free environments; groups of them (in all 23 Borans and 10 grades) were then challenged by parenteral or intranasal inoculation of virulent virus or by contact exposure to reacting cattle. Nasal excretion of virus was studied daily over the 10-to 14-day period following challenge, and simultaneous attempts were made to detect viraemia. The neutralizing antibody response was followed at 6-month intervals over the whole post-vaccination period and then daily for 10 days and at longer intervals to 3 weeks after challenge. All 33 animals which were exposed by various routes failed to react clinically and a rinderpest viraemia was never detected. No transmission of virus from the vaccinates to susceptible in-contact controls occurred within 14 or more days, from the 20 animals which could be so tested. Clearcut serological responses to challenge were seen in six cattle (four Borans and two grades) which were challenged after 7 years or more; these reactions were all delayed to the 9th or 10th days, i.e. they were not typically 'anamnestic'. These results are discussed in relation to mass vaccination campaigns for the control of rinderpest and from the comparative viewpoint of measles vaccination in man.

The protection of rabbits against the herpesvirus of malignant catarrhal fever by inactivated vaccines.

Cell-free virulent malignant catarrhal fever virus inactivated by either formalin or acetylethylenimine (AEI) and administered with Freund's complete adjuvant protected rabbits against two consecutive parenteral challenges with cell-free virus. However 'protected' rabbits succumbed within 20 days to later inoculation of infected rabbit lymph node suspensions. Both inactivated vaccines evoked high titres of neutralising and immunofluorescent antibody, but more of the rabbits given the AEI preparation showed pyrexia after the first challenge with cell-free virus. The degree of protection afforded was compared with that induced by inactivated antigens of other herpesviruses which cause lymphoproliferative disorders

A microtitre technique for the assay of malignant catarrhal fever virus and neutralising antibody.

A microtitre technique for the quantal assay of a cell-free strain of malignant catarrhal fever virus was developed, using serially passaged bovine embryonic kidney cells. End-points were determined after 12 days' incubation and the mean titre recorded for a single virus stock stored at -70 degrees C over a six-month period was 10(5.5) +/- 0.2 (SD). In neutralisation tests serum/virus mixtures were best held at 37 degrees C for 1 h in microtitre trays before the addition of cells; assays were highly reproducible, figures of 10(1.5) +/- 0.2 being obtained for a single reference serum.

Antibody response in cattle and rabbits to early antigens of malignant catarrhal fever virus in cultured cells.

Cytosine arabinoside (Ara-C) inhibited the production of late antigens and of infectious virus in monolayers of bovine kidney cells infected with the high-passage, WC-11 strain of malignant catarrhal fever virus. Early antigens were not affected. Using hyperimmune and acute-phase sera from cattle and rabbits in indirect immunofluorescence tests, it was shown that Ara-C treated cultures contained two early antigens; one was diffuse and distributed throughout the cells, the other was particulate and intranuclear. Antibody to early antigens developed later and attained lower titres in infected animals, especially rabbits; only hyperimmune sera reacted with the diffuse early antigen.

Vaccination against diseases associated with herpesvirus infections in animals: a review.

An account is presented of the development and use of herpesvirus vaccines in domestic animals, with particular reference to those viruses causing cytolytic rather than oncogenic infections. The chief infections covered are Aujeszky's disease (AD or pseudorabies), infectious bovine rhinotracheitis (IBR) and equine rhinopneumonitis (equine abortion; EHV-1). Others mentioned are feline viral rhinotracheitis and malignant catarrhal fever of cattle. Both live-modified and inactivated vaccines are widely used or under development for ADV, IBR and EHV-1. Live vaccines are generally regarded as successful in some circumstances but have major drawbacks, both in regard to safety or immunogenicity for the individual animal or to efforts to control and eliminate the viruses from whole populations. Difficulties remian in the preparation and use of inactivated vaccines, which still suffer from many of the epidemiological objections raised against modified live vaccines.

Experimental transplacental transmission of porcine cytomegalovirus.

