Detection of foot-and-mouth disease virus infection in vaccinated cattle

The aim of this study was to evaluate the value of commercially available kits for the detection of foot-and-mouth disease (FMD) virus infection in vaccinated cattle. The cattle were vaccinated with a commercial aqueous FMD vaccine type A24 and subsequently challenged 28 days post vaccination with homologous FMD virus. Seven of eight animals were protected from clinical disease and all became carriers. They were bled sequentially for up to 130 days post infection and samples of sera were tested with three ELISA kits: CHEKIT FMD-3ABC, Ceditest FMDV-NS and SVANOIR FMDV 3ABC-Ab ELISA. The Ceditest kit appears to be relatively higher sensitive than the others. When examined with this ELISA, all cattle developed of FMDV nonstructural proteins (NSPs) antibodies and remained positive throughout the period of the experiment. The response of antibodies against 3ABC antigen delayed in two cattle challenged with FMDV A24 virus. One of the cattle reacted negatively in Svanoir ELISA kit and sera from two animals were found negative in CHEKIT ELISA. It can be concluded that all tested kits can be a promising tool for FMD control and eradication campaigns in situation where emergency vaccination was applied.     

Presence of antibodies to non-structural proteins of foot-and-mouth disease virus in repeatedly vaccinated cattle

For the purpose of removing infected animals by detecting humoral immune responses to non-structural proteins of the foot-and-mouth disease (FMD) virus, antibodies induced by contaminated residual non-structural proteins contained in less pure FMD vaccine can be problematic for serological screening. The aim of the present study was to measure the possible presence of antibodies against these non-structural proteins in repeatedly vaccinated calves and beef cattle. Five imported FMD vaccines were examined using two commercial ELISA kits, UBI FMDV NS EIA and Ceditest FMDV-NS, for serological testing. After five doses of vaccination, the serum of one calf tested positive, and two vaccines induced a significant increase in anti-3ABC antibodies in calves. This finding demonstrated that a positive reaction to non-structural proteins due to impurities in the FMD vaccine was detectable using commercial tests. A low percentage of field sera sampled from beef cattle in Kinmen also tested positive, but the key factor resulting in the positive reactions could not be positively identified based on our data.     

Cedivac-FMD can be used according to a marker vaccine principle

In this study, we investigated whether Cedivac-FMD, an emergency vaccine against foot-and-mouth disease (FMD), is suitable for use conjointly with a screening program intended to confirm freedom from disease in vaccinated herds based on evidence of virus replication in vaccinates. Different sets of sera were tested using the Ceditest FMDV-NS ELISA for the detection of antibodies against non-structural proteins (NSPs) of FMD virus. During a vaccine safety study, serum samples were collected from 10 calves, 10 lambs and 10 piglets following administration of a double dose and a repeat dose of high payload trivalent Cedivac-FMD vaccine. All serum samples collected both 2 weeks following the administration of a double dose as well as those collected 2 weeks after the single dose booster (given 2 weeks after the double dose) were negative in the Ceditest FMDV-NS ELISA. In a series of vaccine potency experiments, serum samples were collected from 70 vaccinated cattle prior to and following exposure to infectious, homologous FMD virus. When testing cattle sera collected 4 weeks after vaccination with a regular dose of monovalent >6 PD(50) vaccines, 1 of 70 animals tested positive in the NSP antibody ELISA. After infection with FMD virus, antibodies to NSP were detected in 59 of 70 vaccinated cattle and 27 of 28 non-vaccinated control animals within 7 days. Cedivac-FMD vaccines do not induce NSP antibodies in cattle, pigs or sheep following administration of a double dose or a repeat dose. FMD-exposed animals can be detected in a vaccinated group within 7-14 days. Because Cedivac-FMD does not induce NSP antibodies, the principle of 'marker vaccine' applies.     

