Influence of vaccination on Aujeszky's disease virus and disease transmission

Parenteral vaccination of fattening pigs with either modified live or inactivated Aujeszky's disease virus did not prevent infection with field strain virus or the development of clinical disease. The duration and severity of the clinical syndrome was, however, reduced and vaccinated pigs did not suffer the severe weight loss and high mortality experienced by non-vaccinated pigs in the acute phase of disease. The range of tissues in which challenge virus replication took place was more restricted in vaccinated animals and the concentration of virus in infected tissues was reduced. Vaccination shortened the duration of field virus excretion and carriage in the tonsil. Replication of modified live vaccine virus was restricted to the site of inoculation in the neck and associated lymph nodes for two days after vaccination and it was not excreted by vaccinated pigs. Attempts to infect pigs by feeding them tissues taken from non-vaccinated or vaccinated pigs soon after challenge infection were unsuccessful.     

A comparison of the efficacy of two commercial Aujeszky's disease vaccines with glycoprotein-I deletion in pigs

Two commercial Aujeszky's disease vaccines, a modified killed vaccine and a sub-unit vaccine, both carrying a deletion of glycoprotein-I, were evaluated in pigs. Each vaccine was administered to two groups of four pigs, twice at 4-week intervals, with two pigs held as unvaccinated controls. All pigs were challenged with a New Zealand field isolate of Aujeszky's disease virus 3 weeks after the second vaccination. The results indicate that the sub-unit vaccine was able to protect pigs against clinical Aujeszky's disease much better than the pigs vaccinated with the modified killed vaccine when challenged with a virulent virus. However, the amount and the duration of virulent virus excretion following challenge was greater with the sub-unit vaccine than the modified killed vaccine. Pigs vaccinated with the sub-unit vaccine were shown to be latently infected following challenge. Latent infection was demonstrated by excretion of Aujeszky's disease virus from the nasal cavity after dexamethasone treatment and seroconversion of a sentinel in contact pigs to Aujeszky's disease virus.     

Oronasal challenge of fattening pigs after vaccination with an inactivated Aujeszky's disease vaccine

Groups of pigs from vaccinated and unvaccinated sows were vaccinated once or twice between the ages of eight and 20 weeks with a commercial inactivated, oil adjuvanted Aujeszky's disease virus vaccine. Pigs were challenged by the oronasal route when 22 to 27 weeks old. Pigs from unvaccinated sows developed neutralising antibodies after vaccination but no seroconversion was detected in eight-week-old pigs or in 80 per cent of 15-week-old pigs from vaccinated sows. Challenge resulted in severe disease and weight loss in control pigs. In vaccinated animals the duration and severity of clinical signs and the amount of weight lost decreased with increasing serum neutralisation titres. The results indicate that parenteral vaccination at weaning with the vaccine described will not protect pigs at slaughter age against infection and disease, particularly if they were born from seropositive mothers.     

Potency of an experimental DNA vaccine against Aujeszky's disease in pigs

Intradermal vaccination with plasmid DNA encoding envelope glycoprotein C (gC) of pseudorabies virus (PrV) conferred protection of pigs against Aujeszky's disease when challenged with strain 75V19, but proved to be inadequate for protection against the highly virulent strain NIA-3. To improve the performance of the DNA vaccine, animals were vaccinated intradermally with a combination of plasmids expressing PrV glycoproteins gB, gC, gD, or gE under control of the major immediate-early promotor/enhancer of human cytomegalovirus. 12.5 microg per plasmid were used per immunization of 5-week old piglets which were injected three times at biweekly intervals. Five out of six animals survived a lethal challenge with strain NIA-3 without exhibiting central nervous signs, whereas all the control animals succumbed to the disease. This result shows the increased protection afforded by administration of the plasmid mixture over vaccination with a gC expressing plasmid alone. A comparative trial was performed using commercially available inactivated and modified-live vaccines and a mixture of plasmids expressing gB, gC, and gD. gE was omitted to conform with current eradication strategies based on gE-deleted vaccines. All six animals vaccinated with the live vaccine survived the lethal NIA-3 challenge without showing severe clinical signs. In contrast, five of six animals immunized with the inactivated vaccine died, as did two non-vaccinated controls. In this test, three of six animals vaccinated with the DNA vaccine survived without severe clinical signs, whereas three succumbed to the disease. Comparing weight reduction and virus excretion, the DNA vaccine also ranged between the inactivated and modified-live vaccines. Thus, administration of DNA constructs expressing different PrV glycoproteins was superior to an adjuvanted inactivated vaccine but less effective than an attenuated live vaccine in protection of pigs against PrV infection. Our data suggest a potential use of DNA vaccination in circumstances which do not allow administration of live attenuated vaccines.     

