Classical Swine Fever Virus Antibody Research Articles

The role of porcine reproductive and respiratory syndrome virus as a risk factor in the outbreak of porcine epidemic diarrhea in immunized swine herds

The aim of the study was to investigate the relation between porcine epidemic diarrhea (PED) and porcine reproductive and respiratory syndrome virus (PRRSV). Here, we constructed a dynamic clinical experimental model in which sows were infected with PRRSV. The stability of the experimental model was surveyed by detecting the levels of antibodies against classical swine fever virus (CSFV) or PRRSV, respectively. In both group A (infected group) and group B (healthy group), the CSFV antibody level was not significantly different (P > 0.05). In group B, the PRRSV antibody level was also not significantly different (P > 0.05) at the three times of testing. The incidence of porcine epidemic diarrhea virus (PEDV) in piglets and PEDV-carrying rate of sows was detected by real-time fluorescent RT-PCR. In group A, PEDV infection of piglets and the PEDV-carrying rate of sows were significantly higher than in group B (P < 0.01) and the PEDV strains were within the same group according to the phylogenetic analysis based on the complete S gene. The results suggested that PRRSV was a risk factor in the outbreak of PED in immunized swine herds. Besides that, the experimental model was stable and no interference factors affected the results of the study.

An investigation of classical swine fever virus seroprevalence and risk factors in pigs in Timor-Leste

Classical swine fever virus (CSFV) is a highly infectious pathogen of pigs and believed to be a major constraint to pig production in Timor-Leste. The Ministry of Agriculture and Fisheries conducts vaccination campaigns in an attempt to control clinical disease, however, there is no empirical data available concerning the seroprevalence and distribution of CSFV in Timor-Leste. To help address this knowledge deficit, a cross-sectional study to determine seroprevalence was conducted in the three districts that border Indonesia. Data on farmer- and pig-level factors were also collected to look at their impact on CSFV serological status. Overall, true CSFV seroprevalence was estimated at 34.4%. Seroprevalence estimates varied widely between and within districts, subdistricts, and villages. Older pigs and pigs that had been vaccinated for CSFV were more likely to test positive for CSFV antibody. Pigs owned by farmers that experienced the sudden death of pigs in the 12 months prior to the survey were more likely to test positive for CSFV antibody, while pigs that had been sick in the previous three months were less likely to test positive for CSFV antibody. The final multivariable model accounted for a large amount of variation in the data, however, much of this variation was explained by the random effects with less than one percent of the variation explained by the fixed effects. This work further supports the need for a collaborative approach to whole-island CSFV control between West Timor, Indonesia and Timor-Leste. Further work is needed to better understand the risk factors for CSFV serological status in order to allocate resources for control. As CSFV is now endemic in Timor-Leste research involving a combination of serology, antigen detection and in-depth investigation of suspect cases over a period of time may be required.

Results of Serological and Molecular Analysis of African and Classical Swine Fever in the Population of Wild Boars in Ukraine

Abstract The results of the study of lymphoid organs and sera of wild boars for the presence of DNA of African swine fever (ASF) virus and RNA of classical swine fever (CSF) virus are presented, as well as the results of a serological examination for the presence of ASF and CSF virus antibodies. The study was conducted in Ukraine between 2008 and 2013. Biological samples were obtained from wild boars shot during the hunting season, and were examined by real-time PCR and ELISA. In total, 5759 sera were tested for CSF virus antibodies and 4856 for ASF virus antibodies by ELISA. Samples of lymphoid organs totalling 1129 were examined by PCR for the detection of CSF virus RNA and 8102 such samples were examined for the detection of ASF virus DNA. CSF virus antibodies were detected in 6.56% of wild boar sera. RNA of CSF virus was also identified in 1 out of 1129 samples tested. ASF virus antibodies or DNA in lymphoid organ samples were not detected.

