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Epidemiological basis useful for the control of foot-and-mouth disease.

Although known for many years, foot-and-mouth disease is still able to represent a real threat to many farming economies in the world. The recent 2001 Western European epizootics linked to O PanAsia virus strain can illustrate the fact that many questions are still unanswered in the field of foot-and-mouth epidemiology. It also demonstrates that the increase in international trade, including livestock, animal products and animal food, means an increase in the probability of transmitting, through the same way, some animal diseases, foot-and-mouth included. In our economies, a rapid identification of the virus and a fast elimination of infected, contaminated and even some contact animals are still the key factors to react in front of such a disease.

Animal Husbandry↗

A survey to investigate movements off sheep and cattle farms in New Zealand, with reference to the potential transmission of foot-and-mouth disease.

AIM: To quantify the numbers and extent of movements off sheep and cattle farms in New Zealand, in order to construct more realistic simulation models to investigate how infectious diseases such as foot-and-mouth disease (FMD) might spread. METHODS: Farmers from 500 randomly selected farms, comprising 100 from each of the following sectors, viz beef, dairy, grazing/dairy heifer rearing, sheep, and mixed sheep and beef, were asked to fill in diaries in which they recorded the movements of all animals, products, people, vehicles and equipment coming on to or leaving their farms during two separate 3-week periods, representing relatively 'busy' and 'quiet' times of the year with respect to livestock movements. Where possible, the destination of each movement was identified and geo-coded, to allow the distance travelled to be calculated. Each movement was then classified according to the risk of transfer of FMD virus (FMDV), should the disease have been present on the study farm at the time of the movement. The data were then analysed to establish movement frequencies and distributions of distances travelled, by the different pastoral livestock sectors. RESULTS: Two hundred and seventeen farmers returned one or more diaries. One hundred and ninety-three farmers completed a Busy-period diary, recording a total of 12,052 movements off their farms, a crude average of 62.4 per 3-week period, or 2.97 per day. Of these, 4.0% involved the transport of livestock, equating to 0.12 livestock consignments per day. In contrast, 186 Quiet-period diaries were returned, recording a total of 10,885 movements off, representing a crude average of 58.5 during the 3-week period, or 2.78 per day. Of these, 2.1% involved livestock, equating to 0.06 livestock consignments per day. The mean and median distances travelled during the Busy periods were 30.9 km and 13.1 km, respectively (range 0-1,167 km). In comparison, the mean and median distances travelled during Quiet periods were 41.3 and 14 km, respectively (range 0.4-1,203 km). CONCLUSIONS: People, vehicles, livestock and other items can travel off pastoral livestock farms in New Zealand to other farms either directly or via saleyards over extensive distances. This has implications for the potential spread of infectious diseases such as FMD. Movement parameters intended for use in the InterSpread Plus inter-farm simulation model of FMD were established, which will facilitate the prediction of likely spread and efficacy of controls in the unlikely event of a real-life outbreak.

Animals↗

[Combating foot and mouth disease in Brazil: private sector participation].

The fact that cattle herds in Brazil are in such good health and that the meat on the domestic and international markets is safe and of high quality is largely due to the campaign to combat foot and mouth disease and to the improvements that this campaign has made in the organisation of the Brazilian bovine production industry. Launched in 1965, the campaign set itself ambitious objectives, but thanks to the direct participation of producer organisations it received the necessary support and funding. The results are reflected in the growing number of states--fifteen to date--that are internationally recognised as free from foot and mouth disease with vaccination. Thanks to the active participation of producers and of the entire production chain in foot and mouth disease eradication efforts, Brazil has risen to become a leading player in the international beef trade.

Animals↗

Synthetic vaccine against foot-and-mouth disease based on a palmitoyl derivative of the VP1 protein 135-159 fragment of the A22 virus strain.

The peptide Palm2 135-159, a dipalmitoyl derivative of the 135-159 fragment of VP1 protein of the foot-and-mouth disease virus strain A22 was synthesized. In the experiments on mice, guinea pigs and sheep Palm2 135-159 possesses greater immunogenic and protective activity than the nonacylated 135-159 peptide. The synthetic vaccine against foot-and-mouth disease for use in sheep was developed on the basis of the lipopeptide. Synthetic polymethylsiloxane oil was found to be a suitable adjuvant for this vaccine. The dependencies of protective and immunogenic effects from the dose of peptide were studied. The vaccine was found to be stable to storage for 1 year at 18 degrees C. It was shown that the synthetic vaccine provides 1 year protection of sheep against foot-and-mouth disease after a single administration. The vaccine is allowed for veterinary use in Russia.

Animals↗

Overview of foot and mouth disease in southern Africa.

