Classical Swine Fever Vaccine Research Articles

Detection of classical swine fever vaccine virus in blood and tissue samples of pigs vaccinated either with a conventional C-strain vaccine or a modified live marker vaccine

Attenuated live classical swine fever (CSF) viruses are the most efficacious vaccines against the disease. However, little is known about the distribution and detection of CSF vaccine viruses in the host. We therefore compared the new recombinant attenuated marker vaccine virus CP7_E2alf with the conventional C-strain vaccine concerning virus isolation, antigen-, and genome-detection in different samples within the first 42 days post-vaccination (p.v.). Leukocytes and several organs such as tonsils, lymph nodes, spleen, thymus, parotis and kidney were also tested using highly sensitive real-time reverse transcription-polymerase chain reaction (RT-PCR) techniques. It was demonstrated that vaccine virus could be detected by live animal sampling only in a few leukocytes samples at very low titres and genome copy numbers within the first 14 days after immunisation. Vaccine virus could also be isolated from individual tonsil samples within the first 6 days after vaccine application. In contrast, vaccine virus genomes were consistently detected in the tonsils up to day 42 by real-time RT-PCR. Distribution, amount of virus and viral genome levels were similar for both tested vaccines. In conclusion, blood samples could be the sample material of choice for detecting CSF wild type virus infection even in vaccinated animals after more than 14 days p.v., while tonsil sampling provided appropriate material for long-term detection of both tested CSF vaccine viruses using real-time RT-PCR methods.

Evaluación del diagnóstico de la peste porcina clásica y del rendimiento de la vacuna

Rapid and accurate diagnosis is of the utmost importance in the control of epizootic diseases such as classical swine fever (CSF), and efficacious vaccination can be used as a supporting tool. While most of the recently developed CSF vaccines and diagnostic kits are mostly validated according to World Organisation for Animal Health (OIE) standards, not all of the well-established traditional vaccines and diagnostic tests were subject to these validation procedures and requirements. In this report, data were compiled on performance and validation of CSF diagnostic tests and vaccines. In addition, current strategies for differentiating infected from vaccinated animals are reviewed, as is information on the control of CSF in wildlife. Evaluation data on diagnostic tests were kindly provided by National Reference Laboratories for CSF in various European countries.

Epidemiological survey of viral diseases of pigs in the Mekong delta of Vietnam between 1999 and 2003

In the Mekong delta, backyard pig rearing plays an integral role in recycling nutrients in farming systems and generating valuable cash income. However, development has been hampered by fatal epizootics of piglets and reproductive failure of sows. Diseases are named by symptoms and blindly treated with antibiotics. As antibiotics are often ineffectual, involvement of viral diseases are suspected. To identify the causative agent, we first sero-surveyed porcine reproductive and respiratory syndrome (PRRS) and pseudorabies with 478 sera from non-vaccinated pigs collected from backyard farms, state farms and slaughterhouses in Can Tho province between 1999 and 2002. Antibodies for PRRS were first detected in 2002 in backyard farms and at high prevalence in state farms with increased piglet mortality. A few backyard breeder pigs had antibodies for pseudorabies in 2000 and 2002. With compulsory classical swine fever (CSF) vaccination, we examined the relationship between vaccination and antibodies in 70 serum samples. Seventy-nine percent of vaccinated breeders had CSF antibodies—higher than expected with irregular vaccination. Since circulation of CSF virus was suspected, isolation was attempted at 10 farms with fatal epizootics between 2002 and 2003. The viruses were detected at all farms and clustered within genogroup 2, despite vaccines corresponding to genogroup 1. This study demonstrated virologically/serologically the existence of PRRS, pseudorabies and CSF viruses in the Mekong delta of Vietnam. We also identified CSF as a cause of piglet mortality that disastrously affected backyard farming. Vaccine standardization and proper instructions are needed to simplify diagnosis and complement established simultaneous vaccination of sows with piglets.

