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high virulence of african swine fever virus caucasus isolate in european wild boars of all ages. 201222469497
a serosurvey for brucella suis, classical swine fever virus, porcine circovirus type 2, and pseudorabies virus in feral swine (sus scrofa) of eastern north carolina.as feral swine (sus scrofa) populations expand their range and the opportunity for feral swine hunting increases, there is increased potential for disease transmission that may impact humans, domestic swine, and wildlife. from september 2007 to march 2010, in 13 north carolina, usa, counties and at howell woods environmental learning center, we conducted a serosurvey of feral swine for brucella suis, pseudorabies virus (prv), and classical swine fever virus (csfv); the samples obtained at howell ...201222493123
classical swine fever virus p7 protein is a viroporin involved in virulence in swine.the nonstructural protein p7 of classical swine fever virus (csfv) is a small hydrophobic polypeptide with an apparent molecular mass of 6 to 7 kda. the protein contains two hydrophobic stretches of amino acids interrupted by a short charged segment that are predicted to form transmembrane helices and a cytosolic loop, respectively. using reverse genetics, partial in-frame deletions of p7 were deleterious for virus growth, demonstrating that csfv p7 function is critical for virus production in c ...201222496228
novel approach for the generation of recombinant african swine fever virus from a field isolate using gfp expression and 5-bromo-2'-deoxyuridine selection.generation of african swine fever virus (asfv) recombinants has so far relied mainly on the manipulation of virus strains which had been adapted to growth in cell culture, since field isolates do not usually replicate efficiently in established cell lines. using wild boar lung cells (wsl) which allow for propagation of asfv field isolates, a novel approach for the generation of recombinant asfv directly from field isolates was developed which includes the integration into the viral thymidine kin ...201222498259
baculovirus expression and antigenic characterization of classical swine fever virus e2 proteins.genes encoding a major structural glycoprotein, e2, of classical swine fever viruses (csfv) brescia (subgroup 1.2), paderborn (subgroup 2.1) and kanagawa (subgroup 3.4) were constructed by removing the transmembrane domain and adding a c-terminal 6 histidine (his) tag. all the e2 constructs were efficiently expressed in a baculovirus system as 53-kda glycosylated proteins that were identified in western blots by their reaction with anti-his and csfv-specific antibodies. these proteins were used ...201322510427
characterization of c-strain "riems" tav-epitope escape variants obtained through selective antibody pressure in cell culture.classical swine fever virus (csfv) c-strain "riems" escape variants generated under selective antibody pressure with monoclonal antibodies and a peptide-specific antiserum in cell culture were investigated. candidates with up to three amino acid exchanges in the immunodominant and highly conserved linear tav-epitope of the e2-glycoprotein, and additional mutations in the envelope proteins erns and e1, were characterized both in vitro and in vivo.it was further demonstrated, that intramuscular im ...201222515281
prevalence of and risk factors associated with viral and bacterial pathogens in farmed european wild boar.the aim of this study was to estimate in farmed european wild boars the prevalence of and risk factors associated with a range of common porcine viral and bacterial infections, namely, porcine parvovirus (ppv), porcine circovirus type 2 (pcv2), swine influenza virus (siv), aujeszky's disease virus (adv), classical swine fever virus (csfv), swine vesicular disease virus (svdv), coronavirus causing transmissible gastroenteritis (tgev), porcine reproductive and respiratory syndrome virus (prrsv), m ...201222516920
tick-borne viruses in europe.the aim of this review is to present briefly background information on 27 tick-borne viruses ("tiboviruses") that have been detected in europe, viz flaviviruses tick-borne encephalitis (tbev), louping-ill (liv), tyuleniy (tyuv), and meaban (meav); orthobunyaviruses bahig (bahv) and matruh (mtrv); phleboviruses grand arbaud (gav), ponteves (ptvv), uukuniemi (uukv), zaliv terpeniya (ztv), and st. abb's head (sahv); nairoviruses soldado (solv), puffin island (piv), avalon (avav), clo mor (cmv), cri ...201222526290
neoplastic transformation of t lymphocytes through transgenic expression of a virus host modification protein.virus host evasion genes are ready-made tools for gene manipulation and therapy. in this work we have assessed the impact in vivo of the evasion gene a238l of the african swine fever virus, a gene which inhibits transcription mediated by both nf-κb and nfat. the a238l gene has been selectively expressed in mouse t lymphocytes using tissue specific promoter, enhancer and locus control region sequences for cd2. the resulting two independently derived transgenic mice expressed the transgene and dev ...201222558084
development of multiplex pcr for simultaneous detection of six swine dna and rna viruses.uniplex and multiplex reverse transcription-polymerase chain reaction (rt-pcr) and pcr protocols were developed and evaluated subsequently for its effectiveness in detecting simultaneously single and mixed infections in swine. specific primers for three dna viruses and three rna viruses, including classical swine fever virus (csfv), porcine reproductive and respiratory syndrome virus (prrsv), japanese encephalitis virus (jev), porcine circovirus type 2 (pcv2), porcine pseudorabies virus (prv) an ...201222575688
positive selection pressure on the b/c domains of the e2-gene of classical swine fever virus in endemic areas under c-strain vaccination.in cuba, classical swine fever (csf) has become an endemic disease with several outbreaks each year, despite the implemented vaccination program. interestingly, a trend towards a milder presentation of the disease has been observed among the animals during the last years. this study aimed to assess positive selection pressure acting on partial e2 gene of csf viruses to gain insights into the mechanisms governing virulence and the driving forces of classical swine fever virus (csfv) evolution in ...201222580241
diagnosis of porcine teschovirus encephalomyelitis in the republic of haiti.in february and march 2009, approximately 1,500 backyard pigs of variable age became sick, and approximately 700 of them died or were euthanized in the lower artibonite valley and the lower plateau of the republic of haiti. the main clinical sign was posterior ataxia followed by paresis and/or paralysis on the second or third day of illness. no gross lesions were observed at postmortem examinations. the morbidity and mortality were approximately 60% and 40%, respectively. diagnostic samples (who ...201222604774
african swine fever virus strain georgia 2007/1 in ornithodoros erraticus ticks. 022607706
early pathogenesis of classical swine fever virus (csfv) strains in danish pigs.host-virus interactions play an important role for the clinical outcome of classical swine fever virus (csfv) infections in pigs. strain virulence, host characteristics and environment are all factors that markedly influence disease severity. we tested csfv strains of varying virulence in an experimental set-up, reducing the influence of host and environmental factors. thus, weaner pigs were inoculated with one of 4 csfv strains in order to compare the pathogenesis for a 3-week-period after infe ...201222608103