Direct coating of culture medium from cells secreting classical swine fever virus E2 antigen on ELISA plates for detection of E2-specific antibodies

The envelope glycoprotein E2 of classical swine fever virus (CSFV) is widely used as a marker for measuring vaccine efficacy and antibody titer. The glycosylation profile of E2 may affect the immunogenicity of the vaccine and the timing of re-vaccination. In this study, a human embryonic kidney cell line was used to secrete fully-glycosylated CSFV E2, which was then coated onto ELISA plates without purification or adjustment.The resulting E2-secreting medium-direct-coating (E2-mDc) ELISA was successfully used to measure anti-E2 antibody titers in vaccinated and field pig sera samples. Compared with a virus neutralization test (as standard), the E2-mDc ELISA was found to be more accurate (90%) than a commercial CSFV antibody diagnostic kit (62%). In conclusion, the mammalian cell-secreted antigen can provide cheap, accurate and effective assays for vaccine efficacy and disease diagnoses.

Effects of antibacterial peptide on humoral immunity in weaned piglets

The aim of this study was to investigate the effects of dietary antimicrobial peptide (AMP) supplementation on the serum IgG, IgM, IgA, classical swine fever antibody (CSF-Ab), and total complement (CH50) levels in weaned piglets. Ninety weaned piglets were randomly allotted to five groups for 28-day study. Blood samples were collected at 32, 39, 46, and 53 days of age. The results showed that AMP as dietary additive increased the levels of IgG, IgM, IgA, CSF-Ab, CH50 among the whole experimental period, and the effects of AMP supplementation were dose-dependent. The study showed that supplementing weaned piglets diets with AMP enhanced the humoral immune responses of weaned piglets by improving the levels of classical swine fever virus antibody, CH50, and immunoglobulins. We suggest the dosage of AMP was 500 mg/kg.

Evaluation of classical swine fever virus antibody detection assays with an emphasis on the differentiation of infected from vaccinated animals

The aim of this study was to evaluate the general characteristics of commercially available enzyme-linked immunosorbent assays (ELISAs) to detect antibody against classical swine fever (CSF), as well as to assess their potential use as accompanying marker tests able to differentiate infected from vaccinated animals (DIVA). The Chekit* CSF-Sero and the HerdChek* CSFV Ab, both of which detect antibodies against the E2 protein of classical swine fever virus (CSFV), had the highest sensitivity. Both tests were practicable and showed good reproducibility. Comparable sensitivity was shown by the Chekit* CSF-Marker, an Erns ELISA. However, this test does not allow differentiation between antibodies directed against ruminant pestiviruses and those against CSFV. Therefore, it is not suitable for use with the chimeric marker vaccines tested. The PrioCHECK CSFV Erns was the only ELISA suitable for use in DIVA with marker vaccines containing Erns proteins from ruminant pestiviruses. However, this test was less sensitive and selective than the E2-ELISAs and cannot be recommended.

Detection of antibodies against classical swine fever virus in fecal samples from wild boar

Classical swine fever (CSF) is a contagious viral disease that affects pigs. Wild boars can play an important epidemiological role in CSF outbreaks. In the past decades, studies conducted in many countries have reported that the CSF virus (CSFV) may persist in wild boar populations. The existence of CSFV in the free-ranging wild boar populations was indirectly confirmed by determining the prevalence of antibodies against CSFV in the serum of hunted wild boars. However, analyzing sero-prevalence in hunted wild boars to study the risk of CSF outbreaks is difficult due to insufficient number of samples, limitation of hunting area and biased age distribution of hunted wild boars. To improve this survey method, we collected feces of wild boars from their habitat and tested them using CSFV antibody enzyme-linked immunosorbent assay (ELISA) and CSF virus neutralization (VN) test. In this study, ELISA was found to be highly sensitive for detecting antibodies against CSFV in fecal samples. Most of doubtful or positive results obtained in CSFV ELISA were confirmed by VN tests. Despite the high coincidence rate of antibody-positive samples between CSFV ELISA and VN test, the possibility of false positive reaction should be considered. In the regional distribution, a fact that antibody-positive fecal and serum samples were found in geographically close area was shown. Hence, presence of antibodies in fecal samples may provide vital information regarding the risk of CSF outbreaks in wild boar groups in geographical proximity.

Tissues Expression, Polymorphisms Identification of FcRn Gene and Its Relationship with Serum Classical Swine Fever Virus Antibody Level in Pigs.

