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Muscle samples (20 g) from 2025 pig carcases from Aujeszky's disease-free holdings were collected at the slaughterhouse. The samples were frozen and thawed to obtain meat juice, which was then analysed by three ELISA-gE test kits in parallel, to assess their specificity. After two cycles of freezing and thawing, 2.2 per cent of the samples were dry. Three times more of the samples from the sow carcases than from the finisher carcases yielded insufficient juice (< 220 microl). To validate the results of the specificity study, the sensitivity of the test kits was evaluated on 45 samples from gE-seropositive sows. On the basis of the results from 1879 samples, the specificity of the ELISA-gE kits was between 0.995 and 1.000, depending on the classification of the doubtful results. In the case of a positive or doubtful result, it proved useful to repeat the test on the same sample, in order to limit the number of false positive results.

A commercial ELISA test to detect serum anti-gE antibodies to Aujeszky's disease virus was adapted for use with muscle exudates. The muscle samples were taken from the diaphragm of pig carcasses at the slaughterhouse. Three hundred and eighty-nine pairs of samples of serum and muscle exudate were compared to determine the possibility of using muscle exudate samples in a programme to control Aujeszky's disease. Taking the serum samples as the reference, the individual sensitivity of the test was 93.2 per cent and the individual specificity was 98.3 per cent. The concentration of antibodies in the muscle exudates was on average 20 times lower than that in the serum samples.

An attenuated glycoprotein I-negative (gI-)/thymidine kinase-negative (TK-) constructed vaccine was used to eradicate Aujeszky's disease virus from a large farrow-to-finish herd in Sweden. The herd had had problems every year for seven years and two attempts to eradicate the virus without vaccination had failed. At the start of the vaccination programme 86 per cent of the 396 breeding animals were seropositive to the virus. In spite of evidence of virus circulation in the fattening units, no fatteners were vaccinated. The breeding stock was vaccinated every four months and monitored serologically. Seropositive sows and boars were culled at an economic rate. During the programme, four breeding animals seroconverted to gI. Another seven animals which seroconverted to gI were suspected to have been infected shortly before the first test and vaccination. When all the seropositive breeding animals had been culled, the fattening units were sampled and no seropositive animals were found. The herd was declared gI-negative 39 months after the start of the programme. Monitoring of the herd for another four years, until all the vaccinated animals had been culled, showed that the herd remained free from Aujeszky's disease virus.

An outbreak of Aujeszky's disease occurred in a flock of 160 ewes which had been shorn and housed adjacent to pigs which were excreting Aujeszky's disease virus. The 29 affected sheep developed either the classical 'mad itch' signs associated with Aujeszky's disease in ruminants or signs of encephalitis. Aujeszky's disease virus was isolated from the central nervous system of the affected ewes. Five farm cats also became ill and died. Histological lesions of viral encephalitis were observed in one of the cats and Aujeszky's disease virus was isolated from its brain.

An intensive vaccination programme with the glycoprotein I (gI) and thymidine kinase-deleted vaccine strain 783 was applied on all the pig farms in a region with a high pig density. To monitor the spread of Aujeszky's disease virus within breeding herds in that region, all the breeding stock in nine herds were examined for antibodies to gI six times at intervals of four months. The prevalence of gI-seropositive sows decreased greatly in all nine herds. The mean percentage of gI-seropositive sows decreased from 56.4 per cent (range 80.0 to 13.6 per cent) at the start, to 20.3 per cent (range 29.1 to 1.6 per cent) after two years. Nevertheless, seroconversions to gI were detected in all the herds, and in six out of the nine breeding herds even during the second year of the study. The intensive regional vaccination apparently did not completely prevent Aujeszky's disease virus infections within these herds. The source of the virus responsible for these infections was not identified. However, because in most herds only a few sows seroconverted, the virus either circulated at a low level within the herds, or its introduction or reactivation did not lead to an extensive spread of the virus.

Pigs were vaccinated by scarification or intramuscular injection with a swinepox virus-Aujeszky's disease (pseudorabies) recombinant (rSPV-AD) constructed by inserting the linked Aujeszky's disease virus genes coding for glycoproteins gp50 and gp63, attached to a vaccinia virus p7.5 promoter, into the thymidine kinase gene of swinepox virus. By 21 days after vaccination, 90 and 100 per cent of the animals vaccinated by scarification or intramuscular injection, respectively, had developed serum neutralising antibodies to Aujeszky's disease virus. Upon challenge with virulent virus, significantly fewer vaccinated pigs developed clinical Aujeszky's disease, nasal shedding of challenge virus was markedly reduced, and the vaccinated groups of pigs maintained or gained weight during the week after challenge whereas the unvaccinated control group lost weight. No transmission of rSPV-AD to in-contact controls was detected during the three weeks before challenge. In a second experiment, serum neutralising antibodies to Aujeszky's disease virus persisted for 150 days after the pigs were vaccinated with rSPV-AD by scarification or intramuscular injection and all the pigs showed an anamnestic response when they were revaccinated.

