Aujeszky's disease (pseudorabies) virus detection in cerebrospinal fluid in experimentally infected pigs

The presence of Aujeszky's disease virus in cerebrospinal fluid of experimentally infected pigs was studied using the techniques of virus isolation and PCR. Pigs, some of which were previously vaccinated against Aujeszky's disease, were inoculated with different doses of the Aujeszky's disease NIA-3 strain. At the time of death or sacrifice, a sample of cerebrospinal fluid was taken and tested for the presence of virus using the mentioned techniques. Virus was isolated only from one sample, while it was detected by PCR in most of them. The higher sensitivity of the PCR technique and the possible presence of antiviral antibodies in the cerebrospinal fluid are reasons that can be argued to explain this fact. By PCR, the virus was detected more efficiently when digested cerebrospinal fluid cells were used as DNA source than when using whole cerebrospinal fluid, suggesting that the virus could be cell-associated. Aujeszky's disease virus could not be detected by PCR in pigs which survived the acute phase of the infection and were euthanased at 8 weeks post-inoculation, when they were latently infected. This indicated that the cerebrospinal fluid is not an adequate sample for the diagnosis of latency. Since Aujeszky's disease virus was detected from most of the tested samples, we believe that this could be an adequate procedure for the quick diagnosis of Aujeszky's disease.     

Patterns of seroconversion to Aujeszky's disease virus in unvaccinated chronically infected Swedish weaner pig-producing herds

This study was carried out in nine unvaccinated Swedish weaner pig-producing herds, ranging in size from 20 to 134 breeding animals, which had experienced a previous outbreak of Aujeszky's disease (AD) and in which seropositive sows were still present. The objective was to quantify the seroconversion rate to Aujeszky's disease virus (ADV) in breeding animals and to find out whether unvaccinated herds could become free from ADV by using only seronegative animals for replacement. Serum samples were initially obtained from all adult pigs in the herds. Animals testing negative, as well as replacement animals, were subsequently tested every second or third month and the herds were followed for 10-28 months. The herd owners were advised to keep seropositive and seronegative animals separate whenever possible and to implement sanitary measures against the reintroduction of the virus into the herds. All herds experienced long consecutive periods (> or = 4 months, median 9 months) without seroconversion. Three herds became free from ADV during the serological study; in two of these herds no seroconversion was observed. In those seven herds where seroconversion occurred, between 9 and 86% of the susceptible pigs became infected. The reinfection was indicative of the reactivation of latent virus in all herds but two, where the reintroduction of the virus was most likely. The pattern of seroconversion was either sporadic, involving not more than three animals at a time, or epidemic, involving a large proportion of the susceptible animals in the herd. Severe clinical outbreaks hit the two largest herds and these only became ADV-free after the conclusion of the study when vaccine was used, which reduced the incidence of seroconversion to zero. The results from the present study show that it is possible for smaller herds to achieve freedom from ADV without any radical control methods, provided that only ADV-free replacement animals are used and the virus is not reintroduced. Moreover, the finding that long periods can elapse without transmission suggests that when infection is discovered in a herd, the risk of massive spread is not necessarily imminent.     

Prevalence of herds with young sows seropositive to pseudorabies (Aujeszky's disease) in northern Belgium

In Belgium, pseudorabies in swine has been the subject of a mandatory eradication programme since 1993. From December 1995 to February 1996, a survey was conducted in the five provinces of northern Belgium to estimate the provincial pseudorabies virus (PRV) herd seroprevalence. Seven hundred and twenty randomly selected herds were included in this survey. To detect recently infected animals, only young sows were sampled. The results show that 44% of these herds had an important number of PRV-seropositive young sows. The highest herd seroprevalence was observed in West Flanders (68%), followed by Antwerp (60%), East Flanders (43%), Limburg (18%), and Flemish Brabant (8%). Assuming a diagnostic test sensitivity and specificity of 95% and 99%, respectively, and a true PRV within-herd prevalence of 43%, the overall true PRV herd prevalence was estimated to be 35%. A logistic multiple-regression revealed that the presence of finishing pigs was associated with a two-fold increase in odds of a herd being seropositive (odds ratio (OR)=2.07, 95% confidence interval (CI)=1.31–3.26); a breeding herd size ≥70 sows was associated with a four-fold increase in odds of a herd being seropositive (OR=4.09, 95% CI=2.18–7.67); a pig density in the municipality of ≥455 pigs/km² was associated with a 10-fold increase in odds of a herd being seropositive (OR=9.68, 95% CI=5.17–18.12). No association was detected between the PRV herd seroprevalence and purchase policy of breeding pigs (purchased gilts, or use of homebred gilts only).     

