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establishment of persistent infection with foot-and-mouth disease virus in bhk-21 cells.abstract:201121492421
biosecurity and fmd transmission. 201121493485
isolation, identification and complete genome sequence analysis of a strain of foot-and-mouth disease virus serotype asia1 from pigs in southwest of china.abstract: backgroud: foot-and-mouth disease virus (fmdv) serotype asia1 generally infects cattle and sheep, while its infection of pigs is rarely reported. in 2005-2007, fmd outbreaks caused by asia1 type occurred in many regions of china, as well as some parts of east asia countries. during the outbreaks, there was not any report that pigs were found to be clinically infected. results: in this study, a strain of fmdv that isolated from pigs was identified as serotype asia1, and designated as "a ...201121496298
evidence of activation and suppression during the early immune response to foot-and-mouth disease virus.foot-and-mouth disease virus causes a serious disease of livestock species, threatening free global trade and food security. the disease spreads rapidly between animals, and to ensure a window of opportunity for such spread, the virus has evolved multiple mechanisms to subvert the early immune response. the cycle of infection in the individual animal is very short, infection is initiated, disseminated throughout the body and infectious virus produced in <7 days. foot-and-mouth disease virus has ...201121501424
development and validation of a prokaryotically expressed foot-and-mouth disease virus non-structural protein 2c'3ab-based immunochromatographic strip to differentiate between infected and vaccinated animals.abstract:201121513550
rna structural domains in non-coding regions of foot-and-mouth disease virus genome trigger innate immunity in porcine cells and mice.induction of type i interferons (ifn-α/β) in response to viral infection is a crucial step leading to the antiviral state in the host. viruses produce dsrna during their replication cycle that is sensed as non-self by host cells through different receptors. a signalling cascade is then activated to block viral replication and spread. foot-and-mouth disease virus (fmdv) is a picornavirus, highly sensitive to ifn, causing one of the world's most important animal diseases. in this study, we showed ...201121525336
an experimental infection in pigs using a foot-and-mouth disease virus isolated from the 2010 epidemic in japan.in this study, we carried out an experimental infection in pigs using a foot-and-mouth disease virus isolated from the 2010 epidemic in japan to analyze clinical manifestation, antibody response and virus sedding patterns in pigs. we found that the virus was virulent in pigs, producing a synchronous disease in the inoculated pigs and efficient spread to direct contact pigs. these results are useful for epidemiologically investigating the 2010 epidemic in japan and improving the measures for cont ...201121532260
identification of the role of rig-i, mda-5 and tlr3 in sensing rna viruses in porcine epithelial cells using lentivirus-driven rna interference.pathogen recognition receptors are essential for antiviral host immune responses. these specialized receptors detect conserved viral compounds and induce type i interferons (ifn) and pro-inflammatory cytokines. here we evaluated the contribution of rig-i, mda-5 and tlr3 to the recognition of classical swine fever (csfv), foot-and-mouth disease virus (fmdv), vesicular stomatitis virus (vsv) and influenza a virus (iav) to ifn-β responses in the porcine epithelial cell line pk-15. to this end, we i ...201121539869
development of one-step multiplex rt-pcr method for simultaneous detection and differentiation of foot-and-mouth disease virus serotypes o, a, and asia 1 circulating in vietnam.a one-step multiplex rt-pcr method using new primers was developed for the simultaneous detection and differentiation of vietnamese fmdv serotypes o, a, and asia 1 directly from clinical samples. the rt-pcr method used a cocktail of one universal minus-sense primer designed in the 2b gene and three serotype-specific plus-sense primers designed in the hypervariable regions of the capsid vp1 coding gene of fmdv. these serotype-specific primer pairs amplified 658, 535, and 427bp pcr products corres ...201121550367
relationship between clinical signs and transmission of an infectious disease and the implications for control.control of many infectious diseases relies on the detection of clinical cases and the isolation, removal, or treatment of cases and their contacts. the success of such "reactive" strategies is influenced by the fraction of transmission occurring before signs appear. we performed experimental studies of foot-and-mouth disease transmission in cattle and estimated this fraction at less than half the value expected from detecting virus in body fluids, the standard proxy measure of infectiousness. th ...201121551063
[construction and expression of a coxsackievirus a16 vp1 gene plasmid which delivered by live attenuated salmonella.]aim: to develop a coxsackievirus a16 (cox a16) vp1 gene plasmid which delivered by live attenuated salmonella. methods: the plasmid which expressed vp1 protein of coxa16 was constructed by gene recombination. cellular expression was assessed by western bloten analysis. then the recombinant attenuated salmonella which harboring the plasmid were constructed by electro transformation. results: coxa16 vp1 gene sequence was inserted into a eukaryotic expression plasmid. vp1 protein was detected in th ...201121557905
foot and mouth disease virus polyepitope protein produced in bacteria and plants induces protective immunity in guinea pigs.the goal of this project was to develop an alternative foot and mouth disease (fmd) vaccine candidate based on a recombinant protein consisting of efficient viral epitopes. a recombinant gene was designed that encodes b-cell epitopes of proteins vp1 and vp4 and t-cell epitopes of proteins 2c and 3d. the polyepitope protein (h-pe) was produced in e. coli bacteria or in n. benthamiana plants using a phytovirus expression system. the methods of extraction and purification of h-pe proteins from bact ...201121568869
analysis of the acute phase responses of serum amyloid a, haptoglobin and type 1 interferon in cattle experimentally infected with foot-and-mouth disease virus serotype o.abstract: a series of challenge experiments were performed in order to investigate the acute phase responses to foot-and-mouth disease virus (fmdv) infection in cattle and possible implications for the development of persistently infected "carriers". the host response to infection was investigated through measurements of the concentrations of the acute phase proteins (apps) serum amyloid a (saa) and haptoglobin (hp), as well as the bioactivity of type 1 interferon (ifn) in serum of infected anim ...201121592356
phylogenetic analysis of foot-and-mouth disease virus type o circulating in the andean region of south america during 2002-2008.at present, foot-and-mouth disease (fmd) has been successfully controlled in most territories of south america, where only ecuador and venezuela remain as endemic countries. in this context, the precise characterization of circulating viruses is of utmost importance. this work describes the first molecular epidemiology study performed with the complete vp(1)-coding region of 114 field isolates of fmd virus (fmdv) type o, collected in the andean countries mainly during 2002-2008. sequences were a ...201121601999