Determination of genotoxicity of classical swine fever vaccine in vitro by cytogenetic and comet tests

Chromosome damage in lymphocyte cultures induced by live virus vaccine against classical swine fever (CSF) has been observed in previous studies. In vivo cytogenetic tests were made with several doses of vaccines used in Argentina to control the disease. These studies have shown that genotoxic effects increased with dose. In the present study, two different in vitro assays were performed by recording the frequency of cells with chromosome alterations and by assessing the ability of the vaccine to damage DNA, using the single cell gel microelectrophoretic assay (comet test). Frequencies of cells with chromosomal alterations increased significantly when compared with controls and were dose (microl/ml) dependent: 0 = 1.23, 5 = 2.29, 10 = 5.42 and 20 = 11.71%. In the comet assay the variables measured, tail length (TL) and tail moment (TM), also increased. For control cultures TL was 2.32 microm, whereas with concentrations of 20 and 100 microl/ml TL were 12.47 and 42.3 microm, respectively. TM of control cultures was 0.18, whereas with vaccine concentrations of 20 and 100 microl/ml TM were 5.52 and 24.52, respectively. Comet frequency distributions differed significantly among treatments. These results agree with previous in vivo observations. Regarding CSF pathogeny, our results support a direct effect of CSF vaccinal virus on lymphocyte DNA. Genotoxicity of CSF vaccine was corroborated in vitro at the cytogenetic and molecular levels.

Negative impact of porcine reproductive and respiratory syndrome virus infection on the efficacy of classical swine fever vaccine

Recent findings suggest that porcine reproductive and respiratory syndrome virus (PRRSV) possesses immunomodulatory properties. To investigate the effect of PRRSV infection on classical swine fever (CSF) vaccine efficacy, 17-day-old pigs were divided into five groups. The experimental group was infected with a Thai PRRSV (US genotype) a week before CSF vaccination and challenged with a virulent CSF virus (CSFV) 3 weeks following vaccination. The control groups received no PRRSV infection, no CSF vaccination, no CSF challenge, or in combination were included. The results demonstrated that PRRSV infection significantly inhibited host immune response that resulted in vaccination failure in the subsequent CSFV exposure. Following CSF challenge, the PRRSV-infected, vaccinated pigs exhibited clinical, virological and pathological features resembled to those of the non-vaccinated groups. The findings indicated that CSF immunization during an acute phase of PRRSV infection could result in vaccination failure.

The kinetics of cytokine production and CD25 expression by porcine lymphocyte subpopulations following exposure to classical swine fever virus (CSFV)

Surface expression of IL-2R-alpha (CD25) is widely used to identify activated lymphocyte populations, while interferon-gamma (IFN-γ) levels have been shown to be a good indicator of cell-mediated immunity (CMI) in pigs. To investigate the relationship between these two parameters, we developed an intracellular cytokine-staining assay and studied the kinetics of cytokine (IFN-γ and interleukin-10, IL-10) production relative to CD25 expression in porcine lymphocyte subpopulations, following immunization with a classical swine fever (CSF) vaccine. The number of activated memory T cells (CD4 +CD8 +CD25 + cells) increased slightly in the peripheral blood mononuclear cell (PBMC) population soon after vaccination, then diminished within a few weeks. The number of activated cytotoxic T cells (CD4 −CD8 +CD25 + cells) peaked approximately 2 weeks after the memory population. Although the number of IFN-γ producing cells detected in this experiment was relatively low, the CD4 +CD8 + T cells were major IFN-γ producers in the PBMCs throughout the experiment. In another experiment, CSF-vaccinated pigs were challenged with a virulent classical swine fever virus (CSFV), and the kinetics of CD25 expression and cytokine productions were monitored. Following exposure to the virus, the number of IFN-γ producing cells in the PBMCs increased markedly in both the vaccinated and unvaccinated groups. The CD4 −CD8 + cells were major IFN-γ producing cells in vaccinated pigs, while both CD4 +CD8 + and CD4 −CD8 + populations contributed to the IFN-γ production in the control group. Interestingly, the enhanced IFN-γ production was not associated with the upregulation of CD25 expression following the CSFV challenge. In addition, exposure to the virulent CSFV significantly increased interleukin-10 production by the CD4 −CD8 + populations in PBMCs of the unvaccinated pigs. Taken together, our results indicated that CD25 expression and IFN-γ production were not tightly associated in porcine lymphocytes. In addition, the CD4 −CD8 + lymphocytes of the PBMCs played a major role in cytokine productions following the CSFV challenge.