Effect of experimental infection with pseudorabies (Aujeszky's disease) virus on pigs with maternal immunity from vaccinated sows.

Live-virus and inactivated-virus vaccines were used to immunize sows against pseudorabies (Aujeszky's disease) virus. To test the efficacy of the vaccination, 53 pigs of different ages were taken from the 1st and the 2nd litters of vaccinated sows and placed separately in isolation units. The pigs were challenge exposed with virulent pseudorabies virus and examined for clinical signs, virus excretion, and serologic reaction. The challenge inoculum caused severe nervous or respiratory signs of disease in 12 of the 13 control pigs, with a mortality of 76%. The pigs from the 1st litters of sows vaccinated with the live-virus vaccine did not become sick, whereas 2 of the 9 pigs (22%) from the 2nd litters had clinical signs and died of pseudorabies. All pigs from sows vaccinated with the inactivated-virus vaccine remained healthy. The results of virus isolation from oronasal swabs, combined with the serotest results, indicated that challenge exposure of all except 1 of the pigs resulted in a subclinical infection with the formation of active immunity.     

Extent and duration of virulent virus excretion upon challenge of pigs vaccinated with different glycoprotein-deleted Aujeszky's disease vaccines

Different deleted Aujeszky's disease vaccines were compared for their ability to induce an immunity which suppresses virus excretion optimally upon infection. Groups of pigs were vaccinated once with attenuated deleted Aujeszky's disease vaccine (gI, gX or gp63 negative), suspended in phosphate buffered saline. Two additional groups were vaccinated with a gI deleted vaccine virus suspended in an oil-in-water emulsion. Other groups were vaccinated twice with gI deleted inactivated vaccines. The three control groups included were: pigs immune after infection, unvaccinated pigs and pigs receiving vaccine without known deletion in the envelope. Experimental challenge took place 3 or 4 weeks after the only or the last vaccination. The number of excreting pigs, the duration of excretion and the virus titers excreted, were determined for all the groups. All the pigs vaccinated with glycoprotein deletion vaccines suspended in phosphate buffered saline, excreted virus for 2 to 6 days after challenge. A 100 to 1000 fold reduction in excreted virus titers was obtained in vaccinated pigs compared to unvaccinated ones. Some vaccines suppressed virus excretion better than others, but no correlation could be made between the type of deletion (gI, gX or gp63) and the degree of reduction in virus excretion. Similar results were obtained with two applications of inactivated vaccines. The lowest number of excreting pigs, the lowest duration of excretion and the lowest titers were obtained in groups vaccinated with the attenuated vaccine suspended in an oil-in-water emulsion. No vaccine suppressed virus excretion totally.     

Induction of antibodies to glycoprotein I in pigs exposed to different doses of a mildly virulent strain of Aujeszky's disease virus

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.     

Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism

The efficacy of four different commercial live vaccines (vaccines A, B, C, and D) against the infectious laryngotracheitis virus (ILTV) was assessed in specific-pathogen-free (SPF) chickens. SPF chickens were vaccinated intraocularly at 6 wk old with ILTV live vaccines and were challenged intratracheally with the N91B01 strain of virulent Korean ILTV 2 wk after vaccination. The immunity against ILTV live vaccines was assessed by the incidence of latent infection by the challenge virus in the chickens' tracheas and trigeminal ganglia, the reisolation rate of the challenge virus, and the clinical signs in the chickens challenged with the N91B01 strain of ILTV. The latent infection in chickens was assessed by nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Our data showed that the clinical signs and challenge virus isolation were negative in all chickens receiving four difference commercial ILTV live vaccines. The viral DNA of the vaccine strain, but not that of the challenge virus, was detected in chickens vaccinated with vaccine A by nested PCR-RFLP. The viral DNAs of both the vaccine and challenge strains were detected from chickens vaccinated with vaccines B, C, and D. This study showed that only vaccine A can protect chickens from latent infection with the field virulent ILTV. We speculate that the efficacy of infectious laryngotracheitis live vaccines to protect chickens from latent infection with virulent ILTVs can be assessed by nested PCR-RFLP analysis.     

A novel concept for the control of Aujeszky's disease: experiences in two vaccinated pig herds

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)     

gIII antibody responses in pigs following vaccination and challenge to Aujeszky's disease.

Serological responses to a genetically engineered Aujeszky's disease "marker" vaccine (dl gIII + dl tk) were monitored using a blocking-ELISA (B-ELISA), a serum neutralisation test (SNT) and an indirect ELISA (I-ELISA). The B-ELISA is capable of differentiating pigs vaccinated with the above vaccine from natural infection. The SNT and the I-ELISA indicated that the pigs responded to vaccination and challenge. All three tests showed that the controls and the in-contact pigs always reacted negative for antibodies. The B-ELISA was able to detect pigs challenged with a field isolate 24 days post-challenge. These pigs remained positive until 110 days post-challenge when last tested. These findings indicate that the B-ELISA could be used successfully with this vaccine in a control eradication programme. This trial also shows that the vaccine virus did not spread to the in-contact pigs and also the vaccinated and challenged pigs did not transmit the disease to other susceptible pigs when they were introduced 14 days after challenge.     

Safety of an Aujeszky's disease vaccine based on deletion mutant strain 783 which does not express thymidine kinase and glycoprotein I

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 of the ability of a TK-negative and gI/gE-negative pseudorabies virus (PRV) mutant inoculated by different routes to protect pigs against PRV infection

The capacity of a TK-negative (TK-) and gI/gE-negative (gI/gE-) pseudorabies virus (PRV) mutant to protect pigs against Aujeszky's disease carried out by experimental infection with a virulent PRV strain, was tested. There were three groups, each of four susceptible pigs which were inoculated twice by two different schedules. Group 1 received the modified virus by the intradermal (first inoculation)-intramuscular (second inoculation) routes; group 2 was treated by the intranasal (first inoculation)-intramuscular (second inoculation) routes. The third group was left untreated as the control. All of the pigs were challenged intranasally with a virulent PRV strain and they were subsequently injected with dexamethasone. Two pigs in each group were necropsied on days 5 and 15 after dexamethasone inoculation. The challenge exposure resulted in mild clinical signs, increase in growth and a shorter period of virus shedding in vaccinated pigs, whereas the control group showed severe signs of Aujeszky's disease. No difference in the titre of the virulent virus which was excreted by pigs of all three groups, was observed and all animals seroconverted. Both the mutant strain and the wild-type virus established a latent infection although only the latter was reactivated and shed. Slight lesions were observed in target tissues of the vaccinated animals and no significant differences were detected between the two inoculation schedules.     