withdrawn: development of an rt-pcr-rflp assay for the detection and differentiation of wild-type and vaccine strains of classical swine fever virus.this article has been withdrawn at the request of the author(s). the publisher apologizes for any inconvenience this may cause. the full elsevier policy on article withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.201222609180
real-time analysis of the interaction of a multiple-epitope peptide with antibodies against classical swine fever virus using surface plasmon resonance.the e2 envelope glycoprotein is the major immunodominant protein of classical swine fever virus (csfv), and can induce neutralizing antibodies and protective immune responses in infected swine. we developed a tandem-repeat multiple-epitope recombinant protein that contains two copies of each of the regions of e2 spanned by residues 693-704, 770-780, and 826-843, coupled by two copies of the region spanned by residues 1446-1460 of the csfv nonstructural protein ns2-3. the chemically synthesized g ...201222634031
[quantitative detection of anti-classical swine fever virus sirna expression in cells by stem-loop rt-qpcr].rna interference (rnai) is a promising technology in development of specific antiviral therapy, but the quantitative detection of small interfering rna (sirna) expressed in vivo is the main challenge to assess its antiviral effect. in order to detect the sirna molecules (sin1 and sin2) particularly expressed in cells to inhibit the replication of classical swine fever virus (csfv), serial specific stem-loop primers were designed and synthesized. two of them (slp-n1-6 and slp-n2-8) were selected ...201222667106
genetic typing of classical swine fever virus isolates from china.the e2 genes of 73 classical swine fever virus (csfv) originated from csf suspected cases in different regions of china were genetically characterized and compared with reference csf viruses. all chinese viruses that characterized were segregated into two major groups and subdivided into four subgroups. most of isolates (61.6%) belonged to group 2 and were further divided into three subgroups: subgroup 2.1, 2.2 and 2.3. subgroup 2.1 was the largest subgroup which contained 46.6% of isolates, whi ...201322672483
selection of classical swine fever virus with enhanced pathogenicity reveals synergistic virulence determinants in e2 and ns4b.classical swine fever virus (csfv) is the causative agent of classical swine fever (csf), a highly contagious disease of pigs. there are numerous csfv strains that differ in virulence, resulting in clinical disease with different degrees of severity. low-virulent and moderately virulent isolates cause a mild and often chronic disease, while highly virulent isolates cause an acute and mostly lethal hemorrhagic fever. the live attenuated vaccine strain gpe(-) was produced by multiple passages of t ...201222674973
modulation of translation initiation efficiency in classical swine fever virus.modulation of translation initiation efficiency on classical swine fever virus (csfv) rna can be achieved by targeted mutations within the internal ribosome entry site (ires). in this study, cdnas corresponding to the wild-type (wt) or mutant forms of the ires of csfv strain paderborn were amplified and inserted into dicistronic reporter plasmids encoding fluc and rluc under the control of a t7 promoter. the mutations were within domains ii, iiid(1), and iiif of the ires. the plasmids were trans ...201222674994
improved strategy for phylogenetic analysis of classical swine fever virus based on full-length e2 encoding sequences.molecular epidemiology has proven to be an essential tool in the control of classical swine fever (csf) and its use has significantly increased during the past two decades. phylogenetic analysis is a prerequisite for virus tracing and thus allows implementing more effective control measures. so far, fragments of the 5´ntr (150 nucleotides, nt) and the e2 gene (190 nt) have frequently been used for phylogenetic analyses. the short sequence lengths represent a limiting factor for differentiation o ...201222676246
classical swine fever virus ns5a protein localizes to endoplasmic reticulum and induces oxidative stress in vascular endothelial cells.classical swine fever virus (csfv) causes a severe disease of pigs that is characterized by hemorrhage, disseminated intravascular coagulation, and leucopenia. until now, the role of the nonstructural protein 5a (ns5a) produced by csfv in the pathogenesis of csf is not well known. in this study, we investigated the function of csfv ns5a by examining its role in the induction of oxidative stress and related intracellular events. stable swine umbilical vein endothelial cell lines expressing csfv n ...201222718084
african swine fever virus uses macropinocytosis to enter host cells.african swine fever (asf) is caused by a large and highly pathogenic dna virus, african swine fever virus (asfv), which provokes severe economic losses and expansion threats. presently, no specific protection or vaccine against asf is available, despite the high hazard that the continued occurrence of the disease in sub-saharan africa, the recent outbreak in the caucasus in 2007, and the potential dissemination to neighboring countries, represents. although virus entry is a remarkable target for ...201222719252
identification of conformational epitopes and antigen-specific residues at the d/a domains and the extramembrane c-terminal region of e2 glycoprotein of classical swine fever virus.envelope glycoprotein e2 of classical swine fever virus (csfv) is the major antigen that induces neutralizing antibodies in infected pigs. previous studies revealed that both conformation-dependent and linear epitopes are most present within domains b/c/d/a in the n-terminal half of e2. however, studies of antigenicity beyond the b/c domains remain limited. this study revealed that conformational epitopes were present on the d/a domains as well as the proximal c-terminal of e2, since the mutatio ...201222727685
a real-time pcr to detect and analyze virulent emcv loads in sows and piglets.a real-time polymerase chain reaction with sybr green was developed for the detection and quantification of encephalomyocarditis virus (emcv) in porcine tissues; the method uses two primers specific for the 3d gene. the detection limit of this assay was 22 gene copies/reaction, equivalent to 0.001 tcid(50)/ml. the assay was linear over a 10(7) dilution range of template concentrations and was specific for emcv; it did not amplify other porcine pathogens (porcine circovirus 2, porcine reproductiv ...201222752806
the atf6 branch of unfolded protein response and apoptosis are activated to promote african swine fever virus infection.african swine fever virus (asfv) infection induces apoptosis in the infected cell; however, the consequences of this activation on virus replication have not been defined. in order to identify the role of apoptosis in asfv infection, we analyzed caspase induction during the infection and the impact of caspase inhibition on viral production. caspases 3, 9 and 12 were activated from 16 h post-infection, but not caspase 8. indeed, caspase 3 activation during the early stages of the infection appear ...201222764100
