Stimulation by heterotypic antigens of foot-and-mouth disease virus antibodies in vaccinated cattle

Immunisation of cattle with foot-and-mouth disease virus failed to raise a level of antibody that provides protection against heterotypic challenge. Further the 12S substructure, produced from the 146S particle, was ineffective in providing protection against challenge by homotypic virus. These findings suggest considerable antigenic differences in the virus serotypes and between the virus and its substructure. Inoculation of homologous 12S and heterologous 1246S and 12S antigens into vaccinated cattle, however, revealed antigenic relationship between different serotypes, and between serotypes and their substructures.     

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.     

Detection of carriers of foot-and-mouth disease virus among vaccinated cattle

To investigate and optimise detection of carriers, we vaccinated 15 calves with an inactivated vaccine based on foot-and-mouth disease virus (FMDV) A Turkey strain and challenged them and two further non-vaccinated calves with the homologous virus four weeks later. To determine transmission to a sensitive animal, we put a sentinel calf among the infected cattle from 60 days post-infection until the end of the experiment at 609 days post-infection. Samples were tested for the presence of FMDV, viral genome, specific IgA antibodies, antibodies against FMDV non-structural (NS) proteins or neutralising antibodies. Virus and viral genome was intermittently isolated from probang samples and the number of isolations decreased over time. During the first 100 days significantly more samples were positive by RT-PCR than by virus isolation (VI), whereas, late after infection more samples were positive by virus isolation. All the inoculated cattle developed high titres of neutralising antibodies that remained high during the entire experiment. An IgA antibody response was intermittently detected in the oropharyngeal fluid of 14 of the 17 calves, while all of them developed detectable levels of antibodies to NS proteins of FMDV in serum, which declined slowly beyond 34 days post-infection. Nevertheless, at 609 days after inoculation, 10 cattle (60%) were still positive by NS ELISA. Of the 17 cattle in our experiment, 16 became carriers. Despite frequent reallocation between a different pair of infected cattle no transmission to the sentinel calf occurred. It remained negative in all assays during the entire experiment. The results of this experiment show that the NS ELISA is currently the most sensitive method to detect carriers in a vaccinated cattle population.     

4种ELISA检测奶牛O型口蹄疫免疫抗体的比较试验

口蹄疫(FMD)是由口蹄疫病毒引起的以感染偶蹄动物为主的急性、热性、高度传染性疫病,世界动物卫生组织(OIE)将其列为必须报告的动物疫病.我国规定为一类动物疫病.我国奶牛口蹄疫的防控,以免疫接种措施为核心,实行强制免疫政策,要求O型和Asia Ⅰ型疫苗免疫密度必须达100%,通过牛群整体免疫水平的提高来预防口蹄疫的感染和流行[1].     

Transmission of foot-and-mouth disease by vaccinated cattle following natural challenge

Cattle vaccinated with a conventional monovalent type O1 foot-and-mouth disease (FMD) vaccine were challenged between four and 21 days after vaccination by short-term exposure to homologous airborne virus produced by pigs. Transmission was then assessed by housing susceptible cattle with the vaccinated animals and testing and observing all the animals for signs of infection and clinical disease. All 18 cattle vaccinated three weeks before challenge resisted clinical disease and although four contracted subclinical infection, there was no transmission to susceptible cattle in contact. One of the two groups of cattle vaccinated two weeks previously transmitted subclinical infection, but not disease, to susceptible animals housed with them from day 0 after challenge. Subclinical infection was manifested by a transient viraemia which was not followed by a detectable circulating antibody response. Shorter periods (seven or four days) from vaccination to challenge resulted in transmission of disease from clinically normal vaccinated to in-contact animals in one of two experiments. The severe challenge presented by the diseased in-contact animals than overwhelmed the immunity of the vaccinated animals. The results indicate that during emergency vaccination programmes it is advisable to vaccinate all FMD-susceptible animals within the vaccination zone and that at the outer boundary of the zone vaccinated animals should be kept separated from unvaccinated animals for at least three weeks.     

