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PubMed1992

Characterization and immune response of a protein produced by a cDNA clone of foot and mouth disease virus, type Asia 1 63/72.

A 0.9 kb double stranded cDNA of foot and mouth disease virus (FMDV) Type Asia 1, 63/72 was cloned in an expression vector, pUR222. A protein of 38 kd was produced by the clone which reacted with the antibodies raised against the virus. A 20 kd protein which may be derived from the 38 kd protein contained the antigenic epitopes of the protein VP1 of the virus. Injection of 10-20 micrograms of the partially purified 38 and 20 kd proteins or a lysate of cells containing 240 micrograms of the proteins elicited high titers of FMDV specific antibodies in guinea pigs and cattle respectively. Also, at these concentrations, the proteins protected 5 of 8 guinea pigs and 3 of 8 cattle when challenged with a virulent virus.

Animals↗
PubMed1994

Validation of an inhibition ELISA using a monoclonal antibody for foot-and-mouth disease (FMD) primary diagnosis.

An inhibition ELISA (IH-ELISA) test for foot-and-mouth disease virus (FMDV) was validated using 106 epithelial samples from suspected cases of FMD in Argentina submitted to the Argentine National Diagnostics Laboratory (GELAB) over a period of 12 months and examined in parallel with the complement fixation test (CFT). IH-ELISA was found to be more sensitive, detecting 25% (26 samples) more FMDV positives than the CFT in original suspensions of field samples. The effect of storage conditions on 12S stability was examined. Plates stored at 4 degrees C blocked with 1% ovalbumin and plates stored at -20 degrees C with or without blocking buffer could be used for at least 90 days. When various brands of polystyrene plates were compared for 12ps adsorption it was found that those microplates of higher binding capacity were more efficient.

Animals↗
PubMed1988

Immunization against foot-and-mouth disease with synthetic peptides representing the C-terminal region of VP1.

Foot-and-mouth disease virus challenge experiments in guinea-pigs and immunoassays with a range of peptides equivalent to either or both of the sequences 141 to 158 and 200 to 213 of VP1 showed the most effective structure, in terms of protection, to be one in which both 'sites' were present with a minimum of additional amino acids. An 80 residue peptide comprising amino acids 134 to 213 was considerably less effective than 40 or 45 residue peptides. The major site for the induction of protection was deduced to be in the region 141 to 158. Thus, protection with the 40 or 45 residue peptide did not appear to be due to the presence of antibody directed solely to the 200 to 213 sequence. Finally, induction of antibody to the latter site appeared to be dependent on both the size of the peptide and the disposition of 'sites' within it.

Animals↗
PubMed1994

Haptoglobin response of cattle infected with foot-and-mouth disease virus.

Haptoglobin, a major bovine acute phase protein, was evaluated as a marker of the primary replication of foot-and-mouth disease virus in 12 naturally infected cattle from which blood was collected daily. An acute phase response, as measured by an increase in serum haptoglobin concentration and the presence of fever, was not detected during the previraemic stage of disease, but there was a significant increase in serum haptoglobin after the onset of viraemia. It occurred on the same day as the first appearance of clinical signs in three of the cattle, one day later in four and two days later in the other five cattle.

Animals↗
PubMed1996

Isotype-specific antibody responses to foot-and-mouth disease virus in sera and secretions of "carrier' and "non-carrier' cattle.

Isotype-specific antibody responses to foot-and-mouth disease virus (FMDV) were measured in the sera and upper respiratory tract secretions of vaccinated and susceptible cattle challenged with FMDV by direct contact or by intranasal inoculation. A comparison was made between cattle that eliminated FMDV and those that developed and maintained a persistent infection. Serological and mucosal antibody responses were detected in all animals after challenge. IgA and IgM were detected before the development of IgG1 and IgG2 responses. IgM was not detected in vaccinated cattle. Challenge with FMDV elicited a prolonged biphasic secretory antibody response in FMDV "carrier' animals only. The response was detected as FMDV-specific IgA in both mucosal secretions and serum samples, which gained statistical significance (P < 0.05) by 5 weeks after challenge. This observation could represent the basis of a test to differentiate vaccinated and/or recovered convalescent cattle from FMDV "carriers'.