development of a suspension microarray for the genotyping of african swine fever virus targeting the snps in the c-terminal end of the p72 gene region of the genome.african swine fever virus (asfv) causes one of the most dreaded transboundary animal diseases (tads) in suidae. african swine fever (asf) often causes high rates of morbidity and mortality, which can reach 100% in domestic swine. to date, serological diagnosis has the drawback of not being able to differentiate variants of this virus. previous studies have identified the 22 genotypes based on sequence variation in the c-terminal region of the p72 gene, which has become the standard for categoriz ...201322776009
outbreaks of highly pathogenic porcine reproductive and respiratory syndrome in jiangxi province, china.in 2007, herds of pigs in jiangxi province, china experienced outbreaks of a severe form of suspected porcine reproductive and respiratory syndrome (prrs) characterized by high fever, high morbidity and mortality in animals of different ages. 152 swine sera and 42 tissues (consisting of liver, lung, lymph node and kidney) from five herds of pigs were collected. pigs were diagnosed as infected with a highly pathogenic form of the prrs virus (prrsv) based on elisa and reverse transcriptase polymer ...201222784793
african swine fever virus excretion patterns in persistently infected animals: a quantitative approach.the continuing circulation of african swine fever (asf) in russia and in the trans-caucasian countries has led to increased efforts in characterizing the epidemiology of asf. for a better insight in epidemiology, quantitative data on virus excretion is required. until now, excretion data has mainly focused on the initial stages of the disease. in our study we have studied asf virus (asfv) excretion dynamics in persistently infected animals. for this purpose, virus excretion through different rou ...201222795261
classical swine fever virus ns5a regulates viral rna replication through binding to ns5b and 3'utr.in this report, classical swine fever virus (csfv) ns5a inhibit viral rna replication when its concentration reached and surpassed the level of ns5b. three amino acid fragments of csfv ns5a, 137-172, 224-268 and 390-414 individually were shown to be essential to ns5b binding. the former two fragments were independently necessary for regulation of viral rna replication and correlated with ns5b and 3'utr binding activity. we also found that amino acids w143, v145, p227, t246, p257, k399, t401, e40 ...201222795973
production and characterization of monoclonal antibodies against a local isolate of classical swine fever virus.monoclonal antibodies (mabs) against a classical swine fever virus (csfv; subgenogroup 1:1) isolate from assam, india were produced and characterized. four fusions of myeloma cells (sp2/0ag) were made with spleenocytes of 8-10 weeks old balb/c mice immunized with the viral antigen. several hybridoma clones secreting antibodies to the virus were obtained after four fusions, but five hybridoma clones secreting antibody specific to the virus could be stabilized. all the mabs belong to the igg2a iso ...201222822524
detection of antibodies against classical swine fever virus in fecal samples from wild boar.classical swine fever (csf) is a contagious viral disease that affects pigs. wild boars can play an important epidemiological role in csf outbreaks. in the past decades, studies conducted in many countries have reported that the csf virus (csfv) may persist in wild boar populations. the existence of csfv in the free-ranging wild boar populations was indirectly confirmed by determining the prevalence of antibodies against csfv in the serum of hunted wild boars. however, analyzing sero-prevalence ...201222841406
new insights into the antigenic structure of the glycoprotein e(rns) of classical swine fever virus by epitope mapping.the e(rns) glycoprotein of classical swine fever virus (csfv) has been studied in detail concerning biochemical and functional properties, whereas less is known about its antigenic structure. in order to define epitopes recognized by csfv-specific antibodies, the binding sites of seven e(rns)-specific monoclonal antibodies were investigated. mapping experiments using chimeric e(rns) proteins, site-directed mutagenesis and an overlapping peptide library identified one antigenic region located bet ...201222868042
two newly developed e(rns)-based elisas allow the differentiation of classical swine fever virus-infected from marker-vaccinated animals and the discrimination of pestivirus antibodies.new generations of classical swine fever virus (csfv) marker vaccines have recently been developed in order to make emergency vaccination in case of a csf outbreak more feasible. however, the application of a marker vaccine is dependent on the availability of an accompanying discriminatory test allowing differentiation of infected from vaccinated animals (diva). cp7_e2alf, the most promising live marker vaccine candidate currently available, is a genetically modified bovine viral diarrhea virus ...201322902189
classical swine fever virus isolates from cuba form a new subgenotype 1.4.identification and classification of classical swine fever virus (csfv) on the basis of nucleotide sequencing and phylogenetic analysis have become an important tool to study the epidemiology and to control csf disease. according to phylogenetic analyses of short sequences from the 5'nontranslated region (150 nt) and the e2 (190 nt), most csfv isolates from south and central america have been assorted to the subgenotypes 1.1 and 1.3, while csfv isolates from cuba have been allocated to subgenoty ...201322902191
sequential deletion of genes from the african swine fever virus genome using the cre/loxp recombination system.a method has been established to sequentially delete combinations of genes from the asfv genome to test the effect on virus replication and host responses to infection. initially the asfv genes mgf505 2r and mgf505 3r and a truncated mgf360 9l gene were deleted from the genome of the tissue-culture adapted asfv strain ba71v and replaced with bacteriophage loxp sequences flanking the beta-glucuronidase (gus) marker gene to create recombinant virus vδmgf-gus. subsequently the gus gene was removed ...201222902236
prevalence of classical swine fever virus in domestic pigs in south korea: 1999-2011.the major policy for eradication of classical swine fever (csf) in south korea has focused on the implementation of compulsory vaccination of the susceptible pig population. a vaccine strain of csf virus, the lom strain, is used to maintain high herd seroconversion, a practice complementary to the 'stamping-out policy' and restriction of animal movement during disease outbreaks. to survey for the prevalence of csf in domestic pigs in south korea over the past 13 years (1999-2011), we tested 4 19 ...201322925439
in vitro infection with classical swine fever virus inhibits the transcription of immune response genes.classical swine fever virus (csfv) can evade the immune response and establish chronic infection under natural and experimental conditions. some genes related to antigen processing and presentation and to cytokine regulation are known to be involved in this response, but the precise mechanism through which each gene responds to csfv infection remains unclear.201222925563