Neonatal Fc receptor (FcRn) gene encodes a receptor that binds the Fc region of monomeric immunoglobulin G (IgG) and is responsible for IgG transport and stabilization. In this report, the 8,900 bp porcine FcRn genomic DNA structure was identified and putative FcRn protein included 356 amino acids. Alignment and phylogenetic analysis of the porcine FcRn amino acid sequences with their homologies of other species showed high identity. Tissues expression of FcRn mRNA was detected by real time quantitative polymerase chain reaction (Q-PCR), the results revealed FcRn expressed widely in ten analyzed tissues. One single nucleotide polymorphism (SNP) (HQ026019:g.8526 C>T) in exon6 region of porcine FcRn gene was demonstrated by DNA sequencing analysis. A further analysis of SNP genotypes associated with serum Classical Swine Fever Virus antibody (anti-CSFV) concentration was performed in three pig populations including Large White, Landrace and Songliao Black pig (a Chinese indigenous breed). Our results of statistical analysis showed that the SNP had a highly significant association with the level of anti-CSFV antibody (At d 20; At d 35) in serum (p = 0.008; p = 0.0001). Investigation of expression and polymorphisms of the porcine FcRn gene will help us in further understanding the molecular basis of the antibody regulation pathway in the porcine immune response. All these results indicate that FcRn gene might be regarded as a molecular marker for genetic selection of anti-CSFV antibody level in pig disease resistance breeding programmes.

Prevalence of Classical Swine Fever Virus in India: A 6-Year Study (2004-2010)

A study was undertaken regarding the prevalence of classical swine fever virus (CSFV) antibodies and antigens in sera and suspected tissue samples of domestic pigs. The samples were received between January 2004 and September 2010. A total of 594 serum samples from 12 states and 287 tissue samples from 13 states of India were tested using commercial enzyme-linked immunosorbent assay (ELISA) kits. The mean prevalence of CSFV antibodies in suspected sera was 63.3% (376/594), whereas 76.7% (220/287) of the suspected samples were found to contain CSFV antigen. The high prevalence of CSFV antibodies suggests that the disease is endemic in the country. This baseline data will be of use in the formulation of control and eradication programmes.

Investigation of a pig herd with animals seropositive for classical swine fever and where porcine circovirus-associated disease had been diagnosed

AIM: To investigate the cause of classical swine fever (CSF) virus-seropositive animals in a nucleus pig-breeding herd in New Zealand, where porcine circovirus-associated disease had been diagnosed. CASE HISTORY AND CLINICAL FINDINGS: An exotic disease investigation was undertaken to exclude CSF and porcine reproductive and respiratory syndrome (PRRS) on a nucleus pig-breeding herd comprising approximately 300 breeding sows, 1,000 weaners, and 650 grower pigs. The herd was experiencing poor reproductive performance in sows, and breeding records showed a declining farrowing rate attributable to a single manager. The growing pigs (10–15 weeks old) were experiencing respiratory disease and wasting, and the mortality rate by pen varied between 9 and 20%. Post-mortem changes in affected grower pigs were consistent with circovirus-associated diseases. DIAGNOSTIC TESTING: Serological screening using an IDEXX-ELISA gave negative results for PRRS virus antibodies, but two grower pigs and one sow tested positive for CSF virus antibodies. These three seropositive animals remained positive to CSF virus, using three commercial ELISA test kits, over 27 weeks. A newly developed virus neutralisation test (VNT), using a New Zealand isolate of border disease (BD) virus, demonstrated that the seropositive pig sera had higher antibody titres to BD virus than to bovine viral diarrhoea (BVD) virus and CSF virus. PCR performed on tonsil, kidney, ileum and spleen gave negative results for CSF virus, and histopathology on lymph nodes, intestine, lung, kidney, liver and brain showed no evidence of the disease. Virus isolation performed on a number of samples was negative. CLINICAL RELEVANCE: The seropositive samples for CSF virus found in this investigation were likely to be a cross reaction to a pestivirus other than CSF virus. The finding of a possible endemic pestivirus capable of being transmitted between sheep and pigs on this farm may explain findings from previous serological survey work in New Zealand, and supports experience elsewhere, where BD virus was found to be the predominant ruminant pestivirus infecting pigs. The results show that pestivirus cross reactivity can result in unexpectedly high titres, and that testing with a full set of (local) pestiviruses is necessary to reach the correct conclusion. The investigation has direct relevance where pig herds with a low seroprevalence are encountered during surveillance for CSF.

