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detection of classical swine fever virus in semen of infected boars.during the classical swine fever (csf) epidemic in 1997 in the eu member states germany, italy, spain and the netherlands, boars in an artificial insemination (ai) centre were found to be infected with csf virus. this raised a question of epidemiological importance which could not be answered immediately. can csf virus be shed by semen of infected boars and what conclusions concerning the risk of spreading csf infection by semen can be drawn. experimental studies were conducted to answer this qu ...200011042404
the genetic basis for cytopathogenicity of pestiviruses.two biotypes of pestiviruses, cytopathogenic (cp) and noncp viruses, can be distinguished by their effects on tissue culture cells. identification of cp bovine viral diarrhea virus (bvdv) has been frequently reported since antigenically closely related noncp and cp bvdv can be isolated from cattle with fatal mucosal disease (md) and are called a virus pair. in contrast to the bvdv system, only few cp border disease virus (bdv) and cp classical swine fever virus (csfv) strains have been described ...200011042405
comparative immunohistopathology in pigs infected with highly virulent or less virulent strains of hog cholera virus.eight pigs were inoculated subcutaneously with a highly virulent hog cholera virus (hcv) strain ald. the infected pigs developed severe illness and became moribund on postinoculation day (pid) 7 or pid 10. histologic lesions were characterized by severe generalized vasculitis, necrosis of lymphocytes, and encephalitis. hcv antigen was detected in crypt tonsilar epithelial cells, macrophages, and reticular endothelial cells of lymphoid tissues. antigen localization corresponded well with histolog ...200011055862
classical swine fever: essex ban to stay in place. 200011083043
laboratory diagnosis, epizootiology, and efficacy of marker vaccines in classical swine fever: a review.detection of classical swine fever virus (csfv) can be achieved by a range of assays of which the most commonly used are: immunohistochemical and virus culture techniques. new developments have enabled the detection of viral proteins by enzyme-linked immunosorbent assays (elisas) and the detection of the viral genome by rt- pcr. so far, laboratory findings show that the latter assays may supplement or replace the conventional techniques in the near future. the detection of serum antibody against ...200011087126
transmission of classical swine fever virus by artificial insemination during the 1997-1998 epidemic in the netherlands: a descriptive epidemiological study.in the course of the 1997-1998 csf epidemic in the netherlands, two semen collection centres (scc) became infected. as an eradication strategy for an acute crisis situation, it was concluded that all semen of the boars at the sccs collected and distributed in the risk period of 28 january to 7 march 1997 was potentially contaminated (suspect semen). as a consequence, a total of 1,680 pig herds, mainly located in the southern part of the netherlands, were officially declared csf suspect. the purp ...200011087136
deletions of structural glycoprotein e2 of classical swine fever virus strain alfort/187 resolve a linear epitope of monoclonal antibody wh303 and the minimal n-terminal domain essential for binding immunoglobulin g antibodies of a pig hyperimmune serum.the major structural glycoprotein e2 of classical swine fever virus (csfv) is responsible for eliciting neutralizing antibodies and conferring protective immunity. the current structural model of this protein predicts its surface-exposed region at the n terminus with a short stretch of the c-terminal residues spanning the membrane envelope. in this study, the n-terminal region of 221 amino acids (aa) covering aa 690 to 910 of the csfv strain alfort/187 e2, expressed as a fusion product in escher ...200011090160
sero-surveillance of wild boar in the netherlands, 1996-1999.from 1996 to 1999, blood samples were collected from wild boar shot during the hunting season in crown properties, national parks and the free wildlife belt in the netherlands. sera were screened for the presence of antibodies against classical swine fever virus (csfv), swine vesicular disease virus (svdv), aujeszky's disease virus (adv) and trichinella spiralis. the results of the sero-surveillance system indicate that csfv, svdv and adv are uncommon within the wild boar population. hence, the ...200011107628
the inactivation of foot and mouth disease, aujeszky's disease and classical swine fever viruses in pig slurry.the aim of the study was to investigate the decontamination of pig slurry containing exotic viruses of pigs, foot and mouth disease virus (fmdv), aujeszky's disease virus (adv) and classical swine fever virus (csfv). laboratory-scale decontamination experiments showed that fmdv, adv and csfv were heat inactivated in slurry within 3 min at 67 degrees c, 3 min at 62 degrees c and 3 min at 60 degrees c and in glasgow eagles medium within 5 min at 67 degrees c, 4 min at 65 degrees c and 2 min at 65 ...200011119149