Vaccinology of classical swine fever: from lab to field

There are two types of classical swine fever vaccines available: the classical live and the recently developed E2 subunit vaccines. The live Chinese strain vaccine is the most widely used. After a single vaccination, it confers solid immunity within a few days that appears to persist lifelong. The E2 subunit vaccine induces immunity from approximately 10–14 days after a single vaccination. The immunity may persist for more than a year, but is then not complete. The Chinese strain vaccine may establish a strong herd immunity 1–2 weeks earlier than the E2 vaccine. The ability of the Chinese vaccine strain to prevent congenital infection has not been reported, but the E2 subunit vaccine does not induce complete protection against congenital infection. Immunological mechanisms that underlie the protective immunity are still to be elucidated. Both types of vaccine are considered to be safe. A great advantage of the E2 subunit vaccine is that it allows differentiation of infected pigs from vaccinated pigs and is referred to as a DIVA vaccine. However, the companion diagnostic E rns ELISA to actually make that differentiation should be improved. Many approaches to develop novel vaccines have been described, but none of these is likely to result in a new DIVA vaccine reaching the market in the next 5–10 years. Countries where classical swine fever is endemic can best control the infection by systematic vaccination campaigns, accompanied by the normal diagnostic procedures and control measures. Oral vaccination of wild boar may contribute to lowering the incidence of classical swine fever, and consequently diminishing the threat of virus introduction into domestic pigs. Free countries should not vaccinate and should be highly alert to rapidly diagnose any new outbreak. Once a new introduction of classical swine fever virus in dense pig areas has been confirmed, an emergency vaccination programme should be immediately instituted, for maximum benefit. The question is whether the time is ripe to seriously consider global eradication of classical swine fever virus.

Infection of porcine reproductive and respiratory syndrome virus suppresses the antibody response to classical swine fever virus vaccination

The effects of porcine reproductive and respiratory syndrome virus (PRRSV) infection on the antibody response to vaccination with classical swine fever (CSF) vaccine in piglets were investigated. Piglets were inoculated intranasally with the PRRSV BJ-4 strain, and then vaccinated intramuscularly with CSF vaccine 2 days later. Control animals were either vaccinated with CSF vaccine or infected with PRRSV alone. Time course of the antibody response against either PRRSV or CSF vaccine was measured. In animals infected with PRRSV prior to CSF vaccination, the antibody response to CSF vaccine was significantly lower than that observed in animals treated with CSF vaccination alone, especially during 3–5 weeks after vaccination. At the same time, no significant difference in serum PRRSV antibody level was observed between the animals treated with both PRRSV and CSF vaccine and the animals treated with PRRSV alone. These results suggested that the reduced antibody response against CSF vaccine observed in PRRSV-infected animals might be related to the higher prevalence of CSF in PRRSV prevalent farms in China.

The influence of maternal immunity on the efficacy of a classical swine fever vaccine against classical swine fever virus, genogroup 2.2, infection

In Thailand, where vaccination is routinely employed, there has been an increased incidence of chronic classical swine fever (CSF) outbreaks during the past decade. The major causative virus has been identified to be the moderate virulence, classical swine fever virus (CSFV) of the genogroup 2.2. An investigation was made into the efficacy of a CSF vaccine against this genogroup 2.2 challenge. Five-week-old pigs, grouped by their level of passive antibody titer were immunized with lapinized Chinese-strain CSF vaccine and challenged with CSFV genogroup 2.2, 13 days after vaccination. The group containing passive titers of lower than 64 at the time of immunization, had significantly higher number of CSFV-specific IFN-γ secreting cells and was completely protected against the challenge. Interestingly, both cellular and antibody responses were inhibited in the pigs with the higher passive titer. Furthermore, following challenge, CSFV could be isolated from 50% of the pigs in this group. It was demonstrated that the CSF vaccine could induce complete protection in pigs, provided that the maternal derived titer at the time of vaccination was lower than 64. The result implied that an increase in CSFV outbreaks might be due to the inappropriate timing of vaccination as well as the nature of the CSFV genogroup 2.2.

The effect of Trypanosoma evansi infection on pig performance and vaccination against classical swine fever.

