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1.
J Gen Virol ; 89(Pt 3): 660-666, 2008 Mar.
Article in English | MEDLINE | ID: mdl-18272756

ABSTRACT

A virus was isolated from aborted dromedary (Camelus dromedarius) fetuses during an abortion storm in Dubai, United Arab Emirates. Laboratory investigations showed the causative agent to be indistinguishable from equine rhinitis A virus (ERAV), a picornavirus. Two pregnant dromedaries experimentally infected with the camel virus isolate both aborted and an identical virus was reisolated from both fetuses, thus confirming the diagnosis. The extremely high prevalence of antibody (>90 %) and the high titres recorded against ERAV in the dromedary herd clearly showed that ERAV does infect dromedaries. Unlike horses, where ERAV targets the upper respiratory tract, in dromedaries the target organ appears to be the genital tract.


Subject(s)
Aphthovirus/pathogenicity , Camelus/virology , Disease Outbreaks , Fetus/virology , Picornaviridae Infections/veterinary , Abortion, Veterinary/epidemiology , Abortion, Veterinary/virology , Animals , Aphthovirus/classification , Aphthovirus/genetics , Aphthovirus/isolation & purification , Base Sequence , Female , Molecular Sequence Data , Phylogeny , Picornaviridae Infections/epidemiology , Picornaviridae Infections/virology , Pregnancy , Pregnancy Complications, Infectious/veterinary , Pregnancy Complications, Infectious/virology , Sequence Analysis, DNA
2.
J Gen Virol ; 84(Pt 6): 1607-1612, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12771431

ABSTRACT

Equine rhinitis A virus (ERAV) is an important respiratory pathogen of horses and is of additional interest because of its close relationship and common classification with foot-and-mouth disease virus (FMDV). As is the case with FMDV, the VP1 capsid protein of ERAV has been shown to be a target of neutralizing antibodies. In FMDV VP1, such antibodies commonly recognize linear epitopes present in the betaG-betaH loop region. To map linear B cell epitopes in ERAV VP1, overlapping fragments spanning its length were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins. These fusion proteins were tested for reactivity with sera from ERAV-infected horses and with polyclonal sera from ERAV-immunized rabbits and mice. Regions at the N- and C-termini as well as the betaE-betaF and the betaG-betaH loop regions contained B cell epitopes that elicited antibodies in the natural host. GST fusion proteins of these regions also elicited antibodies following immunization of rabbits and mice, which, in general, strongly recognized native ERAV VP1 but which were non-neutralizing. It is concluded that the N-terminal region of ERAV VP1, in particular, contains strong B cell epitopes.


Subject(s)
Antigens, Viral , Aphthovirus/immunology , Capsid Proteins/immunology , Amino Acid Sequence , Animals , Antibodies, Viral , Antigens, Viral/genetics , Aphthovirus/genetics , Aphthovirus/pathogenicity , Capsid Proteins/genetics , Epitope Mapping , Horse Diseases/immunology , Horse Diseases/virology , Horses , Molecular Sequence Data , Picornaviridae Infections/immunology , Picornaviridae Infections/veterinary , Picornaviridae Infections/virology , Rabbits , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology
3.
J Gen Virol ; 82(Pt 7): 1703-1711, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11413382

ABSTRACT

Foot-and-mouth disease virus (FMDV) is known to employ the conserved Arg-Gly-Asp (RGD) tripeptide located on the variable betaG-betaH loop of the VP1 capsid protein for binding to cells. Coxsackievirus A9 (CAV9) also carries an RGD sequence, but on a short C-terminal extension of its VP1 and in a different amino acid context. This apparent relationship raised the question of whether insertion of the heterologous CAV9 sequence into FMDV would influence infection by the genetically modified FMDV. Four VP1 mutants were generated by PCR mutagenesis of a full-length FMDV cDNA plasmid. After transfection of BHK-21 cells, viral protein synthesis and virus particle formation could be detected. Two of the four mutants, mV9b and mV9d, could be propagated in BHK-21 cells, but not in CV-1 cells. Both of these mutants contained 17 amino acids of the C terminus of CAV9 VP1. Infection of BHK cells could be specifically inhibited by rabbit immune serum raised against a synthetic peptide representing the amino acid sequence of the C-terminal extension of CAV9 VP1. This demonstrated the direct involvement of the inserted sequence in cell infection. In fact, genetically modified FMDV O(1)K was capable of employing the VP1 C-terminal RGD region of CAV9 for infection of BHK cells. In addition, these results show that, even in cell culture-adapted viruses, the RGD-containing betaG-betaH loop plays an important role in virus infectivity.