The role of spatial factors in the success of an Aujeszky's disease eradication programme in a high pig density area (Northeast Spain, 2003–2007)

We tested the role of several spatial variables on the risk of a sow herd being Aujeszky's disease virus (ADV) seropositive in certain areas of North Eastern Spain and during different periods of the eradication programme. Distance to the nearest slaughterhouse, distance to the nearest conventional road and number of ADV serologically positive sows and ADV serologically positive fattening pigs within different distances (1000, 1500 and 2000 m) of each sow herd, were included in a hierarchical Bayesian binomial model. A variable without spatial characteristics, type of herd (farrow to weaning and farrow to finish), was also included. Presence of positive fattening pigs or positive sows up to a distance of 1500 m of a sow herd increased its risk of being seropositive, although this variable had no effect on the risk when located at distances up to 1000 or 2000 m. The number of seropositive sows increased the risk of a sow herd being ADV seropositive only in the first period of study, when the proportion of serologically positive sow herds was nearly 60%. The spatial pattern of the residuals of the hierarchical Bayesian binomial model (observed versus predicted) was very similar to the observed infection in sow herds in all of the eradication periods, showing that spatial factors might not be the main factors related to the eradication of Aujeszky's disease from sow herds. Other herd-specific risk factors might be much more strongly related to the risk of a sow herd being ADV seropositive.     

Survival of Aujeszky's disease virus in infected pig slurry

It has been demonstrated that after experimental infection of pig slurry from the space under the slatted floor (infection dose of 10(6)PFU per ml), the Aujeszky's disease virus (ADV) survived for 72 hours at the temperature of 15 degrees C and at pH 6.5, but was inactivated after 96 hours. When technologically treated pig slurry from the storage tanks was saturated with water and infected with ADV at the dose of 10(5)PFU per ml, the virus survived for 23 days when kept at 15 degrees C and 4 degrees C and at pH 6.8, but was inactivated under the same conditions after 30 days. When the infective ADV dose in the technologically treated pig slurry in the storage tanks was reduced to 10(4)PFU per ml, the virus survived 16 days at +4 degrees C and pH 7.0 and 8.0 but was inactivated within 23 days after infection.     

Presence of wild type Aujeszky''s disease virus in swine identified as subclinical low-prevalent serological test reactors within qualified virus-negative herds

Subclinical low-prevalent Aujeszky's disease (AD) serological test reactors test reactors are defined as those few swine within a qualified AD virus (ADV)-negative herd that have antibodies to wild type virus. However, clinical signs of the associated diease are not observed in these putatively infected swine or elsewhere in the herd. Twelve such animals, including 7 previously vaccinated with a genetically modified ADV, were identified in Illinois (USA) during a 2.5 year period. The humoral immune responses of the 5 nonvaccinated swine were assessed by an enhanced virus neutralization test and a radioimmunoassay. Anti-ADV antibodies were determined to be present in the serum from 4 of these swine. Attempts to isolate ADV by in vitro and in vivo inculations of cell cultures and weanling mice, respectively, of tonsillar and trigeminal nerve ganglionic tissue preparations from each animal (vaccinated and nonvaccinated) were unsuccessful. Tonsillar and trigeminal nerve ganglionic tissues of each animal were screened for the presecence of wild type and/or vaccine viral genomes by a soluble polymerase chain reaction (PCR) coupled with Southern hybridization. Unique PCR primers were used distinguish between wild type and vaccine viral DNAs. Additional PCR procedure, which amplifiers a portion of the essential and highly conserved viral gp50 gene, also was employed in an effort to detect viral genomes. Wild type viral DNA was found in the tissues from at least 5 of the vaccinated and 3 of the nonvaccinated swine. These results indicate that such animals should be considered as being infected with ADV. Further, these findings emphasize the need to develope highly specific and sensitive antemortem testing methods for accurate assessment of ADV infection in herds containing such subclinical, yet serologically positive, swine.     