Although Trypanosoma evansi is not considered as an important pathogen in pigs, it may interfere with other pathogens or vaccinations by its immunosuppressive nature. In order to determine whether T. evansi alters pig performance and induces immunosuppression in pigs, induction of immune responses by vaccination against classical swine fever (CSF) and by immunization with a control antigen, human serum albumin (HSA), was assessed in T. evansi-infected and non-infected animals. Although T. evansi infection did not have a significant influence on growth performance, feed conversion or PCV, antibody responses against both the test antigen HSA and the CSF vaccine were significantly reduced in T. evansi-infected animals as compared to uninfected animals. Moreover, the reduced response against the CSF vaccine appears to be accompanied by a less well-developed protection against CSF with higher fever responses and leukopenia. This immunosuppression might explain the accounts of poor protection of CSF-vaccinated pigs reported in T. evansi-endemic areas of Vietnam, and suggests that prior treatments with trypanocidal drugs to improve the efficacy of CSF vaccination, may be justified.

Oral immunisation of swine with a classical swine fever vaccine (Chinese strain) and transmission studies in rabbits and sheep

Seven experiments including a total of 47 pigs, 11 wild boars, 26 rabbits, 10 hares and 16 sheep were carried out to assess the efficacy, safety and transmission of the Chinese vaccine strain of the classical swine fever virus (CSFV) administrated by the oral route. Within 3 weeks after oral vaccination, a clear seroconversion occurred in the pigs. Six weeks after vaccination, vaccinated pigs were fully protected against a virulent challenge. The C-strain was not isolated from tonsils, spleen, lymph nodes, thymus, saliva, urine and faeces of pigs within 4 days after oral vaccination. In one experiment, susceptible pigs were placed in direct contact with vaccinated pigs. None of these contact-exposed pigs became serologically positive for CSFV antibodies. It is concluded that the C-strain induces protection in pigs when administrated by the oral route and is not shed by vaccinated pigs. Serum anti-CSFV antibodies developed in seven out of eight wild boars vaccinated by the oral route. No vaccine virus was detected in the spleen and tonsils of these animals. The results in wild boar were in accordance with those obtained in domestic pigs. Sheep did not show any clinical signs after oral vaccination while rabbits had moderate hyperthermia and growth retardation. No clinical response to oral immunisation in hares was detected. At the end of the experiment, no sheep had detectable serum antibodies against CSFV, whereas a few vaccinated rabbits and hares became seropositive. None of the contact-exposed rabbits and hares seroconverted. These data indicate that the C-strain is safe for sheep and as expected, moderately or not pathogenic for rabbits and hares. These efficacy and safety studies on oral vaccination with the C-strain under experimental conditions provide essential information for further studies in wild boars under experimental and field conditions, including assays with baits to control a CSF epidemic.

Classical swine fever virus diagnostics and vaccine production in insect cells

Classical swine fever virus (CSFV; synonym hog cholera virus), bovine viral diarrhoea virus (BVDV), and border disease virus (BDV), members of the Pestivirus genus of the family of Flaviviridae (Francki et al., 1991) are small, enveloped, positive-stranded RNA viruses (Moennig & Plagemann, 1992). The viruses are structurally, antigenically and genetically closely related. BVDV and BDV can infect ruminants and pigs. CSFV infections are restricted to pigs. The envelope of the CSF virion contains three glycoproteins, El (gp51 to gp54), E2 (gp42 to gp46), and E3 (gp31) (Moormann et al., 1990; Thiel et al., 1991). Animals infected with pestiviruses raise antibodies against at least two viral glycoproteins, namely El and E2 (Kwang et al., 1992; Terpstra & Wensvoort, 1988). El is the most immunodominant protein present in the envelope and plays an important role in virus neutralization (Wensvoort et al., 1990). CSFV causes a highly contagious and often fatal disease in pigs. Outbreaks of the disease occur intermittently in several European countries and can cause large economical losses. An active serological screening program followed by “stamping out” of infected herds, is used to control outbreaks of the disease. This policy is not only expensive but is also not sufficient for eradication of classical swine fever in Europe. Vaccination of pigs with a live attenuated CSFV vaccine strain, the C strain, protects pigs from classical swine fever

Simultaneous vaccination of piglets against foot-and-mouth disease and classical swine fever

Six-week-old piglets, born of unvaccinated sows, were vaccinated against foot-and-mouth disease (FMD) with a trivalent, inactivated vaccine containing an adjuvant or vaccinated against classical swine fever (CSF) with a live attenuated vaccine or against both diseases simultaneously at two different sites. The antibody response to the FMD vaccine was not significantly influenced by the simultaneous vaccination against CSF. FMD vaccine administered simulateously with the CSF vaccine produced a significantly higher antibody response to CSF than occurred with CSF vaccination only.