effects of infection of the tick ornithodoros moubata with african swine fever virus.the effects of infection with african swine fever virus (asfv) on adult and nymphal ornithodoros moubata murray (ixodoidea, argasidae) ticks were examined. three groups of ticks were used, an uninfected control group, one group infected with the vic t90/1 isolate of asfv and another group infected with the liv 13/33 isolate of asfv. infection with asfv did not affect the oviposition rates of infected ticks when compared with uninfected ticks. there was no difference between infected and uninfect ...200011129698
effects of classical swine fever virus infection on the porcine leukocyte subsets.the effects of classical swine fever (csf) virus infection on the porcine leukocyte subsets were investigated by flow cytometry in acute, chronic and convalescent forms of the disease. the virus antigen could be first detected in the monocytes on postinfection (p.i.) day 10 while in the lymphocytes on p.i. day 13. it could be established that the ratio of cd6+ cells decreased until p.i. day 6, but afterwards it started to increase and reached different values. the cd4+cd8+, the cd8+ and the cd6- ...200011402673
biochemical requirements of virus wrapping by the endoplasmic reticulum: involvement of atp and endoplasmic reticulum calcium store during envelopment of african swine fever virus.enwrapment by membrane cisternae has emerged recently as a mechanism of envelopment for large enveloped dna viruses, such as herpesviruses, poxviruses, and african swine fever (asf) virus. for both asf virus and the poxviruses, wrapping is a multistage process initiated by the recruitment of capsid proteins onto membrane cisternae of the endoplasmic reticulum (er) or associated er-golgi intermediate membrane compartments. capsid assembly induces progressive bending of membrane cisternae into the ...200010666244
[serosurveillance of notifiable veterinary diseases in wild boar in the netherlands].during the hunting season 1996-1999, blood samples were collected from wild boar shot in the netherlands. sera were screened for presence of antibodies against classical swine fever virus (csfv), swine vesicular disease virus (svdv), aujeszky's disease virus (adv), and trichinella spiralis. the results indicate that csfv, svdv, and adv are uncommon in the wild boar population. therefore, it seems that csfv, svdv, and adv infection in the wild boar population is not an important reservoir in the ...200010666784
effects of chlorine, iodine, and quaternary ammonium compound disinfectants on several exotic disease viruses.the effects of three representative disinfectants, chlorine (sodium hypochlorite), iodine (potassium tetraglicine triiodide), and quaternary ammonium compound (didecyldimethylammonium chloride), on several exotic disease viruses were examined. the viruses used were four enveloped viruses (vesicular stomatitis virus, african swine fever virus, equine viral arteritis virus, and porcine reproductive and respiratory syndrome virus) and two non-enveloped viruses (swine vesicular disease virus (svdv) ...200010676896
within-farm spread of classical swine fever virus--a blueprint for a stochastic simulation model.a stochastic simulation model to investigate the transmission of classical swine fever (csf) virus within an infected farm is described. the model is structured according to the processes that occur within and between management groups (pig units or houses). it uses the individual pig as the unit of interest and estimates the number of animals in the states 'susceptible', 'infected', 'infectious', and 'removed' for each day of the disease incident. probabilities are assigned to the transitions b ...200010682386
protection of pigs against classical swine fever with dna-delivered gp55.classical swine fever virus causes significant mortality and morbidity in commercial piggeries in many countries in europe and asia. the protective antigen, gp55, is highly conformation-dependent and thus killed virus or bacterially produced proteins are not protective. this report demonstrates that dna vaccination with the gene encoding gp55 can provide protective immunity with inoculation of two doses of 25 microg dna or a single shot of 200 microg. furthermore, the dna can be delivered intram ...200010699343
[laboratory findings during the classic swine fever epidemic of 1997-1998].the results of the laboratory tests carried out by the institute for animal science and health (id-lelystad), the netherlands, on samples collected during the classical swine fever (csf) epidemic 1997-1998 are summarized in this article. the relevance of the different laboratory tests and various samples collected on the eradication of csf during an outbreak is evaluated.200010705606
