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Results for “Classical Swine Fever”

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PubMed1996

Evaluation of a gp 55 (E2) recombinant-based ELISA for the detection of antibodies induced by classical swine fever virus.

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies against classical swine fever virus (CSFV) based on the competition of the serum antibodies with a CSFV-specific monoclonal antibody directed against the viral glycoprotein gp 55 (E2), has been evaluated. A total of 553 sera obtained from pigs experimentally infected in groups with different pestiviruses were included in this study. The ELISA was applied to a group of sera collected from pigs prior to pestivirus inoculation and therefore expected to have no detectable CSFV neutralizing antibodies. The specificity of the ELISA was calculated to range between 93% and 98%. The sensitivity of the ELISA was determined by testing predominantly sera of pigs exposed to CSFV and exhibiting CSFV neutralizing antibodies. When the cut-off level was reduced by 10%, the average sensitivity increased from 60% to 70% for the groups of sera tested. The ELISA was able to discriminate between CSFV and bovine viral diarrhoea virus (BVDV) induced antibodies.

Animals↗

PubMed2006

Classical swine fever virus replicon particles lacking the Erns gene: a potential marker vaccine for intradermal application.

Classical swine fever virus replicon particles (CSF-VRP) deficient for E(rns) were evaluated as a non-transmissible marker vaccine. A cDNA clone of CSFV strain Alfort/187 was used to obtain a replication-competent mutant genome (replicon) lacking the sequence encoding the 227 amino acids of the glycoprotein E(rns) (A187delE(rns)). For packaging of A187delE(rns) into virus particles, porcine kidney cell lines constitutively expressing E(rns) of CSFV were established. The rescued VRP were infectious in cell culture but did not yield infectious progeny virus. Single intradermal vaccination of two pigs with 10(7) TCID(50) of VRP A187delE(rns) elicited neutralizing antibodies, anti-E2 antibodies, and cellular immune responses determined by an increase of IFN-gamma producing cells. No anti-E(rns) antibodies were detected in the vaccinees confirming that this vaccine represents a negative marker vaccine allowing differentiation between infected and vaccinated animals. The two pigs were protected against lethal challenge with the highly virulent CSFV strain Eystrup. In contrast, oral immunization resulted in only partial protection, and neither CSFV-specific antibodies nor stimulated T-cells were found before challenge. These data represent a good basis for more extended vaccination/challenge trials including larger numbers of animals as well as more thorough analysis of virus shedding using sentinel animals to monitor horizontal spread of the challenge virus.

Animals↗

PubMed2004

Vaccination against classical swine fever virus: limitations and new strategies.

The most widely used vaccines for the control of classical swine fever (CSF) in countries where it is endemic are live attenuated virus strains, which are highly efficacious, inducing virtually complete protection against challenge with pathogenic virus. In the European Union (EU), the combination of prophylactic mass vaccination and culling of infected pigs in endemic regions has made it possible to almost eradicate the disease. However, it is not possible to discriminate between infected and vaccinated animals, thus hampering disease control measures that rely on serology. Therefore, vaccination was banned at the end of 1990 before the internal common market was established in the EU. Vaccination is allowed only in severe emergencies. In addition, there are strict restrictions on the international trade in pig products from countries using vaccination. To circumvent these problems, marker vaccines which allow differentiation of infected from vaccinated animals (DIVA) have been developed. There are several approaches, ranging from protective peptides, single expressed proteins, naked DNA and chimeric viruses. To date, two subunit vaccines based on the E2 glycoprotein are commercially available and have been tested extensively for their efficacy. The accompanying discriminatory tests are based on an ELISA detecting another viral glycoprotein, the E(rns). The subunit vaccines were found to be less efficacious than live attenuated vaccines. In addition, the currently available discriminatory tests do not provide high enough specificity and sensitivity. Although there is an urgent need for more advanced marker vaccines and better discriminatory tests, the development of new DIVA vaccines against CSF is hampered by the small market potential for these products.

Animals↗

PubMed2000

Development of a classical swine fever subunit marker vaccine and companion diagnostic test.