Animals↗
PubMed2004

Making a vaccinate-to-live policy a reality in foot-and-mouth disease.

Public opinion and the availability of new technologies are making the use of 'stamping- out' an increasingly unattractive option as the method of first choice for foot-and-mouth disease (FMD) control in FMD-free countries or zones seeking to control incursion of disease. There is therefore increasing pressure to adopt a 'vaccinate-to-live' policy in these circumstances. For a successful vaccinate-to-live policy, veterinary services need access to appropriate, licensed vaccines; to have adequate contingency plans to ensure that they can deliver the required vaccine, where and when it is needed; and to have developed an 'exit strategy' that enables recognition of freedom from disease as quickly as possible. This paper discusses progress towards these requirements and the problems that still need to be addressed before a vaccinate-to-live policy can become the option of first resort.

Animals↗
PubMed2004

An investigation into the source and spread of foot and mouth disease virus from a wildlife conservancy in Zimbabwe.

African buffalo were introduced into a wildlife conservancy in the southeast of Zimbabwe in an effortto increase the conservancy's economic viability, which is primarily based on eco-tourism. The buffalo were infected with SAT serotypes (SAT-1, SAT-2 and SAT-3) of foot and mouth disease (FMD) virus, and in order to isolate the conservancy and prevent the transmission of FMD to adjacent populations of domestic livestock, the conservancy was surrounded by a double-fence system, 1.8 m in height. The intention was to prevent the movement of both wildlife and domestic animals across the perimeter. However, two years after the buffalo were introduced, FMD occurred in cattle farmed just outside of the conservancy. Using serological and molecular diagnostic tests, epidemiological investigations showed that it was most likely that antelope (impala or kudu), infected through contact with the buffalo herd within the conservancy, had jumped over the fence and transmitted the virus to the cattle.

Animals↗
PubMed2000

An integrated model to predict the atmospheric spread of foot-and-mouth disease virus.

The application of a computer model called Rimpuff for simulating the airborne spread of foot-and-mouth disease (FMD) is described. Rimpuff is more sophisticated and accurate than other FMD simulation models previously described. It can be run on a desktop computer and performs analyses very quickly. It can be linked to a geographical information system and so the information generated can be integrated with geographical and demographical data for display in a format that can be easily assimilated and transmitted electronically. The system was validated using historical data from outbreaks of FMD in France and the UK in 1981, and from Denmark and the former German Democratic Republic (GDR) in 1982. A very good fit was obtained between the direction of the plumes of virus simulated by the model and the spread of disease from France to the UK in 1981. Although cattle in the UK were infected during the episode, the concentrations of airborne virus in the plumes simulated by the model were beneath the infectivity threshold for cattle. It was concluded from the analysis that the number of pigs infected in France, and therefore the source concentration of airborne virus, was probably much higher than was recorded at the time of the outbreaks. Analysis of the Denmark/GDR episode pointed to the possibility that the source of virus for the 1982 epidemic in Denmark could have been one or more unreported outbreaks involving pigs in the former GDR.

Animal Husbandry↗
PubMed1996

Pathogenesis of wild-type and leaderless foot-and-mouth disease virus in cattle.

Four calves were experimentally infected via aerosol with foot-and-mouth disease virus. Two were infected with a wild-type virus derived from a full-length infectious clone (A12-IC), and two were infected with a clone-derived virus lacking the leader gene (A12-LLV2), with euthanasia and tissue collection at 24 and 72 h postexposure (hpe). Clinical disease was apparent only in the animal given A12-IC and euthanized at 72 hpe. In situ hybridization revealed that the animal infected with A12-IC and euthanized at 24 hpe had abundant viral nucleic acid in the lung, present in clusters of positive cells in the respiratory bronchiolar epithelium and associated subepithelial regions. At 72 hpe in the A12-IC-infected calf, viral nucleic acid in the lung was present in interstitial areas, and in addition, viral nucleic acid was detectable in epithelial tissues around histologically apparent vesicles. In animals infected with A12-LLV2, viral nucleic acid was detectable in the lung at both 24 and 72 hpe, but staining revealed a more localized distribution with less nucleic acid than was found in animals given A12-IC. Therefore, it appears that after aerosol exposure to A12-IC, early replication is in the region of the lung, with subsequent dissemination to distal sites. In comparison, the A12-LLV2 virus is much less widely disseminated in the lung at 24 hpe, with no lesions or virus detectable in secondary sites at 72 hpe. The greatly reduced pathogenicity of A12-LLV2 may make it an excellent candidate for a modified live viral vaccine.