control of foot-and-mouth disease. 201121602410
high cleavage efficiency of a 2a peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice.when expression of more than one gene is required in cells, bicistronic or multicistronic expression vectors have been used. among various strategies employed to construct bicistronic or multicistronic vectors, an internal ribosomal entry site (ires) has been widely used. due to the large size and difference in expression levels between genes before and after ires, however, a new strategy was required to replace ires. a self-cleaving 2a peptide could be a good candidate to replace ires because o ...201121602908
comparative complete genome analysis of indian type a foot-and-mouth disease virus field isolates.comparative complete genome analysis of 17 serotype a indian field isolates representing different genotypes and sub-lineages is presented in this report. overall 79% of amino acids were invariant in the coding region. chunk deletion of nucleotide was observed in s and l fragment of 5'-utr. more variability which is comparable to that of capsid coding region was found in l and 3a region. functional motifs and residues critical for virus biology were conserved most. polyprotein cleavage sites acc ...201121604149
[diversity of receptor recognition site of type asial foot-and-mouth disease virus].foot-and-mouth disease virus (fmdv) initiates infection by binding to integrin receptors via a highly conserved arg-gly-asp (rgd) sequence found in the g-h loop of the structural protein vp1. however, fmdv is an rna virus, which harbors the evolutionary potential to render the rgd motif dispensable upon changes in constant environment. we studied mutation of rgd motif upon short-time passages fmdv asial/js/china/2005 field strain in different host.201121604558
the importance of fmdv localisation in lymphoid tissue.foot-and-mouth disease virus, a highly contagious pathogen that can cause lameness, low weight and decreased milk production, is a scourge of agricultural livestock around the world. although the acute phase of infection is rarely fatal, infection may persist in animals that have apparently recovered, creating a viral reservoir that some fear could contribute to the spread of disease. we have used an array of molecular techniques to search for traces of virus in tissues from the mouths and throa ...201121616546
rapid screening swine foot-and-mouth disease virus using micro-elisa system.in order to tackle both regional and global foot-and-mouth disease virus (fmdv) epdimics, we hereby develop a rapid microfluidic thermal lens microscopic method to screen swine type o fmdv with good efficiency. the scheme has great merits in terms of field portability, sample volume, assay time, analytical sensitivity, and test reproducibility.201121617821
multiplex rt-pcr detection and microarray typing of vesicular disease viruses.a vesicular disease multiplex reverse transcription (rt)-pcr with an accompanying microarray assay was developed for simultaneous detection and typing of foot-and-mouth disease virus (fmdv) and vesicular stomatitis virus (vsv), and for the detection of swine vesicular disease virus (svdv) and vesicular exanthema of swine virus (vesv). the multiplex rt-pcr successfully detected viral rna from a collection of 49 strains of vesicular viruses, including multiple strains from all seven serotypes of f ...201121620898
genes controlling vaccine responses and disease resistance to respiratory viral pathogens in cattle.farm animals remain at risk of endemic, exotic and newly emerging viruses. vaccination is often promoted as the best possible solution, and yet for many pathogens, either there are no appropriate vaccines or those that are available are far from ideal. a complementary approach to disease control may be to identify genes and chromosomal regions that underlie genetic variation in disease resistance and response to vaccination. however, identification of the causal polymorphisms is not straightforw ...201121621277
outbreak of hand, foot and mouth disease caused by coxsackie a16 virus in a childcare centre in croatia, february to march 2011. 201121632018
virus inactivation by salt (nacl) and phosphate supplemented salt in a 3d collagen matrix model for natural sausage casings.due to possible presence and spread of contagious animal viruses via natural sausage casings the international trade in these food products is subject to veterinary and public health requirements. in order to manage these restrictions we determined the effect of casing preservation on four highly contagious viruses for livestock: foot-and-mouth-disease virus (fmdv), classical swine fever virus (csfv), swine vesicular disease virus (svdv) and african swine fever virus (asfv). we used an in vitro ...201121632134
validation of a recombinant integrin αvβ6/monoclonal antibody based antigen elisa for the diagnosis of foot-and-mouth disease.a sandwich elisa using recombinant integrin αvβ6 as a capture ligand and serotype-specific monoclonal antibodies (mabs) as detecting reagents has been compared with a polyclonal antibody based elisa (using type-specific rabbit antibodies as capture and guinea pig antibodies as detectors), which is employed routinely at the fao world reference laboratory for foot-and-mouth disease (fmd), for the identification and serotyping of fmd virus (fmdv). the study used cell culture grown antigens (1351 fm ...201121635921
enhancement of astragalus polysaccharide on the immune responses in pigs inoculated with foot-and-mouth disease virus vaccine.the effects of astragalus polysaccharides (aps) on the immune response in pigs immunized with foot-and-mouth disease virus (fmdv) vaccine were investigated. fifteen pigs were randomly divided into five groups. four groups were vaccinated with a fmdv inactivated vaccine. pigs in three experimental groups were administered varying doses of aps (aps1, 5mg/kg; aps2, 10mg/kg; aps3, 20mg/kg). the influence of aps on the number of cd3(+)cd4(-)cd8(+) cytotoxic t cells, cd3(+)cd4(+)cd8(+) t helper memory ...201121640133
reducing animal experimentation in foot-and-mouth disease vaccine potency tests.the world organisation for animal health (oie) terrestrial manual and the european pharmacopoeia (ep) still prescribe live challenge experiments for foot-and-mouth disease virus (fmdv) immunogenicity and vaccine potency tests. however, the ep allows for other validated tests for the latter, and specifically in vitro tests if a "satisfactory pass level" has been determined; serological replacements are also currently in use in south america. much research has therefore focused on validating both ...201121640777
[expression and utilization of 3ab nonstructural protein of foot-and-mouth disease virus in escherichia coli].to develop a sensitive and specific elisa for detection of antibodies to the nonstructural protein of fmdv. we cloned and expressed fmdv nonstructural protein 3ab in escherichia coli expression system. the recombinant protein 3ab was purified with ni-nta hisbind resins and characterized by western blotting. an indirect elisa based on purified protein 3ab as a coating antigen was established. the specificity and sensitivity of this assay were evaluated by comparison with a commercial 3abc-elisa k ...201121648221