The development of a classical swine fever (CSF) subunit marker vaccine, based on viral envelope glycoprotein E2, and a companion diagnostic test, based on a second viral envelope glycoprotein E(RNS), will be described. Important properties of the vaccine, such as onset and duration of immunity, and prevention of horizontal and vertical transmission of virus were evaluated. A single dose of the vaccine protected pigs against clinical signs of CSF, following intranasal challenge with 100LD(50) of virulent classical swine fever virus (CSFV) at 2 weeks after vaccination. However, challenge virus transmission to unvaccinated sentinels was not always completely inhibited at this time point. From 3 weeks up to 6 months after vaccination, pigs were protected against clinical signs of CSF, and no longer transmitted challenge virus to unvaccinated sentinels. In contrast, unvaccinated control pigs died within 2 weeks after challenge. We also evaluated transmission of challenge virus in a setup enabling determination of the reproduction ratio (R value) of the virus. In such an experiment, transmission of challenge virus is determined in a fully vaccinated population at different time points after vaccination. Pigs challenged at 1 week after immunization died of CSF, whereas the vaccinated sentinels became infected, seroconverted for E(RNS) antibodies, but survived. At 2 weeks after vaccination, the challenged pigs seroconverted for E(RNS) antibodies, but none of the vaccinated sentinels did. Thus, at 1 week after vaccination, R1, and at 2 weeks, R=0, implying no control or control of an outbreak, respectively. Vertical transmission of CSFV to the immune-incompetent fetus may lead to the birth of highly viraemic, persistently infected piglets which are one of the major sources of virus spread. Protection against transplacental transmission of CSFV in vaccinated sows was, therefore, tested in once and twice vaccinated sows. Only one out of nine once-vaccinated sows transmitted challenge virus to the fetus, whereas none of the nine twice-vaccinated sows did. Finally, our data show that the E(RNS) test detects CSFV-specific antibodies in vaccinated or unvaccinated pigs as early as 14 days after infection with a virulent CSF strain. This indicates that the E2 vaccine and companion test fully comply with the marker vaccine concept. This concept implies the possibility of detecting infected animals within a vaccinated population.

Animals↗

PubMed2001

Duration of the protection of an E2 subunit marker vaccine against classical swine fever after a single vaccination.

The period during which pigs are protected after vaccination is important for the successful usage of a marker vaccine against classical swine fever virus (CSFV) in an eradication programme. In four animal experiments with different vaccination-challenge intervals we determined the duration of protection of an E2 subunit marker vaccine in pigs after a single vaccination. Unvaccinated pigs were included in each group to detect transmission of the challenge virus. Three groups of six pigs were vaccinated once and subsequently inoculated with the virulent CSFV strain Brescia after a vaccination-challenge interval of 3, 51/2, 6 or 13 months. All vaccinated pigs, 16 out of 18, with neutralising antibodies against CSFV at the moment of challenge, 3, 51/2, 6 or 13 months later, survived, whereas unvaccinated control pigs died from acute CSF or were killed being moribund. A proportion of the vaccinated pigs did however develop fever or cytopenia after challenge and two vaccinated pigs were viremic after challenge. Virus transmission of vaccinated and challenged pigs to unvaccinated sentinel pigs did not occur in groups of pigs which were challenged 3 or 6 months after a single vaccination. Two out of eight vaccinated pigs that were found negative for CSFV neutralising antibody at 13 months after vaccination died after subsequent challenge. The findings in this study demonstrate that pigs can be protected against a lethal challenge of CSFV for up to 13 months after a single vaccination with an E2 subunit marker vaccine.

Animals↗

PubMed1994

Classical swine fever: genetic detection and analysis of differences between virus isolates.