efficacy of marker vaccine candidate cp7_e2alf in piglets with maternally derived c-strain antibodies.marker vaccines offer the possibility to differentiate classical swine fever (csf) infected from csf vaccinated animals based on serology and their implementation will ensure free trade with pigs. therefore, new generations of promising marker vaccines have been developed, among them the chimeric vaccine cp7_e2alf. however, in populations previously vaccinated with live attenuated vaccines like the c-strain, passive immunity through maternal antibodies can interfere with efficacy of cp7_e2alf va ...201222939909
comparative evaluation of novel african swine fever virus (asf) antibody detection techniques derived from specific asf viral genotypes with the oie internationally prescribed serological tests.the presence of antibodies against african swine fever (asf), a complex fatal notifiable oie disease of swine, is always indicative of previous infection, since there is no vaccine that is currently used in the field. the early appearance and subsequent long-term persistence of antibodies combined with cost-effectiveness make antibody detection techniques essential in control programmes. recent reports appear to indicate that the serological tests recommended by the oie for asf monitoring are mu ...201322944073
a t-cell epitope on ns3 non-structural protein enhances the b and t cell responses elicited by dendrimeric constructions against csfv in domestic pigs.it has been recently reported by our group that dendrimeric constructs combining b- and t-cell epitopes from classical swine fever virus (csfv) provided partial protection against experimental infection. this research evaluated four newly designed constructions while taking into account our previous work, including the direct implication that a t-cell epitope from the ns3 protein contributes to the generation of the immune response against csfv. to this end, the dendrimeric constructions, includ ...201222959286
first molecular identification and characterization of classical swine fever virus isolates from nepal.classical swine fever (csf) is a major constraint to pig production worldwide, and in many developing countries, the epidemiological status is unknown. here, for the first time, molecular identification and characterization of csfv isolates from two recent outbreaks in nepal are presented. analysis of full-length e2-encoding sequences revealed that these isolates belonged to csfv subgenotype 2.2 and had highest genetic similarity to isolates from india. hence, for csfv, nepal and india should be ...201322975985
viral metagenomics demonstrates that domestic pigs are a potential reservoir for ndumu virus.the rising demand for pork has resulted in a massive expansion of pig production in uganda. this has resulted in increased contact between humans and pigs. pigs can act as reservoirs for emerging infectious diseases. therefore identification of potential zoonotic pathogens is important for public health surveillance. in this study, during a routine general surveillance for african swine fever, domestic pigs from uganda were screened for the presence of rna and dna viruses using a high-throughput ...201223006778
[dynamics of changes in the composition of leukocyte population of peripheral blood during the african swine fever].the comparison of the composition of leukocytes of peripheral blood of healthy and infected swine revealed the quantitative alterations, as well as the appearance of new cells in leukocyte population during african swine fever. it was determined that african swine fever virus induced mass-scale mortality of cells of peripheral blood, especially lymphocytes and neutrophiles. the number of the dead cells reaches 60% of the initial number of all cells at the end of infection. it was also revealed t ...201623012981
african swine fever virus transcription.african swine fever virus (asfv), a large, enveloped, icosahedral dsdna virus, is currently the only known dna-containing arbovirus and the only recognized member of the family asfarviridae. its genome encodes more than 150 open reading frames that are densely distributed, separated by short intergenic regions. asfv gene expression follows a complex temporal programming. four classes of mrnas have been identified by its distinctive accumulation kinetics. gene transcription is coordinated with dn ...201323041356
laboratory methods to study african swine fever virus.we summarize findings of comparative studies in different cells cultures susceptible to asfv infection, through the analysis of virus components and infectious virus particles production, as alternative means to grow field and laboratory asfv strains. we also provide different methods to assay the infectivity of asfv samples and to purify the infective virus particles. finally we describe the general strategy to construct virus deletion mutants that can be engineered to obtain attenuated asfv st ...201323041357
dna vaccination partially protects against african swine fever virus lethal challenge in the absence of antibodies.the lack of available vaccines against african swine fever virus (asfv) means that the evaluation of new immunization strategies is required. here we show that fusion of the extracellular domain of the asfv hemagglutinin (sha) to p54 and p30, two immunodominant structural viral antigens, exponentially improved both the humoral and the cellular responses induced in pigs after dna immunization. however, immunization with the resulting plasmid (pcmv-shapq) did not confer protection against lethal c ...201223049728
pathway analysis in blood cells of pigs infected with classical swine fever virus: comparison of pigs that develop a chronic form of infection or recover.infection of pigs with csfv can lead to either acute disease, resulting in death or recovery, or chronic disease. the mechanisms by which csfv manipulates the pig's first line of defence to establish a chronic infection are poorly understood. therefore, pigs were infected with moderately virulent csfv, and whole blood was collected on a regular basis during a period of 18 days. using whole-genome microarrays, time-dependent changes in gene expression were recorded in blood cells of chronically d ...201323053521
african swine fever virus morphogenesis.this review summarizes recent structural and molecular biology studies related to the morphogenesis of african swine fever virus (asfv). asfv possesses icosahedral morphology and is constituted by four concentric layers: the central nucleoid, the core shell, the inner envelope and the icosahedral capsid. the extracellular virus acquires an external envelope by budding through the plasma membrane. the genes coding for 19 of the 54 structural proteins of the asfv particle are known and the localiz ...201323059353
interactive cellular proteins related to classical swine fever virus non-structure protein 2 by yeast two-hybrid analysis.classical swine fever is caused by the classical swine fever virus (csfv), which has a special affinity to endothelial cells. this fever is characterized by hemorrhage and necrosis of vascular injury. very little information is available on the interaction between vascular endothelial cells and csfv. in the current report, the cdna library of swine umbilical vein endothelial cell (suvec) was constructed by the switching mechanism at 5' end of the rna transcript approach. the yeast two-hybrid (y2 ...201223076522
