Groppelli, Elisabetta; Tuthill, Tobias J.; Rowlands, David J.
Equine rhinitis A virus (ERAV) is genetically closely related to foot-and-mouth disease virus (FMDV), and both are now classified within the genus Aphthovirus of the family Picornaviridae. For disease security reasons, FMDV can be handled only in high-containment facilities, but these constraints do not apply to ERAV, making it an attractive alternative for the study of aphthovirus biology. Here, we show, using immunofluorescence, pharmacological agents, and dominant negative inhibitors, that...
Tobias J Tuthill
Full Text Available Equine rhinitis A virus (ERAV is closely related to foot-and-mouth disease virus (FMDV, belonging to the genus Aphthovirus of the Picornaviridae. How picornaviruses introduce their RNA genome into the cytoplasm of the host cell to initiate replication is unclear since they have no lipid envelope to facilitate fusion with cellular membranes. It has been thought that the dissociation of the FMDV particle into pentameric subunits at acidic pH is the mechanism for genome release during cell entry, but this raises the problem of how transfer across the endosome membrane of the genome might be facilitated. In contrast, most other picornaviruses form 'altered' particle intermediates (not reported for aphthoviruses thought to induce membrane pores through which the genome can be transferred. Here we show that ERAV, like FMDV, dissociates into pentamers at mildly acidic pH but demonstrate that dissociation is preceded by the transient formation of empty 80S particles which have released their genome and may represent novel biologically relevant intermediates in the aphthovirus cell entry process. The crystal structures of the native ERAV virus and a low pH form have been determined via highly efficient crystallization and data collection strategies, required due to low virus yields. ERAV is closely similar to FMDV for VP2, VP3 and part of VP4 but VP1 diverges, to give a particle with a pitted surface, as seen in cardioviruses. The low pH particle has internal structure consistent with it representing a pre-dissociation cell entry intermediate. These results suggest a unified mechanism of picornavirus cell entry.
Grigera, P R; Garcia-Briones, M; Periolo, O; La Torre, J L; Wagner, R R
An oligodeoxynucleotide coding for amino acids 139 through 149 of antigenic site A (ASA) of the VP1 capsid protein of the foot-and-mouth disease virus C3 serotype (FMDV C3) was inserted into three different in-frame sites of the vesicular stomatitis virus New Jersey serotype (VSV-NJ) glycoprotein (G) gene cDNA present in plasmid pKG97 under control of the bacteriophage T7 polymerase promoter. Transfection of these plasmids into CV1 cells coinfected with the T7 polymerase-expressing vaccinia v...
Foot-and-mouth disease virus (FMDV), equine rhinitis A virus (ERAV) and bovine rhinitis B virus (BRBV) comprise the genus Aphthovirus of the Picornaviridae family. Seven genera within this family, Aphthoviruses, Cardioviruses, Erboviruses (ERBV), Kobuviruses, Senecaviruses, Sapeloviruses, and Tescho...
Costa Giomi, M P; Bergmann, I E; Scodeller, E A; Augé de Mello, P; Gomez, I; La Torre, J L
In this paper we report a study of a sample of foot-and-mouth disease virus carrying two polyribocytidylic acid [poly(C)] tracts of different lengths. By plaque purification in tissue culture, we isolated two populations of particles, one carrying the long poly(C) tract and the other carrying only the short homopolymer. The fingerprints of both viruses were indistinguishable from each other and from that of the virus present in the original sample, suggesting that the main difference between the two types of particles is limited to the poly(C) tracts of their genomic RNAs, to the flanking sequences of the poly(C) tract, or to both. In addition, some biological properties of these viruses are reported, such as stability upon serial passages in different cell lines, plaque size, and pathogenicity for cattle. The results indicate that the size of the poly(C) tract is not directly related to the virulence of these viruses. However, the size of the homopolymer could play a role in determining their efficiency of replication, and it appears that the particles with the short poly(C) tract might have some replicative advantage over those carrying the long one. Images PMID:6088803
Costa Giomi, M P; Bergmann, I E; Scodeller, E A; Augé de Mello, P; Gomez, I.; La Torre, J L
In this paper we report a study of a sample of foot-and-mouth disease virus carrying two polyribocytidylic acid [poly(C)] tracts of different lengths. By plaque purification in tissue culture, we isolated two populations of particles, one carrying the long poly(C) tract and the other carrying only the short homopolymer. The fingerprints of both viruses were indistinguishable from each other and from that of the virus present in the original sample, suggesting that the main difference between ...