The prevalence of foot and mouth disease (FMD) in the southern African subcontinent between 1931 and 1990 is summarised, together with the major features of the epidemiology and control of the disease. The author emphasises the role of wildlife, especially African buffalo (Syncerus caffer). A proposal is made for a more structured and co-operative approach to investigating the extent and nature of antigenic variation within the Southern African Territories (SAT) types of FMD virus. Quantification of the economic impact of FMD on the agro-economics of the subcontinent is attempted, and the importance of the social values of rural peoples in this respect is explained.

Africa, Southern↗

[Recent outbreak of foot and mouth disease in Ivory Coast].

An outbreak of foot and mouth disease (FMD) due to SAT2 occurred among cattle, sheep and pigs in Côte-d'Ivoire. The morbidity and mortality were low so vaccination of only high value livestock in intensive production systems was suggested.

Animals↗

Absence of antibodies to foot-and-mouth disease virus in free-ranging roe deer from selected areas of Germany (2001-2002).

Blood samples (n = 223) of free-ranging roe deer (Capreolus capreolus) were collected from selected hunting grounds in Germany between October 2001 and October 2002. Samples originated from Lower Saxony (n = 43) and North-Rhine Westphalia (n = 108) within a 20-km area ("cordon") cordoned off along the border of The Netherlands. This is adjacent to the area of a foot-and-mouth disease outbreak that occurred between 21 March and 22 April 2001 in The Netherlands. Negative control samples were taken from northern Germany (Schleswig-Holstein, n = 72). Two different enzyme-linked immunosorbent assays (ELISAs) were used for the detection of antibodies against foot-and-mouth disease virus (FMDV) serotype O strain Manisa. To confirm ELISA-positive results, a virus neutralization test was performed. All samples tested negative for antibodies against FMDV. These results suggest that FMDV was not transmitted to free-ranging roe deer living in parts of Germany adjacent to the area affected by the 2001 foot-and-mouth disease outbreak in The Netherlands.

Animals↗

[Hand-foot-mouth disease].

A case of hand-foot-and-mouth disease is reported in a 20-year-old female patient. Infection by Coxsackie A 16-virus was diagnosed by serology.

Adult↗

Expression of the Foot-and-Mouth Disease Virus VP1 protein using a replication-competent recombinant canine adenovirus type 2 elicits a humoral antibody response in a porcine model.

To develop a new type vaccine for Foot-and-Mouth Disease (FMD) prevention by using canine adenovirus as vector, the VP1 cDNA of Foot-and-Mouth Disease Virus (FMDV) type O strain China 99 was amplified by RT-PCR and cloned into pEGFP-C1 by replacing the GFP gene with the VP1 cDNA, resulting in an expression plasmid pVP1-C1. The expression cassette of VP1 composed of the CMV promoter, the VP1 gene and the SV40 early mRNA polyadenylation signal was recovered by Nsi I / Mlu I digestion of pVP1-C1 and cloned into the Canine adenovirus type-2 (CAV-2) genome in which E3 region was partly deleted by removing the Ssp I- Ssp I fragment. The recombinant virus (CAV-2-VP1) was obtained by transfecting the recombinant CAV-2-VP1 genome into MDCK cells with Lipofectamine 2000. Immunization trial in pigs with the recombinant virus, CAV-2-VP1, showed that CAV-2-VP1 could stimulate a specific immune response to both FMDV and the vector virus. Immune response to the VP1 and FMDV after VP1 expression was confirmed by ELISA, western blotting analysis and neutralization test. It was indicated that CAV-2 may serve as a vector for FMD vaccine development in pigs.

Adenoviruses, Canine↗

Construction of a chimeric Theiler's murine encephalomyelitis virus containing the leader gene of foot-and-mouth disease virus.

The foot-and-mouth disease virus (FMDV) leader coding region (Lb) was cloned into a full-length cDNA of the DA strain of Theiler's murine encephalomyelitis virus (TMEV) replacing the complete L coding region of TMEV. This construct, pDAFSSC1-Lb, was engineered to contain cleavage sites, at the 3' end of the Lb coding region, for both the FMDV Lb and the TMEV 3C proteases. Transcripts derived from this construct were translated in a cell-free system. Analysis of the translation products showed efficient synthesis and processing of TMEV structural and nonstructural proteins as well as a major band that comigrated with FMDV Lb and was reactive with Lb antiserum. A small plaque virus was recovered from BHK-21 cells transfected with RNA derived from pDAFSSC1-Lb. RT-PCR of RNA isolated from DAFSSC1-Lb virus demonstrated a product corresponding in size and sequence to FMDV Lb. DAFSSC1-Lb virus grew slower than parental virus, DAFSSC1, and to a lower titer. The pattern of viral proteins synthesized in DAFSSC1-Lb virus-infected cells was very similar to the pattern in DAFSSC1 virus-infected cells except that significant amounts of FMDV Lb were produced. In addition, extracts from DAFSSC1-Lb-virus-infected cells cleaved an exogenous source of the translation initiation factor, p220, while DAFSSC1-virus-infected extracts did not. Chimeric viruses that contain coding regions from different picornaviral genera may be valuable tools in investigating the function of particular viral proteins and in studying disease pathogenesis.