Animals↗
PubMed2002

Integrity of GH-loop of foot-and-mouth disease virus during virus inactivation: detection by epitope specific antibodies.

Vaccine against foot-and-mouth disease (FMD) is prepared after inactivating the virus produced in cell culture. Inactivation of the FMD virus (FMDV) was earlier done by formaline. However, several vaccine outbreaks, which occurred in Europe revealed that the formaline treatment is not highly effective for virus inactivation. Subsequently, binary ethyleneimine (BEI) was identified as an effective inactivation reagent for FMDV. However, these chemical reagents are likely to have effect on whole virus particle whose integrity is essential for vaccine potency. Therefore, a need is felt to develop non-chemical methods. We have studied induction of endonucleolytic activity as an alternative method for virus inactivation. This method of inactivation was compared with the chemical methods, and found to be highly effective for virus inactivation. The effects of endonucleolytic activity on the integrity of virus capsid was studied using antibodies raised against recombinant proteins, which elicited antibodies against major epitopes present on the surface of the virus. Further, the effect of the agents on the integrity of the virus capsid was studied by using antigen capture PCR (Ag-RT/PCR) which detects the whole virus. The studies showed that inactivation of the virus by induction of endonucleolytic activity is more effective besides maintaining virus integrity. The effect of various inactivating agents on four serotypes of FMDV has also been studied and found to have varying effects, depending on serotype.

Animals↗
PubMed1966

Inactivation of foot-and-mouth disease virus by interaction of dye and visible light.

The inactivation of foot-and-mouth disease virus was studied by means of the interaction of neutral red, Toluidine Blue, and methylene blue with visible light. The virus, Type A, strain 1, CANEFA of Argentine origin, was grown in tissue culture and tested in the crude and clarified state. Virus and dye were mixed and incubated together at 4 C for 45 min in the dark, or were mixed and immediately exposed to the visible light source without prior incubation together. Mixtures of crude virus and dye, under any of the experimental conditions used, did not inactivate more than 1 to 2 logs of viral infectivity when held in the dark or when exposed to light during a period of 45 min. Complete inactivation of virus was achieved when clarified virus and dye were mixed and immediately exposed to the visible light source for 15 min. Prior incubation of clarified virus and dye permitted inactivation by methylene blue only, whereas no incubation prior to exposure resulted in three of the dyes contributing to inactivation. A concentration of 6 mug of neutral red, Toluidine Blue, methylene blue, and crystal violet was used per milliliter of virus suspension. Crystal violet was not a good viral inactivator under the conditions of the experimentation. Inactive virus induced the formation of neutralizing antibodies in adult chickens and mice. The antibody titer stimulated by the antigen treated with methylene blue and visible light was probably significant.

Animals↗
PubMed2002

Genetic heterogeneity in the foot-and-mouth disease virus Leader and 3C proteinases.