Prevalence of seroreagents to FMDV in the cattle population in Poland: results of 9-year monitoring studies

The aim of this study was to evaluate the occurrence of antibodies to foot-and-mouth disease virus (FMDV) in sera of cattle in Poland. The examinations were performed using the virus neutralization (VN) test and ELISA methods: liquid-phase blocking ELISA (LPBE) and 3ABC-ELISA. During 1993-2001, about 681,000 samples of sera collected from animals held on the territory of Poland were tested. Of about 600,000 sera taken from animals exported to the European Union, 963 samples (0.16%) were found to be positive to FMDV types A, O and/or C. During 1996-2001 out of 85,000 sera tested as part of the national serological surveillance program for FMD, the FMDV antibodies were recorded in 51 (0.06%) samples. Persistence of FMD antibodies was observed in sera of cattle from the region around Zduńska Wola, which had been vaccinated annually during 1985-1985 with trivalent FMD vaccine. The results of the serological studies of 550 animals from this region indicates the presence of FMDV antibodies in sera of 240 (44%) cattle. A half-life of maternal antibodies in sera of calves born to seropositive dams was estimated; the highest level of FMDV antibodies was detected in sera taken from new-born calves aged 5-10 days. The level of FMDV antibodies in beestings of dams was highest during the first 10 hours after parturition; after 24 hours a significant decrease (3-5 times) was found and in two weeks post parturition FMDV antibodies were undetectable in the milk. It was established that all LPBE/VN positive sera of cattle exported from Poland, from the vaccination zone around Zduńska Wola as well as those tested as part of the national serological surveillance program for FMD, were taken exclusively from vaccinated animals or calves born to vaccinated dams.     

Comparison of sensitivity and specificity in three commercial foot-and-mouth disease virus non-structural protein ELISA kits with swine sera in Taiwan

Three commercialized ELISA kits for the detection of antibodies to the non-structural proteins (NSPs) of FMD virus were compared, using sera from uninfected, vaccinated, challenged and naturally infected pigs. The kinetics of the antibody response to NSPs was compared on sequential serum samples in swine from challenge studies and outbreaks. The results showed that ELISA A (UBI) and ELISA B (CEDI) had better sensitivity than that of the 3ABC recombinant protein-based ELISA C (Chekit). The peak for detection of antibodies to NSPs in ELISA C was significantly delayed in sera from natural infection and challenged swine as compared to the ELISA A and B. The sensitivity of the three ELISAs gradually declined during the 6-month post-infection as antibodies to NSP decline. ELISA kits A and B detected NSP antibody in 50% of challenged pigs by the 9-10th-day and 7-8th-day post-challenge, respectively. ELISA B and C had better specificity than ELISA A on sequential serum samples obtained from swine immunized with a type O FMD vaccine commercially available in Taiwan. Antibody to NSPs before vaccination was not detected in swine not exposed to FMD virus, however, antibody to NSPs was found in sera of some pigs after vaccination. All assays had significantly lower specificity when testing sera from repeatedly vaccinated sows and finishers in 1997 that were tested after the 1997 FMD outbreak. However, when testing sera from repeatedly vaccinated sows or finishers in 2003-2004, the specificity for ELISAs A, B and C were significantly better than those in 1997. This effect was less marked for ELISA A. The ELISA B was the best test in terms of the highest sensitivity and specificity and the lowest reactivity with residual NSP in vaccinates.     

A Comparison of Methods for Measuring the Antibody Response in Mice and Cattle following Vaccination against Food and Mouth Disease

We present a comparison of methods for evaluating the potency of foot and mouth disease vaccine in the laboratory. The anti-FMDV antibodies (Ab) in vaccinated mice were tested by liquid phase (lp) ELISA, solid phase (sp) ELISA and virus neutralization (VN), and were compared with the Ab titres detected by lpELISA, which is the official test in Argentina for testing the potency of FMD vaccines and protection against a virulent challenge in cattle. The results demonstrated that it is possible to relate the Ab levels induced in vaccinated mice with both the Ab and protective responses elicited in cattle. Furthermore, it was found that the anti-FMDV Ab titres in mice detected by lpELISA 14 days after vaccination should be an accurate parameter for predicting the results of the challenge test in cattle. Thus, this test in mice appears to be an inexpensive and rapid alternative for testing FMD vaccines in cattle.     