Animals↗

Detection and characterization of functional T-cell epitopes on the structural proteins VP2, VP3, and VP4 of foot and mouth disease virus O1 campos.

Foot and mouth disease virus (FMDV) is the cause of a widespread infectious disease affecting cloven-hoofed animals. It is controlled by vaccination with immune-inactivated virus grown in tissue culture. However, peptide vaccines represent a safer alternative to the current virus-inactivated immunogens. Their design requires the identification and evaluation of the sequences recognized by T- and B-lymphocytes. Four structural proteins, VP1, VP2, VP3, and VP4, comprise the viral capsid of the FMDV, but only VP1 has been extensively studied regarding the existence of relevant T-cell epitopes. Here, we utilize a murine model to present a functional T-cell epitope mapping on the complete sequences of VP2, VP3, and VP4 of FMDV O1 Campos. We used two in vitro assays to describe 13 amino acid sequences, each one of them including at least one T-cell epitope. The in vivo T-cell helper function of these sequences was studied in an adoptive cell-transfer assay in mice. Immunization experiments with a fusion peptide containing one of the sequences characterized were also done comparing the helper activity of this sequence with other T-cell epitopes included in the major immunogenic region of VP1.

Adoptive Transfer↗

Multiple sites of recombination within the RNA genome of foot-and-mouth disease virus.

Recombinant foot-and-mouth disease viruses were isolated from cells infected with a mixture of temperature-sensitive (ts) mutants belonging to different subtype strains. In order to select for recombination events in many different regions of the genome, crosses were performed between various pairs of mutants, with ts mutations in different regions of the genome. ts+ progeny were analysed by electrofocusing virus-induced proteins and RNase T1 fingerprinting of their RNA. All but 5 out of 43 independent isolates, from nine crosses, proved to have recombinant RNA genomes. Maps of these genomes, based on a knowledge of the locations of the unique oligonucleotides, were constructed. Most could be interpreted as being the products of single genetic cross-overs, although three recombinants were formed by two cross-overs each. Cross-overs in at least twelve distinct regions of the genome were identified. This evidence of a large number of recombination sites suggests that RNA recombination in picornaviruses is a general, as opposed to a site-specific, phenomenon.

Aphthovirus↗

Expression in Escherichia coli and purification of biologically active L proteinase of foot-and-mouth disease virus.

The foot-and-mouth disease virus (FMDV) Lb gene was cloned into bacterial expression vectors under the control of a T7 RNA polymerase promoter. The Lb protein was expressed in both an in vitro transcription-translation system and in Escherichia coli. In vitro expression of a construct containing the Lb gene fused to a portion of the VP4 and 3D genes demonstrated cis cleavage activity that could be blocked by the thiol protease inhibitor E-64. Lb expressed in E. coli was purified from the soluble fraction by metal chelation chromatography. Purified Lb had trans cleavage activity at the L/P1 junction and cleaved the p220 component of the cap-binding protein complex.

Aphthovirus↗

Characterization of an acid-resistant mutant of foot-and-mouth disease virus.

A foot-and-mouth disease virus mutant which is stable at pH 6.4 has been isolated from a virus of serotype A. In contrast to the parent (P) virus, which gave a mixture of large and small plaques in BHK21 cells and in a bovine kidney cell line, the acid-resistant (AR) virus gave small plaques which did not increase markedly in size after 24 hr. The infectivity titer of the acid-resistant virus was about 100-fold lower in suckling mice than in BHK21 cells, whether the inoculation was made intraperitoneally or intracerebrally, whereas the parent virus gave similar titers in both systems. Furthermore, in mice the AR virus reached its end point two to three times more slowly. The diameter of the AR virus was almost 20% less than that of the P virus and it had a more distinct topography, but the two viruses cosedimented in sucrose gradients. However, the buoyant density in CsCl of the AR virus was slightly lower (1.42 compared with 1.43 g/cc) in coruns. The RNAs and capsid proteins of the two viruses gave similar profiles in sucrose gradients and by SDS-PAGE, respectively. However, isoelectric focusing of the capsid proteins revealed considerable differences between the two viruses. Whereas the P virus gave four protein bands, corresponding to VP1-VP4, the AR virus gave one band for VP4, two for VP3, two for VP2, and four for VP1. Sequence analysis of the genes coding for the capsid protein regions of the two viruses showed four changes (one silent), resulting in an Ala-3-->Ser substitution in VP1 and Glu-131-->Lys and Asp-133-->Ser substitutions in VP2.