Full Text Available FR108942 AY593808 Aphthovirus internal ribosome entry site (IRES) internal_ribosome_entry_site F ... oot-and-mouth disease virus C4 ... RF00210 Rfam v8.1 FR108942.jpg FR108942.png FR1089 ...
Hato, S.V.; Sorgeloos, F.; Ricour, C.; Zoll, J.; Melchers, W.J.G.; Michiels, T.; Kuppeveld, F.J.M. van
Picornaviruses encompass a large family of RNA viruses. Some picornaviruses possess a leader (L) protein at the N-terminus of their polyprotein. The L proteins of encephalomyocarditis virus, a cardiovirus, and foot-and-mouth disease virus (FMDV), an aphthovirus, are both dispensable for replication
Our recent study has shown that bovine rhinovirus type 2 (BRV2), a new member of the Aphthovirus genus, shares many motifs and sequence similarities with foot-and-mouth disease virus (FMDV). Despite low sequence conservation (36percent amino acid identity) and N- and C-terminus folding differences,...
Foot-and-mouth disease virus (FMDV) is the type species of the Aphthovirus genus, of the family Picornaviridae. Infection of cells with positive-strand RNA viruses results in a rearrangement of intracellular membranes into viral replication complexes. However, the origin of these membranes remains u...
Bovine Rhinitis B Viruses (BRBV) are picornaviruses responsible for mild respiratory infection of cattle and probably the least characterized member of the Aphthoviruses. BRBV is the closest relative known to Foot and Mouth Disease virus (FMDV) with around a 43 percent identical polyprotein sequenc...
Hahn, H.; Palmenberg, A C
Sixteen substitution mutations of the conserved DvExNPGP sequence, implicated in cardiovirus and aphthovirus primary polyprotein cleavage, were created in encephalomyocarditis virus cDNA, expressed, and characterized for processing activity. Nearly all the mutations severely decreased the efficiency of the primary cleavage reaction during cell-free synthesis of viral precursors, indicating a stringent requirement for the natural sequence in this processing event. When representative mutations...
This bachelor's thesis called "Control measures for an outbreak of foot and mouth disease at the point of origin and its surroundings" deals with problems concerning FMD disease occurrence in Chlebov livestock holding belonging to Reprogen JSC. FMD is extremely dangerous and highly contagious disease causing high fever that affects especially cloven-hoofed animals. A viral genus Aphthovirus is the originator, causing blisters and aphthae on intestinal tract, limbs and hairless skin, other sym...
Skern, Tim; Fita, Ignacio; Guarné, Alba
The leader (L) proteinases of aphthoviruses (foot-and-mouth disease viruses) and equine rhinovirus serotypes 1 and 2 cleave themselves from the growing polyprotein. This cleavage occurs intramolecularly between the C terminus of the L proteinases and the N terminus of the subsequent protein VP4. The foot-and-mouth disease virus enzyme has been shown, in addition, to cleave at least one cellular protein, the eukaryotic initiation factor 4G. Mechanistically, inhibitor studies and sequence analy...
