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PubMed1988

Serological and biochemical analysis of some recent type A foot-and-mouth disease virus isolates from the Middle East.

In 1986 and 1987 foot-and-mouth disease virus (FMDV) serotype A was isolated from outbreaks of disease in Saudi Arabia and Iran. Selected virus isolates were antigenically distinct from the prototype A22 virus strain (A22/Iraq/64), but were serologically related to each other. However, polyacrylamide gel electrophoresis showed that whilst the respective Saudi Arabian structural polypeptides were homogeneous, those from an Iran isolate were distinct. Direct sequencing of part of the P-1D (VP1) gene demonstrated considerable difference in nucleotide homology between the two groups of viruses; the Saudi Arabian viruses were closely related to each other but only distantly related to both the A22 prototype virus strain and the Iranian virus isolate. The latter viruses were only slightly more closely related to each other. Thus there appeared to be at least two distinct FMDV type A variants co-circulating in the Middle East, both of which differed considerably from the classical A22 subtype.

Animals↗
PubMed1994

A modified liquid phase (LP) blocking ELISA used to assess type O foot-and-mouth disease virus antigenic variation in Thailand.

A selection of type O foot-and-mouth disease (FMD) viruses isolated in Thailand between 1986 and 1989 were compared to the reference viruses O1 Thailand 1960 (O BKK/60) and O Nakorn Pathom 1965 (O NPT/65) using a liquid-phase blocking ELISA (LP ELISA) to derive serum titres and associated r values. Interpolation techniques were used to increase the precision for estimation of r values through a more accurate estimation of serum titres at predicted equivalent levels of antigen input. Mean r values were 0.45 (for 56 field viruses) relative to O BKK/60 reference virus and 0.56 (for 51 field viruses) relative to O NPT/65. While only two viruses showed considerable difference (r < 0.20) to a reference virus (O BKK/60), 41% and 31% gave r values less than 0.4 for O BKK/60 and O NPT/65 respectively. This indicated antigenic differences between reference and field viruses which may result in a reduction in vaccine efficacy.

Animals↗
PubMed2005

Immunogenicity of the epitope of the foot-and-mouth disease virus fused with a hepatitis B core protein as expressed in transgenic tobacco.

A novel plant-based vaccine protecting against foot-and-mouth disease (FMD) was developed by inserting the VP21 epitope into the internal region of the hepatitis B virus core antigen gene (HBcAg). The specific sequence of the VP21 epitope is located within the VP1 capsid protein of the FMD virus (FMDV). It spans 21 amino acids located between positions 140 and 160 of the G-H loop. The fusion gene, HBCVP, was inserted into the plant binary vector pBI121 and then transformed into tobacco (Nicotiana tabacum) plants via Agrobacterium tumefaciens strain LBA 4404. The presence of HBCVP in the tobacco genome was confirmed by polymerase chain reaction (PCR); its transcription was verified by reverse transcription-PCR; and the recombinant protein expression was confirmed by Western blot analysis. The results of immunologic microscopic observation demonstrated that recombinant fusion protein HBCVP can form a virus-like particle (VLP) structure in transgenic tobacco leaves. Mice, immunized intraperitoneally with a soluble crude extract of transgenic tobacco leaves, were found to produce specific antibody responses to both HBcAg and FMDV VP1. A virus challenge demonstrated that the immunized mice were highly protected against virulent FMD. This work describes a new way to develop an FMD vaccine from plants that will aid the development of new vaccines using HBcAg fused to the conserved epitopes of other pathogenic antigens.

Agrobacterium tumefaciens↗
PubMed1998

Protection of swine by live and inactivated vaccines prepared from a leader proteinase-deficient serotype A12 foot-and-mouth disease virus.

Previously, we demonstrated that a genetically engineered variant of foot-and-mouth disease virus (FMDV) serotype A12 lacking the leader proteinase-coding region (A12-LLV2) was attenuated and induced an immune response that partially protected cattle from FMD. In this study, A12-LLV2 was tested in swine as a live or chemically inactivated vaccine. Animals vaccinated with chemically inactivated A12-LLV2 or wild-type (WT) virus in oil adjuvant developed high levels of neutralizing antibodies and were protected from FMD upon challenge. Animals vaccinated with live A12-LLV2 did not exhibit signs of FMD, did not spread virus to other animals, developed a neutralizing antibody response and antibodies to nonstructural protein 3D, and were partially protected from FMD. Animals given a similar dose of chemically inactivated A12-LLV2 in the absence of adjuvant developed a poor immune response and were not protected from FMD, indicating that limited replication was responsible for the improved immune response found in animals vaccinated with live A12-LLV2. The results demonstrate the potential of A12-LLV2 as a live-attenuated vaccine as well as a safe source of antigen for chemically inactivated vaccines.