Animals↗

Evaluation of three 'ready to formulate' oil adjuvants for foot-and-mouth disease vaccine production.

Foot-and-mouth disease virus (FMDV) type OR(2)/75, grown on BHK 21 clone 13 cell monolayers, was inactivated with formalin. The virus was clarified and was either concentrated with 8% polyethylene glycol 6000 (PEG) or used in its untreated form for the preparation of oil adjuvant vaccines. The oil adjuvants used in this study were Montanide ISA 206 (which renders a water-in-oil-in-water (w/o/w) type of emulsion), Montanide ISA 57 and Montanide ISA 50V (both of which render water-in-oil (w/o) type of emulsions). The vaccines were tested on guinea pigs and calves. The results indicated that vaccines emulsified using Montanide ISA 57 elicited the best protective immune response in the animals, followed by those emulsified with Montanide ISA 50V and Montanide ISA 206. It was also found that vaccines formulated with virus concentrated using 8% polyethylene glycol (PEG) were more immunogenic than the vaccines formulated with the untreated harvest virus.

Adjuvants, Immunologic↗

The sub-type classification of strains of foot-and-mouth disease virus.

Sixteen foot-and-mouth disease virus (FMDV) strains of type SAT 1 were compared in complement-fixation tests. With the test used, the range of antigenic variation within a type appeared to be greater than previously described. The concept of a sub-type group within which all strains are more closely related to each other than to any strain outside the group was not supported. Considering the group of strains studied, it is suggested that the classification of strains is best achieved by moninating a reference strain for each sub-type. Others are classified as related strains in one or more sub-type groups according to their relationships with the reference strains.

Animals↗

Recognition of the initiation codon for protein synthesis in foot-and-mouth disease virus RNA.

Foot-and-mouth disease virus (FMDV) RNA utilizes two in-frame initiation codons to produce two precursor proteins with identical carboxy termini. The 5' untranslated region (5'UTR) directs the ribosome to internal sequences without the need for a cap structure as used in host mRNAs. The FMDV 5'UTR was cloned upstream of the reporter gene chloramphenicol acetyltransferase (CAT) in order to study the selection of initiation site and to facilitate quantification of the translation products. After in vitro transcription with T7 RNA polymerase and translation in rabbit reticulocyte lysate, the two CAT products, resulting from initiation from the two initiation codons, were quantified. The downstream initiator AUG (AUGLb) was selected more efficiently in the wild-type 5'UTR. In truncated RNA, the upstream initiation site (AUGLab) was more efficiently utilized than in the wild-type 5'UTR. Protein synthesis initiation factors were added to translation assays to determine whether these factors influenced initiation site selection. Addition of eIF-2 and of eIF-2B changed the selection process for both types of RNA. These factors induced a 2.5-fold higher usage of the upstream AUGLab for wild-type and 5'UTR-truncated RNA. A change in mRNA concentration also induced a change in the usage of initiation codons; however, the effect of eIF-2 was measured over a broad range of mRNA concentrations. In conclusion, eIF-2 mediates the recognition of the initiation codon during both cap-dependent and internal ribosome entry site-dependent initiation.

Animals↗

Evidence for the role of His-142 of protein 1C in the acid-induced disassembly of foot-and-mouth disease virus capsids.

Foot-and-mouth disease virus (FMDV) capsids are inherently labile under mildly acidic conditions, dissociating to pentamers at pH values in the region of 6.5, with the release of protein 1A and the viral RNA. This acid-induced disassembly is thought to be required for the entry of the virus genome into the host cell. Previous work has highlighted a histidine-alpha-helix charge-dipole interaction at the twofold axes of symmetry between pentamers and has suggested that this interaction plays a role in acid-induced disassembly. The validity of this theory has now been tested by converting the implicated residue, His-142 of protein 1C, to Arg, Phe and Asp. The effects of such changes were studied by using a previously described vaccinia virus expression system, in which synthesis and processing of FMDV capsid proteins results in the self-assembly of capsids. In agreement with the histidine-alpha-helix charge-dipole theory, assembly in the arginine mutant was found to be greatly reduced, while capsids of the aspartic acid mutant were considerably more stable under acidic conditions than the wild-type. Aberrant but acid-stable complexes were obtained in the phenylalanine mutant.

Animals↗