african swine fever virus infection in ornithodoros ticks.african swine fever virus (asfv) is an arbovirus which is vectored by soft ticks of the ornithodoros spp. and in the sylvatic cycle infects wart hogs and bush pigs. asfv infection of domestic swine causes a high mortality disease. on the other hand, asfv infection of the tick can result in a high-titered and persistent infection depending upon the asfv isolate and the tick combination. recently, morphological, classical virology (titration) and recombinant asfv have been used to study the cellul ...201323085123
co-expression of erns and e2 genes of classical swine fever virus by replication-defective recombinant adenovirus completely protects pigs against virulent challenge with classical swine fever virus.the objective of this study was to construct a recombinant adenovirus for future csfv vaccines used in the pig industry for the reduction of losses involved in csf outbreaks. the erns and e2 genes of classical swine fever virus (csfv), which encode the two main protective glycoproteins from the "shimen" strain of csfv, were combined and inserted into the replication-defective human adenovirus type-5 and named the rad-erns-e2. nine pigs were randomly assigned to three treatment groups (three pigs ...201323092714
development and evaluation of indirect enzyme-linked immunosorbent assay for a screening test to detect antibodies against classical swine fever virus.an indirect enzyme-linked immunosorbent assay (elisa) was developed for a screening test to detect antibodies against classical swine fever virus (csfv). viral glycoproteins, which were purified from swine kidney cells infected with csfv ald/a76 strain by the immunoaffinity purification using monoclonal antibody against e2 protein, were adsorbed on a microtiter plate as the antigen for the antibody detection. each antibody titer of serum sample was expressed as a sample per positive value calcul ...201223094583
strain-specific monoclonal antibodies to the e2 protein of classical swine fever virus, paderborn strain.monoclonal antibodies (mabs) against the e2 protein of classical swine fever virus (csfv) are useful for diagnosis and strain characterization. a purified, baculovirus-expressed csfv e2 protein from the paderborn strain was formulated with a saponin adjuvant and successfully used to induce an antigen-specific immune response in mice. after cell fusion a panel, designated f92g, of 12 mouse hybridomas (5-2, 11-1, 14-1, 25-2, 28-2, 31-1, 34-1, 35-2, 37-3, 38-2, 39-1, 41-1) producing csfv-e2 specifi ...201223098300
epidemiology of african swine fever virus.african swine fever virus used to occur primarily in africa. there had been occasional incursions into europe or america which apart from the endemic situation on the island of sardinia always had been successfully controlled. but following an introduction of the virus in 2007, it now has expanded its geographical distribution into caucasus and eastern europe where it has not been controlled, to date. african swine fever affects domestic and wild pig species, and can involve tick vectors. the ab ...201323123296
african swine fever virus serodiagnosis: a general review with a focus on the analyses of african serum samples.african swine fever (asf) is an infectious disease that causes heavy mortality in domestic pigs. at present there is no vaccine against asf, and eradication in countries where the disease is endemic is based only on competent diagnosis programs and the sacrifice of infected animals. due to the presence of natural attenuated strains, certain infection conditions may result in reduced mortality. in these situations, the disease can be diagnosed by detection of specific antibodies. the use of class ...201323131491
virological diagnosis of african swine fever--comparative study of available tests.the rapid and reliable detection of african swine fever virus (asfv) is essential both for timely implementation of control measures to prevent the spread of disease, and to differentiate african swine fever (asf) from other pig disease with similar clinical presentations. many virological tests are currently available for the detection of asfv (live virus), antigen and genome, including virus isolation, elisa, fluorescent antibody, polymerase chain reaction (pcr) and isothermal assays. in recen ...201323131492
endosomal maturation, rab7 gtpase and phosphoinositides in african swine fever virus entry.here we analyzed the dependence of african swine fever virus (asfv) infection on the integrity of the endosomal pathway. using confocal immunofluorescence with antibodies against viral capsid proteins, we found colocalization of incoming viral particles with early endosomes (ee) during the first minutes of infection. conversely, viral capsid protein was not detected in acidic late endosomal compartments, multivesicular bodies (mvbs), late endosomes (les) or lysosomes (ly). using an antibody agai ...201223133661
the survey of porcine teschoviruses in field samples in china with a universal rapid probe real-time rt-pcr assay.a real-time reverse transcription polymerase chain reaction (rt-pcr) based on taqman was established and evaluated for quantitative detection of porcine teschoviruses (ptvs). a pair of primers and a taqman probe targeting on the highly conserved sequence of the 5'-untranslated region (5'-utr) of one to 11 serotypes of ptv were designed. standard plasmid dna containing pcr amplification of the 5'-utr were constructed and used to develop the real-time rt-pcr. the results indicated that the real-ti ...201323138413
african swine fever virus eradication in africa.african swine fever was reported in domestic pigs in 26 african countries during the period 2009-2011. the virus exists in an ancient sylvatic cycle between warthogs (phacochoerus africanus) and argasid ticks of the ornithodoros moubata complex in many of the countries reporting outbreaks and in two further countries in the region. eradication of the virus from the countries in eastern and southern africa where the classic sylvatic cycle occurs is clearly not an option. however, the virus has be ...201323142552
african swine fever virus replication and genomics.african swine fever virus (asfv) is a large icosahedral dna virus which replicates predominantly in the cytoplasm of infected cells. the asfv double-stranded dna genome varies in length from about 170 to 193 kbp depending on the isolate and contains between 150 and 167 open reading frames. these are closely spaced and read from both dna strands. the virus genome termini are covalently closed by imperfectly base-paired hairpin loops that are present in two forms that are complimentary and inverte ...201323142553
comprehensive evaluation of the adenovirus/alphavirus-replicon chimeric vector-based vaccine radv-sfv-e2 against classical swine fever.classical swine fever (csf) is an economically important, highly contagious swine disease caused by classical swine fever virus (csfv). marker vaccines and companion serological diagnostic tests are thought to be a promising strategy for future control and eradication of csf. previously, we have demonstrated that an adenovirus-vectored semliki forest virus replicon construct expressing the e2 glycoprotein from csfv, radv-sfv-e2, induced sterile immunity against a lethal csfv challenge. in this s ...201323153441