classical swine fever virus e(rns) deletion mutants: trans-complementation and potential use as nontransmissible, modified, live-attenuated marker vaccines.an sk6 cell line (sk6c26) which constitutively expressed the glycoprotein e(rns) of classical swine fever virus (csfv) was used to rescue csfv e(rns) deletion mutants based on the infectious copy of csfv strain c. the biochemical properties of e(rns) from this cell line were indistinguishable from those of csfv e(rns). two e(rns) deletion mutants were constructed, virus flc23 and virus flc22. virus flc23 encoded only the utmost n- and c-terminal amino acids of e(rns) (deletion of 215 amino acids ...200010708411
recombinant classical swine fever (csf) viruses derived from the chinese vaccine strain (c-strain) of csf virus retain their avirulent and immunogenic characteristics.two recombinant classical swine fever (csf) viruses (flc2, flc3) transcribed from a dna copy of the genome of the chinese (c) strain, a csf virus vaccine strain, were characterized in vivo in rabbits and pigs. rabbits were inoculated intravenously with flc2 or flc3, the parent c-strain virus, a biologically cloned c-strain or csf virus strain brescia (c.1.1.1). after 24-96 h fever was detected in the rabbits inoculated with the different c-strain viruses. apart from those in the control group, a ...200010738091
transient classical swine fever virus infection in wild boar piglets partially protected by maternal antibodies.an experimental study was conducted to investigate the clinical course of classical swine fever (csf) in wild boar piglets partially protected by maternal antibodies. five healthy wild boar piglets with a low serum titre of colostral antibodies against csf virus were challenged with virulent csf virus at the age of three months. apart of reduced food intake and diarrhoea no major clinical symptoms were noticed after challenge. these signs were seen during the second and third week of infection, ...200010743336
an arid family protein binds to the african swine fever virus encoded ubiquitin conjugating enzyme, ubcv1.the nh(2)-terminal end of a protein, named smcp, which contains an arid (a/t rich interaction domain) dna binding domain and is similar to the mammalian smcy/smcx proteins and retinoblastoma binding protein 2, was shown to bind the african swine fever virus encoded ubiquitin conjugating enzyme (ubcv1) using the yeast two hybrid system and in in vitro binding assays. antisera raised against the smcp protein were used to show that the protein is present in the cell nucleus. immunofluorescence show ...200010760505
[synthesis and immunochemical properties of the recombinant major surface glycoprotein e2 of the classical swine fever virus].recombinant e2 protein from vaccine strain of classical swine fever virus (csfv) and from scfv virulent strain shimen was synthesized in sf-21 and high-five cell culture with baculovirus as the expressing vector. for secretion, hydrophobic c-terminal transmembrane domain was removed and n-terminal signal polypeptide of 38 amino acids was added. maximum accumulation of recombinant products in sf-21 cells was observed after 48 h and in medium 96 h after infection with recombinant baculovirus. in h ...200010765548
[use of recombinant protein e2 of the classical swine fever virus for immunization of animals].recombinant major surface glycoprotein e2 from virulent shimen strain of classical swine fever virus (csfv) has been tested for immunogenicity in animal immunization experiments. immunization of 3-month-old piglets with 200 micrograms of recombinant protein protected the animals from lethal challenge with virulent csfv strain. csfv-specific antibody detection test based on competitive elisa has been developed using the recombinant e2 protein. the test can evaluate specific antibody levels after ...200010765549
introduction to classical swine fever: virus, disease and control policy.classical swine fever virus is a spherical enveloped particle of about 40-60 nm in diameter with a single stranded rna genome of about 12,300 bases with positive polarity, classified as a pestivirus within the family flaviviridae. natural hosts are domestic and wild pigs. the virus causes one of the most severe diseases in pigs world wide with grave economic consequences. the clinical picture of classical swine fever is variable, depending on the age of the affected animals and viral virulence. ...200010785320
structure and presentation of a world wide web database of csf virus isolates held at the eu reference laboratory.a computerized database was generated with the epidemiological data of more than 600 csf virus strains and isolates kept in the eu reference laboratory for classical swine fever in hanover. in addition, as sequence data from defined regions of the genome are increasingly being used for genetic typing of new isolates and are thus being published, it was decided to integrate them into the database. in order to make the epidemiological and the sequence data available to other laboratories through t ...200010785323
genetic typing of classical swine fever virus.three regions of the classical swine fever virus (csfv) genome that have been widely sequenced were compared with respect to their ability to discriminate between isolates and to segregate viruses into genetic groups. sequence data-sets were assembled for 55 csfvs comprising 150 nucleotides of the 5' non-translated region, 190 nucleotides of the e2 envelope glycoprotein gene and 409 nucleotides of the ns5b polymerase gene. phylogenetic analysis of each data-set revealed similar groups and subgro ...200010785324
