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inactivation of foot-and-mouth disease virus in various bovine tissues used for the production of natural sausage casings.bovine intestines, bladders and oesophagus are used for the production of natural casings ("beef casings") as edible sausage containers. derived from cattle experimentally infected with fmdv (initial dosage 10(4) tcid(50)/ml, strain a iran 97), these beef casings were treated with sodium chloride (nacl) or phosphate supplemented salt (p-salt). in addition, different in-vitro experiments using beef casings were done on a small scale with other fmdv strains (a turkey 06, c-oberbayern and o(1) mani ...201122137684
the infectivity and pathogenicity of a foot-and-mouth disease virus persistent infection strain from oesophageal-pharyngeal fluid of a chinese cattle in 2010.abstract: background: foot-and mouth disease (fmd) is an acute, febrile, and contagious vesicular disease affecting cloven-hoofed animals. some animals may become persistent infected carriers when they contact fmd virus (fmdv), and persistent infected animals are a dangerous factor to cause fmd outbreak. findings: 300 op (oesophageal-pharyngeal) fluid samples were collected from cattle without clinic symptom after one month fmd circulated in 2010 in china. a fmdv strain was isolated when a posi ...201122166050
genetic characterization of vaccine and field strains of serotype a foot-and-mouth disease virus from india.extreme antigenic and genetic heterogeneity of serotype a foot-and-mouth disease virus (fmdv) population has resulted in change of vaccine strains in india twice in the last decade. in such a situation, complete characterization of the vaccine strains is imperative. with regard to the frequent outbreaks of this disease, fmdv field strains are also of interest. therefore three vaccine strains and two field strains of type a fmdv from india were completely sequenced and the obtained sequences were ...201122149500
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
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
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
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
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
molecular characterization of amino acid deletion in vp1 (1d) protein and novel amino acid substitutions in 3d polymerase protein of foot and mouth disease virus subtype a/iran87.the nucleotide sequence of the vp1 (1d) and partial 3d polymerase (3d(pol)) coding regions of the foot and mouth disease virus (fmdv) vaccine strain a/iran87, a highly passaged isolate (~150 passages), was determined and aligned with previously published fmdv serotype a sequences. overall analysis of the amino acid substitutions revealed that the partial 3d(pol) coding region contained four amino acid alterations. amino acid sequence comparison of the vp1 coding region of the field isolates reve ...201122122902
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
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
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
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
detection and genetic characterization of foot-and-mouth disease viruses in samples from clinically healthy animals in endemic settings.a total of 1501 oral swab samples from pakistan, afghanistan and tajikistan were collected from clinically healthy animals between july 2008 and august 2009 and assayed for the presence of foot-and-mouth disease virus (fmdv) rna. the oral swab samples from two (of four) live animal markets in pakistan (n = 245), one (of three) live animal market in afghanistan (n=61) and both the live animal markets in tajikistan (n=120) all tested negative. however, 2 of 129 (∼2%) samples from gondal and 11 of ...201122212855
tissue culture independent transformation of the forage crop sunnhemp (crotalaria juncea l.): an easy method towards generation of transgenics.a transformation system which is free of in vitro plant regeneration following agrobacterium infection is established for the forage legume, sunnhemp (crotalaria juncea l.) where in the entire embryo axis of the germinating seed was used as the target tissue for transformation. after standardization of transformation conditions, the cotyledonary node of the embryo axis was infected with agrobacterium host lba 4404 harboring the recombinant vector pcambia 2301. the bivalent 1d gene of the two maj ...201123573040
[early steps of picornavirus infection].picornaviridae is a large family of viruses that cause a variety of infectious diseases in humans and animals. it includes important viruses such as poliovirus, hepatisis a virus and foot and mouth disease virus. early steps of infection play important roles in determining the host range and the target organs for each virus. here, i review the recent advances in the studies of cellular receptors for picornaviruses, mechanisms of cell entry and viral uncoating.201122916565
immune response in goats to different payloads of fmdv monovalent vaccine: protection against virulent challenge and development of carrier status.the relationship of foot-and-mouth disease virus antigen payload and number of dose of vaccine conferring protection against virus challenge in goats was studied. goats vaccinated with oil adjuvant foot-and-mouth disease vaccines containing different antigen payloads with or without booster resisted virulent challenge at 21 days post-vaccination or 7 days after booster respectively. however, localized sub-clinical infection was observed in two vaccinated goats on 35 days post-challenge. rna coul ...201122282634