Two pairs of oligonucleotide primers were designed that specifically amplified regions of the classical swine fever virus genome. These products, corresponding to a 671 bp portion of the genes encoding the E1 and E2 (gp33 and gp55) proteins and a 1090 bp portion of the putative polymerase gene, were amplified from eight virus isolates which had been responsible for a series of classical swine fever outbreaks in Italy involving both domestic pigs and wild boar. For each virus the fragments were partially sequenced to give 475 bp of the E1/E2 glycoprotein and 212 bp of the putative polymerase gene sequences. The data from each set of fragments were compared with one another and with reference strains. This allowed us confidently to assign most of the viruses to one of three subgroups. An analysis of the same viruses with a panel of monoclonal antibodies was much less informative. The subgrouping of the isolates suggested that, in this region of Italy, there had been at least two separate introductions of classical swine fever over a 7 year period and that virus had been transmitted between domestic pigs and wild boar. A consensus nucleotide sequence derived from the glycoprotein fragments of all the viruses examined revealed conservation at the wobble position of some codons.

Animals↗

PubMed2000

Recombinant classical swine fever (CSF) viruses derived from the Chinese vaccine strain (C-strain) of CSF virus retain their avirulent and immunogenic characteristics.

Two recombinant classical swine fever (CSF) viruses (Flc2, Flc3) transcribed from a DNA copy of the genome of the Chinese (C) strain, a CSF virus vaccine strain, were characterized in vivo in rabbits and pigs. Rabbits were inoculated intravenously with Flc2 or Flc3, the parent C-strain virus, a biologically cloned C-strain or CSF virus strain Brescia (C.1.1.1). After 24-96 h fever was detected in the rabbits inoculated with the different C-strain viruses. Apart from those in the control group, all the C-strain inoculated rabbits had developed CSF virus neutralizing antibodies 4 weeks later and were protected against a parent C-strain challenge. In the second experiment, pigs were inoculated with the parent C-strain or recombinant C-strain virus (Flc2 or Flc3) and then challenged after 4 weeks with the virulent CSF virus strain Brescia. None of the pigs showed clinical signs of classical swine fever after vaccination or challenge, whereas the control pigs developed clinical signs typical for acute CSF. Pigs inoculated with the different C-strain viruses were not viremic after inoculation or challenge, and CSF virus neutralizing antibodies were detected from day 14 onwards. The results from both experiments demonstrated that the two recombinant viruses had retained the biological and immunogenic properties of the parent C-strain in rabbits and pigs. We conclude that the full-length cDNA of the C-strain can serve as a matrix for further development of a live recombinant CSF virus marker vaccine.

Animals↗

PubMed1995

Cytopathogenicity of classical swine fever virus caused by defective interfering particles.

For three independent cytopathogenic isolates of classical swine fever virus, defective RNAs were found in infected cells in addition to full-length viral genomes. These RNAs represent the genomes of typical defective interfering (DI) particles because of strict dependence on a complementing helper virus and interference with the replication of the helper virus. Analysis of the DI genomes revealed internal deletions of 4,764 nucleotides encompassing the complete structural protein-coding region of the virus and two flanking nonstructural genes. Plaque isolation and RNA transfection experiments showed that the DI particles are responsible for the cytopathic effect caused by these classical swine fever virus isolates.

Base Sequence↗

PubMed1988

Classical swine fever: virulence and tissue distribution of a 1986 English isolate in pigs.

Following the recurrence of classical swine fever in the United Kingdom in 1986, a virus isolated from a single outbreak was studied. A major factor in the spread of this disease is considered to be the presence of infectious virus in tissues taken from animals at certain stages of infection, although their condition may escape detection by routine inspection either before or after slaughter. Intranasal inoculation of the isolate into eight-week-old pigs reproduced the acute form of the disease. The pigs were killed or died between seven and 25 days after inoculation. The virus concentration was determined in a wide range of tissues taken at different stages of infection. Infectious virus was present at high concentrations in all the tissues taken and at all stages of infection. Any porcine tissue is therefore a potential source of infection even when it is taken either before the animal displays detectable signs of disease of after it develops serum neutralising antibodies.

Animals↗

PubMed1995

A serological study on infection patterns, control and persistence of classical swine fever in infected farms in the Philippines.