Development and validation of a 3ABC indirect ELISA for differentiation of foot-and-mouth disease virus infected from vaccinated animals

Non-structural protein (NSP) 3ABC antibody is considered to be the most reliable indicator of present or past infection with foot-and-mouth disease virus (FMDV) in vaccinated animals. An indirect ELISA was established, using purified His-tagged 3ABC fusion protein as antigen, for detection of the antibody response to FMDV NSP 3ABC in different animal species. The method was validated by simultaneous detection of the early antibody responses to NSP and structural protein (SP) in FMDV Asia 1 infected animals. The performance of the method was also validated by detection of antibody in reference sera from the FMD World Reference Laboratory (WRL) in Pirbright, UK, and comparison with two commercial NSP ELISA kits. The results showed that the antibody response to SP developed more quickly than that to NSP 3ABC in FMDV infected animals. In contact-infected cattle, the antibody response to NSP 3ABC was significantly delayed compared with that to SP antibody. The early antibody responses to SP and NSP 3ABC in FMDV inoculated cattle and contact-infected or inoculated sheep and pigs were generally consistent. In pigs, 3ABC antibody was linked to the presence of clinical signs; however, in sheep, subclinical infection was detected by the development of 3ABC antibodies. Therefore, the antibody responses to 3ABC varied between host species. Eight out of 10 positive serum samples from FMD WRL were tested to be positive at cutoff value of 0.2. The rate of agreement with the ceditest FMDV-NS and the UBI NSP ELISA were 98.05% (302/308) and 93.2% (287/308), respectively. The prevalence of 3ABC antibodies reached 71.4% in some diseased cattle herds. The further work is required to evaluation the performance of this method in different animal species and different field situations.     

Two newly developed Erns-based ELISAs allow the differentiation of Classical Swine Fever virus-infected from marker-vaccinated animals and the discrimination of pestivirus antibodies

New generations of Classical Swine Fever virus (CSFV) marker vaccines have recently been developed in order to make emergency vaccination in case of a CSF outbreak more feasible. However, the application of a marker vaccine is dependent on the availability of an accompanying discriminatory test allowing differentiation of infected from vaccinated animals (DIVA). CP7_E2alf, the most promising live marker vaccine candidate currently available, is a genetically modified Bovine Viral Diarrhea virus expressing the E2 glycoprotein of CSFV strain Alfort/187. The DIVA principle going along with CP7_E2alf is based on the detection of CSFV E^rns-specific antibodies that are raised in the host upon CSFV infection but not after vaccination with the marker vaccine. The aim of this study was to develop novel DIVA tests to be used in combination with CP7_E2alf. Two indirect ELISAs (one for screening, the other one for confirmation purposes) using bacterially expressed recombinant E^rns proteins were designed and evaluated. Both ELISAs detected CSFV-specific antibodies against a broad range of strains and genotypes, and as early as 10 days after infection. They were able to distinguish CSFV-infected pigs from pigs vaccinated with CP7_E2alf and allowed discrimination of antibodies against ruminant pestiviruses in both, sera from domestic pigs and wild boar. Sensitivity and specificity of the screening ELISA was ≥95%. Thus, the ELISAs represent promising DIVA diagnostic tools, as well as an alternative to traditional pestivirus antibody differentiation by serum neutralization test.     