Serum and colostrum samples taken from 499 sows from five herds of pigs endemically infected and vaccinated against Aujeszky's disease virus, were used to investigate whether colostrum could be used to detect antibodies against glycoprotein I (gI) of the virus. Using serum as the reference, the test applied to colostrum had a sensitivity of 97 per cent and a specificity of 88 per cent. When samples were taken from 50 sows from a gI seronegative vaccinated herd, one colostrum sample was gI-positive, giving a specificity for the test of 98 per cent. The mean gI antibody titres in colostrum were about six times higher than in serum. Samples of colostrum were also taken from 132 sows from eight unvaccinated herds free of Aujeszky's disease virus. All these samples were gI-negative, giving a specificity of 100 per cent. Colostrum samples can be stored for at least six weeks at -20 degrees C without compromising the test results, and the repeatability and reproducibility of the test applied to colostrum were good.

This work describes the evaluation of two commercial ELISA kits for the detection of gI antibodies against Aujeszky's disease. A collection of experimental sera from infected pigs, field sample sera, and sera from pigs vaccinated with seven different modified gI-negative commercial vaccines were used to evaluate each test. Both ELISA kits showed good reproducibility, and specificity, but differences could be appreciated in sensitivity when sera obtained at early stages of infection was analysed. These results also indicated that both kits could be used in conjunction with the seven vaccines evaluated in this study.

The introduction of Aujeszky's disease virus into a herd of pigs usually results in a rapid spread of the virus and a high percentage of pigs become seropositive. However, herd monitoring for the virus occasionally reveals a single seropositive breeding pig, referred to as a single reactor. The seropositive status of single reactors may be due to previous vaccination against Aujeszky's disease, or to exposure to a field strain of the virus, or to a false positive reaction in the serological assay. During a monitoring programme in Minnesota, 30 pig herds with single serological reactors were detected. Twenty-seven of these single reactors from 19 herds were segregated from their herds immunosuppressed with dexamethasone. Aujeszky's disease virus was isolated from four of the 27 pigs. Three of the four herds subsequently had outbreaks of Aujeszky's disease, suggesting that some single reactors were infected with Aujeszky's disease virus and had the potential to spread the virus within and between herds.

The safety of an Aujeszky's disease virus vaccine based on strain 783, a deletion mutant which does not express glycoprotein I and thymidine kinase, was assessed in pigs, calves and sheep. Four-day-old piglets which were inoculated intranasally and intramuscularly with 10(7) plaque forming units (PFU) developed only slight depression and fever. The virus was transmitted to a sentinel piglet. Six weeks after inoculation, the pigs were injected with high doses of corticosteroids in an attempt to reactivate the vaccine virus. The pigs did not shed Aujeszky's disease virus, did not develop a rise in virus neutralising antibody titres and sentinel pigs remained seronegative to Aujeszky's disease virus. Strain 783 was passaged in two series of three- to five-day old piglets, but after the third and fourth passages virus could no longer be recovered. Pregnant sows were inoculated with 10(7) PFU of virus strain 783 around day 35 or on day 85 of pregnancy, and their fetuses and piglets were assayed for Aujeszky's disease virus and antibodies against Aujeszky's disease virus. No evidence was found for transplacental transmission of the virus. Calves and sheep were given 10(7) PFU of virus strain 783 intranasally or intramuscularly; they survived and did not develop clinical signs of Aujeszky's disease. All the sheep and the calves inoculated intramuscularly developed neutralising antibodies to Aujeszky's disease virus.

A study was conducted to examine the usefulness of a glycoprotein I (gI)-ELISA to monitor Aujeszky's disease virus infection in two vaccinated pig herds; the gI-ELISA can differentiate between pigs infected with Aujeszky's disease virus and pigs vaccinated against Aujeszky's disease with gI-negative vaccines. The two herds had been vaccinated with gI-negative vaccines for several years. The first survey, in September 1986, revealed that approximately 10 per cent of the breeding pigs in a large multiplier herd were seropositive for antibodies to gI of Aujeszky's disease virus, and it was decided to try to eliminate the virus from the herd by gI-ELISA testing and culling of gI-seropositive pigs. A one month quarantine period for incoming stock was established, and only gI-seronegative pigs were admitted to the herd. After two rounds of testing and culling the herd appeared to be free of wild-type Aujeszky's disease virus, and neither Aujeszky's disease virus nor antibodies could be detected either in 21 sentinel pigs placed on the farm or in 347 stillborn piglets or piglets that died shortly after birth. The herd probably remained free of Aujeszky's disease virus until the end of the 27-month period of monitoring except for two of 639 breeding pigs that were unexpectedly found to be positive in the gI-ELISA in November 1987. These sows were culled. A second breeding herd was monitored for antibodies to gI of Aujeszky's disease virus for two years. The gI-seropositive sows constituted approximately 30 per cent of the herd's breeding pigs, but they were not culled.(ABSTRACT TRUNCATED AT 250 WORDS)

Four methods used for the detection of antibodies to Aujeszky's disease virus in porcine sera were compared. The serum neutralisation test, countercurrent immunoelectrophoresis test, micro-immunodiffusion test and the enzyme-linked immunosorbent assay (Elisa) were assessed with particular regard to their use in large scale screening of porcine serum samples. The Elisa test was found to be the most suitable on the grounds of sensitivity, speed and cost.