Subject(s)
Aphthovirus/pathogenicity , Enterovirus/pathogenicity , Amino Acid Sequence , Animals , Aphthovirus/genetics , Aphthovirus/metabolism , Capsid/genetics , Capsid Proteins , Cell Line , Enterovirus/chemistry , Humans , Molecular Sequence Data , Mutagenesis , Species Specificity , Transfection , Virulence
5.
Vet Rec ; 148(19): 585-6, 2001 May 12.
Article in English | MEDLINE | ID: mdl-11386443

ABSTRACT

Preliminary results indicate that no evidence has been found to support the spread of FMD virus from the burning of animal carcases on open pyres. This finding is subject to a number of assumptions, and is based on a limited number of case studies.


Subject(s)
Air Microbiology , Aphthovirus/pathogenicity , Foot-and-Mouth Disease/transmission , Incineration , Animals , Aphthovirus/isolation & purification , Cattle , Disease Reservoirs/veterinary , Sheep , Swine
13.
J Virol ; 75(8): 3977-83, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11264387

ABSTRACT

The genetic changes selected during the adaptation of a clonal population of foot-and-mouth disease virus (FMDV) to the guinea pig have been analyzed. FMDV clone C-S8c1 was adapted to the guinea pig by serial passage in the animals until secondary lesions were observed. Analysis of the virus directly recovered from the lesions developed by the animals revealed the selection of variants with two amino acid substitutions in nonstructural proteins, I(248)-->T in 2C and Q(44)-->R in 3A. On further passages, an additional mutation, L(147)-->P, was selected in an important antigenic site located in the G-H loop of capsid protein VP1. The amino acid substitution Q(44)-->R in 3A, either alone or in combination with the replacement I(248)-->T in 2C, was sufficient to give FMDV the ability to produce lesions. This was shown by using infectious transcripts which generated chimeric viruses with the relevant amino acid substitutions. Clinical symptoms produced by the artificial chimeras were similar to those produced by the naturally adapted virus. These results obtained with FMDV imply that one or very few replacements in nonstructural viral proteins, which should be within reach of the mutant spectra of replicating viral quasispecies, may result in adaptation of a virus to a new animal host.


Subject(s)
Adaptation, Physiological/genetics , Amino Acid Substitution/genetics , Aphthovirus/genetics , Aphthovirus/physiology , Guinea Pigs/virology , Mutation/genetics , Viral Nonstructural Proteins/metabolism , Animals , Aphthovirus/classification , Aphthovirus/pathogenicity , Cloning, Molecular , Foot-and-Mouth Disease/pathology , Foot-and-Mouth Disease/virology , Genome, Viral , Male , Phenotype , RNA, Viral/genetics , Selection, Genetic , Viral Nonstructural Proteins/genetics , Virus Replication
14.
J Gen Virol ; 82(Pt 4): 747-755, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11257178

ABSTRACT

Foot-and-mouth disease (FMD) is a highly contagious, economically important virus disease of cloven-hoofed animals. The objective of the present study was to examine the early pathogenesis of FMD in pigs by a quantitative time-course study. Under experimental conditions, recipient pigs were infected by contact with donor pigs affected by FMD. Every 24 h from day 1 to day 4 after exposure, two recipient pigs were selected randomly, killed and necropsied. A range of tissues were analysed by a quantitative TaqMan RT-PCR method and by titration of FMD virus on primary bovine thyroid cells. The titres of virus determined by assay in cell culture and calculated from the quantitative TaqMan data correlated strongly (r>0.9), thereby establishing the validity of the TaqMan calculations. The data indicated that the replication of virus in the lungs contributes only in small part to airborne virus excretion. Sites in the pharynx, trachea and nasal mucosa are probably more important in that regard. The sites of earliest virus infection and possibly replication in recipient pigs appeared to be in the pharynx (soft palate, tonsil and floor of pharynx). The data indicated that FMD virus replication in pigs is rapid and that the majority of virus amplification occurs in the skin. A model for the progression of infection is proposed, indicating initial spread from the pharyngeal region, through regional lymph nodes and via the blood to epithelial cells, resulting in several cycles of virus amplification and spread.