A complex-trapping-blocking (CTB) ELISA, using monoclonal antibodies and detecting specifically antibodies directed against foot-and-mouth disease types A, O and C. II. Application

The complex-trapping-blocking (CTB) enzyme-linked immunosorbent assay (ELISA) was evaluated to detect antibodies directed against foot-and-mouth disease virus (FMDV) strains A10 Holland, O1 BFS, and C1 Detmold. Log10 serum titres of uninfected, unvaccinated cattle (n = 100) were less than 1.80 in the CTB-ELISA. Sera from cattle vaccinated with either monovalent or trivalent vaccines were tested in both the CTB-ELISA and the serum neutralisation test (SNT); titres in both tests correlated positively (P less than 0.001). Titres of sera from cattle, sheep, and pigs vaccinated twice with FMDV A10 Holland also correlated positively in both tests. In another experiment, cattle vaccinated with FMDV strain C1 Detmold were intradermolingually challenged 3 weeks after primary vaccination; at the same time two controls were challenged. At 8 days after challenge, serum titres of the controls were distinctly higher in the CTB-ELISA than in the SNT, whereas serum titres of the vaccinated cattle were equally high in both tests. In potency tests for monovalent vaccines against FMDV strains A10 Holland, O1 BFS or C1 Detmold, serum titres correlated strongly in both tests with protection against the homologous FMDV strain. We concluded that the CTB-ELISA is not only sensitive, but easier to perform and more rapid and reproducible than the SNT. The CTB-ELISA may be useful in evaluating the immune response in cattle during FMD vaccine potency tests.     

Four-layer enzyme immunoassay (EIA) detection of differences in IgG, IgM and IgA antibody response to Aujeszky's disease virus in infected and vaccinated pigs

The use of the four-layer enzyme immunoassay (EIA) for the detection of IgG, IgM and IgA antibodies against Aujeszky's disease virus in blood and oropharyngeal swabs of infected and vaccinated pigs is described. Mean antibody titres obtained using the four-layer EIA were 6.1 and 3829 times higher compared with the indirect enzyme-linked immunosorbent assay (ELISA) and virus neutralization (VN) test, respectively. The VN test detected mainly IgG antibodies, while the IgM antibodies did not react. Using the EIA, the first antiviral antibodies in sera were demonstrated on Days 5-7 after infection or vaccination. Up to the 7th day, demonstrable antibodies were almost exclusively of the IgM class. In infected pigs high titres of IgM antibodies were still detected on Day 18, while in vaccinated animals they were absent by this time. Antibodies of the IgG class appeared in infected pigs sooner (Day 7) than in vaccinated pigs (Day 10) and reached higher mean titres. Antibodies of the IgA class were demonstrable from Day 10 only in samples from infected pigs. Similar antibody dynamics and distribution were detected in oropharyngeal swabs, except that the IgG and IgM titres were roughly 100 times lower than in sera. However, titres of IgA antibodies in oropharyngeal swabs were two times higher than in sera. The greatest differences between both groups of animals were recorded on Day 18; in the infected pigs, IgG, IgM and IgA antibodies were present in sera and oropharyngeal swabs at that time, while in vaccinated pigs only IgG antibodies were demonstrable. The effect of infection and vaccination on the pattern of the immune response as well as the importance of the detection of individual immunoglobulin classes for the specificity of the enzyme immunoassay are discussed.     

Concurrent vaccination of goats with foot and mouth disease (FMD) and peste des petits ruminants (PPR) booster vaccines

Foot and mouth disease (FMD) remains subclinical and self-limiting in small ruminants, but risk of spread of infection to susceptible cohorts is of great epidemiological significance; therefore, small ruminants must be included in vaccination campaigns in FMD endemic regions. Three groups of goats already immunized against peste des petits ruminants (PPR) were vaccinated with FMD and PPR vaccines alone or concurrently. The specific antibody response against three FMD virus strains and PPR virus were evaluated by competitive enzyme-linked immunosorbent assay (cELISA). Goats concurrently vaccinated with PPR + FMD vaccines had significantly (p < 0.05) higher antibody titers to two serotypes of FMD virus at 28, 45, and 60 days post-immunization compared to goats vaccinated with FMD vaccine alone, while goats vaccinated with PPR vaccines alone or PPR + FMD vaccines concurrently showed similar antibody kinetics against PPR virus up till 60 days post-vaccination. Overall, antibody kinetic curves for all three tested strains of FMD virus and PPR virus were similar in vaccinated groups during the course of experiment.     