A killed gI-negative vaccine combined with a gI enzyme-linked immunosorbent assay (ELISA) test was used for the first time in Sweden in an attempt to eradicate Aujeszky's disease from a weaner pig producing herd. The herd had experienced three severe outbreaks of the disease during a 10 year period and at the start of the programme 96 per cent of the herd's 104 breeding animals were seropositive to the Aujeszky's virus. In addition, there was serological evidence of active virus circulation among younger animals. During the programme, all breeding animals were vaccinated every sixth month and replacement animals were tested free of disease and vaccinated before entry into the herd. When the originally seropositive animals had been rotated out of the herd, all breeding animals and a sample of weaner pigs were tested twice at six weeks' interval. No seroconversions to gI had taken place and the herd was declared Aujeszky's disease-free, 22 months after the start of the programme.

The use of two monoclonal antibodies specific for glycoproteins GI and GIII of the pseudorabies virus led to the development of a competitive ELISA which made it possible to differentiate animals infected with pseudorabies virus from animals vaccinated with the strains of the virus Bartha, NAI4 or Norden. A postvaccinal serological response could be detected from three to four weeks after vaccination. After the virulent challenge of these vaccinated pigs an infectious serological response became apparent two weeks after the challenge.

It has recently been shown that the antibody response to glycoprotein I (gI) of Aujeszky's disease virus can be used to distinguish infected from vaccinated pigs. To examine whether pigs exposed to low doses of a mildly virulent strain of Aujeszky's disease virus produce antibody to gI four groups of four pigs were inoculated intranasally with 10, 10(2), 10(3) or 10(4) plaque forming units (PFU) of the Sterksel strain. Two unvaccinated pigs and two pigs vaccinated intranasally with Bartha's K strain, a gI-negative vaccine, were placed in contact with each group. The pigs given 10 PFU and the in-contact pigs in this group did not become infected. The inoculated and the unvaccinated in-contact pigs in the other groups developed mild signs of illness and produced antibody to gI. Four of six vaccinated in-contact pigs that became infected showed neither clinical signs nor virus shedding and still produced antibody to gI. The other two vaccinated pigs appeared to be resistant to contact-challenge. The antibody response to gI persisted for at least seven months. These results support the idea that Aujeszky's disease virus may be eradicated by a programme based on vaccination with gI-negative vaccines, in conjunction with the detection and subsequent removal of gI-antibody positive, infected, pigs.

Twelve herds were investigated where outbreaks of clinical Aujeszky's disease had occurred. Clinical Aujeszky's disease was eliminated from all farms following vaccination. After vaccination was stopped in two of the six fattening herds virus was also apparently eradicated, judging from serological examination. These two herds were the smallest of the six fattening farms and size and the throughput of pigs may have contributed to apparent eradication of virus. In two of six breeding farms where controlled vaccination is still practised unvaccinated gilts and boars were seronegative. In this case possible eradication of infection may have resulted from either no excretion or insufficient production of virus from vaccinates to infect susceptible non-vaccinates. It is postulated that a properly controlled and monitored vaccination and culling programme may result in the eradication of disease and perhaps also infection from breeding herds experiencing Aujeszky's disease.

An indirect micro enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to Aujeszky's disease virus in pigs is described. A control antigen prepared from infected cells was included for each serum tested. Of 243 sera from serologically positive farms, 175 (72 per cent) and 147 (60 per cent) were positive by the ELISA test and microtitre serum neutralisation test, respectively. Failure to include a control antigen for each serum would have resulted in 14 sera (6 per cent) being differently recorded. Results for sera from experimental and field infections indicated that seroconversion was more quickly detected by the ELISA test than the microtitre serum neutralisation test. In addition to greater sensitivity the ELISA test has other advantages over the serum neutralisation test. ELISA is a rapid, cheap test which is not dependent on a continuous supply of cell cultures and which can be readily automated.

The spread of antibody to Aujeszky's disease virus through a susceptible pig herd was monitored after the probable introduction of infection by a recently purchased boar. The infection spread slowly through the herd but no clinical signs of Aujeszky's disease were seen. The strain of virus isolated was designated NIA-6. It has been characterised by a series of experimental infections and extends the known range of virulence of isolates of Aujeszky's disease virus made in Northern Ireland. The strain caused no disease in four-week-old piglets and is therefore less virulent than other isolates from Northern Ireland pigs. However, it killed rabbits and a proportion of experimentally infected two-week-old piglets, which differentiates it from the avirulent bovine isolate (NIA-4).