Animals↗
PubMed2004

Genome comparison of a novel foot-and-mouth disease virus with other FMDV strains.

The genome of a novel foot-and-mouth disease virus, HKN/2002, was 8104 nucleotides (nt) in length (excluding the poly(C) tract and poly(A) tail) and was composed of a 1042-nt 5'-untranslated region (UTR), a 6966-nt open reading frame, and a 93-nt 3'-UTR. Genome sequences of HKN/2002 and other known FMDV strains were compared. The VP1, VP2, and VP3-based neighbor-joining (NJ) trees were divided into distinct clusters according to different serotypes, while other region-based NJ trees exhibited some degree of intercross among serotypes. Mutations in HKN/2002 were revealed, including frequent deletions and insertions in the G-H loop of VP1, and deletion involving 10 amino acid residues in the 3A protein. An evolutionary relationship of HKN/2002 with an Asian FMDV lineage isolated from a Hong Kong swine host in 1970 was postulated. A 43-nt deletion identified in the 5'-UTR of HKN/2002 possibly contributed to the loss of one pseudo-knot domain.

3' Untranslated Regions↗
PubMed1993

Characterization of foot-and-mouth disease virus by monoclonal antibodies.

Monoclonal antibodies (MAbs) were produced against foot-and-mouth disease (FMD) virus types O1 Campos Br1/58, A24 Cruzeiro Br1/55, and C3 Indaial Br1/71, which are the strains used for production of FMD vaccines in the majority of South American countries. Within the library of MAbs produced, a group was selected on the basis of their neutralizing titer in cell culture, protective titer in suckling mice, sensitivity to trypsin, and specificity for virus structural proteins. The MAbs were utilized in an ELISA test format to compare European and South American representative field isolates with vaccine production strains in their r1 relationship as obtained by 50% complement fixation (CF50) with polyclonal antibodies (PAb) and their virus neutralization (VN) relationship obtained with sera from one-time-vaccinated and from revaccinated cattle, respectively. The MAbs selected varied in their reactivity against the different strains and, therefore, enabled us to compare field FMDV strains to those against which the MAbs were produced, with definite advantages over the r1 and VN ratios. Thus, panels of MAb produced with the vaccine strains and appropriately selected are significantly useful for the FMD-control programs because they serve to provide guidance on the immunological coverage provided by the vaccines against FMDV strains circulating in the field. The MAbs are also useful for the differentiation of FMD virus strains.

Animals↗
PubMed2005

Immune response in guinea pigs vaccinated with DNA vaccine of foot-and-mouth disease virus O/China99.

In order to obtain the gene P12X3C of foot-and-mouth disease virus (FMDV O/China99) that includes full length P1, 2A, 3C and part of 2B and 3B, the site mutation strategy was used. The recombinant plasmid pcDNA3.1/P12X3C was transfected into BHK-21 cells. The capsid proteins of FMDV expressed in BHK-21 cells were confirmed by sandwich-ELISA and indirect immunofluorescence test. Then the plasmid pcDNA3.1/P12X3C was administered to guinea pigs intramuscularly, and purified FMDV O/China993D protein expressed in yeast cells was injected together with pcDNA3.1/P12X3C. Anti-FMDV antibodies were detected by indirect ELISA, the T-lymphocyte proliferation response was tested by MTT assay, and neutralizating antibodies titers were analyzed by micro-neutralization assay. The result showed that the plasmid pcDNA3.1/P12X3C was able to express immunocompetent proteins of FMDV in BHK-21 cells. Furthermore, anti-FMDV antibodies were elicited and increased by plasmid pcDNA3.1/P12X3C in the second week after vaccination. Neutralizating antibodies were induced and the T-lymphocyte proliferation response was enhanced after vaccination. In the challenge test, all of guinea pigs vaccinated with pcDNA3.1/P12X3C were fully protected from FMDV challenge. However, the result obtained from animals that were injected with protein 3D together with plasmid pcDNA3.1/P12X3C was not satisfied. In conclusion, the results encouraged further work towards the development of a DNA vaccine against FMDV and provided the basis of research for DNA vaccine.