Foot-and-mouth disease. Infection trial in cattle after a single vaccination

Following an outbreak of FMD caused by an A5 strain in the spring of 1984, ten cattle were vaccinated with samples of the five commercial vaccines used for the vaccination campaign in that year, i.e. two animals per vaccine. Six weeks later the cattle were challenged by contact with animals inoculated with the virus strain isolated from the field outbreak. Seven of the ten cattle became severely ill, exhibiting the typical symptoms of FMD, one animal did not show any clinical symptoms, the remaining two weak ones that might have escaped recognition by the cattlemen. Virus could be recovered from the vaccinated animals from days 2 to 10 following contact with the non-vaccinated infected cattle. It was concluded that a single vaccination does not protect cattle against the isolate.     

Immunization of cattle against Aujeszky's disease with inactivated adjuvant vaccine

The experiments with sheep and young cattle were carried out to test the immunizing efficacy of inactivated adjuvant vaccine against Aujeszky's disease. The vaccine application at doses of 1 ml and 2 ml to lambs at the age of eight to ten months caused the neutralizing antibody production with a significant rise of titres after revaccination. A survival of infection induced with a dose of 10(5.5) TKID50 of virulent virus was recorded in 62.5% of once vaccinated animals and in 87.5% of twice vaccinated animals. When applying different doses of vaccines (from 1 to 10 ml) to young cattle, the antibody reaction level was directly dependent on the inoculum quantity. The double inoculation of animals with vaccines of 2 ml and 5 ml caused the neutralizing antibody production at titres of 1:35, or 1:46. The animals, immunized with the live or inactivated IBR-vaccine possessing high antibody titres against IBR-virus, reacted upon the vaccination with inactivated Aujeszky's vaccine anamnestically, by early production of antibodies in high titres. Metaphylactic vaccination (2 ml of vaccine) of cattle in herds with an acute course days, however earlier during five days from the revaccination when it was carried out in seven days following the first vaccination.     

Assessment using ELISA of the herd immunity levels induced in cattle by foot-and-mouth disease oil vaccines

The development of a liquid-phase blocking sandwich ELISA (LPBE) to measure antibodies (Ab) produced in cattle with the O, A and C foot-and-mouth disease virus (FMDV) types of commercial vaccines used in Argentina is described. The test was specific: 99% of naïve cattle sera (n = 130) gave titres below log₁₀ = 1.2, and none had a titre above log₁₀ = 1.5. Comparative studies with serum neutralization test (SNT) using sera from cattle which received one or more vaccine doses is reported. The overall rank correlation coefficient (Spearman's , r_s) between SNT and LPBE were highly significant (r_s > 0.67, P < 0.0001) for all vaccine strains. LBPE Ab titres on sera collected 90 days post vaccination were compared with results of cattle protection tests by applying a logistic regression. The minimum Ab titres at which 85% and 75% of the cattle were protected for each FMDV type were determined in order to interpret field Ab data in terms of protection. Application of this method allows large scale serological examinations to monitor antibody levels in vaccinated animals as an indirect indicator of the FMD control program status in the field. Its use in the evaluation of commercial batches of FMD vaccine is discussed.     

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

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

Evaluation of safety and efficacy of DNA vaccines against bovine herpesvirus-1 (BoHV-1) in calves

Four DNA vaccines against BoHV-1 were evaluated for their efficacy in calves. Twelve animals were divided into four groups which were injected with four different DNA vaccines: pVAX-tgD (Vaccine A); pVAX-tgD co-immunised with pVAX-48CpG (Vaccine B); pVAX-UbiLacI-tgD-L (Vaccine C); pVAX-UbiLacI-tgD-L co-immunised with pVAX-48CpG (Vaccine D). Three additional calves were given the plasmid vector and served as controls. Ninety days after the first vaccination all calves were challenge infected with BoHV-1.All animals developed a severe form of infections bovine rhinotracheitis. Only the calves given the pVAX-tgD co-immunised with pVAX-48CpG (Vaccine B) developed humoral antibodies against BoHV-1 between 56 and 90 days after the first vaccination, whereas in calves of other groups and in the controls, antibodies appeared only after the infection. In the calves vaccinated with either pVAX-tgD (Vaccine A) or pVAX-tgD combined with pVAX-48CpG (Vaccine B), BoHV-1-specific IFN-γ secreting cells were detected in PBMCs 90 days after the first vaccination and their number increased after challenge exposure. In the other groups the IFN-γ secreting cells were detected after virus infection and at low values.     