self replicating gene vaccine carrying p1-2a gene of fmdv serotype o and its effects on the immune responses of cattle.dna vaccines are considered as alternatives to live attenuated ones for those diseases like foot-and-mouth disease (fmd) where the production and application of live vaccines have been found unsuccessful. however, stability of dna and the quantity of antigen expressed are the major limitation with naked dna vaccines. to address these issues self replicating gene vaccine construct was made for foot-and-mouth disease virus (fmdv) type 'o' and studied. the vector for vaccine construct, designated a ...201123637502
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
development and laboratory evaluation of two lateral flow devices for the detection of vesicular stomatitis virus in clinical samples.two lateral flow devices (lfd) for the detection of vesicular stomatitis (vs) virus (vsv), types indiana (vsv-ind) and new jersey (vsv-nj) were developed using monoclonal antibodies c1 and f25vsvnj-45 to the respective vsv serotypes. the performance of the lfds was evaluated in the laboratory on suspensions of vesicular epithelia and cell culture passage derived supernatants of vsv. the collection of test samples included 105 positive for vsv-ind (92 vesicular epithelial suspensions and 13 cell ...201122230813
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
comparative sequence analysis of representative foot-and-mouth disease virus genomes from southeast asia.foot-and-mouth disease (fmd) is endemic in mainland southeast asia where up to seven genetically distinct viral lineages co-circulate (o/sea/mya-98, o/sea/cam-94, o/me-sa/panasia, o/me-sa/panasia-2, o/cathay, a/asia/sea-97, and serotype asia 1). the aim of this study was to analyse the sequence variability between representative complete genomes for these seven lineages. the genome sequences varied from 8130 to 8192 nucleotides in length and shared nucleotide identities ranging from 91.8 to 78.9 ...201121479678
foot-and-mouth disease in feral swine: susceptibility and transmission.experimental studies of foot-and-mouth disease (fmd) in feral swine are limited, and data for clinical manifestations and disease transmissibility are lacking. in this report, feral and domestic swine were experimentally infected with fmdv (a24-cruzeiro), and susceptibility and virus transmission were studied. feral swine were proved to be highly susceptible to a-24 cruzeiro fmd virus by intradermal inoculation and by contact with infected domestic and feral swine. typical clinical signs in fera ...201121418546
virus survival in slurry: analysis of the stability of foot-and-mouth disease, classical swine fever, bovine viral diarrhoea and swine influenza viruses.farm slurry can be highly contaminated with viral pathogens. the survival of these pathogens within slurry is important since this material is often distributed onto farm land either directly or after heat treatment. there is clearly some risk of spreading pathogens in the early stages of an outbreak of disease before it has been recognized. the survival of foot-and-mouth disease virus, classical swine fever virus, bovine viral diarrhoea virus and swine influenza virus, which belong to three dif ...201122226541
comparison of ires and f2a-based locus-specific multicistronic expression in stable mouse lines.efficient and stoichiometric expression of genes concatenated by bi- or multi-cistronic vectors has become an invaluable tool not only in basic biology to track and visualize proteins in vivo, but also for vaccine development and in the clinics for gene therapy. to adequately compare, in vivo, the effectiveness of two of the currently popular co-expression strategies - the internal ribosome entry site (ires) derived from the picornavirus and the 2a peptide from the foot-and-mouth disease virus ( ...201122216134
direct typing and molecular evolutionary analysis of field samples of foot-and-mouth disease virus collected in viet nam between 2006 and 2007.in this study, we used universal or duplex serotype-specific (o and asia 1) rt-pcr to analyze clinical field samples of foot-and-mouth disease virus (fmdv) or virus isolates collected in viet nam between 2006 and 2007. we found viral serotypes o and asia 1 circulating concurrently during this period. direct sequencing of type-specific rt-pcr products revealed the existence of three different topotypes of serotype o: southeast asia (sea), middle east-south asia (me-sa), and cathay. of these, sea ...201120667669
outbreak investigations and genetic characterization of foot-and-mouth disease virus in ethiopia in 2008/2009.the study was conducted in three regional states of ethiopia: amhara, oromia, and addis ababa from august 2008 to april 2009 with the objectives of identifying the genetic diversity of serotypes and topotypes in ethiopia, and determining the attack rate and associations of potential risk factors with foot-and-mouth disease (fmd) seropositivity. a total of 496 cattle were clinically and serologically examined for presence of specific lesions and nonstructural protein for fmd, respectively. of whi ...201120717724