foot and mouth disease (fmd) virus: quantification of whole virus particles during the vaccine manufacturing process by size exclusion chromatography.foot and mouth disease (fmd) is a highly infectious viral disease that affects cattle, sheep, goats and swine causing severe economic losses worldwide. the efficacy of inactivated vaccines is critically dependent on the integrity of foot and mouth disease virus (fmdv) particles. the recommended method to quantify the active ingredient of vaccines is the 140s quantitative sucrose density gradient analysis. this method has been an immensely valuable tool over the past three decades but it is highl ...201121651939
effective inhibition of foot-and-mouth disease virus (fmdv) replication in vitro by vector-delivered micrornas targeting the 3d gene.foot-and-mouth disease virus (fmdv) causes an economically important and highly contagious disease of cloven-hoofed animals. rnai triggered by small rna molecules, including sirnas and mirnas, offers a new approach for controlling viral infections. there is no report available for fmdv inhibition by vector-delivered mirna, although mirna is believed to have more potential than sirna. in this study, the inhibitory effects of vector-delivered mirnas targeting the 3d gene on fmdv replication were e ...201121663611
immune potential of a novel multiple-epitope vaccine to fmdv type asia 1 in guinea pigs and sheep.to develop a safe and efficient recombinant subunit vaccine to foot-and-mouth disease virus (fmdv) type asia 1 in sheep, a tandem repeated multiple-epitope gene consisting of residues 137-160 and 197-211 of the vp1 gene of fmdv was designed and artificially synthesized. the biologically functional molecule, the ovine igg heavy constant region (oigg) as a protein carrier was introduced for design of the multiple-epitope recombinant vaccine and recombinant expression plasmids pet-30a-re and pet-30 ...201121667339
in silico analysis of ires rnas of foot-and-mouth disease virus and related picornaviruses.foot-and-mouth disease virus (fmdv) uses an internal ribosome entry site (ires), a highly structured segment of its genomic rna, to hijack the translational apparatus of an infected host. computational analysis of 162 type ii picornavirus ires rna sequences yielded secondary structures that included only base pairs supported by comparative or experimental evidence. the deduced helical sections provided the foundation for a hypothetical three-dimensional model of fmdv ires rna. the model was furt ...201121681504
predicting antigenic sites on the foot-and-mouth disease virus capsid of the south african territories (sat) types using virus neutralization data.foot-and-mouth disease virus (fmdv) outer capsid proteins 1b, 1c and 1d contribute to the virus serotype distribution and antigenic variants that exist within each of the seven serotypes. this study presents phylogenetic, genetic and antigenic analyses of south african territories (sat) serotypes prevalent in sub-saharan africa. here, we show that the high levels of genetic diversity in the p1-coding region within the sat serotypes are reflected in the antigenic properties of these viruses and t ...201121697350
[foot and mouth disease virus keeps lurking around the corner]. 201121702179
in-vitro and in-vivo phenotype of type asia 1 foot-and-mouth disease viruses utilizing two non-rgd receptor recognition sites.foot-and-mouth disease virus (fmdv) uses a highly conserved arg-gly-asp (rgd) triplet for attachment to host cells and this motif is believed to be essential for virus viability. previous sequence analyses of the 1d-encoding region of an fmdv field isolate (asia1/js/cha/05) and its two derivatives indicated that two viruses, which contained an arg-asp-asp (rdd) or an arg-ser-asp (rsd) triplet instead of the rgd integrin recognition motif, were generated serendipitously upon short-term evolution ...201121711567
detection of foot-and-mouth disease virus in the breath of infected cattle using a hand-held device to collect aerosols.exhaled air of individual cattle infected experimentally with foot-and-mouth disease virus (fmdv) was sampled to assess the feasibility of a rapid, non-invasive general screening approach for identifying sources of fmdv infection. the air sampler used was a handheld prototype device employing electrostatic particle capture in a microchip chamber of 10-15╬╝l and was shown to effectively capture a high percentage of airborne microorganisms. the particles were eluted subsequently from the chip cham ...201121723882
understanding foot-and-mouth disease virus early pathogenesis and immune responses. 201121733132
structural basis for the biological relevance of the invariant apical stem in ires-mediated translation.rna structure plays a fundamental role in internal initiation of translation. picornavirus internal ribosome entry site (ires) are long, efficient cis-acting elements that recruit the ribosome to internal mrna sites. however, little is known about long-range constraints determining the ires rna structure. here, we sought to investigate the functional and structural relevance of the invariant apical stem of a picornavirus ires. mutation of this apical stem revealed better performance of g:c compa ...201121742761
differential effects of viral vectors on migratory afferent lymph dendritic cells in vitro predicts enhanced immunogenicity in vivo.targeting dendritic cells (dc) is key to driving effective immune responses. lymphatic cannulation provides access to the heterogeneous populations of dc draining peripheral sites in rodents and ruminants. afferent lymph dec-205(+)cd11c(+)sirpa(+) dc were preferentially infected ex vivo with three vaccine viral vectors: replication defective human adenovirus-5 (rhuadv5); modified vaccinia virus ankara (rmva) and fowlpox virus (rfpv) all expressing green fluorescent protein (gfp). the rhuadv5 inf ...201121752909
a brief review on diagnosis of foot-and-mouth disease of livestock: conventional to molecular tools.foot-and-mouth disease (fmd) is one of the highly contagious diseases of domestic animals. effective control of this disease needs sensitive, specific, and quick diagnostic tools at each tier of control strategy. in this paper we have outlined various diagnostic approaches from old to new generation in a nutshell. presently fmd diagnosis is being carried out using techniques such as virus isolation (vi), sandwich-elisa (s-elisa), liquid-phase blocking elisa (lpbe), multiplex-pcr (m-pcr), and ind ...201121776357
translation of viral mrna without active eif2: the case of picornaviruses.previous work by several laboratories has established that translation of picornavirus rna requires active eif2a for translation in cell free systems or after transfection in culture cells. strikingly, we have found that encephalomyocarditis virus protein synthesis at late infection times is resistant to inhibitors that induce the phosphorylation of eif2a whereas translation of encephalomyocarditis virus early during infection is blocked upon inactivation of eif2a by phosphorylation induced by a ...201121779397