In 3 farrow-to-finish farms with enzootic classical swine fever (CSF) all sick pigs were culled and an intensive vaccination program was introduced. Boars and cows were vaccinated every 6 months and piglets at the age of 6 and 8 weeks. The infection was monitored on clinical grounds and by means of serological examination of the farms. Serological results reflected well the clinical situation in the farms. High titers and high percentage of animals with positive titers were observed in infected farms, compared to lower titers in a clinically "free" farm, which were interpreted as titers due to vaccination. Although clinical improvement was noticed, the rigid vaccination regime was not able to control CSF in none of the farms. In all cases, a small percentage of pigs in the finishing stage was still affected. Since these pigs seemed not to have responded to vaccination at 6 and 8 weeks of age, the programme was questioned. Additionally, very high titers were recorded in sows from infected farms, which would automatically extend the period of passive protection in offspring and postpone the age at which piglets could be vaccinated. More intensive programmes, which include also vaccination beyond the age of 8 weeks, were suggested to bring enzootic infections under control.

Aging↗

PubMed2003

Genetic typing of recent classical swine fever virus isolates from Croatia.

During a period of 5 years (1997-2001) several outbreaks of classical swine fever (CSF) were recorded in Croatia. For genetic typing, fragments of 150 nucleotides within the 5'-non-translated region (5'-NTR) and 190 nucleotides within the E2 glycoprotein coding gene of nine field isolates that were derived from domestic pigs and wild boars were used. For better epizootiological understanding, isolates from other European countries were included in the study. The results show that the isolates belong to subgroups 2.1 and 2.3 of CSF virus. Isolates from subgroup 2.1 were collected from domestic pigs during sporadic outbreaks in June 1997 and are genetically closely related. A genomic similarity between these isolates and CSF virus isolates from pigs in other European countries from the same year could also be confirmed. In contrast, the isolate from October 1997 was found to be a member of subgroup 2.3, and is closely related to European CSF virus isolates from outbreaks in the last decade in Western and Central European countries. These results show that two different sources of CSF virus caused outbreaks in Croatia during the same year. Furthermore, a close relationship was found between an isolate from a domestic pig in 1999 and isolates of subgroup 2.3 that originated from Croatian wild boars.

Animals↗

PubMed1999

Experimental infection of slaughter pigs with classical swine fever virus: transmission of the virus, course of the disease and antibody response.

The spread of classical swine fever virus was investigated in an isolation unit containing four pens, each containing six slaughter pigs. One pig in the middle pen of three adjacent pens was inoculated intramuscularly and intranasally with the virus. The fourth pen was located in a separate compartment. The pens were visited in a strict order to study, first, the effect of indirect contact via contaminated clothing and footwear on the spread of the virus to adjacent pens and, secondly, the airborne transmission of the virus between compartments. The pigs were examined and blood samples were taken every other day for 62 days for virological and serological analyses. The virus was highly contagious for the five pigs that were in direct contact with the inoculated pig, but spread to the other pens only after all the pigs in the originally infected pen had become viraemic. The spread of the virus was promoted by contaminated clothing and footwear, but airborne transmission contributed considerably to the spread of the virus within the pighouse. The first clinical signs observed after the virus was introduced into a pen were decreased feed intake, increased mean rectal temperature and apathy. Neither the clinical course of the infection, nor the pattern of seroconversion observed over time, was affected by the differences in the intensity of contact with the virus between the pigs in the different pens.

Animals↗

PubMed2000

Pathogenesis of classical swine fever: renal haemorrhages and erythrodiapedesis.