classical swine fever virus: a ring test to evaluate rt-pcr detection methods.six laboratories participated in an exercise to compare the sensitivity and specificity of rt-pcr tests for the detection of classical swine fever virus (csfv). two sets of coded samples were prepared by serial dilution of positive samples and then distributed to each of the laboratories. one set comprised 34 samples of random primed cdna. these had been synthesised from viral rna representative of seven different genetic subtypes of csfv. the other set comprised 40 clinical samples containing t ...200010785325
survival and inactivation of classical swine fever virus.classical swine fever virus, like many enveloped viruses, may be regarded as moderately fragile. it shows a short but variable survival time in the environment, depending on physical conditions, but may remain viable for prolonged periods in favourable circumstances as found, for example, in stored meat. published information is reviewed on the survival time of the virus under a variety of physical and chemical conditions, including the variable influence of environmental factors. the principles ...200010785326
laboratory experience during the classical swine fever virus epizootic in the netherlands in 1997-1998.from february 1997 till may 1998 the national reference laboratory for classical swine fever (csf) in the netherlands was confronted with millions of samples taken from pigs during an outbreak of csf in a pig dense region. in a limited period major logistic problems needed to be solved regarding the processing of samples and information at the laboratory facilities. in total over 2.3 million samples were examined by different csf diagnostic methods. the majority (approximately 2.1 million) of th ...200010785328
development of a classical swine fever subunit marker vaccine and companion diagnostic test.the development of a classical swine fever (csf) subunit marker vaccine, based on viral envelope glycoprotein e2, and a companion diagnostic test, based on a second viral envelope glycoprotein e(rns), will be described. important properties of the vaccine, such as onset and duration of immunity, and prevention of horizontal and vertical transmission of virus were evaluated. a single dose of the vaccine protected pigs against clinical signs of csf, following intranasal challenge with 100ld(50) of ...200010785329
susceptibility of piglets to rabbit hemorrhagic disease virus following experimental infection.the possibility exists that rabbit hemorrhagic disease virus (rhdv) can be transmitted to swine, through lapinized hog cholera virus (hcv) vaccine. to investigate the infectivity of rhdv in swine, 16 four- to six-week-old piglets were inoculated subcutaneously with rhdv, and samples of liver, lung, spleen, kidney, bile, adrenal gland, tonsil, mesenteric lymph node, thymus, urine, buffy coat, and feces were collected from each of 2 animals on days 0, 1, 2, 3, 5, 7, 14, and 28 post infection. usin ...200010805254
determination of the onset of the herd-immunity induced by the e2 sub-unit vaccine against classical swine fever virus.for a recently developed e2 subunit vaccine against classical swine fever (csf), the reduction in transmission, at different moments after vaccination, was assessed by animal experiments and statistical calculations. two experiments were performed to estimate the reproduction ratio r. experiment 1 consisted of three groups and experiment 2 of two groups each of 10 pigs. in four of these groups, all pigs were vaccinated intramuscularly with the vaccine. the pigs in the fifth group remained unvacc ...200010618535
vaccination with a single dose of a recombinant porcine adenovirus expressing the classical swine fever virus gp55 (e2) gene protects pigs against classical swine fever.a recombinant porcine adenovirus (rpav) with the gp55 (e2) gene from the classical swine fever virus (csfv) 'weybridge' strain inserted into the right hand end of the pav serotype 3 (pav3) genome was constructed. expression of gp55 was directed by the major late promoter and tri-partite leader sequences located and cloned from pav3. no compensatory deletions of pav dna sequences were made. vaccination of outbred pigs with a single dose of the recombinant virus (rpav-gp55) resulted in complete pr ...200010590324
evaluation of vaccine-induced maternal immunity against classical swine fever.the vaccine-induced maternal immunity against classical swine fever (csf) was investigated in this study. eight sows were vaccinated with the chinese strain of classical swine fever virus (csfv). the length of time between vaccination and farrowing was 167-217 days. milk samples from the front, middle and back udder sections and blood samples were taken from the sows on days 3 and 14 after farrowing. blood samples were obtained from the piglets at the age of 3, 6 and 10 weeks. the antibody level ...200111402686