modelling foot-and-mouth disease virus dynamics in oral epithelium to help identify the determinants of lysis.foot-and-mouth disease virus (fmdv) causes an economically important disease of cloven-hoofed livestock; of interest here is the difference in lytic behaviour that is observed in bovine epithelium. on the skin around the feet and tongue, the virus rapidly replicates, killing cells, and resulting in growing lesions, before eventually being cleared by the immune response. in contrast, there is usually minimal lysis in the soft palate, but virus may persist in tissue long after the animal has recov ...201120725794
multiple invasions of o1 fmdv serotype into israel revealed by genetic analysis of vp1 genes of israeli's isolates from 1989 to 2007.foot-and-mouth disease virus (fmdv), one of the most dangerous viruses affecting cloven-hoofed animals, comprises seven serotypes that do not mutually cross-protect, with a total of about 80 subtypes. the middle east is an fmd-endemic region, with repeated fmd outbreaks and in spite of its compulsory vaccination policy in israel, outbreaks occur repeatedly. in order to compare the israeli isolates, the complete viral vp1 genes of representative viruses isolated during the major outbreaks from 19 ...201120702050
introduction of tag epitopes in the inter-aug region of foot and mouth disease virus: effect on the l protein.foot-and-mouth disease virus (fmdv) initiates translation from two in-frame aug codons producing two forms of the leader (l) proteinase, lab (starting at the first aug) and lb (starting at second aug). in a previous study, we have demonstrated that a cdna-derived mutant fmdv (a24-l1123) containing a 57-nucleotide transposon (tn) insertion between the two aug initiation codons (inter-aug region) was completely attenuated in cattle, suggesting that this region is involved in viral pathogenesis. to ...201120849893
meta-analysis on the efficacy of foot-and-mouth disease emergency vaccination.the objectives of this study were to provide a summary quantification of the efficacy of fmd emergency vaccination based on a systematic review and a meta-analysis of available literature, and to further discuss the suitability of this review and meta-analysis to summarize and further interpret the results. peer-reviewed, symposium, and unpublished studies were considered in the analysis. clinical protection and virological protection against fmd were used as parameters to assess the efficacy of ...201120869130
phylogenetic structure of serotype a foot-and-mouth disease virus: global diversity and the indian perspective.global epidemiological analysis is vital for implementing progressive regional foot-and-mouth disease control programmes. here, we have generated vp1 region sequences for 55 indian type a outbreak strains and have included complete vp1 sequences from 46 other countries to obtain a comprehensive global phylogeographical impression. a total of 26 regional genotypes within three continental topotypes, based on a 15% nucleotide divergence cut-off criterion, could be identified. these genotypes corre ...201121228130
the current state of vaccines used in the field for foot and mouth disease virus in china. 201121162615
capsid proteins from field strains of foot-and-mouth disease virus confer a pathogenic phenotype in cattle on an attenuated, cell-culture-adapted virus.chimeric foot-and-mouth disease viruses (fmdvs) have been generated from plasmids containing full-length fmdv cdnas and characterized. the parental virus cdna was derived from the cell-culture-adapted o1kaufbeuren b64 (o1k b64) strain. chimeric viruses, containing capsid coding sequences derived from the o/ukg/34/2001 or a/turkey 2/2006 field viruses, were constructed using the backbone from the o1k b64 cdna, and viable viruses (o1k/o-ukg and o1k/a-tur, respectively) were successfully rescued in ...201121270284
gm-csf and il-2 as adjuvant enhance the immune effect of protein vaccine against foot-and-mouth disease.cytokines as molecular adjuvant play a critical role in differentiation of effector t cell subsets and in determination of the magnitude of the response after vaccination. in this study, we investigated the effects of gm-csf and il-2 as adjuvant on the immune responses of vp1 recombinant protein as a model antigen for foot and mouth disease.201121214955
effect of thiomersal on dissociation of intact (146s) foot-and-mouth disease virions into 12s particles as assessed by novel elisas specific for either 146s or 12s particles.intact (146s) foot-and-mouth disease virions (fmdvs) can dissociate into specific (12s) viral capsid degradation products. using two single-domain antibody fragments that bind specifically to either 146s or 12s particles we developed two elisas for the quantification of these particles in fmdv antigen preparations used for vaccine manufacturing. only o serotype strains are detected in the 146s specific elisa whereas strains of most serotypes are detected in the 12s specific elisa. however, the 1 ...201121316500
epidemic and economic impacts of delayed detection of foot-and-mouth disease: a case study of a simulated outbreak in california.the epidemic and economic impacts of foot-and-mouth disease virus (fmdv) spread and control were examined by using epidemic simulation and economic (epinomic) optimization models. the simulated index herd was a ≥2,000 cow dairy located in california. simulated disease spread was limited to california; however, economic impact was assessed throughout the united states and included international trade effects. five index case detection delays were examined, which ranged from 7 to 22 days. the simu ...201121217024