The Leader and 3C proteinases of foot-and-mouth disease virus (FMDV) are responsible for almost all the proteolytic processing events of the viral polyprotein precursor. Investigation into the genetic heterogeneity of the regions encoding these proteins from isolates of six FMDV serotypes revealed the 3C proteinase to be more conserved than the Leader proteinase. Maximum likelihood analysis indicated similar phylogenetic groupings for the non-structural protein coding regions of both the Leader and 3C. These groupings were different from the structural VP1 protein coding region which, as shown previously, grouped according to serotype. Two distinct clades were apparent for both the Leader and 3C coding regions: one comprising of serotypes A, O and C together with SAT (South African Territories) isolates from eastern Africa. The other clade consisted of SAT isolates originating from southern Africa. Only one virus isolate, obtained from a buffalo in Uganda, did not conform to this phylogenetic pattern. This SAT 1 virus grouped with types A, O and C in the Leader analysis, but with the southern African SAT types in the 3C analysis, implicating intertypic recombination. The leader proteinases of southern African SAT type isolates differed from those present in European type isolates, particularly in the self-processing region. A three-dimensional structure was modeled for the Leader proteinase of one of the SAT type viruses, ZIM/7/83/2, and compared with the previously elucidated crystal structure of O(1)Kaufbeuren Leader proteinase. The active sites of the two leaders were found to superimpose closely, despite the observed sequence variation between the two molecules. Comparison of the 3C proteinase P1 cleavage sites suggested that the FMDV 3C proteinase may possess a broader substrate specificity, as observed in hepatitis A virus 3C proteinase.

3C Viral Proteases↗
PubMed2005

Analysis of foot-and-mouth disease virus internalization events in cultured cells.

It has been demonstrated that foot-and-mouth disease virus (FMDV) can utilize at least four members of the alpha(V) subgroup of the integrin family of receptors in vitro. The virus interacts with these receptors via a highly conserved arginine-glycine-aspartic acid amino acid sequence motif located within the betaG-betaH loop of VP1. While there have been extensive studies of virus-receptor interactions at the cell surface, our understanding of the events during viral entry into the infected cell is still not clear. We have utilized confocal microscopy to analyze the entry of two FMDV serotypes (types A and O) after interaction with integrin receptors at the cell surface. In cell cultures expressing both the alphaVbeta3 and alphaVbeta6 integrins, virus adsorbed to the cells at 4 degrees C appears to colocalize almost exclusively with the alphaVbeta6 integrin. Upon shifting the infected cells to 37 degrees C, FMDV capsid proteins were detected within 15 min after the temperature shift, in association with the integrin in vesicular structures that were positive for a marker of clathrin-mediated endocytosis. In contrast, virus did not colocalize with a marker for caveola-mediated endocytosis. Virus remained associated with the integrin until about 1 h after the temperature shift, when viral proteins appeared around the perinuclear region of the cell. By 15 min after the temperature shift, viral proteins were seen colocalizing with a marker for early endosomes, while no colocalization with late endosomal markers was observed. In the presence of monensin, which raises the pH of endocytic vesicles and has been shown to inhibit FMDV replication, viral proteins were not released from the recycling endosome structures. Viral proteins were not observed associated with the endoplasmic reticulum or the Golgi. These data indicate that FMDV utilizes the clathrin-mediated endocytosis pathway to infect the cells and that viral replication begins due to acidification of endocytic vesicles, causing the breakdown of the viral capsid structure and release of the genome by an as-yet-unidentified mechanism.

Antibodies, Viral↗
PubMed2005

A decision-support system for real-time risk assessment of airborne spread of the foot-and-mouth disease virus.

OBJECTIVES: [corrected] The application of epidemic models during the first days following the confirmation of a virus outbreak should significantly contribute to minimize its costs. Here we describe the first version of a decision-support system for the calculation of the airborne spread of a virus and its application to foot-and-mouth disease (FMD). The goal is to provide geographical maps depicting infection risk for various animal species to support the national health authorities. METHODS: The major tool of the decision-support system is a specific epidemic (or atmospheric) model: A so-called Gaussian dispersion model to calculate 3-dimensional virus plumes. Additional tools providing input data and visualizing the output are: A veterinary data base of geo-referenced premises, a geographical information system (GIS), and, as an external part running at the National Weather Service, a numerical weather prediction (NWP) model. To demonstrate the features of the decision-support system a pilot study in Styria, Austria, has been performed simulating an artificial FMD outbreak. RESULTS: One result of this simulation experiment is the determination of neighboring premises at which animals are at risk to be infected. Particular attention has been turned to cattle, sheep and swine. Using actual hourly NWP data from April 25, 2003, and a source of ten swine excreting a virus, cattle have been estimated to be at risk downwind 1,000-12,000 m, sheep 200-1,300 m, and swines 70-330 m. CONCLUSIONS: A system for real-time risk assessment of the airborne spread of a virus, applied to FMD, was introduced. Due to the forcing of the Gaussian dispersion model with NWP data, it is designed to run in both analysis and forecast mode. The system was applied for the first time during the Austrian real-time exercise on FMD, instructed by the European Union, in November 2004.