Prevalence of Antibodies to Selected Viral and Bacterial Pathogens in Wild Boar (Sus scrofa) in Campania Region, Italy

Serum samples were collected from wild boars (Sus scrofa) harvested during the 2005-2006 hunting season in Campania, southern Italy. Samples were tested for antibodies to Leptospira interrogan, Brucella spp., Salmonella spp., Aujeszky disease virus (ADV), porcine reproductive and respiratory stress syndrome virus (PRRSV), porcine parvovirus (PPV), classical swine fever virus (CSFV), and swine vesicular disease virus (SVDV). Of the 342 serum samples tested, 15 (4.4%) were seropositive to Brucella spp., nine (2.6%) were seropositive to L. interrogans, 66 (19.3%) were seropositive for Salmonella spp., 105 (30.7%) were seropositive for ADV, 27 (7.9%) were seropositive for PPV, and 129 (37.7%) were seropositive for PRRSV. All sera tested seronegative for SVDV and CSFV antibodies. These results, recorded for the first time in Campania, support the hypothesis that wild boar are reservoirs of certain infectious agents, but some infections in wild boars originate from their domestic counterparts.

Antigens containing TAVSPTTLR tandem repeats could be used in assaying antibodies to Classical swine fever virus

Classical swine fever virus (CSFV) causes a highly contagious and often fatal viral disease in pigs. The highly conserved epitope TAVSPTTLR on the glycoprotein E2 was shown to be suitable for differentiation of CSFV from other pestiviruses. In this study, we found that swine CSFV antisera contained TAVSPTTLR-related rather than TAVSPTTLR-specific CSFV antibodies. The CSFV antisera reacted only to some extent with a synthetic TAVSPTTLR-containing peptide, but inhibited the binding of TAVSPTTLR-specific antibodies to the viral antigen. Since chimeric antigens containing TAVSPTTLR tetramers or hexamers were recognized by the swine CSFV antisera, such antigens could be potentially applied to the detection of CSFV antibodies. These results might be helpful in designing a TAVSPTTLR epitope-based CSFV vaccine and a corresponding serological test.

Characterization of porcine MMP-2 and its association with immune traits

Matrix metalloproteinase-2 (MMP-2) plays important roles in inflammation and immunity besides its basic role in degrading and remodelling extracellular matrix (ECM). The expression of MMP-2 is up-regulated in many human as well as animal models of inflammatory and immune diseases. In this study, we cloned the 5′-upstream sequence, 3′-downstream sequence as well as other missed genomic sequences of porcine MMP-2, the genomic structure and the promotor sequence were analyzed and found to share high similarity with those of human MMP-2. Porcine MMP-2 was assigned to SSC6p14–p15, and closely linked to microsatellite SW1108 (53cR, LOD score 7.59) by IMpRH panel. Real-time PCR analysis revealed that the expression of porcine MMP-2 was remarkably different in diverse tissues, a high level expression was observed in the testis and uterus, relatively low expression in other tissues. Allele frequencies determination in different pig breeds and association study were performed on the selected SNP and indel. The results showed that the SNP AcyI in exon 12 was significantly associated with white blood cell count (WBC) of neonate piglets at 0 day ( P = 0.0079), and classical swine fever virus antibody level (CSFV-AB) of pigs at 17 days ( P = 0.0461), the indel MspI in intron 4 had remarkable correlation with mean corpuscular hemoglobin (MCH) of pigs at 17 days ( P < 0.0001).