[differential diagnosis of classical swine fever and border disease: seroepidemiological investigation of a pestivirus infection on a mixed sheep and swine farm].during recent years neutralizing antibodies against border disease virus (bdv) were found repeatedly in german pig herds. consequently there was a demand for a differential diagnostic system. a permanent sheep cell line and bdv reference strain moredun were chosen and were applied in a could be used case study. a pestivirus could be isolated from piglets on a mixed farm and was characterised as 'non-classical swine fever' (csf) by using monoclonal antibodies. due to a csf suspicion the pig herd ...200111417380
atypical cilia in the bronchiolar epithelium of pigs experimentally infected with hog cholera virus.to study the effect of hog cholera virus on the epithelial cells of the bronchiolar mucosa, 12 pigs were inoculated with a highly virulent strain. immunohistochemical and ultrastructural examination of the ciliated epithelial cells demonstrated an increase in the number of atypical cilia. the latter showed alterations in the microtubular pattern, possibly resulting from viral interference with the normal metabolism of the epithelial cells.200111428186
transovarial transmission of african swine fever virus in the argasid tick ornithodoros moubata.the aim of this study was to determine filial infection prevalence of experimentally infected colony ornithodoros moubata walton (ixodoidea: argasidae) ticks for african swine fever virus (asfv). three groups of ticks were used: an uninfected control group, one group orally infected with the vic t90/1 isolate and another group orally infected with the liv 13/33 isolate of asfv. the results show that filial infection prevalences were not constant but were highly variable between egg batches from ...200111434547
african swine fever virus structural protein pe120r is essential for virus transport from assembly sites to plasma membrane but not for infectivity.this report examines the role of african swine fever virus (asfv) structural protein pe120r in virus replication. immunoelectron microscopy revealed that protein pe120r localizes at the surface of the intracellular virions. consistent with this, coimmunoprecipitation assays showed that protein pe120r binds to the major capsid protein p72. moreover, it was found that, in cells infected with an asfv recombinant that inducibly expresses protein p72, the incorporation of pe120r into the virus partic ...200111435554
classical swine fever: pathogenesis of glomerular damage and immunocharacterization of immunocomplex deposits.twenty-six pigs were inoculated with a virulent isolate (quillota strain) of classical swine fever (hog cholera) virus to determine the chronological development of lesions in the renal glomeruli and the pathogenesis of glomerular damage and immunocomplex deposition. the study included the use of histopathological, ultrastructural and immunohistochemical (detection of viral antigen gp55, myeloid-histiocyte antigen, igm, igg and c1q) techniques. the main changes in glomerular structure were obser ...200111437500
classical swine fever: morphological and morphometrical study of pulmonary intravascular macrophages.to gain further insight into the pathogenesis of classical swine fever (csf), the changes induced by hog cholera (hc) virus in pulmonary intravascular macrophages (pims) were examined. twelve pigs were inoculated by the intramuscular route with a virulent strain of hc virus (quillota strain) and killed in groups of three at 4, 7, 10 and 14 days post-inoculation. immunohistochemical and ultrastructural examination revealed hc virus infection in endothelial cells, pims, and interstitial and alveol ...200111437510
[african swine fever virus: achievements over the last decade of the 20th century].complete nucleotide sequence of african swine fever (asf) virus genome was determined in 1993-1999. deletion mutants with low virulence for pigs were obtained. genes of structural (p72, p54, p12, cleavage products pp220 and pp60, hemagglutinin) and nonstructural (p32) proteins were mapped. the significance of different proteins in virus adsorption and resistance to challenge was elucidated, their location in infected cell and virion was determined. lipid composition of the virus was studied. a p ...200111449798
[genetic typing of german isolates of classical swine fever virus].during the last decade several outbreaks of classical swine fever (csf) occurred in germany in domestic pigs and in wild boar, respectively. two major epidemics which also affected other eu member states were recorded. to support epidemiological investigations genetic typing was applied and virus isolates originating from different outbreaks in germany were assigned to groups and virus types. two genomic regions were selected for the phylogenetic analysis, namely 150 nucleotides from the 5' non- ...200111449911