Subject(s)
Aphthovirus/pathogenicity , Foot-and-Mouth Disease/virology , Reverse Transcriptase Polymerase Chain Reaction , Animals , Aphthovirus/genetics , Aphthovirus/isolation & purification , Cell Culture Techniques , Lung/virology , Pharynx/virology , RNA, Viral/analysis , Swine , Viremia/virology , Virus Replication
15.
J Gen Virol ; 82(Pt 1): 93-101, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11125162

ABSTRACT

The 3' noncoding region (NCR) of the genomic picornaviral RNA is believed to contain major cis-acting signals required for negative-strand RNA synthesis. The 3' NCR of foot-and-mouth disease virus (FMDV) was studied in the context of a full-length infectious clone in which the genetic element was deleted or exchanged for the equivalent region of a distantly related swine picornavirus, swine vesicular disease virus (SVDV). Deletion of the 3' NCR, while maintaining the intact poly(A) tail as well as its replacement for the SVDV counterpart, abrogated virus replication in susceptible cells as determined by infectivity and Northern blot assays. Nevertheless, the presence of the SVDV sequence allowed the synthesis of low amounts of chimeric viral RNA at extended times post-transfection as compared to RNAs harbouring the 3' NCR deletion. The failure to recover viable viruses or revertants after several passages on susceptible cells suggests that the presence of specific sequences contained within the FMDV 3' NCR is essential to complete a full replication cycle and that FMDV and SVDV 3' NCRs are not functionally interchangeable.


Subject(s)
3' Untranslated Regions/genetics , Aphthovirus/genetics , Aphthovirus/pathogenicity , Enterovirus/genetics , Animals , Animals, Suckling , Blotting, Northern , Cell Line , Chimera , Gene Deletion , Mice , RNA, Viral/analysis , RNA, Viral/genetics , Swine , Virulence , Virus Replication
16.
J Virol ; 74(18): 8316-23, 2000 Sep.
Article in English | MEDLINE | ID: mdl-10954530

ABSTRACT

Passage of foot-and-mouth disease virus (FMDV) in cell culture in the presence of the mutagenic base analog 5-fluorouracil or 5-azacytidine resulted in decreases of infectivity and occasional extinction of the virus. Low viral loads and low viral fitness enhanced the frequency of extinction events; this finding was shown with a number of closely related FMDV clones and populations differing by up to 10(6)-fold in relative fitness in infections involving either single or multiple passages in the absence or presence of the chemical mutagens. The mutagenic treatments resulted in increases of 2- to 6.4-fold in mutation frequency and up to 3-fold in mutant spectrum complexity. The largest increase observed corresponded to the 3D (polymerase)-coding region, which is highly conserved in nonmutagenized FMDV populations. As a result, nucleotide sequence heterogeneity for the 3D-coding region became very similar to that for the variable VP1-coding region in FMDVs multiply passaged in the presence of chemical mutagens. The results suggest that strategies to combine reductions of viral load and viral fitness could be effectively associated with extinction mutagenesis as a potential new antiviral strategy.


Subject(s)
Aphthovirus/genetics , Viral Load , Amino Acid Substitution , Animals , Antigens, Viral/genetics , Aphthovirus/pathogenicity , Azacitidine/pharmacology , Capsid/genetics , Capsid Proteins , Cell Line , Cell Survival , Cells, Cultured , Cricetinae , Fluorouracil/pharmacology , Mutagenesis , Mutagens/pharmacology , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, RNA , Viral Nonstructural Proteins/genetics
17.
Can J Vet Res ; 64(2): 130-3, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10805253

ABSTRACT

Four calves were exposed via aerosol to 1 of 2 strains of foot-and-mouth disease virus. Two animals received virus derived from an infectious clone virus (A12-IC) and 2 received virus derived from the same clone but which lacked the leader coding region (A12-LLV2) that codes for a protein responsible for turning off host protein synthesis. Animals were euthanized at 24 and 72 h post exposure. Cattle receiving A12-IC had a rapid course of disease with more virus in tissues while A12-LLV2-infected cattle did not develop clinical signs of disease. Formalin-fixed, paraffin-embedded tissue sections were probed with digoxigenin-labeled riboprobes corresponding to the coding sequence for bovine interferon (IFN) alpha and IFNbeta. Staining for IFNalpha mRNA was noted in mononuclear cells of the lungs of all animals and in respiratory lymph nodes of cattle receiving A12-IC. Staining for IFNbeta mRNA was confined to bronchiolar epithelium and present only in the animals infected with A12-IC. Inability of the A12-LLV2 virus to achieve levels of spread seen with A12-IC may be related to translation of IFNalpha in A12-LLV2-infected cells, which renders adjacent cells less susceptible to productive infection.