Investigation of an outbreak of foot-and-mouth disease in vaccinated dairy cattle in Thailand

SUMMARY An outbreak of foot-and-mouth disease that spread between two related vaccinated dairy herds was investigated. Although the cattle were of similar vaccination status, in one herd there was high morbidity, whereas in the other there was considerably lower morbidity. The relationship between the vaccine virus and the outbreak virus was expressed as an r value determined by the two-dimensional neutralisation test. Bovine serum homologous to the vaccine virus indicated a close antigenic relationship between the vaccine virus and the outbreak virus (r = 0.61). The source of the outbreak virus was not determined. The investigation suggested a requirement for close contact between stock for foot-and-mouth disease to spread in a tropical environment, in contrast to the capacity of the disease to spread considerable distances by aerosol transmission in temperate climates.     

Comparable sensitivity and specificity in three commercially available ELISAs to differentiate between cattle infected with or vaccinated against foot-and-mouth disease virus

Three commercially available ELISAs for the detection of antibodies to the non-structural proteins of foot-and-mouth disease virus (FMDV) were evaluated, using sera from uninfected, vaccinated, infected, inoculated, first vaccinated and subsequently infected, and first vaccinated and subsequently inoculated cattle. We compared antibody kinetics to non-structural proteins, sensitivity, and specificity. One of the ELISAs had a higher sensitivity and much lower specificity than the other two, therefore we established standardised cutoff values for the compared assays using receiver operated characteristic (ROC) curves. Using the standardised cutoff values, all three ELISAs produced comparable results with respect to sensitivity and specificity. Antibody development to non-structural proteins after infection and after vaccination/infection was not significantly different. Development of antibodies, however, both neutralising and directed to non-structural proteins, was significantly delayed after intranasal inoculation as compared to intradermolingual infection. Based on results of sera obtained after vaccination and experimental infection all three assays can be used for testing sera collected between 4 weeks and 6 months after infection. More information is needed on the prevalence of positive reactors in a situation where emergency vaccination has been used and FMD transmission was still observed.     

牛口蹄疫病毒VP2结构蛋白抗体间接ELISA方法的建立

为建立牛口蹄疫(FMD)抗体的检测方法,本研究将口蹄疫病毒(FMDV)的VP2基因,通过pPROEXTM HTb表达载体在大肠杆菌DH5α中表达,获得大小为35ku的重组VP2蛋白(rVP2),western blot证实rVP2可与FMDV5种血清型的牛阳性血清发生特异性反应。以纯化复性的rVP2为抗原建立了FMDVrVP2间接ELISA方法。重复性试验证实批内、批间变异系数均小于10%。特异性交叉试验表明,该抗原不与常见的其他7种牛病阳性血清发生交叉反应。检测非免疫无口蹄疫国家牛阴性血清的特异性为100%;检测感染血清敏感性为97.3%;检测O-AsiaⅠ的二价苗免疫牛血清,与4种商品化试剂盒比较,其符合率分别为69.0%、95.0%、90.4%和86.8%。实验结果表明建立的ELISA方法可以用于口蹄疫感染和免疫抗体检测。     

The airborne dispersal of foot-and-mouth disease virus from vaccinated and recovered pigs, cattle and sheep after exposure to infection.

Foot-and-mouth disease virus was detected during two periods in the air of looseboxes which housed susceptible, vaccinated or recovered pigs, cattle or sheep exposed to infection. The first was 30 min to 22 h after exposure and occurred in all animals. The second was two to seven days after exposure and occurred with those susceptible and vaccinated animals which developed clinical lesions, and with vaccinated and recovered pigs and sheep, which did not develop clinical lesions. Vaccination of animals before exposure resulted in less or no virus being detected. The virus during the first period was attributed to virus trapped on the animal during exposure, and the virus during the second period to limited multiplication in the respiratory tract. Control of movement for two weeks after contact with infection is suggested as a means of preventing spread of foot-and-mouth disease in areas that contain vaccinated animals.     

Residual foot-and-mouth disease virus antibodies in French cattle and sheep six years after the vaccination ban

A serological survey was carried out on French cattle to establish a reference pattern of residual vaccine antibodies and non-specific reactions against the foot-and-mouth disease virus 6 years after the ban on vaccination and in the absence of any foot-and-mouth disease outbreak. Most of the multi-vaccinated cattle still displayed high titres of antibodies and up to 50% of those which had received a single injection still had antibodies. Non-specific reactors were also recorded among animals born during and after 1991. Most of them displayed low titres close to the threshold. Sheep were also tested and, as for cattle, 4.6% of non-specific reactors were recorded, with titres close to the threshold for two-thirds of them. As part of these animals have been resampled and retested, sera revealed negative confirming that these animals are true non-specific reactors. Serological testing as a mean of FMD control should take these facts into account.     