基于SYBR Ⅰ实时荧光定量PCR对猪流行性腹泻病毒野毒和疫苗弱毒鉴别诊断方法的建立

本研究参考GenBank中登录的猪流行性腹泻病毒(Porcine epidemic diarrhea virus,PEDV)野毒株和弱毒疫苗株(CV777弱毒疫苗株)在高度保守ORF3基因核苷酸序列的差异,设计一对特异性荧光定量引物,分别建立基于SYBRⅠ实时荧光定量RT-PCR方法。结合熔解曲线分析可见,其野毒株和弱毒疫苗株熔解温度(Tm)分别为(81.84±0.17)℃和(83.16±0.14)℃,扩增产物的熔解曲线分析均只出现1个单特异峰,对传染性胃肠炎病毒、猪轮状病毒、猪细小病毒、猪流感病毒、猪繁殖与呼吸综合征病毒、猪伪狂犬病毒、猪瘟病毒均检测不到荧光信号。结合熔解曲线可直接鉴别猪群中PEDV的感染情况和程度,可对免疫猪群PEDV野毒感染和疫苗免疫做出快速准确的鉴别诊断,尤其是对PEDV弱毒疫苗免疫后仍爆发PEDV野毒感染的研究更有临床意义。     

Vaccination of sheep against Aujeszky's disease with a gI-deleted mutant]

The use of gl deleted live vaccines against Aujeszky's disease (AD) facilitates to differentiate vaccinated from field-virus infected animals. In this study different modes of vaccination were tried to find out how sheep can be protected from a lethal infection with ADV. It could clearly be demonstrated that Aujeszky disease virus (ADV) is spread by horizontal transmission from infected pigs to sheep. The nasal discharges of infected pigs contained a maximum of 10(8.75)TCID50/g mucus at days 3 and 4 p.i. and those of the contact-pigs 10(8.5)TCID50/g mucus at days 6 and 7 after contact. Non-vaccinated contact sheep were infected horizontally by the pigs. The highest titres ranged from 10(6.25) to 10(7.5)TCID50/g mucus. These animals were sacrificed at day 5 p.i. exhibiting acute symptoms of AD. The nasal discharge of vaccinated sheep contained much lower amounts of ADV (maximum: 10(4.25)TCID50/g mucus). All surviving animals had developed antibodies. Following challenge with the ADV-strain NIA3, no febrile response or virus-shedding was observed in sheep vaccinated 2x s.c. or 2x i.m. with a gl deleted live vaccine, whereas sheep, vaccinated only 1x i.m. (4 out of 4 animals) or 1x i.m. (3 out of 4 animals) or 1x i.n. and 1x i.m. (1 out of 4 animals) had to be sacrificed after showing acute symptoms of AD. In conclusion it can be stated that a double parental vaccination with a gl deleted live vaccine protects sheep against a field-virus AD infection.     

Studies on immunisation of pigs with the Bartha strain of Aujeszky's disease virus.

The K strain of Aujeszky's disease virus (ADV) grown in Vero cells was used to vaccinate pigs. Following intramuscular inoculation, the pigs remained healthy, no vaccine virus was excreted and virus could be detected only at the inoculation site. One inoculation gave good protection against challenge with a virulent strain of ADV, and the amount of virulent ADV excreted was geatly curtailed. Following vaccination only low leads of serum neutralizing antibody were detected (geometric mean titre 1/2), but three weeks after challenge very high levels were found (GMT 1/1773). Intranasal vaccination gave similar results. There was minimal excretion of vaccine virus. The clinical reaction on challenge was less severe than in the intramuscularly challenged group, although lower antibody levels were detected three wekks following challenge (GMT 1/483). A field trial, using this strain given subcutaneously, indicated that one inoculation of this vaccine is effective.     

Field experiments to evaluate the feasibility of eradication of Aujeszky's disease virus by different vaccination schedules.

In the present report, the extent of the reduction in Aujeszky's disease virus (ADV) dissemination achieved when pigs were intensively vaccinated with gI-deleted vaccines under field circumstances, was examined. On widely dispersed breeding-fattening farms, a gI-negative status was most rapidly obtained and the rate of new waves of infections was lowest when the attenuated Bartha strain was administered to both the sows and the fatteners. It was more difficult not only to reach but also to keep a gI-negative status on farms on which the sows were vaccinated with an inactivated vaccine and the fatteners with the attenuated Bartha strain or when the fattening pigs were not vaccinated at all. In a densely populated area, 9 of the 17 farms had gI-positive fatteners at the start of the intensive vaccination programme in which the attenuated Bartha strain was given to both the sows and the fatteners. Antibodies were not detected in the sera of the fatteners of each farm at some time during the experiments, but the fatteners on 7 of the 18 farms still showed antibodies against gI after 20 months of vaccination. At the end of the experiment, the percentage of fatteners with antibodies on these farms was markedly reduced compared with the percentage at the start of the experiment. Therefore, elimination of field virus may be feasible if intensive vaccination is carried out over a sufficiently long period of time. However, the high rate of reinfections experienced either due to reintroduction of the virus or to recrudescence should be a warning against too much optimism, particularly in regions with a dense swine population.     