Subject(s)
Aphthovirus/pathogenicity , Foot-and-Mouth Disease/immunology , Interferon-alpha/biosynthesis , Interferon-beta/biosynthesis , Animals , Cattle , Cloning, Molecular , In Situ Hybridization/veterinary , Interferon-alpha/analysis , Interferon-beta/analysis , RNA, Messenger/analysis
18.
J Virol ; 74(2): 987-91, 2000 Jan.
Article in English | MEDLINE | ID: mdl-10623761

ABSTRACT

In 1997, a devastating outbreak of foot-and-mouth disease (FMD) in Taiwan was caused by a serotype O virus (referred to here as OTai) with atypical virulence. It produced high morbidity and mortality in swine but did not affect cattle. We have defined the genetic basis of the species specificity of OTai by evaluating the properties of genetically engineered chimeric viruses created from OTai and a bovine-virulent FMD virus. These studies have shown that an altered nonstructural protein, 3A, is a primary determinant of restricted growth on bovine cells in vitro and significantly contributes to bovine attenuation of OTai in vivo.


Subject(s)
Aphthovirus/pathogenicity , Disease Outbreaks , Foot-and-Mouth Disease/virology , Amino Acid Sequence , Animals , Aphthovirus/genetics , Cattle , Cell Line , Cricetinae , Foot-and-Mouth Disease/epidemiology , Genome, Viral , Molecular Sequence Data , Swine , Taiwan/epidemiology , Viral Nonstructural Proteins/genetics , Virulence/genetics
19.
Arch Virol ; 144(5): 905-19, 1999.
Article in English | MEDLINE | ID: mdl-10416374

ABSTRACT

Chaperonin 10 of M. tuberculosis conferred partial or total protection against generalized foot-and-mouth disease (FMD) in guinea-pigs challenged with O1 Lausanne FMD virus. Chaperonin 10-immunized animals mounted an antibody response to the protein, one epitope of which was found in the C-terminal half. A similar recognition pattern was observed in FMD-convalescent guinea-pigs, swine and cattle. Anti-chaperonin 10 sera showed antiviral activity against FMDV-infected BHK-21 cells. There was strong evidence that early after infection these cells actively secrete their histones and that antisera to the chaperonin recognize them. The same antisera reacted with purified histones in immunoblotting. Most important, exogenously added histones abrogated the anti-viral activity of the antiserum and an anti-histone monoclonal antibody had strong antiviral activity against FMDV-infected BHK-21 cells. These results are consistent with previous reports on displacement of histones from the nuclear compartment and immune recognition of self-histones after viral infections. On the whole, they indicate that M. tuberculosis chaperonin 10 enables the immune system to react against early abnormalities of virus-infected cells; this is accomplished by antibody cross-reacting with histones released during virus infection.


Subject(s)
Aphthovirus/immunology , Chaperonin 10/therapeutic use , Foot-and-Mouth Disease/immunology , Foot-and-Mouth Disease/prevention & control , Mycobacterium tuberculosis , Amino Acid Sequence , Animals , Antibodies, Viral/immunology , Antibody Formation , Aphthovirus/pathogenicity , Cattle , Cell Line , Chaperonin 10/chemistry , Chaperonin 10/immunology , Cricetinae , Guinea Pigs , Histones/biosynthesis , Histones/immunology , Molecular Sequence Data , Mycobacterium tuberculosis/immunology , Peptide Fragments/chemistry , Peptide Fragments/immunology , Peptide Fragments/therapeutic use , Swine
20.
Vet J ; 158(1): 6-13, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10409411

ABSTRACT

Despite representing the largest part of the world's foot-and-mouth disease (FMD)-susceptible domestic livestock, sheep and goats have generally been neglected with regard to their epidemiological role. This is partly due to the often inapparent nature of the disease in these hosts. Nevertheless, their ability to become carriers represents a reservoir for further infection and spread of disease, and so trade of live sheep and goats present a major risk of entry of FMD to disease-free countries. Research and epidemiological studies continue to be necessary in order both to prevent the entry of the virus and to assist in control should the disease reoccur. This review concentrates primarily on more recent studies relating to sheep and goats and, in particular, considers the importance of these hosts in the overall epidemiology of FMD.


Subject(s)
Aphthovirus/pathogenicity , Disease Reservoirs , Foot-and-Mouth Disease/epidemiology , Goat Diseases/epidemiology , Sheep Diseases/epidemiology , Animals , Antibodies, Viral/blood , Disease Reservoirs/veterinary , European Union , Foot-and-Mouth Disease/transmission , Goat Diseases/transmission , Goat Diseases/virology , Goats , Immunization/veterinary , Sheep , Sheep Diseases/transmission , Sheep Diseases/virology , Viral Vaccines/therapeutic use
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