Transient classical swine fever virus infection in wild boar piglets partially protected by maternal antibodies

An experimental study was conducted to investigate the clinical course of classical swine fever (CSF) in wild boar piglets partially protected by maternal antibodies. Five healthy wild boar piglets with a low serum titre of colostral antibodies against CSF virus were challenged with virulent CSF virus at the age of three months. Apart of reduced food intake and diarrhoea no major clinical symptoms were noticed after challenge. These signs were seen during the second and third week of infection, afterwards the piglets recovered completely. CSF virus could be re-isolated from blood samples taken on day 12 and day 19 post challenge. From blood samples taken later on and from the organ material taken at post mortem examinations no CSF virus could be isolated anymore. It can be concluded that the presence of maternal antibodies influences the clinical course of CSF in terms that the outcome is rather transient than lethal. Such wild boar could play a crucial role in the spread of CSF virus and might contribute to the maintenance of long lasting epizootics.     

Genetic differentiation of infected from vaccinated animals after implementation of an emergency vaccination strategy against classical swine fever in wild boar

Oral emergency vaccination against classical swine fever is a powerful tool to control disease outbreaks among European wild boar and thus to safeguard domestic pigs in affected regions. In the past, when virus detection was mainly done using virus isolation in cell culture or antigen enzyme-linked immunosorbent assays, modified live vaccine strains like C-strain "Riems", were barely detectable after oral vaccination campaigns. Nowadays, the use of highly sensitive molecular techniques has given rise to an increase in vaccine virus detections. This was also the case during the 2009 outbreak among German wild boar and the subsequent vaccination campaigns. To guarantee a rapid differentiation of truly infected from C-strain vaccinated animals, a combination of differentiating multiplex rRT-PCR assays with partial sequencing was implemented. Here, we report on the rational and use of this approach and the lessons learned during execution. It was shown that positive results in the recently developed vaccine strain (genotype) specific rRT-PCR assay can be taken as almost evidentiary whereas negative results should be confirmed by partial sequencing. Thus, combination of multiplex rRT-PCR assays as a first line differentiation with partial sequencing can be recommended for a genetic DIVA strategy in areas with oral vaccination against classical swine fever in wild boars.     

Efficacy of lyophilised C-strain vaccine after oral immunisation of domestic pigs and wild boar against classical swine fever: first results

The aim of this study was to evaluate the efficacy of lyophilised C-strain vaccine in domestic pigs and wild boar after oral application. A new spherical bait form (diameter 3 cm) containing lyophilised vaccine virus and the recent vaccine baits were used for animal experiments. Four vaccination groups were established in experiment 1 (group 1: recent liquid bait vaccine; group 2: spherical baits containing one dose of the lyophilised vaccine; groups 3 (domestic pigs) and 4 (wild boar): spherical baits containing two doses of the lyophilised vaccine) and two groups in experiment 2 (group 1: recent liquid bait vaccine; group 2: spherical baits with two doses of the lyophilised vaccine). Challenge was carried out with the highly virulent virus strain "Alfort 187" (using 100 TCID50 in the first and 1.000 TCID50 in the second experiment). Our results showed that the animals vaccinated with lyophilised C-strain vaccine developed high neutralising antibody titres comparable to those obtained after vaccination with the recent bait vaccine. All pigs which picked up the baits remained healthy after challenge. Neither clinical symptoms nor viremia or virus shedding were observed after infection except in one pig (group 2, experiment 2) which had not consumed the vaccine bait. The surviving domestic pigs and wild boar were tested negative for CSFV and viral RNA at the end of the study. This result demonstrates that lyophilised vaccine may become an effective vaccine formulation for oral immunisation of wild boar against CSF in the near future.     

Two outbreaks of classical swine fever in wild boar in France

In 2002 and 2003, two successive outbreaks of classical swine fever were declared in wild boar in northern France. The first was in Moselle, near the town of Thionville and the border with Luxembourg, and the second was in the northern Vosges area, near the German border. The outbreaks were investigated by serological and virological diagnosis of dead or shot animals. Hunting restrictions were applied to limit the spread of the outbreaks. The virus was detected eight times between April and July 2002 in the Thionville area, an area well delimited by natural or artificial barriers such as rivers or highways. Cooperation between the authorities concerned was good, and hunting restrictions were applied for one year. No virus was detected after July 2002 and the Thionville outbreak was officially considered over in March 2005. In the northern Vosges the situation was different, with no barriers to animal movements, continuous forest, difficulties in establishing hunting restrictions in this huge area, and the circulation of the virus in Germany close to the frontier. Virus of a different strain from that isolated in the Thionville outbreak was still being isolated in the northern Vosges in 2004, and owing to the failure of the hunting restrictions, the French health authorities decided to vaccinate wild boar.     