Immune Effect of Inactivated Vaccines against Foot-and-Mouth Disease and Their Interference on Differential Diagnosis

Vaccination with inactivated vaccine is an important measure to prevent and control foot-and-mouth disease (FMD), however, the immune effect and antigenic purity of inactivated vaccines are two major concerns for the establishment and evaluation of FMD free zones with vaccination. In this study, four groups of FMD type O and type A bivalent inactivated vaccines from 3 FMD vaccine manufacturers (designated as A, B and C) were selected to inoculate healthy juvenile cattle of FMD free. All cattle were immunized 3 or 4 times at a 1-month interval. Serum samples were collected before and after 1 month of every vaccination to determine the level of antibody to structural protein and non-structural protein. Results：(1) The qualified rates of antibody to structural protein: in group a1 (vaccine from company A, different batches), the antibody qualified rate could reach 100% for type O and type A, respectively after each vaccination. In group a2 (vaccines from company A, same batch), the antibody qualified rates were 36.7%, 98.3% and 100% for type O, and 15%, 86.7% and 100% for type A after the first to the third vaccination, respectively. In group b (vaccine from company B, same batch), the antibody qualified rates were 18.3%, 97% and 100% for type O, and 1.7%, 45% and 53.3% for type A after the first to the third vaccination, respectively. In group c (vaccines from company C, same batch), the antibody qualified rates were 26.7%, 96.7% and 100% for type O, and 21.7%, 71.7% and 100% for type A after the first to the third vaccination, respectively. (2) Antibody positive rate to non-structural protein 3ABC (confirmed with second ELISA test): In group a1, the positive rates were 0.7%, 1.4%, 9.5% and 4.8% after the first to the fourth vaccination, respectively; In group a2 and c, no 3ABC antibody-positive animal was detected after 3 repeated vaccination; In Group b, only one animal with a positive rate of 0.6% was detected after the third vaccination. The antibody qualified rates to the structural protein of FMDV in 3 of the 4 groups were far less than 70% after the primary vaccination, however, those were increased significantly after boost and repeated vaccination. The antigen purity of vaccines in three groups (a2, b and c) can meet the requirement of OIE standard on the FMD vaccine, however, the seroconversion to 3ABC antibody was obvious in animals from group a1 after repeated vaccination, which would cause some extent of interference to differential diagnosis. Also, a combination of a primary screening test and a confirmatory ELISA test can further improve the accuracy of differential diagnosis. This study provides an important scientific basis to make a rational program for establishment and evaluation of FMD free zone with vaccination.     

口蹄疫灭活疫苗的免疫效果及其对鉴别诊断的干扰

灭活疫苗免疫是防控口蹄疫的重要措施,但是灭活疫苗的免疫效果及其对感染与免疫鉴别诊断的干扰一直是口蹄疫免疫无疫区建设评估需要明确的重要问题。本研究中选择了3个企业（代号A、B与C）的4组口蹄疫O型与A型二价灭活疫苗,分别免疫口蹄疫抗体阴性健康未成年牛,免疫3～4次,测定免疫前后结构蛋白和非结构蛋白抗体的应答水平。结果显示：（1）结构蛋白抗体合格率：a1组（A企业多批次疫苗）4次免疫后O型和A型均为100%;a2组（A企业同批次疫苗）一~三免O型为36.7%、98.3%与100%,A型为15%、86.7%与100%;b组（B企业疫苗）一~三免O型为18.3%、97%与100%,A型为1.7%、45%与53.3%;c组（C企业疫苗）一~三免O型为26.7%、96.7%与100%,A型为21.7%、71.7%与100%。（2）非结构蛋白3ABC抗体阳性率（两种方法复核结果）：a1组一~四免分别为0.7%、1.4%、9.5%与4.8%;a2组和c组三次免疫均未检测到阳性;b组仅三免后阳性率为0.6%。3组灭活疫苗首次免疫牛的抗体合格率远不及70%,但加强免疫后抗体合格率均显著提高;非结构蛋白抗体检测结果表明有3组疫苗的抗原纯净度符合OIE的要求,但a1组灭活疫苗免疫后,仍然对感染与免疫鉴别诊断存在干扰;采用两种非结构蛋白抗体检测方法进行复核检验,可以提高感染与免疫鉴别诊断的准确性。本研究为口蹄疫免疫无疫评价方案的制定提供了科学依据。     