Six serologically negative sows were infected by intranasal instillation of porcine cytomegalovirus (PCMV) between 31 and 85 days of pregnacy. Four sows showed an afebrile anorexia and lethargy 14-25 days after infection and all 6 developed significant increases in indirect immunofluorescent (IIF) antibody titres within 35 days. Virus was recovered from nasal and/or cervical swabs from 2 sows during life and from lung macrophage cultures after death. At term the sows were killed and their fetuses harvested by caesarean section. The number of mummified and stillborn fetuses increased from 4/63 in 6 previous litters to 18/60 in the 6 present litters. Nine of 43 fetuses born alive were reared in isolators for up to 6 weeks but the majority were killed for examination on the day of birth. Virus was isolated from 16 piglets from 4 of the 6 litters examined; it was isolated most frequently from lungs and liver but also from spleen, kidney, brain and nasal mucosa. Unsuckled day-old pigs had insignificant IIF titres, irrespective of whether they were excreting virus or not. The 5 congenital excretors which were reared all died within 7 days but no death occurred among their 4 litter-mates. Post-natal infection of 2 of these piglets reared in contact with congenitally infected pigs was suggested by the recovery of virus from nasal swabs 17 and 27 days after birth and the subsequent rise in IIF titre to 1/256 by day 42.

Cytomegalovirus excretion in gnotobiotic pigs.

Germ-free piglets were infected intranasally with porcine cytomegalovirus (PCMV) at 1 day (group A) or 3 weeks of age (group B). Viraemia and virus excretion by the nasal, pharyngeal and conjunctival routes was studied up to the time of death or to 12 weeks. Virus was also sought in tissues at death or at slaughter, as well as in a few urine samples. Viraemia was detected in group A between days 5 and 19 after infection and in group B between days 14 and 16 inclusive. The chief route of virus excretion was the nasal mucosa, followed by the pharynx and conjunctiva; the maximal duration of excretion by these routes was 32, 25 and 14 days for pigs of group A and 9, 7 and 4 days for group B. The quantity of virus was also greater in the former group, of which died of generalized PCMV infection. A viruria was demonstrated in 2 animals. Antibody detectable in indirect immunofluorescence (IIF) tests appeared towards the end of the third week, reaching maximal titres at 5 to 7 weeks after infection. The mean peak titre of antibody in group B was lower than in group A. Corticosteroid treatment at days 56--62 after infection resulted in some recrudescence of virus excretion, accompanied in group B by about a twofold increase in IIF antibody. PCMV was isolated in cultures of lung macrophages from 4 of 7 animals killed at about 12 weeks after inoculation.

The behaviour of porcine cytomegalovirus in commercial pig herds.

A longitudinal, virological and serological study of pigs in two herds with respiratory disease showed that infection by porcine cytomegalovirus (PCMV) was universal in both. Virus excretion usually began when piglets were between 3 and 6 weeks of age and reached a maximum between 5 and 8 weeks; it was usually no longer detectable at 11-12 weeks. Antibody demonstrable in indirect immunofluorescence (IIF) tests was present to moderate or high titre in all piglets at 2-3 weeks. This was presumed to be maternal in origin as it declined in titre between 2-3 and 5-6 weeks. After this fall the majority of piglets showed seroconversion as a result of virus infection. One group of 12 pigs in which infection occurred earlier than usual showed a very poor antibody response, which, nevertheless, persisted through to week 27. The findings are discussed with relation to porcine atrophic rhinitis and cytomegalovirus infection in other species.

Relationship of porcine cytomegalovirus and B bronchiseptica to atrophic rhinitis in gnotobiotic piglets.

Both porcine cytomegalovirus (PCMV) and Bordetella bronchiseptica produced rhinitis and pneumonia when inoculated intranasally into young gnotobiotic pigs. With PCMV the nasal lesions were confirmed to the lamina propria, while Bordetella produced atrophy of the turbinate bones and hyperplasia and degeneration of the nasal epithelium. Some exacerbation of the lesions was observed in the nasal mucosa of pigs given both agents, but the degree of bone atrophy was not increased.