Long-term studies on maternal immunity for Aujeszky's disease and classical swine fever in wild boar piglets

The aim of the studies was to fathom the duration and the role of maternal immunity for Aujeszky's disease (AD) and classical swine fever (CSF) in wild boar offspring. In one experiment, two wild boar sows were infected with a low pathogenic pseudorabies virus (PRV) in 1999. A total of 51 offspring was born between 1999 and 2002 and was monitored for PRV maternal antibodies. In a second experiment, the maternal immunity for CSF was analysed. Therefore, a sow was orally vaccinated against CSF using vaccine baits containing the live-attenuated C-strain vaccine. The vaccination took place in January 1999. The sow gave birth to four piglets in 2001 and to two piglets in 2002. With respect to maternal immunity for AD, some piglets reacted positive in the ELISA up to 27-week post-partum while in the neutralization test antibodies were detected up to 15-week post-partum. The calculated half-life of neutralizing antibodies was 21 days. Regarding CSF, the neutralization titres of maternal antibodies dropped continuously reaching values of < or =10 ND50 20-week post-partum. After the 12th week post-partum, most of the sera reacted negative in the ELISA. However, after the third month, low levels of neutralization titres were still detectable. The results are discussed with respect to the epidemiology and control of both diseases in wild boar populations.     

Protection of gruntlings against classical swine fever virus-infection after oral vaccination of sows with C-strain vaccine

The objective of this study was to investigate the maternal protection of gruntlings derived from wild sows vaccinated orally against classical swine fever (CSF) using C-strain vaccine. Three vaccinated sows and one unvaccinated control sow were included. Challenge infection of the progeny was carried out either intranasally or by contact at the beginning of the third month of life (61-65 days post-natum). Whereas, two of three litters had maternal antibodies, the progeny of one vaccinated sow was seronegative at challenge. The progeny of the control sow, which was challenged by contact infection, developed moderate clinical signs except for one animal which became ill and died. Two gruntlings derived from the vaccinated sows also died of CSF, although one of them had a relatively high maternal antibody titre (128 ND(50)). The transient infection and partial virus shedding observed in a small number of gruntlings with maternal antibodies and the fact that one animal with maternal antibodies became ill and died confirm the incomplete maternal protection at this age. The reason for this incomplete protection is discussed. As none of the surviving gruntlings could be shown to carry CSFV or viral RNA at the end of the experiment (36 or 70 d.p.i.), it may be concluded that these animals do not represent a potential CSFV reservoir.     

Evaluation of vaccine-induced maternal immunity against classical swine fever

The vaccine-induced maternal immunity against classical swine fever (CSF) was investigated in this study. Eight sows were vaccinated with the Chinese strain of classical swine fever virus (CSFV). The length of time between vaccination and farrowing was 167-217 days. Milk samples from the front, middle and back udder sections and blood samples were taken from the sows on days 3 and 14 after farrowing. Blood samples were obtained from the piglets at the age of 3, 6 and 10 weeks. The antibody level of the milk was examined by ELISA, and that of blood samples by the virus neutralization (VN) test as well. In all 3-week-old piglets and in 80% of the 6-week-old animals the neutralizing antibody level reached the titre of 1:40. In none of the 10-week-old piglets did the titre exceed the value of 1:20, but in about 25% of the piglets it reached 1:20; the half of these piglets came from two litters. In none of the piglets did the antibody level reach the negative threshold in the ELISA test during the study. No significant differences were found between the udder sections in milk antibody level by ELISA.     

Oral immunisation of wild boar against classical swine fever: evaluation of the first field study in Germany

The effectiveness of oral immunisation of wild boar against classical swine fever (CSF) was studied in a field trial in Lower Saxony for two years, from 1993 to 1995. This field study was performed in an area of ca. 270 km(2)50% of young boars did not feed on vaccine baits nor become immunised. Therefore, an intensive hunting of this age group is a necessary adjunct to the use of oral vaccination. After the third immunisation period, no virus was detected in the areas where oral immunisation took place.     