the complete nucleotide sequences of serotype 'o' and 'a' indian vaccine strains of foot-and-mouth disease virus.the primary genomic structure of indian foot-and-mouth disease virus (fmdv) type a and o vaccine strains was determined, and sequences coding for individual proteins were compared with those of several complete genomic sequences. the clustering based on the whole genome was reflected by that observed with vp1 alone. the indian vaccine strains clustered with currently circulating regional strains, and type a and type o vaccine seeds showed close nucleotide similarity with a22iraq and o1manisa, re ...201121779906
enterovirus 71 and coxsackievirus a16 3c proteases: binding to rupintrivir and their substrate, and anti-hfmd drug design.enterovirus 71 (ev71) and coxsackievirus a16 (cva16) are the major causative agents of hand foot and mouth disease (hfmd) which is prevalent in asia. thus far there are no prophylactic or therapeutic measures against hfmd. the 3c proteases from ev71 and cva16 play important roles in viral replication and are therefore ideal drug targets. by using biochemical, mutational and structural approaches, we broadly characterized both proteases. a series of high-resolution structures of the free or subst ...201121795339
chemical disinfection of high-consequence transboundary animal disease viruses on nonporous surfaces.disinfection is a critical part of the response to transboundary animal disease virus (tadv) outbreaks by inactivating viruses on fomites to help control infection. to model the inactivation of tadv on fomites, we tested selected chemicals to inactivate foot and mouth disease virus (fmdv), african swine fever virus (asfv), and classical swine fever virus (csfv) dried on steel and plastic surfaces. for each of these viruses, we observed a 2 to 3 log reduction of infectivity due to drying alone. w ...201121798759
proteomics analysis of porcine serum proteins by lc-ms/ms after foot-and- mouth disease virus (fmdv) infection.to analyze serum proteomics differences between normal and foot and mouth disease virus (fmdv)-infected piglets, an analytical method based on liquid chromatography with tandem mass spectrometry (lc-ms/ms) was used. samples of venous blood were collected before and after fmdv infection and high abundance serum albumin was removed using a commercial kit. after trypsin digestion, serum samples were processed with lc-ms/ms. proteins were identified by peptide mass fingerprinting. we found that apol ...201121799299
the codon usage model of the context flanking each cleavage site in the polyprotein of foot-and-mouth disease virus.to investigate the codon usage pattern of the contexts flanking 11 cleavage sites of foot-and-mouth disease virus (fmdv) polyprotein, the codon usage model of the corresponding codon position and the synonymous codon usage in the target contexts of 66 strains were characterized by two simple methods based on the relative synonymous codon usage value. the synonymous codons usage pattern was also compared between this virus and two species of hosts (cattle and domestic pig). it is indicated that f ...201121801856
combining livestock trade patterns with phylogenetics to help understand the spread of foot and mouth disease in sub-saharan africa, the middle east and southeast asia.international trade in animals and their products is recognised as a primary determinant of the global epidemiology of transboundary diseases such as foot and mouth disease (fmd). as well as causing serious production losses, fmd is highly contagious, being transmitted through multiple routes and hosts, which makes it one of the most important diseases affecting trade in livestock. its occurrence has dramatic consequences for the agricultural economy of a normally disease-free country, as well a ...201121809754
expression of foot and mouth disease virus nonstructural polyprotein 3abc with inactive 3c(pro) in escherichia coli.nonstructural 3abc protein of foot and mouth disease virus (fmdv) was widely used to differentiate vaccinated from natural fmdv-infected animals. 3abc is a polyprotein which is auto-processed to 3a, three copies of 3b and 3c(pro) by 3c(pro) protease. the 3abc gene was cloned and expressed in escherichia coli as native or mutated 3abc (mu3abc) forms. cysteine residues 142 and 163 of the catalytic triad within the 3c(pro) of mu3abc were changed to serine and glycine, respectively, to inhibit its p ...201121810474
mechanisms governing the selection of translation initiation sites on foot-and-mouth disease virus rna.translation initiation dependent on the foot-and-mouth disease virus internal ribosome entry site (ires) occurs at two sites (lab and lb), 84 nt apart. in vitro translation of an mrna comprising the ires and lab-lb intervening segment fused to a cat reporter has been used to study the parameters influencing the ratio of the two products and the combined product yield, as measures of relative initiation site usage and productive ribosome recruitment, respectively. with wild-type mrna ~40% of init ...201121813609
evolutionary analysis of serotype a foot-and-mouth disease viruses circulating in pakistan and afghanistan during 2002-2009.foot-and-mouth disease (fmd) is endemic in pakistan and afghanistan. three different serotypes of the virus, namely o, a and asia-1, are responsible for the outbreaks of this disease in these countries. in the present study, the nucleotide (nt) coding sequences for the vp1 capsid protein (69 samples) or for all four capsid proteins (p1, 7 representative samples) of the serotype a fmd viruses circulating in pakistan and afghanistan were determined. phylogenetic analysis of the fmdv vp1 coding seq ...201121813704
a dna vaccine encoding foot-and-mouth disease virus b and t-cell epitopes targeted to class ii swine leukocyte antigens protects pigs against viral challenge.development of efficient and safer vaccines against foot-and-mouth disease virus (fmdv) is a must. previous results obtained in our laboratory have demonstrated that dna vaccines encoding b and t cell epitopes from type c fmdv, efficiently controlled virus replication in mice, while they did not protect against fmdv challenge in pigs, one of the fmdv natural hosts. the main finding of this work is the ability to improve the protection afforded in swine using a new dna-vaccine prototype (pcmv-apc ...201121820470
Comparative study of codon substitution patterns in foot-and-mouth disease virus (serotype O).We compared genetic variations in the VP1 gene of foot-and-mouth disease viruses (FMDVs) isolated since 2000 according to the region of the world from. We analyzed relative synonymous codon usage (RSCU) and phylogenetic relationship between geographical regions, and calculated the genetic substitution patterns between Korean isolate and those from other countries. We calculated the ratios of synonymously substituted codons (SSC) to all observed substitutions and developed a new analytical parame ...201121825834
recombinant vp1, an akt inhibitor, suppresses progression of hepatocellular carcinoma by inducing apoptosis and modulation of ccl2 production.the application of viral elements in tumor therapy is one facet of cancer research. recombinant capsid protein vp1 (rvp1) of foot-and-mouth disease virus has previously been demonstrated to induce apoptosis in cancer cell lines. here, we aim to further investigate its apoptotic mechanism and possible anti-metastatic effect in murine models of hepatocellular carcinoma (hcc), one of the most common human cancers worldwide.201121826248