Humoral Response to 2 Inactivated Bluetongue Virus Serotype‐8 Vaccines in South American Camelids

Background: Bluetongue virus serotype 8 (BTV‐8) has caused disease in domestic ruminants in several countries of northern Europe since 2006. In 2008 a mass‐vaccination program was launched in most affected countries using whole virus inactivated vaccines. Objective: To evaluate 2 inactivated vaccines (Bovilis BTV 8; BTVPUR AlSap8) for immunogenicity and safety against BTV‐8 in South American camelids (SAC) in a field trial. Animals: Forty‐two SAC (25 Alpacas, 17 Llamas) aged between 1 and 16 years. Methods: The animals were vaccinated twice at intervals of 21 days. They were observed clinically for adverse local, systemic, or both reactions throughout the trial. Blood samples collected on days 0, 14, 21, 43, and 156 after vaccination were tested for the presence of BTV‐8 virus by real time‐polymerase chain reaction and of specific antibodies by competitive ELISA and a serum neutralization test. Results: All vaccinated animals developed antibodies to BTV‐8 after the 2nd administration of the vaccine. No adverse effects were observed except for moderate local swellings at the injection site, which disappeared within 21 days. Slightly increased body temperatures were only observed in the first 2 days after vaccination. The BTV was not detected in any of the samples analyzed. Conclusions and Clinical Importance: The administration of the 2 inactivated commercial vaccines was safe and induced seroconversion against BTV‐8 in all vaccinated animals. The results of this study suggest that 2 doses injected 3 weeks apart is a suitable vaccination regimen for SAC.     

Developments in diagnostic techniques for differentiating infection from vaccination in foot-and-mouth disease

Foot-and-mouth disease (FMD) is a highly contagious and economically significant disease of cattle, pigs, sheep, goats and wild ruminant species. The FMD virus genome encodes a unique polyprotein from which the different viral polypeptides are cleaved by viral proteases, including eight different non-structural proteins (NSPs). Both structural and non-structural antigens induce the production of antibodies in infected animals. In contrast, vaccinated animals which have not been exposed to replicating virus will develop antibodies only to the viral antigens in the inactivated material. Vaccination against FMD is a key element in the control of the disease in addition to slaughter and movement restrictions. However, countries that vaccinate in the event of an outbreak will have to re-establish their FMD free status to the satisfaction of their trading partners.Because currently available vaccines stimulate the production of antibodies indistinguishable from those produced by infected animals in response to live virus and because vaccinated animals can be infected and become carriers of FMD virus, efforts have been made to develop diagnostic test that can differentiate vaccinated animals from those that are convalescent and from those that have been vaccinated and become carriers following subsequent contact with live virus. Currently the detection of antibodies to non-structural protein's (NSPs) is the preferred diagnostic method to distinguish virus infected, carrier, animals from vaccinated animals. However this is currently only possible at the herd level because of the great variability in the initiation, specificity and duration of the immune response in individual animals to the NSPs shown in many studies. Considerable effort and attention is now being directed toward the development of new methods and techniques for the rapid and accurate detection of anti-NSP antibodies, harmonization and standardization of current diagnostic techniques, as well as the production of defined reagents.