Influence of vaccination route on the efficacy of Aujeszky's disease deletion-mutant vaccine.

In order to compare the effect of the route of immunization on the efficacy of a modified live Aujeszky's disease (AD) vaccine, which had deletions in both thymidine kinase (TK-) and glycoprotein gIII genes (gpIII-), 20 six-week-old pigs were vaccinated by either the intramuscular (IM) (n = 10) or subcutaneous (SC) (n = 10) route. All the animals, including five non-vaccinated control animals, were challenged with virulent AD virus 22 days after vaccination. Four of five non-vaccinated animals died within 12 days after challenge. Although none of vaccinated animals died, three of animals in the SC group exhibited clinical signs, and average daily gains in the SC group were depressed. The animals in the IM group were not found to shed challenge virus, but those in the SC group shed the virus up to 9 days. Virus neutralizing antibody titers in the vaccinated animals were low or non-detectable by 21 days after vaccination. A glycoprotein gII (gpII) screening ELISA detected gpII antibody in all animals in the IM group. While, only 30% of animals in the SC group were positive by the same test. The results of this study indicate that TK-, gpIII modified live AD virus vaccine is effective against challenge with virulent AD virus; however, vaccination by the SC route reduced vaccine efficacy in comparison with IM route.     

Intranasal vaccination of pigs against Aujeszky's disease: comparison with one or two doses of attenuated vaccines in pigs with high maternal antibody titres

Ten-week-old pigs with high levels of maternally derived antibody (MDA) against Aujeszky's disease virus (ADV) were given either a single intranasal vaccination or one or two doses (with an interval of three weeks) of commercially available attenuated ADV vaccines intramuscularly. The pigs did not produce a clear neutralising antibody response to ADV. However, pigs vaccinated intranasally and pigs given two doses of attenuated ADV vaccines were protected against intranasal challenge with virulent ADV two months after the first vaccination. Pigs given one parenteral dose of attenuated ADV vaccine were insufficiently protected. Protection was shown by shorter periods of growth arrest and fever and a greater reduction of virulent virus shedding after challenge in vaccinated pigs than in unvaccinated control pigs. Although intranasal vaccination conferred protection comparable to two parenteral doses of attenuated vaccines, it reduced shedding of virulent virus much more effectively. These results, together with those of other studies, show that intranasal vaccination confers better protection against Aujeszky's disease in pigs with MDA than parenteral vaccination. However, the efficacy of intranasal vaccination also decreases with increasing levels of MDA at the time of vaccination.     

Protective effects of vaccination with bovine leukemia virus (BLV) Tax DNA against BLV infection in sheep

A DNA vaccination trial was performed on sheep to determine whether vaccination with bovine leukemia virus (BLV) transactivator Tax DNA is effective against BLV infection. Immunization was carried out with cationic liposomes containing the Tax-expressing plasmid DNA and subsequently all sheep were challenged with BLV. BLV titers in peripheral blood mononuclear cell (PBMC) determined by syncytium formation assay and BLV provirus load detected by genomic PCR analysis showed higher levels of virus titers in control sheep than those in Tax-vaccinated sheep. Higher levels of IFN-gamma mRNA expression have been demonstrated in vaccinated sheep after the challenge. These results suggested that Th1 type immune response induced by Tax DNA vaccine inhibited BLV propagation in vaccinated sheep at the early phase of infection.