detection of foot-and-mouth disease virus rna in pharyngeal epithelium biopsy samples obtained from infected cattle: investigation of possible sites of virus replication and persistence.foot-and-mouth disease (fmd) is a highly contagious viral infection of significant financial importance to the export and trade of agricultural products. the occurrence of persistently infected "carriers" of fmd-virus (fmdv) in ruminant species adds further complications to disease control. there have been significant discrepancies in reports regarding the pathogenesis of fmdv infection in cattle with specific emphasis on the anatomical sites involved in early and persistent virus replication. i ...201121831538
reanalysis of the start of the uk 1967 to 1968 foot-and-mouth disease epidemic to calculate airborne transmission probabilities.the aims of this study were to statistically reassess the likelihood that windborne spread of foot-and-mouth disease (fmd) virus (fmdv) occurred at the start of the uk 1967 to 1968 fmd epidemic at oswestry, shropshire, and to derive dose-response probability of infection curves for farms exposed to airborne fmdv. to enable this, data on all farms present in 1967 in the parishes near oswestry were assembled. cases were infected premises whose date of appearance of first clinical signs was within ...201121846685
develope monoclonal antibody against foot-and-mouth disease virus a type.in order to develop an anti-fmdv a type monoclonal antibody (mab), babl/c mice were immunized with fmdv a type. monoclonal antibodies (mabs) 7b11 and 8h4 against foot-and-mouth disease virus (fmdv) serotype a were produced by fusing sp2/0 myeloma cells with splenocyte from the mouse immunized with a/av88. the microneutralization titer of the mabs 7b11 and 8h4 were 1024 and 512, respectively. both mabs contain kappa light chains, the mabs were igg1. in order to define the mabs binding epitopes, t ...201121847759
recombinant non-structural polyprotein 3ab-based serodiagnostic strategy for fmd surveillance in bovines irrespective of vaccination.in india, the proportion of bovines vaccinated against foot-and-mouth disease (fmd) is increasing since the implementation of the government supported 'fmd control programme', and non-structural protein (nsp)-based serological assays for discriminating between antibodies induced by infection or vaccination (diva) could be useful. the fmd virus nsp 3ab was expressed in a prokaryotic system and an indirect elisa (r3ab(3) i-elisa) was developed and validated as a screening assay for detecting virus ...201121864578
Modulation of Cytokine mRNA Expression in Pharyngeal Epithelial Samples obtained from Cattle Infected with Foot-and-Mouth Disease Virus.A novel technique of endoscopical collection of small tissue samples was used to obtain sequential tissue samples from the dorsal soft palate (DSP) of individual cattle infected with foot-and-mouth disease virus (FMDV) at different phases of the infection. Levels of mRNA encoding interferon (IFN)-a and IFN-ß as well as tumour necrosis factor (TNF)-a were measured in these samples by quantitative reverse transcriptase polymerase chain reaction. Expression of IFN-ß mRNA was significantly down-regu ...201121868026
cpg oligodeoxynucleotide and montanide isa 206 adjuvant combination augments the immune responses of a recombinant fmdv vaccine in cattle.foot-and-mouth disease (fmd) is a highly contagious disease of cloven-hoofed animals. to prevent the spread of fmdv, inactivated virus vaccines are used to immunize animals in developing countries. however, there are safety concerns. in addition, it is difficult to distinguish the vaccinated animals from those naturally infected ones. in our lab, we have developed a recombinant fmdv vaccine named a7. a7 contained multiple b cell and t cell epitopes, which reside in a capsid protein (vp1) of fmdv ...201121872635
Role of macrophages in early protective immune responses induced by two vaccines against foot and mouth disease.Foot and Mouth Disease (FMD) is an acute disease of cloven-hoofed species. We studied the protection and early immune response induced in the murine model by vaccines formulated with inactivated virus and two different adjuvants. The presence of IMS12802PR or ISA206VG adjuvants yielded protection against viral challenge at early times post vaccination and induced FMDV-specific, but non neutralizing, antibody titers. In vivo macrophage depletion in vaccinated mice severely decreased the protectio ...201121878353
Development of a serotype colloidal gold strip using monoclonal antibody for rapid detection type Asia1 foot-and-mouth disease.In this study, we developed a rapid, one step colloid gold strip (CGS) capable of specifically detecting type Asia1 foot-and-mouth disease virus (FMDV). We have produced two monoclonal antibodies (mAb) to type Asia1 FMD (named 1B8 and 5E2). On the test strip, the purified 1B8 labelled with the colloidal gold was used as the detector, and the purified 5E2 and goat anti-mouse antibodies were wrapped onto nitrocellulose (NC) membranes as the test and the control line, respectively. The rapid colloi ...201121880157
impact of gene vector design on the control of recombinant monoclonal antibody production by chinese hamster ovary cells.in this study, we systematically compare two vector design strategies for recombinant monoclonal antibody (mab) synthesis by chinese hamster ovary (cho) cells; a dual open reading frame (orf) expression vector utilizing separate cytomegalovirus (cmv) promoters to drive heavy chain (hc) and light chain (lc) expression independently, and a single orf vector design employing a single cmv promoter to drive hc and lc polypeptide expression joined by a foot and mouth disease virus f2a polypeptide self ...201121882365
An overview on ELISA techniques for FMD.FMD is one of the major causes of economic loss of cloven-hoofed animals in the world today. The assessment of dominant genotype/lineage and prevalent trends and confirmation the presence of infection or vaccination not only provides scientific basis and first-hand information for appropriate control measure but also for disease eradication and regaining FMD free status following an outbreak. Although different biological and serological approaches are still applied to study this disease, ELISA ...201121888667
vesicular stomatitis virus glycoprotein g carrying a tandem dimer of foot and mouth disease virus antigenic site a can be used as dna and peptide vaccine for cattle.effective foot and mouth disease virus (fmdv) peptide vaccines for cattle have two major constraints: resemblance of one or more of the multiple conformations of the major vp1 antigenic sites to induce neutralizing antibodies, and stimulation of t cells despite the variable bovine-mhc polymorphism. to overcome these limitations, a chimeric antigen was developed, using vesicular stomatitis virus glycoprotein (vsv-g) as carrier protein of an in tandem-dimer of fmdv antigenic site a (asa), the majo ...201121889542