Rai, Devendra K.; Lawrence, Paul; Pauszek, Steve J.; Piccone, Maria E.; Knowles, Nick J.; Rieder, Elizabeth
Bovine rhinitis viruses (BRVs) cause mild respiratory disease of cattle. In this study, a near full-length genome sequence of a virus named RS3X (formerly classified as bovine rhinovirus type 1), isolated from infected cattle from the UK in the 1960s, was obtained and analyzed. Compared to other closely related Aphthoviruses, major differences were detected in the leader protease (Lpro), P1, 2B, and 3A proteins. Phylogenetic analysis revealed that RS3X was a member of the species bovine rhinitis A virus (BRAV). Using different codon-based and branch-site selection models for Aphthoviruses, including BRAV RS3X and foot-and-mouth disease virus, we observed no clear evidence for genomic regions undergoing positive selection. However, within each of the BRV species, multiple sites under positive selection were detected. The results also suggest that the probability (determined by Recombination Detection Program) for recombination events between BRVs and other Aphthoviruses, including foot-and-mouth disease virus was not significant. In contrast, within BRVs, the probability of recombination increases. The data reported here provide genetic information to assist in the identification of diagnostic signatures and research tools for BRAV. PMID:27081310
Rai, Devendra K.; Paul Lawrence; Steve J. Pauszek; Piccone, Maria E.; Knowles, Nick J.; Elizabeth Rieder
Bovine rhinitis viruses (BRVs) cause mild respiratory disease of cattle. In this study, a near full-length genome sequence of a virus named RS3X (formerly classified as bovine rhinovirus type 1), isolated from infected cattle from the UK in the 1960s, was obtained and analyzed. Compared to other closely related Aphthoviruses, major differences were detected in the leader protease (Lpro), P1, 2B, and 3A proteins. Phylogenetic analysis revealed that RS3X was a member of the species bovine rhini...
Gullberg, Maria; Polacek, Charlotta; Bøtner, Anette;
Foot-and-mouth disease virus (FMDV) is responsible for one of the most economically important diseases of farm animals (estimated annual costs are about US$10 billion globally). The virus is the prototypic Aphthovirus within the family Picornaviridae and has a positive sense RNA genome (ca. 8.3kb...... these into susceptible cells it is possible to rescue specifically altered FMDVs. We have used this approach to generate modified viruses that have particular properties; these studies can assist in the development of improved and safer vaccines to protect against FMDV. For example, we have made changes to the leader (L...... “self-tagged” virus particles that contain the VP1-2A precursor (Gullberg et al., 2013). This approach works for two of the most common FMDV serotypes (O and A) and offers the possibility of a single approach to purifying virus particles from different serotypes using reagents targeted to the conserved...
Devendra K. Rai
Full Text Available Bovine Rhinitis B Virus (BRBV is a picornavirus responsible for mild respiratory infection of cattle. It is probably the least characterized among the aphthoviruses. BRBV is the closest relative known to Foot and Mouth Disease virus (FMDV with a ~43% identical polyprotein sequence and as much as 67% identical sequence for the RNA dependent RNA polymerase (RdRp, which is also known as 3D polymerase (3Dpol. In the present study we carried out phylogenetic analysis, structure based sequence alignment and prediction of three-dimensional structure of BRBV 3Dpol using a combination of different computational tools. Model structures of BRBV 3Dpol were verified for their stereochemical quality and accuracy. The BRBV 3Dpol structure predicted by SWISS-MODEL exhibited highest scores in terms of stereochemical quality and accuracy, which were in the range of 2Å resolution crystal structures. The active site, nucleic acid binding site and overall structure were observed to be in agreement with the crystal structure of unliganded as well as template/primer (T/P, nucleotide tri-phosphate (NTP and pyrophosphate (PPi bound FMDV 3Dpol (PDB, 1U09 and 2E9Z. The closest proximity of BRBV and FMDV 3Dpol as compared to human rhinovirus type 16 (HRV-16 and rabbit hemorrhagic disease virus (RHDV 3Dpols is also substantiated by phylogeny analysis and root-mean square deviation (RMSD between C-α traces of the polymerase structures. The absence of positively charged α-helix at C terminal, significant differences in non-covalent interactions especially salt bridges and CH-pi interactions around T/P channel of BRBV 3Dpol compared to FMDV 3Dpol, indicate that despite a very high homology to FMDV 3Dpol, BRBV 3Dpol may adopt a different mechanism for handling its substrates and adapting to physiological requirements. Our findings will be valuable in the