Air Microbiology↗
PubMed2026

Genetic Variation and Evolutionary Characteristics of Coxsackievirus B1: F3 Subtype Associated With Hand, Foot and Mouth Disease in China.

Coxsackievirus B1 (CV-B1) is primarily associated with meningitis but can also cause localized outbreaks of hand, foot, and mouth disease (HFMD). This study analyzed the genetic diversity of the VP1 gene in 39 strains of the CVB1 virus isolated from HFMD children across 15 provinces in China between 2010 and 2024, as well as 179 strains from 17 countries. Based on the average nucleotide difference of VP1 gene, we classified CVB1 virus into six genotypes A to F, Notably, genotype F is newly classified. Since 2010, genotype F guadually replaced genotype E as the dominant genotype in China and has further subdivided into three subtypes: F1, F2, and F3, with F3 being the most prevalent subtype in China currently. We specifically study the mild and severe cases within the F3 subtype. Temperature-sensitivity experiments revealed no differences between mild and severe cases of the F3 subtype, and they all belong to temperature-sensitive strains. Interestingly, we found that mild cases of the F3 subtype did not involve recombination, whereas all severe cases of the F3 subtype showed recombination with Coxsackievirus B4 (CVB4). CVB4 has consistently been the primary pathogen responsible for severe neonatal illnesses, suggesting that recombination between the F3 subtype and CVB4 may be associated with the development of severe HFMD. These findings provide fundamental scientific data for further investigation into the epidemiology and genetic characteristics of variants of Coxsackievirus B1 in China.

Humans↗
PubMed2005

Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus.

One step TaqMan real-time reverse transcription polymerase chain reaction (R/T RT-PCR) using a set of primers/probes was developed for the detection of foot-and-mouth disease (FMD) virus. The gene-specific probes labeled fluorogen for the internal ribosomal entry site, Leader sequence and 2B regions were used to detect FMD virus (FMDV). This assay specifically detected FMDV both in cell culture preparations and clinical samples, and was capable of distinguishing FMD from other viral diseases similar to clinical signs (swine vesicular disease, vesicular stomatitis and bovine viral diarrhea). This assay was shown to be 1000-fold more sensitive than the conventional RT-PCR method. The detection limits of this assay was 1 TCID(50)/ml of the FMDV RNA concentration. Quantification was obtained by a standard curves plotting threshold cycle values versus known infectivity titer. The assay was sensitive, specific and rapid enough to detect FMDV RNA genome in probang samples. As such, the described method is reliable and provides faster disease diagnostics than the conventional RT-PCR procedure to detect FMDV.

Animals↗
PubMed2004

Extent of reduction of foot-and-mouth disease virus RNA load in oesophageal-pharyngeal fluid after peak levels may be a critical determinant of virus persistence in infected cattle.

To investigate whether foot-and-mouth disease virus (FMDV) RNA loads in oesophageal-pharyngeal fluid (OP-fluid) in the early course of infection is related to the outcome of virus persistence, viral RNA in OP-fluid samples from cattle experimentally infected with FMDV type O was quantitatively analysed by using a quantitative real-time RT-PCR. Viral RNA was detected within 24 h post-infection (p.i.) in all infected animals. Rapid virus replication led to peak levels of viral RNA load by 30-53 h p.i., and then the load declined at various rates. In some animals (n=12, so-called non-carriers) viral RNA became undetectable between 7 and 18 days p.i. In contrast, in persistently infected animals (n=12, so-called carriers) viral RNA persisted in OP-fluid samples at detectable levels beyond 28 days p.i. Analysis of early viral decay/clearance and virus clearance half-life in OP-fluid samples showed that the extent of reduction of viral RNA in OP-fluid samples immediately following peak levels is a critical determinant of the outcome of FMDV persistence.

Animals↗