Prevalence of antibodies to bovine virus diarrhoea-mucosal disease virus in Tanzanian cattle

A serological survey to determine the prevalence of antibodies to bovine virus diarrhoea-mucosal disease (BVD-MD) virus was conducted on 419 bovine serum samples originating from 18 of 20 regions (except Mwanza and Shinyanga) of the Tanzania mainland. The sera were a small proportion of samples collected for the appraisal of immune response to rinderpest vaccination. The survey indicated that the virus is prevalent in cattle populations and approximately 12% of sera tested contained demonstrable neutralising antibodies against BVD-MD virus.     

Detection of foot-and-mouth disease virus infected cattle using infrared thermography

In this study, infrared thermography (IRT) was assessed as a means of detecting foot-and-mouth disease virus (FMDV)-infected cattle before and after the development of clinical signs. Preliminary IRT imaging demonstrated that foot temperatures increased in FMDV-infected animals. The maximum foot temperatures of healthy (n = 53), directly inoculated (DI) (n = 12), contact (CT) (n = 6), and vaccine trial (VT) (n = 21) cattle were measured over the course of FMD infection. A cut-off value was established at 34.4 °C (sensitivity = 61.1%, specificity = 87.7%) with the aim of detecting FMDV-infected animals both before and after clinical signs were observed. Seven of 12 (58%) DI and 3/6 (50%) CT animals showed maximum foot temperatures exceeding the 34.4 °C cut-off before the development of foot vesicles. In contrast, only 5/21 (24%) VT animals displayed pre-clinical foot temperatures above this cut-off possibly indicating partial vaccine protection of this group. These results show IRT as a promising screening technology to quickly identify potentially infected animals for confirmatory diagnostic testing during FMD outbreaks. Further evaluation of this technology is needed to determine the value of IRT in detecting animals with mild clinical signs or sub-clinical infections.     

Estimation of the transmission of foot-and-mouth disease virus from infected sheep to cattle

The quantitative role of sheep in the transmission of foot-and-mouth disease virus (FMDV) is not well known. To estimate the role of sheep in the transmission of FMDV, a direct contact transmission experiment with 10 groups of animals each consisting of 2 infected lambs and 1 contact calf was performed. Secretions and excretions (oral swabs, blood, urine, faeces and probang samples) from all animals were tested for the presence of FMDV by virus isolation (VI) and/or RT-PCR. Serum was tested for the presence of antibodies against FMDV. To estimate FMDV transmission, the VI, RT-PCR and serology results were used. The partial reproduction ratio R₀^p i.e. the average number of new infections caused by one infected sheep introduced into a population of susceptible cattle, was estimated using either data of the whole infection chain of the experimental epidemics (the transient state method) or the final sizes of the experimental epidemics (the final size method). Using the transient state method, R₀^p was estimated as 1.0 (95% CI 0.2 - 6.0) using virus isolation results and 1.4 (95% CI 0.3 - 8.0) using RT-PCR results. Using the final size method, R₀^p was estimated as 0.9 (95% CI 0.2 - 3.0). Finally, R₀^p was compared to the R₀’s obtained in previous transmission studies with sheep or cattle only. This comparison showed that the infectivity of sheep is lower than that of cattle and that sheep and cattle are similarly susceptible to FMD. These results indicate that in a mixed population of sheep and cattle, sheep play a more limited role in the transmission of FMDV than cattle.     

Single dilution ELISA for detection of serum antibody to foot-and-mouth disease virus in cattle

A single dilution blocking ELISA was developed and evaluated for measuring serum antibody to foot-and-mouth disease virus (FMDV). Basic parameters of the assay were established and a positive-negative threshold determined from testing 176 specific antibody negative sera from Australian cattle. Sera collected from immunised animals in Thailand were tested by ELISA and virus-neutralisation (VN) tests and the results compared. A positive correlation between ELISA and VN titres was recorded for each of the 3 FMDV serotypes endemic in Thailand, with the overall correlation coefficient being r = 0.8990. A positive correlation for each of the serotypes was also found between ELISA titre and the degree of blocking (percentage inhibition) of each test serum at a dilution of 1:16, with the overall correlation being r = 0.8704. This simplified ELISA was sensitive, specific and gave reproducible results, and had the potential to test quickly and efficiently a considerable number of sera.