a virally encoded chaperone specialized for folding of the major capsid protein of african swine fever virus.it is generally believed that cellular chaperones facilitate the folding of virus capsid proteins, or that capsid proteins fold spontaneously. here we show that p73, the major capsid protein of african swine fever virus (asfv) failed to fold and aggregated when expressed alone in cells. this demonstrated that cellular chaperones were unable to aid the folding of p73 and suggested that asfv may encode a chaperone. an 80-kda protein encoded by asfv, termed the capsid-associated protein (cap) 80, b ...200111461995
isolation of a non-haemadsorbing, non-cytopathic strain of african swine fever virus in madagascar.african swine fever (asf) suspected clinically in madagascar (1998-9) was confirmed by polymerase chain reaction (pcr) and nucleotide sequencing, following virus isolation. no haemadsorption or cytopathic effect could be detected following leukocyte inoculation, but viral growth in cells was confirmed by pcr. detection of asf virus genome was carried out by amplification of a highly conserved region coding for the p72 protein. nucleotide sequencing of the amplicon revealed 99.2% nucleotide ident ...200111467803
application of competitive enzyme-linked immunosorbent assay for the serologic diagnosis of classical swine fever virus infection.a competitive enzyme-linked immunosorbent assay (c-elisa), based on a truncated e2 recombinant protein of the alfort/187 strain of classical swine fever virus (csfv) and a specific monoclonal antibody m1669, was evaluated using 2,000 sera from clinically healthy pigs in canada (a csfv-free country) and sera from experimentally infected pigs. the relative specificity and sensitivity of the c-elisa were 100% and 86%, respectively, at a cutoff of 25% inhibition using negative and positive pig sera, ...200111478613
characterization of pseudotype vsv possessing hcv envelope proteins.the genome of hepatitis c virus (hcv) encodes two envelope glycoproteins (e1 and e2), which are thought to be responsible for receptor binding and membrane fusion resulting in virus penetration. to investigate cell surface determinants important for hcv infection, we used a recombinant vesicular stomatitis virus (vsv) in which the glycoprotein gene was replaced with a reporter gene encoding green fluorescent protein (gfp) and produced hcv-vsv pseudotypes possessing chimeric hcv e1 or e2 glycopro ...200111485395
evidence for the presence of two novel pestivirus species.the genus pestivirus of the family flaviviridae comprises four species, namely bovine viral diarrhea virus-1 (bvdv-1), bvdv-2, border disease virus (bdv), and classical swine fever virus (csfv). comparative analyses of partial sequences have suggested that pestivirus isolates from giraffe (giraffe-1) and reindeer (reindeer-1) are distinct from the established species (becher et al., virology 262, 64--71, 1999). in this study, we report the complete genomic sequences of pestivirus strains giraffe ...200111485413
novel virulence and host range genes of african swine fever virus.current work is beginning to reveal the complex mechanisms by which african swine fever virus interacts with its swine and tick hosts. this work includes the identification of novel viral genes that mediate virulence and host range, and influence important cellular regulatory pathways.200111495811
characterization of classical swine fever virus associated with defective interfering particles containing a cytopathogenic subgenomic rna isolated from wild boar.classical swine fever virus (csfv) strain wb82, isolated from a wild boar in 1982, induced a distinct cytopathic effect (cpe) in primary swine testicle cell culture and in most of the porcine cell lines. this strain of csfv was found to be composed of two biotypes. cytopathogenic (cp) csfv, as a minor population, and noncytopathogenic (noncp) csfv, as a major population. the noncp csfv (designated strain wb82/e+) was obtained by biological cloning, and it showed the exaltation of newcastle disea ...200111503902
classical swine fever (csf): a historical review of research and vaccine production on the isle of riems.a review on classical swine fever (csf) research and vaccine production is given about four historical periods (1924-1948, 1949-1969, 1970-1991, since 1992). similar as to research on foot and mouth disease, applied topics as diagnosis, pathogenesis, epidemiology and control represented the csf research over many years. the development of vaccines and application procedures, e.g. oral and aerogenic immunisation and combined vaccines for large pig farms were the prominent investigations between 1 ...200111505796
oral immunisation against classical swine fever (csf): onset and duration of immunity.in an experimental study, onset and duration of immunity after oral immunisation of pigs with a classical swine fever (csf) live virus vaccine based on the strain "c" has been evaluated. sixteen weaner piglets (group 1) were orally instilled by syringe with the content of one vaccine bait whereas eighteen piglets (group 2) were fed with one bait. six unvaccinated piglets represented the control group (group 3). the pigs having 2, 4, 6 and 10 days post vaccination (p.v.) were challenged with the ...200111506924