developing vaccines against foot-and-mouth disease and some other exotic viral diseases of livestock.vaccines remain the main tool for the control of livestock viral diseases that pose a serious threat to animal and occasionally human health, reduce food security, distort trade in animals and their products, and undermine agricultural development in poor countries. globalization and climate change increase the likelihood for new patterns of emergence and spread of livestock viruses. conventionally attenuated and killed virus products have had spectacular success, and recent examples include the ...201121893540
Factors associated with within-herd transmission of serotype A foot-and-mouth disease virus in cattle, during the 2001 outbreak in Argentina: a protective effect of vaccination.Argentina suffered an extensive foot-and-mouth disease (FMD) epidemic between July 2000 and January 2002, 3 months after obtaining the official FMD-free without vaccination status conferred by the World Organization for Animal Health. This is one of the largest FMD epidemics controlled by implementation of a systematic mass vaccination campaign in an FMD-free country. In 2000, 124 herds were reported as FMD positive, 2394 herds in 2001 and one in January 2002; the total number of cattle herds in ...201121896154
Identification of H-2d restricted T cell epitope of foot-and-mouth disease virus structural protein VP1.Foot-and-mouth disease (FMD) is a highly contagious and devastating disease affecting livestock that causes significant financial losses. Therefore, safer and more effective vaccines are required against Foot-and-mouth disease virus(FMDV). The purpose of this study is to screen and identify an H-2d restricted T cell epitope from the virus structural protein VP1, which is present with FMD. We therefore provide a method and basis for studying a specific FMDV T cell epitope.201121896206
Lentviral-mediated RNAi to inhibit target gene expression of the porcine integrin av subunit, the FMDV receptor, and against FMDV infection in PK-15 cells.shRNA targeting the integrin av subunit, which is the foot-and-mouth disease virus (FMDV) receptor, plays a key role in virus attachment to susceptible cells. We constructed a RNAi lentiviral vector, iav pLenti6/BLOCK -iT™, which expressed siRNA targeting the FMDV receptor, the porcine integrin av subunit, on PK-15 cells. We also produced a lentiviral stock, established an iav-PK-15 cell line, evaluated the gene silencing efficiency of mRNA using real-time qRT-PCR, integrand av expression by ind ...201121899738
characterization of foot-and-mouth disease virus from outbreaks in ecuador during 2009-2010 and cross-protection studies with the vaccine strain in use in the region.during the years 2009 and 2010 relevant epidemic waves of foot-and-mouth disease (fmd) serotype o occurred in ecuador, representing a great drawback for the last stages of the ongoing eradication program in south america. this study describes the molecular and antigenic characterizations of 29 isolates collected from various regions in the country and their relationship to the vaccine strain. the phylogenetic tree derived from sequences spanning the complete vp(1) protein showed that, despite th ...201121911020
Tempo and mode of inhibitor-mutagen antiviral therapies: a multidisciplinary approach.The continuous emergence of drug-resistant viruses is a major obstacle for the successful treatment of viral infections, thus representing a persistent spur to the search for new therapeutic strategies. Among them, multidrug treatments are currently at the forefront of pharmaceutical, clinical, and computational investigation. Still, there are many unknowns in the way that different drugs interact among themselves and with the pathogen that they aim to control. Inspired by experimental studies w ...201121911373
lethal mutagenesis of foot-and-mouth disease virus involves shifts in sequence space.lethal mutagenesis or virus transition into error catastrophe is an antiviral strategy that aims at extinguishing a virus by increasing the viral mutation rates during replication. the molecular basis of lethal mutagenesis is largely unknown. previous studies showed that a critical substitution in the foot-and-mouth disease virus (fmdv) polymerase was sufficient to allow the virus to escape extinction through modulation of the transition types induced by the purine nucleoside analogue ribavirin. ...201121917974
Use of inactivated Escherichia coli enterotoxins to enhance respiratory mucosal adjuvanticity during vaccination in swine.In order to augment responses to respiratory vaccines in swine, various adjuvants were intranasally coadministered with a foot-and-mouth disease virus (FMDV) antigen to pigs. Detoxified Escherichia coli enterotoxins LTK63 and LTR72 enhanced antigen-specific mucosal and systemic immunity, demonstrating their efficacy as adjuvants for nonreplicating antigens upon intranasal immunization in swine.201121918121
Suicidal gene therapy in the effective control of primary human hepatocellular carcinoma as monitored by noninvasive bioimaging.Hepatocellular carcinoma (HCC) is usually refractory to the available treatments. For cancer gene therapy purposes, real-time imaging of therapeutic gene expression is of great importance because there are multiple factors that modulate the therapeutic gene expression in a complex tumor microenvironment. As a consequence, multiple doses of therapeutic viral vectors may be required for improved efficacy. In the present study, the luciferase reporter gene and the yeast cytosine deaminase (yCD) gen ...201121918545
Genetic characterization of a new pandemic Southeast Asia topotype strain of serotype O foot-and-mouth disease virus isolated in China during 2010.The full-length nucleotide sequence of the foot-and-mouth disease virus O/BY/CHA/2010 strain, Mya-98 lineage of Southeast Asia (SEA) topotype, was determined and compared with O/HKN/20/2010 and other known FMDV strains. Homology analysis indicated >98.0% nucleotide identity between O/BY/CHA/2010 and the epidemic strains, O/HKN/20/2010, and O/VN/2009. However, with the exception of the VP4, 2A, and 3BCD regions, O/BY/CHA/2010 showed a lower similarity with SEA topotype strains, O/VN/2006, and HLJ ...201121932049
Nanobodies®: new ammunition to battle viruses.In 1989, a new type of antibody was identified, first in the sera of dromedaries and later also in all other species of the Camelidae family. These antibodies do not contain a light chain and also lack the first constant heavy domain. Today it is still unclear what the evolutionary advantage of such heavy chain-only antibodies could be. In sharp contrast, the broad applicability of the isolated variable antigen-binding domains (VHH) was rapidly recognized, especially for the development of thera ...201121939690
Validation of an r3AB1-FMDV-NSP ELISA to distinguish between cattle infected and vaccinated with foot-and-mouth disease virus.Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed livestock which has a drastic economic impact for affected countries. Although FMDV is distributed worldwide, many regional programs have been effective eradicating this agent. In Argentina, as in many other regions of South America, the combination of a systematic vaccination plan, together with an effective detection system capable of differentiating infection from vaccination, has been successful for eradicating this ...201121946290