Serum antibodies directed against classical swine fever virus and other pestiviruses in wild boar (Sus scrofa) in the Republic of Croatia

The presence of serum antibodies directed against classical swine fever (CSF) virus and other pestiviruses among the wild boar (Sus scrofa) population in Croatia was investigated. During 2003, serum samples from 214 wild boars were collected in 10 hunting areas in the continental part of the country.The sera were examined by enzyme immunoassay (ELISA) and in the virus neutralization test (VNT). Out of 214 sera tested 111 (51.87 %) were positive by ELISA and regarding neutralising antibodies, against CSFV 75 (35.05 %) samples were positive. In the VNT with the C-strain (conventional live vaccine strain China) and the strain Uelzen were used. Samples were also tested for neutralizing antibodies against border disease virus (BDV) using the strain 137/4 and against bovine viral diarrhoea virus (BVDV) using the NADL strain. Neutralizing antibodies against the C-strain were detected in 36 sera (16.82 %), against strain Uelzen in 17 sera (7.94 %) and in 22 sera (10.28 %) against both strains. In five sera (2.33 %) neutralizing antibodies against BVDV and BDV were found.     

Adsorption of colostral antibodies against classical swine fever, persistence of maternal antibodies, and effect on response to vaccination in baby pigs.

OBJECTIVE: To determine kinetics of antibody absorption, persistence of antibody concentrations, and influence of titers on vaccination of baby pigs with a vaccine against classical swine fever (CSF). ANIMALS: 15 sows and their litters. PROCEDURE: Farrowings were supervised. Initial time of suckling was recorded. In the first experiment, blood samples were collected at farrowing, 2 and 4 hours after suckling, and hourly until 10 hours after initial suckling. Samples were assayed for CSF antibodies, using a serum neutralizing (SN) test. A second experiment included 33 baby pigs vaccinated as follows: 10 prior to ingestion of colostrum, 18 between 1 and 4 hours after ingestion of colostrum, and 5 at 12 hours after ingestion of colostrum. Fourteen pigs were vaccinated when 7 weeks old, and 15 pigs were not vaccinated. At 10 weeks of age, pigs were challenge-exposed with virulent CSF virus. Blood samples were collected and assayed for CSF antibodies and p125 antigen and p125 antibodies. RESULTS: CSF antibodies were detected in pigs beginning 2 hours after suckling. Colostral antibodies persisted for > 7 weeks (half-life, 79 days). Vaccination of pigs before suckling provided effective protection from severe disease after challenge-exposure. However, vaccination of neonates with antibody titers was not effective, because 19 of 23 (82%) pigs succumbed after challenge-exposure. All pigs vaccinated when 7 weeks old resisted challenge-exposure, whereas all unvaccinated control pigs succumbed. CONCLUSIONS AND CLINICAL RELEVANCE: Vaccination before ingestion of colostrum conferred good protection against CSF in baby pigs. Vaccination of 7-week-old pigs that had decreasing concentrations of passively acquired antibodies was efficacious.     

Monitoring of classical swine fever in wild boar (Sus scrofa) in Slovenia

Classical swine fever (CSF) is a highly contagious multi-systemic haemorrhagic viral disease of pigs. Not only domestic pigs, but also wild boar appear to play a crucial role in the epidemiology of CSF. Spleen (n = 739) and blood coagulum (n = 562) sampled from wild boars (Sus scrofa) shot in 2002, and serum samples from 746 wild boar shot in 2003 and 2004, were tested throughout Slovenia. In 2002, 17 samples were positive on enzyme-linked immunosorbent assay (ELISA) test for antibodies against classical swine fever virus (CSFV). Positive ELISA test was confirmed by a virus neutralization test. All other samples were negative. This is the first report that describes the epidemiology of CSFV from 2002 on, and the monitoring of the wild boar population in Slovenia at present.     

猪瘟单克隆抗体的制备及ACI-ELISA检测猪瘟病毒的研究

本研究用猪瘟石门毒(CSFV-Shimen)免疫BALB/C小鼠,按常规单克隆抗体(McAb)技术方法制作,最终获得4株McAb,分别命名为AC9、CF8、DG5和EC9,4株McAb与基因工程CSFV E2蛋白反应结果表明:AC9、CF8和EC9是抗CS-FV E2蛋白的McAb.用AC9和CF8McAb对CSFV进行抗原捕获间接ELJSA试验(ACI-ELISA),通过一元McAb和二元McAb CAI-ELISA试验的比较,结果表明AC9与CF8两种McAb有协同作用,其捕获CSFV的能力比一元McAb显著提高.方阵试验结果表明:McAb和血清多抗(PcAb)的最佳工作稀释度分别为1:400和1:200.特异性试验和敏感性试验结果显示本法特异性强,敏感性高.最后用ACI-ELISA与PCR对30份病料的检测结果比较,表明ACI-ELISA与PCR检测结果相符.上述结果说明本研究所获得AC9和CF8可用作猪瘟诊断试剂盒的研制,是检测CSFV的有效方法.     

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 simultaneously with the CSF vaccine produced a significantly higher antibody response to CSF than occurred with CSF vaccination only.     

