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PubMed2001

Gene gun-mediate DNA vaccination against foot-and-mouth disease virus.

Foot-and-mouth disease (FMD) is one of the most dangerous diseases of cloven-hoofed animals and is a constant threat in the Middle-East and other regions throughout the world despite intensive vaccination programs. In this work, we describe the ability of FMDV expression constructs to protect pigs from FMDV challenge when used as a vaccine. The construct consists of encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), the entire P1 and 2A together with 3CD sequences, all in the same reading frame. Another plasmid that was tested, carries the serotype O1 (G) VP1, Asia1 VP1 and O1 (G) 3C. Between each of the genes the 3C cleavage sequences were inserted. All constructs carried the cytomegalo virus (CMV) promoter. Using immunofluorescent and immunoblot techniques, we could show the expression and processing of viral proteins. Following the application of FMDV expression constructs into pigs skin by 'Gene Gun', pigs were partially protected from FMDV challenge.

Animals↗

PubMed2005

Short hairpin RNA targeted to the highly conserved 2B nonstructural protein coding region inhibits replication of multiple serotypes of foot-and-mouth disease virus.

Foot-and-mouth disease virus (FMDV) is one of the most contagious agents of animals. Recent disease outbreaks in FMD-free countries have prompted the development of new control strategies that could improve the levels of protection against this virus. We have delivered a plasmid expressing a short hairpin RNA (shRNA) directed against a highly conserved sequence in the 2B nonstructural protein coding region of FMDV RNA to porcine cells. After virus infection, these cells showed a significant reduction in the synthesis of viral RNA and proteins, as well as a decrease in virus yield when compared to control cells. The antiviral effect was sequence specific and not attributable to induction of the interferon pathway. Since FMDV is an antigenically variable virus, we examined the effectiveness of this strategy against multiple serotypes and found that expressed 2B shRNA resulted in efficient silencing of at least 4 FMDV serotypes. Thus, RNA interference may be a potential alternative control strategy to limit the spread of this highly contagious virus in livestock.

Animals↗

PubMed2003

Effects of potassium and chloride on ribosome association with the RNA of foot-and-mouth disease virus.

Foot-and-mouth disease virus (FMDV) and other picornaviruses initiate translation of their polyprotein cap-independently at an internal site of the positive-strand viral RNA. This process is mediated by the internal ribosome entry site (IRES), a highly structured cis-acting RNA element that binds translation initiation factors and ribosomal subunits. During their life cycle, picornaviruses induce proliferation of membrane structures involved in viral replication and an increase in membrane permeability probably facilitating virus progeny release. Here, I analyze the efficiency of association of the ribosomal subunits with the FMDV IRES RNA at elevated salt concentrations. Potassium stimulates FMDV translation, whereas sodium chloride concentrations up to 150 mM neither stimulate nor interfere with FMDV translation. Even high potassium concentrations allow binding of the viral RNA to ribosomes. Chloride stimulates binding of ribosomes to the viral RNA at the stage of 48S initiation complex formation and FMDV translation at concentrations up to 150 mM. Only at elevated concentrations, binding of ribosomal subunits and translation are inhibited by chloride. However, FMDV start site selection is not influenced by potassium salts. These results indicate that the association of the viral RNA with ribosomal subunits is well adapted to high salt conditions that are induced during picornavirus infection.

Animals↗

PubMed1993

Avridine and LPS from Brucella ovis: effect on the memory induced by foot-and-mouth disease virus vaccination in mice.

Foot-and-mouth disease is one of the more economically important diseases among meat-producing biungulate species. In contrast to natural infection, current foot-and-mouth disease virus (FMDV) vaccines, prepared with inactivated virus and adjuvants, elicit short-lived protection. The immunomodulating effect on FMDV vaccines of avridine and lipopolysaccharide of Brucella ovis (LPS) was tested in a murine model. The duration of immunity, protection, stimulation of immunocompetent cells producing a long-lasting secondary response and immunoglobulin (Ig) isotypes were examined. The incorporation of either immunomodulator into aqueous and oil vaccines induced a long-lasting specific antibody response. The neutralizing titres and protection were significantly higher than those observed in animals immunized with control vaccines. Data collected from repopulation assays indicated that the immunomodulators used participate in the activation of immune cell populations involved in long-lasting memory. This resulted in an efficient B-cell secondary response even in the absence of T cells, which were necessary for the stimulatory effect of the immunomodulators in donor mice. Avridine and LPS stimulated IgG1, IgG2a and IgG2b production, which was correlated with the improvement of the protection induced by these vaccines.