Full Text Available Foot-and-mouth disease virus (FMDV is an aphthovirus that belongs to the Picornaviridae family and causes one of the most important animal diseases worldwide. The capacity of other picornaviruses to express foreign antigens has been extensively reported, however, little is known about FMDV. To explore the potential of FMDV as a viral vector, an 11-amino-acid (aa HSV epitope and an 8 aa FLAG epitope were introduced into the C-terminal different regions of 3A protein of FMDV full-length infectious cDNA clone. Recombinant viruses expressing the HSV or FLAG epitope were successfully rescued after transfection of both modified constructs. Immunofluorescence assay, Western blot and sequence analysis showed that the recombinant viruses stably maintained the foreign epitopes even after 11 serial passages in BHK-21 cells. The 3A-tagged viruses shared similar plaque phenotypes and replication kinetics to those of the parental virus. In addition, mice experimentally infected with the epitope-tagged viruses could induce tag-specific antibodies. Our results demonstrate that FMDV can be used effectively as a viral vector for the delivery of foreign tags.
Analysis of the VP1 capsid protein coding region of simian virus (SV) 2, SV16, SV18, SV42, SV44, SV45, and SV49 demonstrates that they are clearly distinct from members of the Enterovirus genus and from members of other existing picornavirus genera. To further characterize this group of viruses and to clarify their classification within the Picornaviridae, we have determined the complete genomic sequence of SV2 (8126 nucleotides). The genome was typical of members of Picornaviridae, encoding a single open reading frame. The putative polyprotein contained typical picornavirus protease cleavage sites, yielding mature proteins homologous to each of the known picornavirus proteins. SV2 contained an amino-terminal extension of the reading frame, which was analogous to the leader protein of members of the Aphthovirus, Cardiovirus, Erbovirus, Kobuvirus, and Teschovirus genera, but there was no significant amino acid homology with any of these known leader proteins. The 2A protein also aligned poorly with the 2A proteins of other picornaviruses. The deduced amino acid sequences of the SV2 structural and nonstructural proteins were related to but phylogenetically distinct from those of enteroviruses and human rhinoviruses. The major distinguishing features of SV2 were the presence of a type 2 internal ribosome entry site in the 5'-NTR, a putative leader protein encoded upstream of the structural proteins, and an unusually large 2A protein. On the basis of the molecular analysis, we propose that SV2, SV16, SV18, SV42, SV44, SV45, SV49, and porcine enterovirus 8 be classified as members of a new genus in Picornaviridae and that SV2 (strain 2383) be designated as the type strain
Full text: Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of farm livestock. The etiological agent, FMD virus (FMDV), is a single-stranded, positive-sense RNA virus belonging to the genus Aphthovirus within the family Picornaviridae. Rapid and accurate confirmation of the presence of FMDV is needed for effective control and eradication of the disease. An on-site detection test would be highly advantageous as the time taken to transport suspect clinical material to a central laboratory can often be lengthy, thus delaying a definitive diagnosis in the event of an outbreak. This study describes the development of a molecular assay for the detection of all seven serotypes of FMDV using novel technology, namely: Linear-After-The- Exponential (LATE)-PCR, for transfer onto a portable, easy-to-use, fully automated sample preparation and RT-PCR instrument. Primers and a mismatch tolerant probe were designed from consensus sequences in the FMDV 3D (RNA polymerase) gene to detect the target and its variants at low temperature. An internal control (IC) was included to validate negative results. After demonstrating that the LATE RT-PCR signal at end-point was proportional to number of target molecules over the range 10 to 1 million copies, the assay was compared with a one-step real-time RT-PCR (rRT-PCR) assay (also targeting the 3D) used routinely by reference laboratories. The LATE RT-PCR assay amplified RNA extracted from multiple strains of all FMDV serotypes. Of the 121 FMDV-positive samples tested, 119 were positive by both rRT-PCR and LATE RT-PCR tests while 118 had tested positive by virus isolation at the time of receipt. Twenty-eight FMDVnegative samples failed to react in all 3 tests. There were no false positive signals with RNA from other vesicular disease-causing viruses. Each FMDV-negative sample generated a signal from the IC, ruling out amplification failures. A dilution series of an FMDV reference strain demonstrated
Ko, K.; Ha, K.; Park, S.; Kim, Y.; Lee, K.