Animals↗
PubMed1990

Isotype responses of infected, virus-vaccinated and peptide-vaccinated cattle to foot-and-mouth disease virus.

An ELISA to measure bovine serum immunoglobulin isotypes (IgG1, IgG2, IgM and IgA) specific for foot-and-mouth disease virus (FMDV) or for synthetic FMDV peptides is described. Sera from cattle infected by FMDV, vaccinated with conventional inactivated virus vaccines or vaccinated with synthetic peptides were examined using this assay. Generally IgG subclasses dominated the antibody responses of all groups after an early IgM response had waned. An exception to this pattern was seen in the case of a group of immature calves given multiple or high doses of synthetic peptide and in which levels of IgM continued to rise until the end of the experimental period. Both infected animals and those vaccinated with inactivated virus mounted antibody responses in which IgG1 titres tended to predominate over those of IgG2. In some infected animals, an early IgG2 response was evident but resolution of lesions and clinical recovery did not occur until IgG1 antibody appeared in the serum some days later. In synthetic-peptide immunized animals the response was more variable but IgG1:IgG2 ratios at 21 days postvaccination were significantly lower than those of virus-vaccinated animals. It is proposed that differences in the isotype profiles induced by conventional FMD vaccines and those resulting from vaccination of cattle with synthetic FMDV peptides may in part account for the lower protective index of peptide-induced antibodies.

Animals↗
PubMed1976

[Testing the activity of anti-foot-and-mouth disease vaccine in cattle: quantitative methods of antigenic extinction].

Two methods of evaluating the activity of the foot-and-mouth disease vaccine are compared, leading to the evaluation of the number of protective doses 50% or potency. These methods differ in sensitivity, precision and equivalence potency-percentage of protection. For the same vaccine the value of the PD50 is always larger than that of Pb. We chose the Pb method using a diluent containing no adjuvant. Standards based on the evaluation of average potency have not been taken into consideration; only those standards based on the evaluation of minimum potency ensure respect of OIE's recommendation. This recommendation: 70% minimum potency with 0.95 probability is not an easier alternative, since the average protection observed corresponds in fact to at least 90% taking into account the biological variability of bovines.

Animals↗
PubMed2005

Antigenic and genetic analyses of foot-and-mouth disease virus type A isolates for selection of candidate vaccine strain reveals emergence of a variant virus that is responsible for most recent outbreaks in India.

Recent reports indicated presence of two antigenic and genetic groups (genotypes VI and VII) of foot-and-mouth disease virus (FMDV) type A in India and are divergent from the vaccine strains. In order to choose suitable field isolate as candidate vaccine strain, anti-sera against representative isolates from both the genotypes and two in-use vaccine strains are tested in neutralization assay. Two candidate vaccine strains from both the genotypes are selected with close antigenic match to the field isolates. From the result it is evident that IND 81/00 (genotypes VII), gave a better antigenic coverage (antigenic relationship (r)-value>0.40 with 79% of isolates of 2002--2003) than IND 258/99 (genotype VI; r-value>0.40 with 42% of 2002--2003 isolates). Phylogenetic analysis based on P1 genomic region placed all the recent isolates (2001--2003) into genotype VII, with emergence of a new variant virus (VIIb--VP3(59)del) having amino acid deletion at an antigenically critical residue (VP3(59)), indicating a major evolutionary jump probably due to immune selection. Though very limited in its extent, this data indicates an apparent dominance of genotype VII over genotype VI and underscores the need to continue further molecular epidemiological investigations to substantiate this finding.

Animals↗
PubMed1974

Fluorescent antibody responses of cases and contacts of hand, foot, and mouth disease.