Thirty pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (hog cholera) virus to establish the chronological occurrence of lesions in the kidney and to determine the mechanism responsible for renal haemorrhages. The study included the use of histopathological, ultrastructural, immunohistochemical (detection of viral antigen gp55, MAC387, lambda chains, CD3 and C1q) and morphometrical techniques (vascular area). Renal interstitial oedema and haemorrhages were detected from 7 days post-inoculation (dpi), associated with a slight interstitial mononuclear infiltrate and evidence of viral infection in macrophages and fibroblasts, and in a small proportion of lymphocytes. Viral infection was not detected in capillary endothelial cells. An intense mononuclear infiltrate, with B cells, T cells and small numbers of macrophages, was detected from 10 dpi. In the final phase of the experiment (14 dpi), slight proliferation and degranulation of mast cells were observed. Increased expression of the C1q component of complement was also detected. A significant increase in vascular area was observed from 7 dpi. These results suggest that haemorrhages observed in the kidneys of pigs inoculated with the Quillota strain resulted from erythrodiapedesis and increased vascular permeability, probably aggravated by mast cell degranulation in the final stage of the experiment. The results suggested that mast cell degranulation was linked to activation of the complement system.

Animals↗

PubMed2003

Classical Swine Fever: pathology of bone marrow.

Twenty pigs were inoculated with a virulent isolate (Quillota strain) of classical swine fever (CSF) virus to determine the chronological development of lesions in bone marrow. Histopathologic, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, CD3 antigen, and FVIII-rag), and morphometric techniques were employed. Viral antigen was detected from 2 days postinfection (dpi) in stromal and haematopoitic cells, and severe atrophy related to apoptosis of haematopoitic cells was observed. Megakaryocytes (MKs) did not show significant changes in number, but there were important qualitative changes including 1) increased numbers of cloud-nuclei MKs, microMKs, apoptotic MKs, and atypical nucleated MKs and 2) decreased number of typical nucleated MKs. Morphometric study of these cells showed a decrease in cytoplasmic area. MK infection was detected from 2 dpi, but in a small percentage of cells. Myeloid cells showed quantitative changes, with an increase in granulocyte numbers. Apoptosis of lymphocytes and viral infection of erythroblasts were also observed. The main changes in stroma were depletion of T lymphocytes in the middle phase of the experiment and macrophages. Viral infection was also observed in these cells. MK lesions suggest dysmegakaryocytopoiesis, which would aggravate the thrombocytopenia already present and could be responsible for it. Granulocyte changes would lead to the appearance of circulating immature forms, whereas lymphocyte apoptosis in bone marrow would contribute to lymphopenia.

Animals↗

PubMedDate not supplied

Ultrastructural glomerular changes in experimental infection with the classical swine fever virus.

Ultrastructural studies of glomerular changes were performed on 16 pigs experimentally infected with a highly virulent strain of the classical swine fever virus. Our observations revealed the thickening of glomerular basement membranes, swelling of endothelial cells and cytoplasmic vacuolization within podocytes containing abundant viral particles. An early viral infection of podocytes was suggested as the cause of selective swelling of the foot processes of these cells with the consequent obliteration of the glomerular urinary spaces. To our knowledge this is the first report of ultrastructural evidence of classical swine fever virions infecting glomerular podocytes.

Animals↗

PubMed2006

Genome comparison of a novel classical swine fever virus isolated in China in 2004 with other CSFV strains.

The genome of a novel classical swine fever virus (CSFV), SWH/CA/2004, isolated from a hog pen in Henan Province, central China, is 12,296 nucleotides (nt) in length. It is composed of a 373-nt 5' terminal non-translated region (NTR), a 11,697-nt open reading frame (ORF) encoding a polyprotein of 3,898 amino acids (aa), and a 226-nt 3'-NTR. Genome comparison of the SWH/CA/2004 isolate (GenBank Accession: DQ127910) with other known CSFV isolates was performed and analyzed. Corresponding segments from SWH/CA/2004 and other reported strains shared 80.4-99.8% identity at the nucleotide level and 89.5-99.8% identity at the amino acid level. From an evolutionary point of view, isolate SWH/CA/2004 is closely related to the highly virulent isolate cF114/CA/2001, with a pairwise distance of 0.013; and distantly related to the moderately virulent isolate GXWZ02/CA/2003, with pairwise distance 0.170. The phylogenetic trees of the full-length genome and the following region E(rns), E1, E2, and NS5B-based neighbor-joining (NJ) method were constructed and approximately divided into different genetic groups according to avirulence, moderate virulence and high virulence, while other region-based NJ trees demonstrated sequence conservation between these groups. The four genomic regions may constitute important criteria for genetic typing of diverse CSFV isolates. Based on these analyses, isolate SWH/CA/2004 was deduced to belong to the highly virulent isolate group. However, SWH/CA/2004 also contains a 14-U deletion in the 3'-NTR that is characteristic of avirulent isolates. These analyses constitute a comprehensive study of the phylogenetics of CSF based on distinct regions of the genome and may provide the basis for future molecular epidemiology research to identify virulent strain outbreaks and trigger implementation of appropriate control measures.