foot-and-mouth disease virus (fmdv) leader proteinase negatively regulates the porcine interferon-λ1 pathway.foot-and-mouth disease is a highly contagious viral disease caused by foot-and-mouth disease virus (fmdv) of wild and domestic cloven-hoofed animals, and causes an economically important disease in the swine industry. in this study, we found that the replication of fmdv in ibrs-2 cells could be significantly inhibited after treatment with the purified recombinant porcine interferon lambda 1 (ifn-λ1), a newly identified type iii interferon. however, fmdv could not activate the ifn-λ1 promoter and ...201121975014
molecular characterization of serotype asia-1 foot-and-mouth disease viruses in pakistan and afghanistan; emergence of a new genetic group and evidence for a novel recombinant virus.foot-and-mouth disease (fmd) is endemic in pakistan and afghanistan. the fmd virus serotypes o, a and asia-1 are responsible for the outbreaks in these countries. diverse strains of fmdv, even within the same serotype, co-circulate. characterization of the viruses in circulation can facilitate appropriate vaccine selection and tracing of outbreaks. the present study characterized foot-and-mouth disease serotype asia-1 viruses circulating in pakistan and afghanistan during the period 1998-2009. p ...201121983559
field application of a recombinant protein-based elisa during the 2010 outbreak of foot-and-mouth disease type a in south korea.a recombinant protein-based enzyme-linked immunosorbent assay (rp elisa) exists for the detection of antibodies to foot-and-mouth disease virus (fmdv) type a. in this study, the efficacy of the rp elisa was compared to that of other current tests by examining sera collected in the field during an fmd type a outbreak in south korea in 2010. the rp elisa detected early antibodies to fmdv with the same sensitivity as the liquid-phase blocking elisa (lpb elisa), identifying fmd farm outbreaks correc ...201222001272
Cattle remain immunocompetent during the acute phase of foot-and-mouth disease virus infection.ABSTRACT: Infection of cattle with foot-and-mouth disease virus (FMDV) results in the development of long-term protective antibody responses. In contrast, inactivated antigen vaccines fail to induce long-term protective immunity. Differences between susceptible species have also been observed during infection with FMDV, with cattle often developing persistent infections whilst pigs develop more severe symptoms and excrete higher levels of virus. This study examined the early immune response to F ...201122014145
inoculation of newborn mice with non-coding regions of foot-and-mouth disease virus rna can induce a rapid, solid and wide-range protection against viral infection.we have recently described the ability of in vitro-transcribed rnas, mimicking structural domains in the 5' and 3' non-coding regions (ncrs) of the foot-and-mouth disease virus (fmdv) genome, to trigger the innate immune response in porcine cultured cells and mice. in this work, the antiviral effect exerted in vivo by these small synthetic non-infectious rna molecules was analyzed extensively. the susceptibility of transfected newborn swiss mice to fmdv challenge was tested using a wide range of ...201122020303
increased efficacy of an adenovirus-vectored foot-and-mouth disease capsid subunit vaccine expressing nonstructural protein 2b is associated with a specific t cell response.we previously demonstrated that an adenovirus-based foot-and-mouth disease virus (fmdv) serotype a24 capsid subunit vaccine, ad5-a24, expressed under the control of a cytomegalovirus promoter (cmv) can protect swine and bovines against homologous challenge, but in a similar approach using swine vaccinated with an ad5-vectored fmdv o1 campos vaccine, ad5-o1c, the animals were only partially protected when challenged at 21 days post-vaccination (dpv). recently, we demonstrated that inclusion of th ...201122027486
foot-and-mouth disease virus particles inactivated with binary ethylenimine are efficiently internalized into cultured cells.conventional foot-and-mouth disease (fmd) vaccines are produced from virus grown in cell culture that is chemically inactivated by using binary ethylenimide (bei). here, we show that bei treatment preserves both the architecture of fmdv particles, as inactivated viral particles showed by electron microscopy characteristics similar to those of infectious virions, as well as the general features of infectious virus internalization. binding of inactivated particles to bhk-21 cells was blocked by pr ...201122027488
role of arginine-56 within the structural protein vp3 of foot-and-mouth disease virus (fmdv) o1 campos in virus virulence.fmdv o1 subtype undergoes antigenic variation under diverse growth conditions. of particular interest is the amino acid variation observed at position 56 within the structural protein vp3. selective pressures influence whether histidine (h) or arginine (r) is present at this position, ultimately influencing in vitro plaque morphology and in vivo pathogenesis in cattle. using reverse genetics techniques, we have constructed fmdv type o1 campos variants differing only at vp3 position 56, possessin ...201222036313
rapid typing of foot-and-mouth disease serotype asia 1 by reverse transcription loop-mediated isothermal amplification.a reverse transcriptase loop-mediated isothermal amplification (rt-lamp) assay was rapidly used to detect serotype asia 1 of foot-and-mouth disease virus (fmdv) within 45 min at 61°c. all fmdv serotype asia 1 reference strains were positive by rt-lamp, while other viruses such as fmdv serotypes o, c, a and classical swine fever virus, swine vesicular disease virus, porcine reproductive and respiratory syndrome virus and japanese encephalitis virus remained negative. furthermore, fmdv sreotype as ...201122040459
colour determination of beef juices as an indicator of beef cooking temperatures.dominant wavelengths and other colour parameters of beef juices were investigated as predictors of maximum internal temperatures attained during beef cooking since beef imported by the usa from south american countries has to be cooked to a temperature compatible with the inactivation of the foot-and-mouth disease virus. although the correlation between these parameters was highly significant, the prediction error was high and similar in magnitude to that obtained with other methods. however, ac ...197922055351
Investigation of serum protein profiles in sheep naturally infected with foot-and-mouth disease virus.This study was carried out to determine serum protein profiles in naturally infected sheep with foot-and-mouth disease virus (FMDV). The study material consisted of twelve healthy and 36 sheep with foot-and-mouth disease (FMD). FMD had been diagnosed on the basis of clinical findings and results of serological examination. Serotypes serologically detected in the FMDV-infected sheep were as follows: O (n = 11), A (n = 8) and mixed infection with serotypes O, A and Asia-1 (n = 17).The total protei ...201122059288
Establishment of a Stable Cell Line Coexpressing Dengue Virus-2 and Green Fluorescent Protein for Screening of Antiviral Compounds.This study aimed to generate a stable cell line harboring subgenomic dengue virus replicon and a green fluorescent gene (DENV/GFP) for a cell-based model to screen anti-DENV compounds. The gene-encoding envelope protein of DENV-2 was deleted and then replaced with fragments of the GFP gene and a foot-and-mouth-disease virus 2A-derived cleavage site. The human cytomegalovirus immediate early and antisense hepatitis delta virus ribozyme sequences were added at the 5'- and 3'-ends. An internal ribo ...201122068705