Paired sera from clinical cases, familial contacts, and school contacts of hand, foot, and mouth disease were tested by indirect immunofluorescence against a strain of coxsackie A16 virus which had been adapted to human fibroblast tissue-cultured cells. All of the 22 clinical cases developed immunoglobulin (Ig) G antibodies, and 11 responded with IgM and 10 with IgA antibodies. Seventeen of 21 familial contacts of cases showed subclinical infection as determined by development of IgG antibodies. Only seven and nine, respectively, of these had demonstrable IgM and IgA antibodies. Thirteen of 16 school classmates of three clinical cases were shown to have experienced subclinical infection by the development of IgG antibodies. Only five had antibodies in the IgM fraction, and three had antibodies in the IgA fraction. A comparison of IgG titers with those obtained by neutralization tests provides further evidence that the indirect fluorescent antibody technique represents a rapid diagnostic procedure for this disease.

Adolescent↗
PubMed2003

Early dissemination of foot-and-mouth disease virus through sheep marketing in February 2001.

The results of epidemiological investigations suggest that livestock on up to 79 premises, spread widely throughout the British Isles, may have been exposed to infection by foot-and-mouth disease (FMD) virus by the movement of infected sheep before the first case of the disease was confirmed at an abattoir in Essex on February 20, 2001. A further 36 premises may have been infected by this route before the national livestock movement ban was imposed on February 23.

Abattoirs↗
PubMed1975

[Determination of the subtype membership of the epizootic foot-and-mouth disease virus by means of the agar gel diffusion test].

Comparative studies were carried out by means of the agar gel-diffusion technique (AGDT) to establish the antigenic identity between the epizootic foot-and-mouth disease strain (the Nch/73 representative strain) and the standard viruses of type A. The epizootic strain was found to belong to the subtype group A5. Discussed is the usefulness of AGDT in determining the antigenic relations between the foot-and-mouth disease viruses.

Aphthovirus↗
PubMed2005

Comparison of immune responses after intra-typic heterologous and homologous vaccination against foot-and-mouth disease virus infection in pigs.

This study compares the immune responses and protection induced by intra-typic heterologous vaccination with that induced by homologous vaccination against challenge with foot-and-mouth disease virus (FMDV). Humoral and cell-mediated immune responses and protection against challenge with FMDV O Taiwan were examined in a non-vaccinated group, a group vaccinated with O Taiwan FMD vaccine and a group vaccinated with O Manisa FMD vaccine. Five pigs from each group were challenged with FMDV type O Taiwan 14 days after vaccination and five other pigs were contact-exposed to the inoculated pigs. Both homologous and heterologous vaccination protected against challenge with FMDV O Taiwan at 2 weeks after vaccination. In the heterologous vaccinated group, cross-neutralizing antibody titres against O Taiwan could be detected although the ratio 'r(1)' was 0.4, which was significantly smaller than the critical r-value. Cell-mediated immune responses were detected after both homologous and heterologous vaccination. Virus-induced in vitro lymphocyte (cross-) proliferation and production of both a Th1-type (IFN-gamma) and a Th2-type (IL-10) cytokine response were demonstrated in cultures of peripheral blood mononuclear cells (PBMC). The findings show that heterologous (emergency) vaccination can prevent clinical disease and shedding of virus. The induction of the cell-mediated immune responses after (heterologous) vaccination needs more research but data on these responses might provide additional tools for both vaccine choice and vaccine development.

Animals↗
PubMed2006

2'-C-methylcytidine as a potent and selective inhibitor of the replication of foot-and-mouth disease virus.

We report on the potent and selective in vitro antiviral activity of 2'-C-methylcytidine (2'-C-MetCyt) against foot-and-mouth disease virus (FMDV). FMDV belongs to the Picornaviridae and has the potential to cause devastating epidemics in livestock. The 50% and 90% effective concentrations (EC50 and EC90) for inhibition of the FMDV-induced cytopathic effect (CPE) formation were 6.4+/-3.8 and 10.8+/-5.4 microM. Comparable EC50 values for inhibition of viral RNA synthesis were observed. Treatment of FMDV-infected BHK-21 cells with 77 microM 2'-C-MetCyt resulted in a (1.6-3.2)x10(3)-fold reduction of infectious virus yield. Time-of-drug addition experiments suggest that 2'-C-MetCyt interacts with viral replication at a time point that coincides with the onset of intracellular viral RNA synthesis. In contrast to emergency vaccination, a potent and selective antiviral agent may provide almost immediate (prophylactic/therapeutic) protection against infection and thus constitute an important alternative/supplementary option to contain outbreaks such as those caused by FMDV.