african swine fever virus controls the host transcription and cellular machinery of protein synthesis.throughout a viral infection, the infected cell reprograms the gene expression pattern in order to establish a satisfactory antiviral response. african swine fever virus (asfv), like other complex dna viruses, sets up a number of strategies to evade the host's defense systems, such as apoptosis, inflammation and immune responses. the capability of the virus to persist in its natural hosts and in domestic pigs, which recover from infection with less virulent isolates, suggests that the virus disp ...201323154157
antibody-mediated neutralization of african swine fever virus: myths and facts.almost all viruses can be neutralized by antibodies. however, there is some controversy about antibody-mediated neutralization of african swine fever virus (asfv) with sera from convalescent pigs and about the protective relevance of antibodies in experimentally vaccinated pigs. at present, there is no vaccine available for this highly lethal and economically relevant virus and all classical attempts to generate a vaccine have been unsuccessful. this failure has been attributed, in part, to what ...201323159730
a comparative analysis on the synonymous codon usage pattern in viral functional genes and their translational initiation region of asfv.the synonymous codon usage pattern of african swine fever virus (asfv), the similarity degree of the synonymous codon usage between this virus and some organisms and the synonymous codon usage bias for the translation initiation region of viral functional genes in the whole genome of asfv have been investigated by some simply statistical analyses. although both gc12% (the gc content at the first and second codon positions) and gc3% (the gc content at the third codon position) of viral functional ...201323161403
particle tracking analysis for the intracellular trafficking of nanoparticles modified with african swine fever virus protein p54-derived peptide.previous studies showed that the cytoplasmic transport of nanoparticles to the nucleus is driven by a vesicular sorting system. artificial approaches for targeting a microtubule-associating motor complex is also a challenge. we describe herein the development of a liposomal nanoparticle, the surface of which is modified with stearylated octa-arginine (str-r8), and a dynein light chain (lc8)-associated peptide derived from an african swine fever virus protein p54 (p54(149-161)) with polyethyleneg ...201323164937
identification and utility of innate immune system evasion mechanisms of asfv.the interferon (ifn) system is an early innate anti-virus host defense mechanism that takes place shortly after entry of the pathogen and long before the onset of adaptive immunity. thus, african swine fever virus (asfv), as an acute and persistent virus in pigs, is predicted to have evolved multiple genes for the manipulation and evasion of interferon. although, asfv is known to interfere with signaling pathways controlling the transcription of cytokines, surprisingly no individual virus gene m ...201323165138
african swine fever virus, tanzania, 2010-2012. 201223171814
classical swine fever virus failed to activate nuclear factor-kappa b signaling pathway both in vitro and in vivo.classical swine fever virus (csfv) is the cause of csf which is a severe disease of pigs, leading to heavy economic losses in many regions of the world. nuclear factor-kappa b (nf-κb) is a critical regulator of innate and adaptive immunity, and commonly activated upon viral infection. in our previous study, we found that csfv could suppress the maturation and modulate the functions of monocyte-derived dendritic cells (mo-dcs) without activating nf-κb pathway. to further prove the effects of csfv ...201223186553
efficient and stable rescue of classical swine fever virus from cloned cdna using an rna polymerase ii system.conventional reverse genetics for classical swine fever virus (csfv) is based on the transfection of permissive cells with either in vitro or intracellularly synthesized rna transcripts from a viral genomic cdna clone. these strategies are complicated, inefficient and time-consuming. this study is aimed to develop an improved reverse genetics method for the direct, rapid and efficient recovery of csfv from cloned cdna. the cdna clone pbrcism was constructed, which harbors the full-length genomic ...201323192173
cellular immunity in asfv responses.african swine fever virus (asfv) infection usually results in an acute haemorrhagic disease with a mortality rate approaching 100% in domestic pigs. however, pigs can survive infection with less-virulent isolates of asfv and may become chronically infected. surviving animals are resistant to challenge with homologous or, in some cases, closely related isolates of the virus indicating that pigs can develop protective immunity against asfv. during asymptomatic, non-virulent asfv infections natural ...201323201582
comparison of african swine fever virus prevalence and risk in two contrasting pig-farming systems in south-west and central kenya.we describe a horizontal survey of african swine fever virus (asfv) prevalence and risk factors associated with virus infection in domestic pigs in two contrasting production systems in kenya. a free range/tethering, low input production system in ndhiwa district of south-western kenya is compared with a medium input stall fed production system in kiambu district of central kenya. analysis of variance (anova) of data derived from cluster analysis showed that number of animals, number of breeding ...201323219357
rna helicase is involved in the expression and replication of classical swine fever virus and interacts with untranslated region.to investigate whether cytoplasmic rna helicase a (rha) influences the expression and replication of classical swine fever virus (csfv), an sirna molecule targeted to rha was transfected into pk-15 cells. the sirna was found to reduce cytoplasmic rha. in csfv subgenomic replicon transfected cells, incubation with the sirnas negatively impacted viral ns3 and rna production. in the csfv infected cells, treatment with the sirna resulted in a significant reduction of viral replication by 65-70%. fur ...201323220337
poly(c)-binding protein 1, a novel n(pro)-interacting protein involved in classical swine fever virus growth.n(pro) is a multifunctional autoprotease unique to pestiviruses. the interacting partners of the n(pro) protein of classical swine fever virus (csfv), a swine pestivirus, have been insufficiently defined. using a yeast two-hybrid screen, we identified poly(c)-binding protein 1 (pcbp1) as a novel interacting partner of the csfv n(pro) protein and confirmed this by coimmunoprecipitation, glutathione s-transferase (gst) pulldown, and confocal assays. knockdown of pcbp1 by small interfering rna supp ...201223221550
a179l, a new viral bcl2 homolog targeting beclin 1 autophagy related protein.autophagy is a relevant cellular defense mechanism that directly eliminates intracellular pathogens and has a crucial role for innate and adaptive immune responses. some viruses have developed tools to counteract this cellular response. a179l, the viral bcl2 homolog of african swine fever virus, interacts with proapoptotic bcl2 family proteins to inhibit apoptosis. here we report that this gene manipulates autophagy by interacting with beclin 1 through its bh3 homology domain. at subcellular lev ...201323228131