detection of foot-and-mouth disease virus rna by reverse transcription loop-mediated isothermal amplification.a reverse transcription loop-mediated isothermal amplification (rt-lamp) assay was developed for foot-and-mouth disease virus (fmdv) rna. the amplification was able to finish in 45 min under isothermal condition at 64°c by employing a set of four primers targeting fmdv 2b. the assay showed higher sensitivity than rt-pcr. no cross reactivity was observed from other rna viruses including classical swine fever virus, swine vesicular disease, porcine reproductive and respiratory syndrome virus, japa ...201122070774
differentiating foot-and-mouth disease virus-infected from vaccinated animals with baculovirus-expressed specific proteins.abstract we had shown in preliminary studies with a small number of animals that antibodies against 2c could be detected in cattle and pigs which had been infected with fmdv but not in animals which had been vaccinated against the disease. to determine whether this test was generally applicable, seta from several hundred animals which had been vaccinated with different products in many countries have been tested in an elisa using baculovirus expressed 2c. our results show that only 1-2% of the s ...199822077289
cattle response to foot-and-mouth disease virus nonstructural proteins as antigens within vaccines produced using different concentrations.abstract four groups of ten nine-month-old nelore heifers were used for this study. each group received one of four foot-and-mouth disease (fmd) trivalent vaccines for the duration of the experiment. the four vaccine formulations (normal, 2x, 4x and 8x) differed in 140s content to determine the serological reactivities to fmd virus (fmdv) nonstructural proteins 2c, 3abc and 3d. vaccination was by the intramuscular administration of vaccine on day 0, 180 and 360. bleedings were done at 30 days po ...199822077290
diagnostic potential of mab-based elisas for antibodies to non-structural proteins of foot-and-mouth disease virus to differentiate infection from vaccination.abstract this paper summarises the development of monoclonal antibody (mab)-based immunoassays measuring antibodies to non-structural proteins of fmdv to differentiate infection from vaccination. of the three non-structural proteins 2c, 3c and 3abc evaluated in this study, the polypeptide 3abc was the most immunogenic. three elisas for the detection of antibodies to 3abc were developed. two assays rely on the competition of test sera against either a anti-3a mab or against antisera to 3abc raise ...199822077292
rt-pcr in foot-and-mouth.summary a rt-pcr assay for the specific detection of rna sequences from foot-and-mouth disease virus (fmdv) has been developed. the procedure permits also the detection of sequences that correlate with established fmdv serotypes. a computer program that allows selection of genotype-specific primers for rt-pcr amplification was used for the identification of fmdv specific sequences for pcr amplification on rna replicase (3d) gene regions. specific, rapid and highly sensitive detection was achieve ...199822077295
detection of foot-and-mouth disease by reverse transcription polymerase chain reaction and virus isolation in contact sheep without clinical signs of foot-and-mouth disease.summary two non-vaccinated sheep were experimentally, infected with fmdv and one day later 4 other sheep were brought in contact. although the contact sheep showed no clinical signs, serology indicated that all sheep became infected. various secretion samples, taken over a period of at least one month, and various tissue samples were examined for the presence of fmdv by rt-pcr and by virus isolation. fmdv was most often found in saliva (mouth swabs), followed by nasal secretion and sera. faecal ...199822077296
Foot and mouth disease leader protease (Lb(pro)): Investigation of prime side specificity allows the synthesis of a potent inhibitor.Foot and mouth disease virus expresses its genetic information as a single polyprotein that is translated from the single-stranded RNA genome. Proteinases contained within the polyprotein then generate the mature viral proteins. The leader protease (Lb(pro)) performs the initial cleavage by freeing itself from the growing polypeptide chain; subsequently, Lb(pro) cleaves the two homologues of the host cell protein eukaryotic initiation factor 4G (eIF4G). We showed that Lb(pro) possesses specific ...201122085639
Normal variation in thermal radiated temperature in cattle: implications for foot-and-mouth disease detection.ABSTRACT:201122104039
intranasal delivery of cationic plga nano/microparticles-loaded fmdv dna vaccine encoding il-6 elicited protective immunity against fmdv challenge.mucosal vaccination has been demonstrated to be an effective means of eliciting protective immunity against aerosol infections of foot and mouth disease virus (fmdv) and various approaches have been used to improve mucosal response to this pathogen. in this study, cationic plga (poly(lactide-co-glycolide)) nano/microparticles were used as an intranasal delivery vehicle as a means administering fmdv dna vaccine encoding the fmdv capsid protein and the bovine il-6 gene as a means of enhancing muco ...201122110686
Inoculation of swine with foot-and-mouth disease SAP-mutant virus induces early protection against disease.Foot-and-mouth disease virus (FMDV) leader proteinase (L(pro)) cleaves itself from the viral polyprotein and cleaves the translation initiation factor eIF4G. As a result, host cell translation is inhibited, affecting the host innate immune response. We have demonstrated that L(pro) is also associated with degradation of nuclear factor ?B (NF-?B), a process that requires L(pro) nuclear localization. Additionally, we reported that disruption of a conserved protein domain within the L(pro) coding s ...201122114339
Rapid detection of foot-and-mouth disease virus using a field-portable nucleic acid extraction and real-time PCR amplification platform.Rapid and accurate field diagnostic tools can be used to support clinical diagnosis during outbreaks of exotic livestock diseases. This study evaluated a mobile PCR amplification platform that performs RNA extraction, real-time reverse-transcription PCR (rRT-PCR) and interpretation of results without operator intervention. Initial studies showed that there was equivalence between the detection limit generated by RNA extracted using the mobile platform and an automated laboratory-based system. In ...201122115952
disinfection of foot-and-mouth disease and african swine fever viruses with citric acid and sodium hypochlorite on birch wood carriers.transboundary animal disease viruses such as foot-and-mouth disease virus (fmdv) and african swine fever virus (asfv) are highly contagious and cause severe morbidity and mortality in livestock. proper disinfection during an outbreak can help prevent virus spread and will shorten the time for contaminated agriculture facilities to return to food production. wood surfaces are prevalent at these locations, but there is no standardized method for porous surface disinfection; commercial disinfectant ...201122115968
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