Adjuvants, Immunologic↗

PubMed2004

Comparison of ELISA for the detection of porcine serum antibodies to non-structural proteins of foot-and-mouth disease virus.

Three foot-and-mouth disease virus non-structural protein antibody detection kits, CHEKIT FMD-3ABC, UBI FMD NS EIA and DVIVR NSP ELISA, were compared in the study. The results showed that the specificity of the kits ranged from 96.7 to 100% in nai;ve pigs and from 93.6 to 98.1% in vaccinated pigs, and that the DVIVR kit had the highest analytical sensitivity. The kappa statistics for the detection of 612 sera were 0.582, 0.447 and 0.658 for CHEKIT/UBI, CHEKIT/DVIVR and UBI/DVIVR, respectively. This study also revealed that measurable non-structural protein specific antibodies in some of infected pigs were sustained either for shorter periods or in intermittent patterns, thus aggravating the difficulties associated with the removal of pre-exposed pigs in the field.

Animals↗

PubMed2002

Dose-dependent responses of sheep inoculated intranasally with a type O foot-and-mouth disease virus.

Unlike foot-and-mouth disease (FMD) in cattle and pigs, which spreads rapidly, resulting in easily detectable foci of clinical infection, the disease in sheep is characterized by restricted transmission, low morbidity and sporadic clinical cases. The study described was designed to investigate whether the ability of sheep to transmit and maintain FMD virus was dose-related. The viral isolate used was known to be associated epidemiologically with rapid fade-out of transmission within sheep flocks. Five separate transmission experiments were performed, with different doses of FMD virus, each experiment containing five intranasally inoculated donor sheep and 10 in-contact recipient sheep. The lowest dose required to cause clinical infection by inoculation (10(4) 50% tissue culture infectious doses; 10(4) TCID50) was also the optimum dose for producing in-contact transmission. Inoculation of donor sheep with higher doses (10(5) and 10(6) TCID50) resulted in reduced transmission, characterized by reduced duration and degree of viraemia and an early humoral and cell-mediated immune response. Principal component analysis was used to interpret the complex interactions of the dose-related responses to infection.

Administration, Intranasal↗

PubMed2002

Further studies to quantify the dose of natural aerosols of foot-and-mouth disease virus for pigs.

Foot-and-mouth disease virus (FMDV) can be spread by a variety of mechanisms, including wind. Simulation models, developed to predict the risk of airborne spread, have played an important part in decision making in some outbreaks. The amount of airborne virus excreted as well as the minimal infectious dose (MID) of FMDV for different species are important determinants of airborne spread. The objective of this study was to obtain data for the O1 Lausanne, O SKR 2000 and O UKG 2001 strains of FMDV to enhance the capability of such models. Pigs were exposed to naturally generated aerosols of the three strains using an experimental design which delivered high doses of the two strains O1 Lausanne and O SKR 2000 over a short period, or of the O UKG 2001 strain over an extended period. The average excretion of the O1 Lausanne strain was 10(6.4) TCID50 per pig per hour. The excretion of the O SKR 2000 strain averaged 10(5.8) and the O UKG 2001 strain 10(6.1) TCID per pig per 24 h. The results show that the previous estimate of 'above' 800 TCID50 as the MID50 for the O1 Lausanne strain is a considerable under-estimate and that the real dose may be as high as 6000 TCID50. A dose of around 650 TCID50 of the O SKR 2000 strain failed to infect any pigs. Thus, the aerosol MID50 for pigs for this isolate is at least 1000 TCID50 and likely to be as high or higher than the O1 Lausanne strain. The exposure of pairs of recipient pigs kept physically separated from donor pigs in a series of rooms to aerosol exposure doses of the O UKG 2001 strain of around 50 TCID50 per min for 24-48 h failed to infect any of eight pigs. Thus, the present experiment confirms our previous findings that pigs, compared to cattle and sheep, are relatively resistant to infection with airborne FMDV.

Aerosols↗

PubMed2026

Evaluation of amplicon-based nanopore sequencing for foot-and-mouth disease viruses in clinical and environmental samples.