Enhancement of Peripheral Blood CD8+T Cells and Classical Swine Fever Antibodies by Dietaryβ-1,3/1,6-glucan Supplementation in Weaned Piglets

The experiment was conducted to evaluate the effect of dietary supplementation with 0, 50, 100 and 200 mg/kg of beta-1,3/1,6-glucan extracted from Saccharomyces cerevisiae on cellular immune responses and classical swine fever virus (CSFV) antibody production of weaned piglets. Thirty-two (Landrace x Large White; 26 days old; 7.05+/-0.25 kg body weight) castrated male piglets were randomly allocated to four treatments, eight pigs per treatment. All piglets were vaccinated with live-attenuated Chinese strain cell vaccines at 26 days of age. The results showed that dietary supplementation with beta-1,3/1,6-glucan could significantly (P<0.01) enhance peripheral blood lymphocyte proliferation activity in weaned piglets after 14 days administration, whereas no significant effect (P>0.05) was observed on day 28. Dietary beta-1,3/1,6-glucan supplementation could significantly enhance CSFV antibody blocking rate (P<0.05) on days 14 and 21 after inoculation. Although dietary beta-1,3/1,6-glucan supplementation did not significantly influence (P>0.05) lymphocyte subgroups on day 14, high levels of beta-1,3/1,6-glucan (100 and 200 mg/kg) increased the percentage of cytotoxic/suppressor lymphocytes (CD8(+)) significantly (P<0.05), decreased the ratio of CD4(+)/CD8(+) significantly (P<0.05) and increased the percentage of total T lymphocytes (CD3(+)) on day 28. These results suggest that beta-1,3/1,6-glucan may have a beneficial effect on the immune system of swine, and dietary beta-1,3/1,6-glucan supplementation had a transient impact on lymphocyte proliferation activity. The CSFV antibody blocking rate and the percentage of peripheral blood CD8(+) T cells could be enhanced by continuous beta-1,3/1,6-glucan supplementation. These findings support the potential application of beta-1,3/1,6-glucan as a prophylactic agent in increasing animal immune functions and controlling CSF disease.

Immunohistochemical detection of B and T lymphocytes in mandibular lymph nodes of experimentally infected piglets with classical swine fever virus

Classical swine fever (CSF) is a highly contagious viral disease of domestic pigs and wild boars with severe consequences on animal welfare, livestock production, trade and national economy. We investigated if colostral antibodies in piglets, originating from sows vaccinated with Chine strain (C-strain) and challenged with CSF virus, affected the distribution of the B and T lymhocites in mandibular lymph nodes. Nineteen 45 days old cross breed pigs of both sexes were divided in three groups. All animals in the first group, originating from sows vaccinated with C-strain vaccine were serologically positive for the presence of specific colostral CSF virus antibodies. The second group consisted of pigs originating from unvaccinated sows serologically negative for CSF virus antibodies. Three healthy pigs serologically negative for the presence of CSF virus antibodies, originating from unvaccinated sows against CSF virus served as a control group. In the group of animals originating from unvaccinated sows against CSF, severe depletion of CD79 positive B lymphocytes was detected. In the group of pigs originating from sows vaccinated with C-strain (CSF virus antibody positive group of pigs), a reduced number of CD79 positive B lymphocytes was detected, but B cells were still present in the periphery of the germinative centres of secondary lymph follicles. There was an increase of the number of CD3 positive cells in the mandibular lymph nodes of pigs with or without maternal antibodies.

Surveillance of Classical Swine Fever Virus Antibody Titers in Sows and their Piglets under Different Vaccination Programs

Surveillance of Classical Swine Fever Virus Antibody Titers in Sows and their Piglets under Different Vaccination Programs

A DNA vaccine expressing the E2 protein of classical swine fever virus elicits T cell responses that can prime for rapid antibody production and confer total protection upon viral challenge