Foot-and-mouth disease (FMD) or hoof-and-mouth disease is a severe plague for animal farming that affects cloven-hoofed animals such as cattle, pigs, sheep, and goats. Since it is highly infectious and can be easily proliferated by infected animals, contaminated equipments, vehicles, clothing, people, and predators. It is widely known that the virus responsible for FMD is a picornavirus, the prototypic member of the genus Aphthovirus. A serious outbreak of foot-and-mouth disease, leading to the stamping out of 3.53 millions of pigs and cattle and the construction of 4,538 burial sites until 15th March, 2011. The build-up of carcass burial should inevitably produce leachate by the decomposition of buried livestock affecting the surround environment such as air, soil, groundwater, and surface water. The most important issues which are currently raised by scientists are groundwater contamination by leachate from the livestock burial sites. This study examined the current status of FMD outbreak occurred in 2010-2011 and the issues of groundwater contamination by leachate from livestock burial sites. The hydrogeochemical, geophysical, and hydrogeological studies were executed to identify and trace groundwater contamination by leachate from livestock burial sites. Generally livestock mortality leachate contains high concentrations of NH3-N, HCO3-, Cl-, SO42-, K+, Na+, P along with relative lesser amounts of iron, calcium, and magnesium. The groundwater chemical data around four burial sites showed high NH3-N, HCO3-, and K+ suggesting the leachate leakage from burial sites. This is also proved by resistivity monitoring survey and tracer tests. The simulation results of leachate dispersion showed the persistent detrimental impacts for groundwater environment for a long time (~50 years). It is need to remove the leachate of burial sites to prevent the dispersion of leachate from livestock burial to groundwater and to monitor the groundwater quality. The most important
Foot and mouth disease (FMD) is a highly infectious and contagious viral disease affecting bovidae (cattle, zebus, domestic buffaloes, yaks), sheep, goats, swine, all wild ruminants and suidae. Camelidae (camels, dromedaries, llamas, vicunas) have low susceptibility. Foot and mouth disease is caused by a RNS virus of the family Picornaviridae, genus Aphthovirus. There are seven immunologically distinct serotypes: A, O, C, SAT1, SAT2, SAT3, Asia 1. Foot and mouth disease causes significant economic loss both to countries who manage it as an endemic disease (with or without vaccination), as well as those FMD free countries which may become infected. The mortality rate is low in adult animals, but often higher in young due to myocarditis. Foot and mouth disease is endemic in parts of Asia, Africa, the Middle East and South America (sporadic outbreaks in free areas). The Office of International Epizootics (OIE), also referred to the World Organization for Animal Health maintains an official list of free countries and zones.1 The OIE Terrestrial Code (Chapter 2.2.10) provides detailed information on the categories of freedom that can be allocated to a country as well as guidelines for the surveillance for foot and mouth disease (Appendix 3.8.7). In short, countries may be completely free of FMD, free with vaccination or infected with foot and mouth disease virus (FMDV). Source of FMDV include incubating and clinically affected animals with virus present in breath, saliva, faeces, urine, milk and semen. In experimental settings virus has been detected in milk several days before the onset of clinical signs2. Additional sources of virus are meat and by-products in which pH has remained above 6.0 as well as persistently infected carrier animals. Carrier animals may include cattle and water buffalo; convalescent animals and exposed vaccinates (virus persists in the oropharynx for up to 30 months in cattle or longer in buffalo, 9 months in sheep). Pigs do not become carriers