bovine plasmacytoid dendritic cells are the major source of type i interferon in response to foot-and-mouth disease virus in vitro and in vivo.type i interferons (alpha/beta interferons [ifn-a/ß]) are the main innate cytokines that are able to induce a cellular antiviral state, thereby limiting viral replication and disease pathology. plasmacytoid dendritic cells (pdcs) play a crucial role in the control of viral infections, especially in response to viruses that have evolved mechanisms to block the type i ifn signal transduction pathway. using density gradient separation and cell sorting, we have highly enriched a population of bovine ...201121307187
transmission of swine pathogens: different means, different needs.there seems to be two main types of pathogens that cause diseases in swine: those that are mainly introduced through direct pig contacts, and those that are often, and in some situations mainly introduced by indirect transmission means. in this review, the mange mite (sarcoptes scabiei), toxigenic pasteurella multocida and brachyspira hyodysenteriae will be used as examples of the first type, and foot and mouth disease virus, mycoplasma hyopneumoniae and porcine reproductive and respiratory synd ...201121241537
influence of mutagenesis and viral load on the sustained low-level replication of an rna virus.lethal mutagenesis is an antiviral strategy that aims to extinguish viruses as a consequence of enhanced mutation rates during virus replication. the molecular mechanisms that underlie virus extinction by mutagenic nucleoside analogues are not well understood. when mutagenic agents and antiviral inhibitors are administered sequentially or in combination, interconnected and often conflicting selective constraints can influence the fate of the virus either towards survival through selection of mut ...201121256131
evaluation of cross-protection between o1 manisa and o1 campos in cattle vaccinated with foot-and-mouth disease virus vaccine incorporating different payloads of inactivated o1 manisa antigen.serology is used to predict vaccine induced protection against challenge with a heterologous strain of the same serotype of foot-and-mouth disease virus (fmdv). to evaluate the accuracy of such predictions, we compared the protection afforded to cattle vaccinated with the o(1) manisa strain of fmdv against challenge with either a homologous (o(1) manisa) or a heterologous strain (o(1) campos). serology by virus neutralization test (vnt) using o(1) manisa antiserum predicted an acceptable protect ...201121236232
molecular epidemiology of foot-and-mouth disease virus serotypes a and o with emphasis on korean isolates: temporal and spatial dynamics.we determined complete 1d gene sequences for one serotype a and seven additional serotype o korean foot-and-mouth disease viruses (fmdv) and then analyzed them together with published sequences for 180 type a and 300 type o isolates from throughout the world using a bayesian coalescent approach. here, korean serotype a virus was linked with those from laos. korean serotype o viruses were divided into three clades and were closely related to isolates from japan, thailand, the uk, france, ireland, ...201121279395
effects of macromolecular crowding on the inhibition of virus assembly and virus-cell receptor recognition.biological fluids contain a very high total concentration of macromolecules that leads to volume exclusion by one molecule to another. theory and experiment have shown that this condition, termed macromolecular crowding, can have significant effects on molecular recognition. however, the influence of molecular crowding on recognition events involving virus particles, and their inhibition by antiviral compounds, is virtually unexplored. among these processes, capsid self-assembly during viral mor ...201121281589
diagnostic performance and application of two commercial cell viability assays in foot-and-mouth disease research.cell-based assays are still used widely in foot-and-mouth disease (fmd) research, despite the existence of a wide variety of molecular techniques. the aim of this study was to validate an automated, quantitative spectrometric reading to replace the time-consuming and subjective microscopic (mic) evaluation of the fmd virus-induced cytopathic effect (cpe). therefore, the diagnostic performance of two commercial cell viability assays (celltiter 96(®) aqueous one solution cell proliferation assay ( ...201121295609
common conformational changes induced in type 2 picornavirus iress by cognate trans-acting factors.type 2 internal ribosomal entry sites (iress) of encephalomyocarditis virus (emcv), foot-and-mouth disease virus (fmdv) and other picornaviruses comprise five major domains h-l. initiation of translation on these iress begins with specific binding of the central domain of initiation factor, eif4g to the j-k domains, which is stimulated by eif4a. eif4g/eif4a then restructure the region of ribosomal attachment on the ires and promote recruitment of ribosomal 43s pre-initiation complexes. in additi ...201121306989