Animals↗
PubMed2006

Reporting of suspect cases of foot-and-mouth-disease during the 2001 epidemic in the UK, and the herd sensitivity and herd specificity of clinical diagnosis.

We described the clinical diagnostic process utilized during the 2001 epidemic of foot-and-mouth-disease in the United Kingdom (UK), and considered it as a series of diagnostic tests. Premises were classified according to these diagnostic-test results and actual disease status, determined by the reference test, which in this case was one or more internationally accepted laboratory tests. The herd-level sensitivity (HSe) and herd-level specificity (HSp) of the clinical diagnostic process were calculated directly, relative to these internationally accepted reference tests. In this process, the first diagnostic test was 'routine monitoring', which resulted in the identification of suspect cases based solely on the clinical observations of farmers or veterinarians. 6762 suspect cases were identified, and the test had a HSe of 97.6% (95% C.I.: 96.7, 98.3) and a HSp of 95.2% (95% C.I.: 95.0, 95.3). Suspect cases were then subject to the second diagnostic test, termed 'declaration', which consisted of a review of a description of the clinical signs by government veterinarians. Premises that tested positive became 'clinical cases'. The HSe of this test was 97.1% (95% C.I.: 96.2, 97.9), and the HSp was 90.9% (95% C.I.: 90.1, 91.6). During the epidemic, these tests were combined and applied in series, with an overall HSe of 94.7% (95% C.I.: 93.5, 95.7) and an overall HSp of 99.6% (95% C.I.: 99.5, 99.6). We also examined the effect of a policy shift that prohibited delaying the diagnosis pending laboratory testing where the animals exhibited equivocal clinical signs.

Animals↗
PubMed1995

Antigenic analysis of SAT 2 serotype foot-and-mouth disease virus isolates from Zimbabwe using monoclonal antibodies.

This paper compares strains of foot-and-mouth disease (FMD) serotype SAT (South African Territories) 2 viruses isolated from Zimbabwe and other African countries using monoclonal antibodies (MAb). A sandwich-ELISA was used to examine the relative binding of anti-SAT 2 MAb to the various viruses. The MAb-binding profiles of viruses isolated from field samples were compared using hierarchical cluster analysis. Viruses were obtained from game animals, mainly African buffalo (Syncerus caffer) which is the natural host and reservoir for SAT serotypes in Africa, and from cattle showing clinical signs of FMD, as well as from animals suspected of carrying the virus subclinically. Some isolates have been adapted for use as vaccine strains. The results showed that most of the Zimbabwe isolates collected between 1989 and 1992 were an antigenically closely-related group. Although differences were observed between Zimbabwe isolates collected between 1989 and 1992 and those collected in 1987, there was no correlation with the different MAb binding patterns within the 1987 group and the epidemiological information received from the field. Similar profiles were observed for many SAT 2 viruses, including viruses isolated over a 50-year period and from geographically distant areas. This indicates an inherent stability in antigenic profiles of SAT 2 viruses. The MAb panel was capable of assessing antigenic variation, since very different profiles were obtained for some isolates. The work also allowed comparison and characterization of anti-type SAT 2 MAb from different laboratories. The findings are discussed with reference to selection of vaccine strains.

Animals↗
PubMed2002

Review of the status of foot and mouth disease and approach to control/eradication in Europe and Central Asia.

The authors describe the situation of foot and mouth disease (FMD) in Europe over the past 70 years and analyse the origin of the disease and methods of control, particularly since preventive vaccination was banned in Europe in the early 1990s. Since then, and until 2001, despite several incursions of the virus, the disease has always been contained and eradicated rapidly. Therefore, the ban on vaccination did not result in an increase of FMD outbreaks. However, the massive outbreak which took place in 2001 in the United Kingdom (UK) with 2,030 outbreaks, raised questions on the policy utilised to date to control the disease in Europe. In future, the utilisation of ring vaccination should be considered as an alternative to mass culling of large numbers of animals. Based on the recent source of introduction of the virus, the authors review the lines of defence which should be reinforced to reduce the risk of further introduction of the disease. The FMD situation in the Commonwealth of Independent States (CIS) is also examined. The situation in the Central Asian Republics and the Caucasian region deteriorated after the collapse of the Soviet Union, despite the continuous effort of Russia to support these countries. International support is needed to prevent FMD from becoming endemic in the region.

Animals↗