Immunization of domestic pigs with a DNA vaccine expressing the complete E2 protein of classical swine fever virus (CSFV) conferred total protection against a severe viral challenge. Immunization with three doses of plasmid pcDNA3.1/E2 elicited a consistent and specific, MHC class II restricted T cell response in the three domestic pigs analyzed, in the absence of detectable anti-CSFV antibodies in serum. Upon challenge specific T cell responses were boosted in the three vaccinated pigs, and a rapid rise in the titers of CSFV neutralizing antibodies was noticed in two of them, which correlated with a total protection. In these two pigs, neither disease symptoms were observed nor was virus detected at any time after CSFV infection. Neutralizing antibody titers were lower in the third vaccine, which developed a mild and transient peak of pyrexia. As expected, similar analyses in three control pigs (injected with the empty vector or PBS) did not reveal the induction of specific T cells or viral antibodies and, upon challenge, animals developed severe symptoms of the disease, including high titers of viremia, hyperthermia and virus spread to different organs. Control pigs developed, also, a marked leucopenia, resulting in SWC3+ (myelomonocytic cells) being the major PBMC population, and a drastic decrease CD3+ T cells. This T cell depletion was prevented in animals immunized with pcDNA3.1/E2. The total protection achieved, in the absence of CSFV antibodies before challenge, supports the relevance in the antiviral response observed of specific T cell responses primed by pcDNA3.1/E2 vaccine, which, upon challenge, led to a rapid induction of neutralizing antibodies. The observation that CSFV antibodies could only be detected in protected animals after viral challenge opens the possibility of exploring the potential of the DNA vaccine approach used to develop marker vaccines against CSF.

Laboratory decision-making during the classical swine fever epidemic of 1997–1998 in The Netherlands

The National Reference Laboratory for classical swine fever (CSF) virus in the Netherlands examined more than two million samples for CSF virus or serum antibody during the CSF epizootic of 1997–1998. The immense amount of samples and the prevalence of border disease (BD) virus and bovine viral diarrhoea (BVD) virus infections in Dutch pig herds necessitated the diagnostic efforts of the laboratory to be focused on generating CSF specific test results throughout the eradication campaign. Detection of 82% of the 429 outbreaks was achieved through the combined use of a direct immunofluorescence and peroxidase assay (FAT/IPA) with samples (tonsils) collected from clinically-suspected pigs. This suggests that in the majority of the outbreaks, the pigs had clinical signs that were recognised by the farmer and/or veterinarians, indicating the presence of CSF virus in a pig herd. A positive diagnosis of 74% of all the tissue samples (tonsils) collected at infected pig holdings was established by FAT. More than 140,000 heparinised blood samples were examined by virus isolation, resulting in the detection of 4.5% of the infected herds. CSF virus was isolated in approximately 29% of all the blood samples collected from pigs at infected or suspected farms. Several serological surveys — each done within a different framework — led to the detection of 13.5% of the total number of outbreaks. The detection of CSF virus antibody in serum was carried out by semi-automated blocking ELISA. Approximately 28.5% of the sera which reacted in the ELISA were classified as CSF virus-neutralising antibody positive and 26.5% as positive for other pestiviruses following the virus neutralisation test (VNT). We concluded that two of the CSF laboratory diagnostic methods described were determinative in the eradication campaign: first, the FAT for the screening of diseased pigs; and second, the ELISA and VNT when millions of predominantly healthy pigs needed to be screened for the presence of CSF serum antibody. Decision-making on the basis of results generated by either method can, however, be seriously hindered when samples are examined from pig herds with a high prevalence of non-CSF pestiviruses.

Serum neutralization as a differential serological test for classical swine fever virus and other pestivirus infections

Serum neutralization tests (SN) were performed against classical swine fever virus (CSFV), bovine viral diarrhea virus (BVDV) and border disease virus (BDV) on samples of swine serum collected for screening of antibodies to CSFV, in order to determine the SN value as a differential serological test. Ninety-nine sera out of a sample of 16,664 were positive for antibodies to pestiviruses in an ELISA test which did not distinguish antibodies to different pestiviruses. When submitted to SN, 81 sera were positive for CSFV antibodies only. In 17 sera, crossreactive antibodies to either CSFV, BVDV or BDV were detected. In most of these sera (13 out of 17) the differences between SN titres against the three viruses were not sufficient to estimate which was the most likely antibody-inducing virus. It was concluded that, for the SN to be useful in such differentiation, it is essential to examine a sample which must include a representative number of sera from the same farm where suspect animals were detected. When isolated serum samples are examined, such as those obtained with the sampling strategy adopted here, the SN may give rise to inconclusive results.

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.