the leader proteinase of foot-and-mouth disease virus negatively regulates the type i interferon pathway by acting as a viral deubiquitinase.the leader proteinase (l(pro)) of foot-and-mouth disease virus (fmdv) is a papain-like proteinase that plays an important role in fmdv pathogenesis. previously, it has been shown that l(pro) is involved in the inhibition of the type i interferon (ifn) response by fmdv. however, the underlying mechanisms remain unclear. here we demonstrate that fmdv lb(pro), a shorter form of l(pro), has deubiquitinating activity. sequence alignment and structural bioinformatics analyses revealed that the catalyt ...201121307201
study on seroprevalence, risk factors, and economic impact of foot-and-mouth disease in borena pastoral and agro-pastoral system, southern ethiopia.cross-sectional serological study and questionnaire survey were conducted in borana pastoral and agro-pastoral area to determine seroprevalence and risk factors associated with foot-and-mouth disease (fmd) infection and to assess community perceptions as to importance of the disease. a multistage random sampling was carried out to select cattle for seroprevalence and households for interviews. totally, 768 sera were collected from 111 herds. the overall individual level seroprevalence of 23.0% ( ...201121249448
picornavirus non-structural proteins as targets for new anti-virals with broad activity.picornaviridae is one of the largest viral families and is composed of 14 genera, six of which include human pathogens. the best known picornaviruses are enteroviruses (including polio, pv, and rhinoviruses), foot-and-mouth disease virus (fmdv), and hepatitis a virus (hav). although infections often are mild, certain strains may cause pandemic outbreaks accompanied with meningitis and/or paralysis. vaccines are available for pv, hav and fmdv. when the oral vaccines are given to immunocompromised ...201121236302
biochemical characterization of enterovirus 71 3d rna polymerase.an unusual enterovirus 71 (ev71) epidemic has begun in china since 2008. ev71 rna polymerases (3d(pol)) showed polymerase activity with an mn(2+). little activity was detected with co(2+), and no activity was detected with mg(2+), ca(2+), cu(2+), ni(2+), cd(2+), or zn(2+). it is a primer-dependent polymerase, and the enzyme functioned with both di- and 10-nucleotide rna primers. dna primer, dt15, increased primer activity, similar to other enterovirus 3d(pol). however, ev71 3d(pol) initiated de ...201121220056
genomic comparison of foot-and-mouth disease virus r strain and its chick-passaged attenuated strain.the present study examined the genomic differences between foot-and-mouth disease virus (fmdv) r strain and its attenuated, chick-passaged (r(304)) strain. eleven pairs of primers were used to amplify the complete genome of fmdv r and r(304) by rt-pcr. each fragment was cloned into pmd18-t vector and sequenced. nucleotide analyses showed that the genome encoding regions of r and r(304) strains open reading frame (orf) were both 6966 nucleotides (nt) in length, encoding 2322 amino acids. one hund ...201121330068
rescue of foot-and-mouth disease viruses that are pathogenic for cattle from preserved viral rna samples.foot and mouth disease is an economically important disease of cloven-hoofed animals including cattle, sheep and pigs. it is caused by a picornavirus, foot-and-mouth disease virus (fmdv), which has a positive sense rna genome which, when introduced into cells, can initiate virus replication.201121298025
a replication analysis of foot-and-mouth disease virus in swine lymphoid tissue might indicate a putative carrier stage in pigs.abstract: foot-and-mouth disease virus (fmvd), one of the most contagious viruses of cloven-hoofed animals, may cause a prolonged, asymptomatic but persistent infection in ruminants, named the "carrier state". however, it remains an open question whether this carrier state occurs in pigs. here we present quantitative analyses of the duration of fmdv rna and infectivity in lymphoid and epithelial tissues in experimentally infected pigs with fmdv c-s8c1. the data indicated that although fmdv rna r ...201121314905
dna immunization of pigs with foot-and-mouth disease virus minigenes: from partial protection to disease exacerbation.despite several attempts to design new vaccines, there are as of yet no available alternatives to the conventional fmdv vaccines. here, we present the divergent results obtained in pigs after immunization with two experimental dna vaccines encoding one b and two t cell fmdv epitopes, either expressed alone (pcmv-btt) or fused to a strong signal peptide (pcmv-spbtt). while all pigs vaccinated with pcmv-spbtt showed both a delay in the disease onset and reduced severity of signs and lesions after ...201121315115
molecular characterization of foot-and-mouth disease virus: implications for disease control in bangladesh.foot-and-mouth disease (fmd) is endemic in bangladesh, and to implement an effective fmd control programme, it is essential to understand the complex epidemiology of the disease. here, we report on the characterization of fmd virus (fmdv) recovered from fmd outbreaks in bangladesh in late 2009. all isolated viruses belonged to the fmdv serotype o. the phylogenetic reconstruction showed that all isolates belonged to the middle east-south asia (me-sa) topotype, but fell into two distinct sublineag ...201121320294