Animals↗

PubMedDate not supplied

Long-term monitoring of classical swine fever in wild boar (Sus scrofa sp.) using serological data.

In the European Community, epizootics of classical swine fever (CSF) in the wild boar (Sus scrofa) are compulsorily monitored because transmission may occur between wild boars and domestic pigs, causing heavy economic losses to the pork industry. The estimation of incidence in populations of wild boars is generally based on viroprevalence. However, viral isolation becomes rare when the incidence is low because the virus cannot be detected for more than a few weeks following infection. On the contrary, seroprevalence is detectable at low incidence levels, because antibodies can be detected for the lifetime of the infected animal. We thus attempted to analyse the long-term evolution of CSF incidence using serological data. The data came from France, where CSF had been monitored from 1992 to 2002, and where the virus has not been detected since 1997. We assumed that the overall seroprevalence would estimate the proportion of immune wild boars, that seroprevalence in juveniles would approximate incidence and that seroprevalence in different age classes would show the evolution of incidence in a given cohort. Spatial and temporal trends of incidence and seroprevalence were explored using logistic modelling and the spatial trend was analysed using polynomial regression. In 1992, incidence peaked in the northern area. After 1993, incidence decreased but remained the highest in the northern area. After 2000, no seropositive juvenile was observed, suggesting the extinction of the epizootic. Our results support the reliability of serological monitoring since it allowed a longer detection of viral transmission and provided more information on the spatio-temporal evolution of incidence than did viral isolation. We advocate that the highest persistence of infection in northeastern France is not independent from infection persistence in Reinland-Pfalz (Germany). Such persistence may be due to favourable local conditions and/or the social organisation of wild boars.

Animals↗

PubMed2003

Neighbourhood infections of classical swine fever during the 1997-1998 epidemic in The Netherlands.

Data of the 1997-1998 epidemic of classical swine fever (CSF) in The Netherlands were analysed in survival analysis to identify risk factors that were associated with the rate of neighbourhood infections. The study population consisted of herds within 1000 m of exclusively one previously infected herd. Dates of virus introduction into herds were drawn randomly from estimated probability distributions per herd of possible weeks of virus introduction. (To confirm the insensitivity of the results for this random data-selection procedure, the procedure was repeated 9 times (resulting in 10 different data sets).) The data set had 906 non-infected and 59 infected neighbour herds, which were distributed over 215 different neighbourhoods. Neighbour herds that never became infected were right-censored at the last date of the infectious period of the infected source herd. Neighbour herds that became empty within the infectious period or within the following 21 days due to preventive depopulation or due to the implemented buying-out programme were right-censored 21 days before the moment of becoming empty. This was done as a correction for the time a herd could be infected without being noticed as such. The median time to identified infection of neighbour herds was 2 weeks, whereas the median time to right censoring of non-infected neighbour herds was 3 weeks. The risk factors, radial distance < or =500 m, cattle present on source herd and increasing herd size of the neighbour herd were associated multivariably with the hazard for neighbour herds to become infected. We did not find an association between time down wind and infection risk for neighbour herds. Radial dispersion of CSFV seemed more important in neighbourhood infections than dispersion along the road on which the infected source herd is situated. The results of this study support the strategy of preventive depopulation in the neighbourhood of an infected herd. Recommendations are presented to adapt the applied control strategy for neighbourhood infections.

Animal Husbandry↗