detection of foot-and-mouth disease serotype o by elisa using a monoclonal antibody.an elisa assay with monoclonal antibody (melisa) was used to type serotype o of foot-and-mouth disease virus (fmdv). all fmdv serotype o reference strains were positive by melisa, while other viruses such as fmdv serotypes asia 1, c, a and classical swine fever virus, swine vesicular disease virus, and porcine reproductive and respiratory syndrome virus remained negative. further, fmdv serotype o positive samples were able to be detected by melisa. this assay may be particularly suitable for dia ...201223244327
the feasibility of a novel bioreactor for vaccine production of classical swine fever virus.the performance of a new type of tide mode culture system was investigated in this study. this novel bioreactor provides two separated stages, liquid and gas, for cell growth requirements. the immobilized cells absorbed the nutrient from medium during the liquid stage and subsequently were exposed directly to fresh air to absorb oxygen during the gas stage. operating with pk15 cells under optimal conditions, we obtained 2.3×10(9) cells in 500ml reactor. it is 30 times higher than the initial ino ...201323261041
african swine fever virus-cell interactions: from virus entry to cell survival.viruses have adapted to evolve complex and dynamic interactions with their host cell. the viral entry mechanism determines viral tropism and pathogenesis. the entry of african swine fever virus (asfv) is dynamin-dependent and clathrin-mediated, but other pathways have been described such as macropinocytosis. during endocytosis, asfv viral particles undergo disassembly in various compartments that the virus passes through en route to the site of replication. this disassembly relies on the acid ph ...201223262167
modulation of chemokine and chemokine receptor expression following infection of porcine macrophages with african swine fever virus.african swine fever virus (asfv) is the only member of the asfarviridae, a large dna virus family which replicates predominantly in the cytoplasm. most isolates cause a fatal haemorrhagic disease in domestic pigs, although some low virulence isolates cause little or no mortality. the modulation of chemokine responses following infection of porcine macrophages with low and high virulence isolates was studied to indicate how this may be involved in the induction of pathogenesis and of effective im ...201223265239
african swine fever in the north caucasus region and the russian federation in years 2007-2012.since the introduction of the virus into the republic of georgia in 2007 african swine fever (asf) has become a large-scale epidemic involving the domestic pig population but wild boars are involved as well. from 2008 to 2009 the asf epidemic affected wild and domestic pigs in all the southern regions of the russian federation (rf). the driving force of the epidemic in its initial stages was direct contact between infected wild boars and between wild boars and traditionally free-ranging domestic ...201223266725
increased viral load and prevalence of torque teno sus virus 2 (ttsuv2) in pigs experimentally infected with classical swine fever virus (csfv).torque teno sus viruses (ttsuvs) are considered non-pathogenic viruses, although lately they have been linked to porcine circovirus diseases, mainly with post weaning multisystemic wasting syndrome (pmws). these associations point out a possible pathogenic role of ttsuvs or, alternatively, that ttsuv replication is up-regulated under disease conditions. in order to further explore the association of ttsuvs with disease occurrence, ttsuvs prevalence and viral load were assessed before and after a ...201223274109
real-time spr characterization of the interactions between multi-epitope proteins and antibodies against classical swine fever virus.the envelope glycoprotein e2 is the major immunodominant protein of the classical swine fever virus and can induce neutralizing antibodies and protective host-immune responses in infected swine. we designed, expressed, and purified multi-epitope protein (gst-bt22) that contains a tandem repeat of the e2 antigenic-determinant residues 693-704, 770-780, and 826-843, each of which is separated by a ggssgg sequence. in the same manner, we also designed, expressed, and purified a second protein (gst- ...201323291170
african swine fever virus organelle rearrangements.like most viruses african swine fever virus (asfv) subsumes the host cell apparatus in order to facilitate its replication. asfv replication is a highly orchestrated process with a least four stages of transcription, immediate-early, early, intermediate and late. as the infective cycle progresses through these stages most if not all of the organelles that comprise a nucleated cell are modified, adapted or in some cases destroyed. the entry of the virus is receptor-mediated, but the precise mecha ...201323291273
genome and molecular characterization of a csfv strain isolated from a csf outbreak in south china.in the present study, the full-length nucleotide sequences of the csfv-gz-2009 strain of classical swine fever virus (csfv) isolated from a hog pen in guangdong province in china was determined. results demonstrated that the genome of csfv-gz-2009 is 12,298 nucleotides (nt) in length, is composed of a 373-nt 5'-untranslated region (utr), has an 11,697-nt open reading frame encoding a polyprotein of 3,898 amino acids, and has a 228-nt 3'-utr. genome comparison of the csfv-gz-2009 isolate (genbank ...201323296074
standardization of pathological investigations in the framework of experimental asfv infections.african swine fever is still one of the major viral diseases of swine for which a commercial vaccine is lacking. for the design and development of such preventive products, researchers involved in african swine fever virus (asfv) vaccinology need standardized challenge protocols and well characterized clinical, pathological and immunological responses of inbreed and outbreed pigs to different viral strains and vaccine-like products. the different approaches used should be assessed by immunologis ...201323313935
detection of african swine fever virus-like sequences in ponds in the mississippi delta through metagenomic sequencing.metagenomic characterization of water virome was performed in four mississippi catfish ponds. although differing considerably from african swine fever virus (asfv), 48 of 446,100 sequences from 12 samples were similar enough to indicate that they represent new members in the family asfarviridae. at present, asfv is the only member of asfarviridae, and this study presents the first indication of a similar virus in north america. at this point, there is no indication that the identified virus(es) ...201323338931
surveillance of bungowannah pestivirus in the upper midwestern usa.pestiviruses, a genetically and antigenically highly diverse group, include one of the most historically significant swine pathogens, that is, classical swine fever virus. in australia, investigations into swine outbreaks characterized by neonatal mortality, stillbirths and mummified foetuses resulted in the discovery of a new pestivirus, bungowannah virus. this finding raised the possibility that bungowannah virus, or a variant thereof, was circulating in swine herds elsewhere in the world. if ...201423343245