absence of heat intolerance (panting) syndrome in foot-and-mouth disease-affected indian cattle (bos indicus) is associated with intact thyroid gland function.foot-and-mouth disease (fmd) is a highly contagious and economically important viral disease with high morbidity and reduced productivity of affected animals. we studied the heat intolerance (hi) (panting) syndrome and the effect of fmd virus (fmdv) infection on thyroid gland function in indian cattle (bos indicus). experimental infection with fmdv asia 1 resulted in a mild form of disease with superficial lesions. heat intolerance syndrome and its signs were not observed among the recovered ani ...201121388520
the importance of being short: the role of rabies virus phosphoprotein isoforms assessed by differential ires translation initiation.the rabies virus (rv) phosphoprotein p is a multifunctional protein involved in viral rna synthesis and in counteracting host innate immune responses. we have previously shown that rv p gene expression levels can be regulated by using picornavirus internal ribosome entry site (ires) elements. here we exploited a particular feature of the foot-and-mouth disease virus (fmdv) ires, namely, preferential initiation at a downstream initiation codon, to address the role of n-terminally truncated rv pho ...201121397980
pan-serotypic detection of foot-and-mouth disease virus using a minor groove binder probe reverse transcription polymerase chain reaction assay.a novel assay for the pan-serotypic detection of foot-and-mouth disease virus (fmdv) was designed using a 5' conjugated minor groove binder (mgb) probe real-time rt-pcr system. this assay targets the 3d region of the fmdv genome and is capable of detecting 20 copies of a transcribed rna standard. the linear range of the test was eight logs from 2 × 10¹ to 2 × 108 copies and amplification time was approximately 2 h. using a panel of 83 rna samples from representative fmdv isolates, the diagnostic ...201121419170
active protein aggregates induced by terminally attached self-assembling peptide elk16 in escherichia coli.in recent years, it has been gradually realized that bacterial inclusion bodies (ibs) could be biologically active. in particular, several proteins including green fluorescent protein, ß-galactosidase, ß-lactamase, alkaline phosphatase, d-amino acid oxidase, polyphosphate kinase 3, maltodextrin phosphorylase, and sialic acid aldolase have been successfully produced as active ibs when fused to an appropriate partner such as the foot-and-mouth disease virus capsid protein vp1, or the human ß-amylo ...201121320350
viral genome segmentation can result from a trade-off between genetic content and particle stability.the evolutionary benefit of viral genome segmentation is a classical, yet unsolved question in evolutionary biology and rna genetics. theoretical studies anticipated that replication of shorter rna segments could provide a replicative advantage over standard size genomes. however, this question has remained elusive to experimentalists because of the lack of a proper viral model system. here we present a study with a stable segmented bipartite rna virus and its ancestor non-segmented counterpart, ...201121437265
rapid and transient activation of +¦+¦ t cells to ifn-+¦ production, nk cell-like killing, and antigen processing during acute virus infection.+¦+¦ t cells are the majority peripheral blood t cells in young cattle. the role of +¦+¦ t cells in innate responses against infection with foot-and-mouth disease virus was analyzed on consecutive 5 d following infection. before infection, bovine wc1(+) +¦+¦ t cells expressed a nonactivated phenotype relative to cd62l, cd45ro, and cd25 expression and did not produce ifn-+¦ ex vivo. additionally, cd335 expression was lacking and no spontaneous target cell lysis could be detected in vitro, althoug ...201121383249
biosecurity and fmd transmission. 201121493485
development of expression vectors based on pepino mosaic virus. 201121396092
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
closed ampoule isothermal microcalorimetry for continuous real-time detection and evaluation of cultured mammalian cell activity and responses.closed ampoule isothermal microcalorimetry (imc) is a simple, powerful, nondestructive, and convenient technique that allows continuous, real-time detection and evaluation of cultured cell activity and responses. at a selected set temperature, imc measures the heat flow between a sample and a heat sink and compares it to the heat-flow between a thermally inactive reference and the heat sink. since heat flow rates are proportional to the rates of chemical reactions and changes of state, imc provi ...201121468980
foot-and-mouth disease virus leader proteinase inhibits dsrna-induced rantes transcription in pk-15 cells.the chemokine rantes (regulated upon activation, normal t-cells expressed and secreted) plays an essential role in inflammation and immune response. infection with wild-type foot-and-mouth disease virus (fmdv) in pk-15 cells strongly inhibits the expression of rantes compared to infection with a genetically engineered mutant lacking the leader protein (l(pro)) coding region. this suggests that l(pro) is involved in rantes regulation. however, the underlying molecular mechanism remains unclear. i ...201121399922
fine mapping of a foot-and-mouth disease virus epitope recognized by serotype-independent monoclonal antibody 4b2.vp2 is a structural protein of the foot-and-mouth disease virus (fmdv). in this study, a fmdv serotype-in-dependent monoclonal antibody (mab), 4b2, was generated. by screening a phage-displayed random 12-peptide library, we found positive phages displaying the consensus motif ettxle (x is any amino acid (aa)), which is highly homologous to (6)ettlle(11) at the n-terminus of the vp2 protein. subsequently, a series of gst-fusion proteins expressing a truncated n-terminus of vp2 were examined by we ...201121369985