Oral immunisation of wild boar against classical swine fever: concluding analysis of the recent field trials in Germany

The recent oral immunisation trials in wild boar against classical swine fever (CSF) in Germany are described and evaluated in summary. After the first field study in Lower Saxony from 1993-1995 further immunisation trials started in Mecklenburg-Western Pomerania, Brandenburg, Lower Saxony, Baden-Württemberg and Saxony-Anhalt. The immunisation strategies and the size of the vaccination zones were different in the individual federal states. In principle, the bait vaccine based on the CSF virus strain "C" were laid out by hand. Later also the aerial distribution was carried out in selected areas of Mecklenburg-Western Pomerania. The application of baits by plane was introduced at the beginning of the immunisation measures in Saxony-Anhalt apart from the manual distribution. Up to now, the field trials show that the oral immunisation can be an additional tool for CSF control by increasing of herd immunity and reduction of the CSFV prevalence. However, the immunisation was not sufficient enough for young boars in the most field studies. Based on the evaluation of the immunisation experiments an improved immunisation procedure is recommended.     

Epidemiology and control of an outbreak of classical swine fever in wild boar in Switzerland

An outbreak of classical swine fever in wild boar in the southern part of Switzerland (Canton of Ticino) was investigated after the implementation of control measures in a defined infected area (the risk zone), and in a surrounding surveillance zone (the non-risk zone). After the disease had been detected, hunting was not allowed in the risk zone for over six months, during which the disease was left to run its course, but hunting was continued in the non-risk zone for one month. After seven months, a hunting strategy targeted at young animals was implemented in both zones. Between May 1998 and January 2000,1294 wild boar were shot or found dead, and diagnostic and biological data were collected and analysed. Only one animal from the non-risk zone was found to be seropositive for antibodies to the virus, whereas 179 of 528 wild boar from the risk zone were virus positive and 162 were seropositive. The proportion of virus-positive animals decreased from 62.7 per cent to zero over one year. During the first hunting season, seropositive animals were found in all age groups, but 12 months later only animals more than one year old had antibodies against the virus.     

Factors critical for successful vaccination against classical swine fever in endemic areas

Classical swine fever (CSF) or hog cholera, caused by the classical swine fever virus (CSFV), is one of the most important viral diseases that cause serious economic loss to the swine industry worldwide. During the past 5 years, several techniques for measuring porcine cell-mediated immunity (CMI) were applied, in conjunction with other conventional techniques, to study factors that influence the induction of CSFV-specific immunity. Information, obtained from a series of experiments, demonstrated cell-mediated immune responses in providing protective immunity against CSF infection. Although it has been confirmed that commercially available modified live CSF vaccines are able to induce complete protection in vaccinated pigs, several factors including maternal immunity, the age of primary vaccination, vaccination protocol and complications caused by other pathogens, can greatly affect the effectiveness of CSF vaccines in the field.     

“猪瘟和非洲猪瘟的综合防治措施”专栏(三) 影响猪瘟流行地区疫苗成功接种的关键因素

猪瘟(Classical Swine Fever,CFS)又称猪霍乱,由猪瘟病毒(CSFV)引起,是能给全球养猪业造成严重经济损失的最重要的病毒性疾病之一.在过去的5年中,多项用于测定猪细胞免疫(Cell-Mediated Immunity,CMI)的技术与其他常规技术一起,被用来研究可影响猪瘟病毒特异性免疫诱导的因素.从一系列试验获得的资料证明,细胞免疫应答可提供抗猪瘟感染的保护性免疫.尽管业已证实商用改良型猪瘟活疫苗能够诱导免疫猪产生完全的保护,但包括母源免疫、首次免疫年龄、免疫接种方案和其他病原体引起的并发症等多种因素,均会严重影响猪瘟疫苗在田闻的免疫效果.     

影响猪瘟流行地区疫苗成功接种的关键因素

猪瘟（Classical Swine Fever,CFS）又称猪霍乱,由猪瘟病毒（CSFV）引起,是能给全球养猪业造成严重经济损失的最重要的病毒性疾病之一。在过去的5年中,多项用于测定猪细胞免疫（Cell-Mediated Immunity,CMI）的技术与其他常规技术一起,被用来研究可影响猪瘟病毒特异性免疫诱导的因素。从一系列试验获得的资料证明,细胞免疫应答可提供抗猪瘟感染的保护性免疫。尽管业已证实商用改良型猪瘟活疫苗能够诱导免疫猪产生完全的保护,但包括母源免疫、首次免疫年龄、免疫接种方案和其他病原体引起的并发症等多种因素,均会严重影响猪瘟疫苗在田间的免疫效果。