hobi-like viruses: an emerging group of pestiviruses.the genus pestivirus is composed of 4 important pathogens of livestock: bovine viral diarrhea virus 1 and 2 (bvdv-1 and bvdv-2), classical swine fever virus (csfv), and border disease virus of sheep (bdv). bvdv are major pathogens of cattle, and infection results in significant economic loss worldwide. a new putative pestivirus species, tentatively called "hobi-like," "bvdv-3," or "atypical pestiviruses," was first identified in europe in fetal bovine serum (fbs) imported from brazil. hobi-like ...201323345268
down-regulation of cellular protein heme oxygenase 1 inhibits proliferation of classical swine fever virus in pk-15 cells.heme oxygenase 1 (ho-1) is an inducible enzyme that exerts potent antioxidant and anti-inflammatory effects, which also plays a critical role in host defenses against microbial, and particularly viral, infections. in our previous study, up-regulation of ho-1 was observed in peripheral blood leukocytes (pbls) by genomic expression profiling, following infection of pigs with virulent classical swine fever virus (csfv), the causative agent of a highly contagious disease threatening global pig indus ...201323357296
african swine fever virus conquers new territories. 201323376880
comparison of pcr methods for detection of classical swine fever virus and other pestiviruses.classical swine fever (csf) is a notifiable, highly contagious disease of swine controlled mainly with costly administrative methods. swine may be infected not only with classical swine fever virus (csfv), but also with other, non porcine, genetically and antigenically related pestiviruses. differentiation of infections with csfv and other pestiviruses is a crucial element of diagnostics. in the present study two real-time pcr methods and conventional one-tube nested pcr for specific detection o ...201223390749
host dna damage response facilitates african swine fever virus infection.studies with different viral infection models on virus interactions with the host cell nucleus have opened new perspectives on our understanding of the molecular basis of these interactions in african swine fever virus (asfv) infection. the present study aims to characterize the host dna damage response (ddr) occurring upon in vitro infection with the asfv-ba71v isolate. we evaluated protein levels during asfv time-course infection, of several signalling cascade factors belonging to ddr pathways ...201323398667
asfv in tanzania: asymptomatic pigs harbor virus of molecular similarity to georgia 2007.in 2011 african swine fever virus (asfv) genome was detected in asymptomatic pigs in field samples in mbeya, tanzania. the aim of this paper is to partly characterize the virus that was harbored in these pigs and furthermore to confirm, by a second sampling, the latest occurrence of asfv in the study area. asfv genome was detected in serum from 10 out of 127 healthy european/crossbreed pigs. asfv dna was polymerase chain reaction (pcr) amplified and sequenced from sera with high viral loads usin ...201323398669
cp7_e2alf oral vaccination confers partial protection against early classical swine fever virus challenge and interferes with pathogeny-related cytokine responses.the conventional c-strain vaccine induces early protection against classical swine fever (csf), but infected animals cannot be distinguished from vaccinated animals. the cp7_e2alf marker vaccine, a pestivirus chimera, could be a suitable substitute for c-strain vaccine to control csf outbreaks. in this study, single oral applications of cp7_e2alf and c-strain vaccines were compared for their efficacy to induce protection against a csf virus (csfv) challenge with the moderately virulent bas-rhin ...201323398967
complete genome sequences of classical swine fever virus isolates belonging to a new subgenotype, 2.1c, from hunan province, china.two isolates of a new classical swine fever virus (csfv) subgenotype, 2.1c (hnly-2011 and hnsd-2012), were recently isolated from pigs in hunan province, china. the most significant difference in the amino acid sequences of the polyproteins from subgenotypes 2.1a and 2.1b is an spa → tpv amino acid substitution at positions 777 and 779 in the e2 protein.201323405338
sequencing approach to analyze the role of quasispecies for classical swine fever.classical swine fever virus (csfv) is a positive-sense rna virus with a high degree of genetic variability among isolates. high diversity is also found in virulence, with strains covering the complete spectrum from avirulent to highly virulent. the underlying genetic determinants are far from being understood. since rna polymerases of rna viruses lack any proof-reading activity, different genome variations called haplotypes, occur during replication. a set of haplotypes is referred to as a viral ...201323415390
approaches to define the viral genetic basis of classical swine fever virus virulence.classical swine fever (csf), a highly contagious disease of pigs caused by the classical swine fever virus (csfv), can lead to important economic losses in the pig industry. numerous csfv isolates with various degrees of virulence have been isolated worldwide, ranging from low virulent strains that do not result in any apparent clinical signs to highly virulent strains that cause a severe peracute hemorrhagic fever with nearly 100% mortality. knowledge of the molecular determinants of csfv virul ...201323415391
in vitro antiviral activity of fluoroquinolones against african swine fever virus.african swine fever (asf) is a viral swine disease against which neither an effective vaccine nor a treatment is available. the antiviral effect of thirty fluoroquinolones on the infectivity of african swine fever virus (asfv) was screened in vitro. there was a severe reduction of the cytopathic effect in asfv-infected vero cells when exposed to six independent fluoroquinolones, or to some of their combinations, from an early phase of infection. moreover, after 7-day treatments, asfv genome coul ...201323415476
bacmam immunization partially protects pigs against sublethal challenge with african swine fever virus.lack of vaccines and efficient control measures complicate the control and eradication of african swine fever (asf). limitations of conventional inactivated and attenuated virus-based vaccines against african swine fever virus (asfv) highlight the need to use new technologies to develop efficient and safe vaccines against this virus. with this aim in mind, in this study we have constructed bacmam-shapq, a baculovirus based vector for gene transfer into mammalian cells, expressing a fusion protei ...201323428670
conformational and thermal stability improvements for the large-scale production of yeast-derived rabbit hemorrhagic disease virus-like particles as multipurpose vaccine.recombinant virus-like particles (vlp) antigenically similar to rabbit hemorrhagic disease virus (rhdv) were recently expressed at high levels inside pichia pastoris cells. based on the potential of rhdv vlp as platform for diverse vaccination purposes we undertook the design, development and scale-up of a production process. conformational and stability issues were addressed to improve process control and optimization. analyses on the structure, morphology and antigenicity of these multimers we ...201323460801
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