a mutant of asia 1 serotype of foot-and-mouth disease virus with the deletion of an important antigenic epitope in the 3a protein.foot-and-mouth disease is a highly contagious viral disease of cloven-hoofed animals. the availability of a vaccine for differentiating infected from vaccinated animals (diva) is crucial for the control and eradication of foot-and-mouth disease virus (fmdv). because traditional inactivated vaccines may contain trace nonstructural proteins interfering with the diva, we hypothesized that mutant fmdv with deletion of immunodominant epitopes may be valuable. our previous study has generated a full-l ...201121358757
integrin sub-unit expression in cell cultures used for the diagnosis of foot-and-mouth disease.the ability to propagate foot-and-mouth disease virus (fmdv) plays an important role in laboratory diagnosis and the production of vaccines to control the spread of the disease. many established cell lines suffer from poor sensitivity for isolating virus from field samples. one possible factor that limits sensitivity to fmdv is the lack of expression of surface integrins, the primary class of cell receptor used by fmdv to initiate infection. in this study we have sequenced cdnas encoding these m ...201121329991
the expressions of hsp70 and ab-crystallin in myocarditis associated with foot-and-mouth disease virus in lambs.this study describes the expression of heat shock protein70 (hsp70) and alpha-basic-crystallin (a-bc) and their association with apoptosis and some related adaptor proteins in the pathogenesis of foot-and-mouth disease virus (fmdv)-induced myocarditis in lambs. hsp70 was generally overexpressed in the myocardial tissues and inflammatory cells of fmdv-induced myocarditis with differential accumulation and localization in same hearts when compared to non-foot-and-mouth disease control hearts. a-bc ...201121368565
a simple and rapid colloidal gold-based immunochromatogarpic strip test for detection of fmdv serotype a.a sandwich format immunochromatographic assay for detecting foot-and-mouth disease virus (fmdv) serotypes was developed. in this rapid test, affinity purified polyclonal antibodies from guinea pigs which were immunized with sucking-mouse adapted fmd virus (a/av88(l) strain) were conjugated to colloidal gold beads and used as the capture antibody, and affinity purified polyclonal antibodies from rabbits which were immunized with cell-culture adapted fmd virus (a/cha/09 strain) were used as detect ...201121331888
experimental evidence for competitive growth advantage of genotype vii over vi: implications for foot-and-mouth disease virus serotype a genotype turnover in nature.in india, systematic genotype replacement has been observed for serotype a foot-and-mouth disease virus. after a decade of co-circulation of genotypes vi and vii, genotype vii emerged as the single dominant genotype since 2001. to derive possible explanations for such epochal evolution dynamics, in vitro intergenotype growth competition experiments involving both co- and superinfection regimes were conducted. coinfection of bhk-21 cells demonstrated abrupt loss in the genotype vi viral load with ...201121338995
a pseudotype baculovirus expressing the capsid protein of foot-and-mouth disease virus and a t-cell immunogen shows enhanced immunogenicity in mice.foot-and-mouth disease (fmd) is a highly contagious disease of livestock which causes severe economic loss in cloven-hoofed animals. vaccination is still a major strategy in developing countries to control fmd. currently, inactivated vaccine of fmdv has been used in many countries with limited success and safety concerns. development of a novel effective vaccine is must.201121342530
development and initial results of a low cost, disposable, point-of-care testing device for pathogen detection.development of small footprint, disposable, fast, and inexpensive devices for pathogen detection in the field and clinic would benefit human and veterinary medicine by allowing evidence-based responses to future out breaks. we designed and tested an integrated nucleic acid extraction and amplification device employing a loop-mediated isothermal amplification (lamp) or reverse transcriptase-lamp assay. our system provides a screening tool with polymerase-chain-reaction-level sensitivity and speci ...201121342806
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
establishment of persistent infection with foot-and-mouth disease virus in bhk-21 cells.abstract:201121492421
development of vaccines toward the global control and eradication of foot-and-mouth disease.foot-and-mouth disease (fmd) is one of the most economically and socially devastating diseases affecting animal agriculture throughout the world. although mortality is usually low in adult animals, millions of animals have been killed in efforts to rapidly control and eradicate fmd. the causing virus, fmd virus (fmdv), is a highly variable rna virus occurring in seven serotypes (a, o, c, asia 1, sat 1, sat 2 and sat 3) and a large number of subtypes. fmdv is one of the most infectious agents kno ...201121434805