Foot-and-mouth disease (FMD) causes severe global economic loss, necessitating rapid viral characterization. Nanopore sequencing provides a simple, real-time workflow suitable for on-site outbreak response, addressing the limitations of conventional methods. In this study, we optimized a previously published amplicon-based protocol and used this method to characterize a diverse range of samples (vesicular fluid, epithelium, serum, nasal/oral swabs, and environmental samples) collected during FMD outbreaks in 2025 in the Republic of Korea. Of the 129 samples collected, we successfully recovered complete genomes from 37 samples and VP1 sequences from 85 samples. Amplifying the S-fragment in isolation and separately barcoding each pool of PCR amplicons markedly improved sequence recovery. Furthermore, sequencing success depended on viral load and sample type. Based on comparisons with real-time RT-PCR results, whole-genome sequence (WGS) recovery exceeded 77.3% at cycle threshold (Ct) values ≤25 across all clinical samples. In the Ct > 30 category, serum samples yielded the highest WGS recovery rates (44.4%). This rate was markedly higher than the success rates observed for epithelium (20.0%) and nasal swabs (9.1%), whereas oral swabs and environmental samples failed to yield any sequences (0%). However, VP1 recovery from environmental samples reached 80% at Ct ≤ 30 (8/10), providing an approach to enable non-invasive monitoring. These findings demonstrate that amplicon-based nanopore sequencing is a practical method for the rapid generation of genomic data during FMD outbreaks.IMPORTANCEAlthough rapid detection and genomic data analysis are crucial for effective foot-and-mouth disease (FMD) control, the collection of these data can be challenging for certain sample types and impacted by reduced viral loads that result from nationwide FMD vaccination. This study provides a practical solution through large-scale evaluation of an optimized amplicon-based nanopore sequencing protocol to enhance the sequencing success rates for both clinical and environmental samples. Using a modified protocol to enhance genome recovery, we demonstrated that sequence data could be retrieved from diverse sample types (even with high real-time RT-PCR cycle threshold values). We identified serum as the most suitable sample, with environmental sample sequencing allowing for non-invasive monitoring during outbreaks. These results support the use of nanopore sequencing for rapid genomic analysis, particularly in outbreak responses, such as rapid surveillance, emergency vaccine selection, and epidemiological monitoring.

Foot-and-Mouth Disease↗

PubMed2002

Global perspective for foot and mouth disease control.

The world distribution of foot and mouth disease (FMD) is almost a mirror image of the global economic structure. In general, industrialised countries are free while the disease is endemic in developing countries. In recent years, several incursions of FMD have been recorded in countries belonging to the Organization for Economic Co-operation and Development (OECD), all of which have been financially and socially costly to eliminate. At the same time, this single disease bars many developing countries from participation in formal trade, both regionally and internationally. However, recent studies have predicted an unprecedented high demand for animal protein, which can only be met through enhanced participation of developing countries in trade in livestock products. Accordingly, globalisation trends will exacerbate the exclusion of poor communities and countries from markets unless a long-term strategy is implemented to progressively build market opportunities for these countries, without placing the livestock of industrialised countries at undue risk from FMD and other major transboundary animal diseases. The authors submit that there is sufficient knowledge of FMD to make an international initiative for the progressive control of FMD a viable objective. Consequently, a four-stage pathway is proposed for developing a global FMD programme. The proposed strategy involves a build-up of the epidemiology and global status of FMD, including establishing an international early warning system, a risk-reduction phase to lower the incidence of FMD in the primary endemic areas and a control phase leading to the creation of zones of assured FMD-freedom. The authors also propose that an international FMD programme be co-ordinated, based on the experience of the Global Rinderpest Eradication Programme, the Hemispheric Plan for the eradication of FMD for the Americas, the South-East Asia Foot and Mouth Disease control and eradication campaign and the European Commission for the Control of FMD.

Animals↗

PubMed1990

The epidemic of foot-and-mouth disease in Saskatchewan, Canada, 1951-1952.

The epidemic of foot-and-mouth disease in Saskatchewan in 1951 and 1952 was studied in order to determine origins of outbreaks and methods of spread. The epidemic was initially considered to be vesicular stomatitis and foot-and-mouth disease was not recognized until February 1952, three months after the initial infection. The reports prepared at that time were reviewed in order to obtain details of the numbers of animals infected and the source and date of infection for the outbreaks. Methods of spread were rated according to their likelihood. The introduction of infection by an immigrant through his clothes as well as by sausage was possible. The sequence of events from the first outbreak to the spread from a feedlot/packing plant and from a dairy farm, which failed to report the disease, were clarified. Methods of spread included movement of animals, animal products and people and the airborne route. Milk delivery and artificial insemination did not result in spread of infection. The quarantine of affected farms reduced spread by animals and deterred visits by people. The original diagnosis of vesicular stomatitis was due to misinterpretation of a lesion in an inoculated horse. Laboratory tests established the presence of foot-and-mouth disease. The limited extent of the epidemic, despite the delay in diagnosis, is attributed to (i) the low density of cattle, (ii) few infected pigs and hence less airborne virus and (iii) absence of waste food feeding and milk collection in addition to the limited quarantine imposed.

Animals↗

PubMed2004

Inhibition of foot-and-mouth disease virus replication by small interfering RNA.