Subclinical Aujeszky's disease virus infection in a pig herd and the characterisation of the strain of virus isolated

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).     

Implications derived from a mathematical model for eradication of pseudorabies virus

Simple mathematical models based on experimental and observational data were applied to evaluate the feasibility of eradicating pseudorabies virus (PRV) regionally by vaccination and to determine which factors can jeopardise eradication. As much as possible, the models were uncomplicated and our conclusions were based on mathematical analysis. For complicated situations, Monte-Carlo simulation was used to support the conclusions. For eradication, it is sufficient that the reproduction ratio R (the number of units infected by one infectious unit) is < 1. However, R can be determined at different scales: at one end the region with the herds as units and at the other end compartments with the pigs as units. Results from modelling within herds showed that contacts between groups within a herd is important whenever R between individuals (R_ind) is 1 in one or more groups. This is the case within finishing herds. In addition, if the R_ind is more than 1 within a herd, the size of the herd determines whether PRV can persist in the herd and determines the duration of persistence. Moreover, when reactivation of PRV in well-vaccinated sows is taken into account, R_ind in sow herds is still less than 1. In sow herds with group-housing systems, it is possible that in those groups R_ind is 1. Results from modelling between herds showed that whether or not R_herd is < 1 in a particular region is determined by two factors: (1) the transmission of infection between nucleus herds and rearing herds through transfer of animals and (2) contacts among finishing herds and among rearing herds. The transmission between herds can be reduced by reduction of the contact rate between herds, reduction of the herd size, and reduction of the transmission within herds.     

Evidence of long distance airborne transmission of Aujeszky's disease (pseudorabies) virus

Aujeszky's disease has been the subject of an eradication campaign in Denmark since 1980. A detailed knowledge of the virus strains present in the country was provided by restriction fragment analyses of older clinical isolates, and of isolates from all the virologically confirmed outbreaks since 1985. The introduction of foreign strains into southern border areas was demonstrated during the winters of 1984/85, 1986/87 and 1987/88. An epizootic during the winter of 1987/88 was shown to correlate with an unusual predominance of southerly winds. Both conventional and specific pathogen free herds became infected. A herd level case-control analysis of the outbreaks during the winter of 1987/88 revealed that there was a positive correlation between the risk of infection and the size of the herd. The observations support the hypothesis of airborne transmission of the disease.     

Survival rate of Suid herpesvirus (SuHV-1, Aujeszky's disease virus, ADV) in composted sewage sludge

Sewage sludge constitutes a source of valuable biogenic raw materials, but it is a carrier of many pathogenic microorganisms and viruses. Subjected to an effective sanitization by means of the process of composting, it is suitable to use in agriculture as fertilizers. The aim of this study was to observe the survival rate of Suid Herpesvirus under the influence of the temperature alone (water bath) as well as in sewage sludge subjected to the process of composting (pile). The samples were taken at different time intervals, and the virus titres were determined. The viruses survived considerably longer under laboratory conditions: at 30 degrees C as long as 21 days, at 40 degrees C - 93 hours, and at 50 degrees C - less than an hour. In the compost pile, in spite of the lack of the thermophylic phase, the total survival time of the viruses ranged from 34 to 44.5 hours, which indicates the vast importance of other physicochemical factors, apart from the temperature, contributing to virus inactivation.     