antiviral activity of bovine type iii interferon against foot-and-mouth disease virus.foot-and-mouth disease (fmd) is one of the most serious threats to the livestock industry. despite the availability of a vaccine, recent outbreaks in disease-free countries have demonstrated that development of novel fmd control strategies is imperative. here we report the identification and characterization of bovine (bo) interferon lambda 3 (ifn-++3), a member of the type iii ifn family. expression of boifn-++3 using a replication-defective human adenovirus type 5 vector (ad5-boifn-++3) yielde ...201121435672
genetic diversity of foot-and-mouth disease virus serotype o in pakistan and afghanistan, 1997-2009.foot-and-mouth disease (fmd) is endemic in pakistan and afghanistan; serotypes o, a and asia-1 of the virus are responsible for the outbreaks in these countries with fmdv type o usually being the most common. in the present study, the nucleotide sequences encoding the fmdv capsid protein vp1 from virus samples were determined. phylogenetic analysis of the serotype o fmd viruses circulating in pakistan and afghanistan between 1997 and 2009 revealed the presence of at least three different lineage ...201121419880
phylogenomics and molecular evolution of foot-and-mouth disease virus.this report describes the use of bayesian methods to analyze polyprotein coding region sequences (n = 217) obtained from genbank to define the genome-wide phylogeny of foot and mouth disease virus (fmdv). the results strongly supported the monophyly of five fmdv serotypes, o, a, asia 1, c, and sat 3, while sequences for the two remaining fmdv serotypes, sat 1 and sat 2 did not separate into entirely distinct clades. the phylogenomic tree revealed three sister-group relationships, serotype o + as ...201121448588
skin as a potential source of infectious foot and mouth disease aerosols.this review examines whether exfoliated, virus-infected animal skin cells could be an important source of infectious foot and mouth disease virus (fmdv) aerosols. infectious material rafting on skin cell aerosols is an established means of transmitting other diseases. the evidence for a similar mechanism for fmdv is: (i) fmdv is trophic for animal skin and fmdv epidermis titres are high, even in macroscopically normal skin; (ii) estimates for fmdv skin cell aerosol emissions appear consistent wi ...201121450741
a highly sensitive detection for foot-and-mouth disease virus by gold nanopariticle improved immuno-pcr.abstract: background: foot-and-mouth disease (fmd) is one of the most contagious of all artiodactyl animal diseases, and its infection has an obvious ability to spread over long distances and to contribute to epidemics in fmd-free areas. a highly sensitive and specfic method is required to detect fmdv. in this study, we evaluated the usefulness of a bio-barcode assay (bca) technique for detecting clinical samples of fmdv. methods: highly sensitive gold nanopariticle (gnp) improved immuno -pcr (g ...201121453461
outbreak of hand, foot and mouth disease caused by coxsackie a16 virus in a childcare centre in croatia, february to march 2011. 201121632018
viewpoint: factors involved in type i interferon responses during porcine virus infections.since type i interferon (ifn-i) is considered a potent antiviral defence mechanism, it is not surprising that during evolution viruses have development of various mechanisms to counteract ifn-i induction or release. despite this, certain virus infections are associated with very high levels of systemic ifn-i. one explanation for this observation is the presence of plasmacytoid dendritic cells (pdc), which are able to produce high levels of ifn-i despite the presence of viral ifn-i antagonists. e ...201121458079
florfenicol impairs the immune responses to vaccination against foot-and-mouth disease in mice.florfenicol is a new type of broad-spectrum antibacterial that has been used in veterinary clinics. it showed immunosuppressive activity on the immune responses to vaccination against foot-and-mouth disease virus (fmdv) serotype o in mice. in the present study, balb/c mice were immunized subcutaneously with fmdv serotype o antigen on days 1 and 14. beginning on day 21, mice were treated with a single daily oral dose of florfenicol (50, 100, and 200?mg/kg) for seven consecutive days. on day 28, b ...201121428714
differentiation of foot-and-mouth disease-infected pigs from vaccinated pigs using antibody-detecting sandwich elisa.the presence of serum antibodies for nonstructural proteins of the foot-and-mouth disease virus (fmdv) can differentiate fmdv-infected animals from vaccinated animals. in this study, a sandwich elisa was developed for rapid detection of the foot-and-mouth disease (fmd) antibodies; it was based on an escherichia coli-expressed, highly conserved region of the 3abc nonstructural protein of the fmdv o/tw/99 strain and a monoclonal antibody derived from the expressed protein. the diagnostic sensitivi ...201121467761
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
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
[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
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
[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
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
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
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
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