Foot-and-mouth disease, caused by foot-and-mouth disease virus (FMDV), is one of the most dangerous diseases of cloven-hoofed animals and is a constant threat to the dairy and beef industries in the Middle East and other regions of the world, despite intensive vaccination programmes. In this work, the ability of specific small interfering (si)RNAs to inhibit virus replication in BHK-21 cells was examined. By using bioinformatic computer programs, all FMDV sequences in public-domain databases were analysed. The analysis revealed three regions of at least 22 bp with 100 % identity in all FMDV entries. From these sequences, three specific siRNA molecules were prepared and used to test the ability of siRNAs to inhibit virus replication. By using real-time quantitative PCR to measure the amount of viral RNA in infected cells, it was shown that virus replication was inhibited in cells that were transfected with siRNAs. When viral titres were examined, 100 % inhibition of growth could be demonstrated in cells transfected with a mixture of all three anti-FMDV siRNAs, compared with control cells transfected with anti-LacZ siRNA.

Animals↗

PubMed1976

An assessment of the current status of foot-and-mouth disease in Nigeria.

Foot-and-mouth disease has been widespread throughout the country in recent years. The disease occurs mainly in cattle though spillover of infections has been observed in sheep and pigs. Virus types A, SAT 2 and SAT 1 are the usual cause of outbreaks. The present field control method is a 6-monthly vaccination schedulae for animals in government ranches, the few semi-commercial ranches and piggeries as well as for other valuable animals and exotic animals of high production quality. Arrangements are under way to institute a strict and more comprehensive control programme against the disease in view of the envisaged expansion of livestock development projects and improved zoo-sanitary status projected for our 3rd National Livestock Development Plan. A national policy of a barrier and frontier vaccination programme supplemented with strategic vaccinations in vulnerable foci is here recommended and discussed on the basis of a survey on the present status of the disease.

Animals↗

PubMed2005

Use of confocal immunofluorescence microscopy to localize viral nonstructural proteins and potential sites of replication in pigs experimentally infected with foot-and-mouth disease virus.

Replication of foot-and-mouth disease virus in infected pig epithelium has been studied by immunofluorescence labeling of the viral nonstructural protein 3ABC and confocal microscopy. The results were correlated with viral RNA copy numbers in tissue samples from adjacent sites determined by reverse transcription-PCR (RT-PCR). Lesion formation was seen in the tongues and coronary band epithelia of infected pigs 2 days after infection. Viral replication was observed in cells of the epithelium of the tongue and coronary band but not in the associated stromal cells. Infected epithelial cells were present in the stratum spinosum, away from the lesion, with small lesions formed above the basement membrane. Viral replication was markedly reduced in tongue epithelium by day 3 postinfection but remained apparent in the coronary band tissue up to 5 days postinfection. These results were confirmed by the RNA copy number determined by RT-PCR.

Animals↗

PubMed1990

Freeze-drying foot-and-mouth disease virus antigens. II. For use in the ELISA.

Live and inactivated preparations of foot-and-mouth disease virus strains 01 BFS 1860 and A22 IRQ 24/64 were freeze-dried in the presence or absence of additive solutions and assessed for their reactivity by ELISA at intervals over a six month storage period at various temperatures and also after reconstitution and subsequent storage with or without glycerination. The type specificity of all antigen preparations was maintained throughout the study period and the potency of antigens, judged by titration in ELISA, remained constant during the freeze-drying procedure and throughout subsequent storage at -20 degrees C and 4 degrees C with or without additives having been made to virus suspensions prior to freeze-drying. This was also the case with antigens reconstituted and stored at either -20 degrees C with glycerol or at 4 degrees C without glycerol. Certain additive solutions were necessary, however, to preserve the activity of antigens stored at the elevated temperature of 37 degrees C. The reactivity of all freeze-dried antigens was not unduly affected in the liquid-phase blocking ELISA using bovine convalescent antisera of each of the seven serotypes of foot-and-mouth disease virus and known negative, non-immune bovine sera. The results suggest that shipment and long-term storage of freeze-dried foot-and-mouth disease virus antigens is possible for use in the ELISA in the absence of refrigeration. This has attractive advantages for reducing both shipment and storage costs of antigens and for the development of ELISA kits for the diagnosis of foot-and-mouth disease virus.

Animals↗

PubMed2002

Anti-3AB antibodies in the Chinese yellow cattle infected by the O/Taiwan/99 foot-and-mouth disease virus.