Variation of Aujeszky's disease viruses in wild swine in USA

In the United States of America, Aujeszky's disease (pseudorabies) has been eradicated from all domestic swine. Some re-emergence of infection occurred as vaccine use diminished. Sporadic outbreaks have also occurred because of the reservoir of infection in feral swine that have spread across the southern two-thirds of the country and Hawaii. In order to be able to understand the origins of re-emerging virus, sequence analysis of variable genes in pseudorabies virus (PRV) has been used to differentiate strains. Most PRV from feral swine can be distinguished from virus circulating in domestic pigs during the national epizootic. However, several feral swine isolates of PRV from south central states are closely related or identical in sequence to strains from domestic pigs. Extensive study by PCR for the presence of virus in the oral cavity of feral pigs disclosed that the viral DNA is distributed widely in tonsils salivary glands, taste buds and even mucosa in the vicinity of tusks. Clearly the virus in feral swine has multiple mechanisms of transmission to insure persistent infection and the threat of re-emergence in domestic swine continues.     

Herd factors affecting the selection and success of intervention strategies in the program for eradication of pseudorabies (Aujeszky's disease) virus from Illinois swine farms

The program for eradication of pseudorabies virus (PrV) from swine herds in Illinois was evaluated with respect to compliance with Livestock Conservation Institute (LCI) guidelines for selection of intervention strategies and for the effectiveness of these interventions under different herd conditions. The sample consisted of 395 swine operations quarantined between 1988 and 1994. These herds were followed until the end of 1996. The association of herd characteristics (number of sows, sow PrV seroprevalence, type of housing, number of PrV-seropositive farms within 1.5 mi) at the time of quarantine with the producer's selection of an initial intervention strategy (vaccination, offspring segregation, test-and-removal, depopulation-and-repopulation) was analyzed using logistic multiple regression. The interaction of herd characteristics with intervention strategies to affect the duration of quarantine was analyzed using multivariable Cox regression.

Factors favoring selection of vaccination were increased herd size, higher sow PrV seroprevalence, and more PrV-seropositive farms within 1.5 mi. Offspring segregation was preferred when sow PrV seroprevalence was higher, and test-and-removal was preferred when seroprevalence was lower. Depopulation-and-repopulation was more likely in outdoor operations. Except for depopulation-and-repopulation, selection of an intervention strategy was in accordance with LCI guidelines.

Vaccination and offspring segregation were associated with longer times under quarantine, and test-and-removal and depopulation-and-repopulation with shorter times. Test-and-removal was more effective in reducing the duration of quarantine when sow PrV seroprevalence was low. Vaccination increased the duration of quarantine less when sow PrV seroprevalence was high. Vaccination increased the duration of quarantine more when there were one or more PrV-seropositive farms within 1.5 mi than when there were no PrV-seropositive farms within 1.5 mi. It is apparent that herd characteristics affect the duration of quarantine and therefore need to be taken into account in the selection of a PrV-intervention strategy.     

A study of outbreaks of Aujeszky's disease in cattle. I. Virological and epidemiological findings.

Twenty-nine outbreaks of Aujeszky’s disease in cattle, involving 54 animals, were studied virologically and epidemiologically. The outbreaks could be divided into two distinct groups, viz., such with pruritus on the anterior (Group I) and such with pruritus on the posterior part of the body (Group II). Though low, the average number of affected animals per outbreak was twice as high in Group I as in Group II. Besides in the tissue of the central nervous system, virus was demonstrated in the oral, pharyngeal, and nasal mucous membranes of eight out of 12 animals from Group I outbreaks, which was thought to be indicative of respiratory infection, and in the vagina of the three animals with perineal pruritus from which such material was examined. It is concluded that the site of pruritus need not be identical with the site of virus entry. The probable routes of infection in the outbreaks examined will be discussed in detail in a subsequent paper.     

A study of outbreaks of Aujeszky's disease in cattle. II. Further investigations on the routes of infection.

The probable routes of infection in 29 outbreaks of Aujeszky’s disease in cattle are discussed in the light of virological and epidemiological data, given in a preceding paper, and of histories of the individual outbreaks given in the present paper. The view was substantiated that anterior pruritus is mostly associated with infection by the respiratory route. In outbreaks with posterior pruritus the route of infection appeared to be a question of a more complex nature. With reference to cases examined later, it is concluded that perineal pruritus is often associated with vaginal infection. In a few cases there was evidence of virus transmission by man. Three cases with pruritus on a hind leg were suggestive of an alimentary infection. In many outbreaks with posterior pruritus, the route of infection remained obscure.