The O/Taiwan/99 foot-and-mouth disease virus (FMDV), a South Asian topotype of serotype O, was introduced into Taiwan in 1999. The Chinese yellow cattle infected by the virus did not develop clinical lesions under experimental and field conditions. A blocking enzyme-linked immunosorbent assay (ELISA) kit with the 3AB antigen, a polypeptide of FMDV non-structural (NS) proteins, was used to evaluate the development and duration of anti-3AB antibodies, proving active viral replication, in the Chinese yellow cattle. The specificity of the assay was 99%, as was established with negative sera from regularly vaccinated and from naïve cattle. The sensitivity tested with sera from naturally infected animals was approximately 64% and it was lower than that obtained by serum neutralization (SN) test. Under experimental infection, the Chinese yellow cattle developed lower anti-3AB antibodies than that developed in other species. Duration of anti-3AB antibodies was traced in two herds of naturally infected animals, indicating that anti-3AB antibodies persisted for approximately 6 months after outbreaks. On the basis of this study, we propose that the Chinese yellow cattle may have natural resistance, which limits viral replication and reduces the development of anti-3AB antibodies.

Animals↗

PubMed2002

Use of molecular assay in diagnosis of hand, foot and mouth disease caused by enterovirus 71 or coxsackievirus A 16.

Hand, foot and mouth disease is a common illness in children and is usually caused by coxsackievirus A 16 and enterovirus 71. It has been noted that enterovirus 71 infection is more severe with significantly greater frequency of serious complications and fatality than coxsackievirus A 16. Therefore, it is important to develop a rapid and specific assay for discriminating coxsackievirus A 16 and enterovirus 71 in hand, foot and mouth disease outbreaks. In this study we designed two sets of RT-PCR primers specific for coxsackievirus A 16 and enterovirus 71. One hundred and eighty-nine viruses were evaluated for this molecular diagnosis assay. Among 110 enterovirus 71 strains, the enterovirus 71 specific primers gave clear signal for 107 clinical enterovirus 71 isolates and three reference enterovirus 71 strains. None of coxsackievirus A 16, other enteroviruses or non-enteroviruses show signal for enterovirus 71-specific primers. On the other hand, among 28 coxsackievirus A 16 strains, the coxsackievirus A 16-specific primers detect 27 clinical isolates and one reference strain but show no cross-reaction with other viruses. The molecular assay developed in this study provides a sensitive and specific way to distinguish coxsackievirus A 16 and enterovirus 71 induced hand, foot and mouth disease, which will be a useful rapid diagnostic method in future outbreaks.

Enterovirus↗

PubMed1984

Serological response of cattle to simultaneous vaccinations against foot-and-mouth disease and haemorrhagic septicaemia.

In Malaysia, where vaccination campaigns against foot-and-mouth disease and haemorrhagic septicaemia are routinely carried out, it was desirable to determine whether it was safe and efficacious to administer both vaccines simultaneously. A trial group of 104 cattle was divided into three groups; group 1 animals received both vaccines simultaneously, group 2 animals received only foot-and-mouth disease vaccine and group 3 animals received only haemorrhagic septicaemia vaccine. The serological response to vaccinations was monitored at 0, 21 and 35 days by the virus neutralisation test for foot-and-mouth disease and the mouse-protection and indirect haemagglutination tests for haemorrhagic septicaemia. The simultaneous administration of the two inactivated vaccines produced no adverse effects and the serological response did not differ from the response to either vaccine given separately, thus indicating that cattle may be safely and effectively vaccinated simultaneously in this way.

Animals↗

PubMed2004

Validation of binary ethyleneimine (BEI) used as an inactivant for foot and mouth disease tissue culture vaccine.

The complete inactivation of Foot and Mouth Disease (FMD) virus is a critical requirement in the production of FMD vaccine to ensure the safety of the product. Binary ethyleneimine (BEI) is an aziridine compound, produced from bromoethylamine hydrobromide (BEA) commonly used for the inactivation of FMD virus during vaccine manufacturing. The validation of BEI, when used as an inactivant, is essential to ensure the quality of the inactivating agent and the validity of the process. In the present study, the inactivation kinetics of Foot and Mouth Disease virus (O, A and Asia-1 serotypes) were determined for different concentrations of BEI (0.4 mM, 0.8 mM, 1.2 mM, 1.6 mM and 2.0 mM). Statistically significant differences in the inactivation kinetics were observed between 0.4 mM and 1.6 mM of BEI. The results indicated that BEI at 1.6 mM was able to inactivate the FMD virus within 8-10 h. Increasing the concentration of BEI beyond 1.6 mM did not appreciably improve the inactivation process. No differences in the inactivation kinetics were found between the various serotypes studied. This study can be used as a guideline for routine procedures for validating the quality of BEA and the inactivation process.

Animals↗