mutagenesis of hepatitis c virus e1 protein affects its membrane-permeabilizing activity.the e1 glycoprotein of hepatitis c virus is a transmembrane glycoprotein with a c-terminal anchor domain. when expressed in escherichia coli, e1 induces a change in membrane permeability that is toxic to the bacterial cell. the c-terminal hydrophobic region (aa 331-383) of e1 is mainly responsible for membrane association and for inducing changes in membrane permeability. these observed changes are similar to those produced in e. coli by influenza virus m2, human immunodeficiency virus gp41 and ...200111514735
evaluation of the potential of dogs, cats and rats to spread classical swine fever virus. 200111548961
a recombinant classical swine fever virus with a marker insertion in the internal ribosome entry site.based on an infectious cdna clone of classical swine fever virus (csfv) strain alfort/187 (ruggli et al., j virol 70, 3478-3487, 1996) a full-length cdna was constructed harbouring a nonviral 44 base insertion in the internal ribosome entry site (ires) within the 5' nontranslated region (5'ntr) of the genome. genome size rna transcribed in vitro served as a positive control in routine rt-pcr used to detect csfv rna in diagnostic material. unexpectedly this rna proved to be infectious upon transf ...200111556403
attenuated lapinized chinese strain of classical swine fever virus: complete nucleotide sequence and character of 3'-noncoding region.the complete nucleotide sequence including precise 5'- and 3'-terminal non-coding regions (ncrs) of the attenuated lapinized chinese strain (hclv) of classical swine fever virus (csfv) was determined from overlapping cdna clones constructed by separated rt-pcr and rapid amplification of cdna ends (race) methods. the genomic rna of the hclv strain consists of 12.310 nucleotides (nts) including 374 nts and 242nts in the 5'- and 3'-ncrs, respectively. it contains one large open reading frame (orf) ...200111556404
classical swine fever (csf) marker vaccine. trial i. challenge studies in weaner pigs.two commercial marker vaccines against classical swine fever virus (csfv) and companion diagnostic tests were examined in 160 conventional pigs. to test the vaccines in a "worst case scenario", group of 10 weaners were vaccinated using a single dose of an e2 (gp55) based vaccine at days -21, -14, -10 or -7, and subsequently challenged at day 0. the challenge virus was csfv 277, originating from a recent outbreak of classical swine fever (csf) in germany. in all groups, only 5 out of 10 pigs were ...200111557151
classical swine fever (csf) marker vaccine. trial ii. challenge study in pregnant sows.the efficacy of two marker vaccines against classical swine fever (csf) was tested in a large scale laboratory trial in several national swine fever laboratories (nsfl) of the eu member states. the vaccines were: bayovac csf marker (vaccine a) from bayer, leverkusen, germany and porcilis pesti (vaccine b) from intervet, boxmeer, the netherlands. at the nsfl of belgium, the netherlands and germany experiments were carried out to examine the ability of the vaccines to prevent transplacental transm ...200111557152
classical swine fever (csf) marker vaccine. trial iii. evaluation of discriminatory elisas.the objective of the marker vaccine trial was to test the two available csf marker vaccines in scenarios which are likely to occur in the field and to evaluate the reliability of the discriminatory tests. the evaluation of the discriminatory tests was of special importance because there is no requirement for formal data concerning their performance by the european medicinal products evaluation agency (emea) in london. emea is responsible for the licensing procedure of the marker vaccines within ...200111557153
interaction of classical swine fever virus with membrane-associated heparan sulfate: role for virus replication in vivo and virulence.passage of native classical swine fever virus (csfv) in cultured swine kidney cells (sk6 cells) selects virus variants that attach to the surface of cells by interaction with membrane-associated heparan sulfate (hs). a ser-to-arg change in the c terminus of envelope glycoprotein e(rns) (amino acid 476 in the open reading frame of csfv) is responsible for selection of these hs-binding virus variants (m. m. hulst, h. g. p. van gennip, and r. j. m. moormann, j. virol. 74:9553-9561, 2000). in this i ...200111559790
african swine fever virus protein p54 interacts with the microtubular motor complex through direct binding to light-chain dynein.dynein is a minus-end-directed microtubule-associated motor protein involved in cargo transport in the cytoplasm. african swine fever virus (asfv), a large dna virus, hijacks the microtubule motor complex cellular transport machinery during virus infection of the cell through direct binding of virus protein p54 to the light chain of cytoplasmic dynein (lc8). interaction of p54 and lc8 occurs both in vitro and in cells, and the two proteins colocalize at the microtubular organizing center during ...200111559815
an e2 sub-unit marker vaccine does not prevent horizontal or vertical transmission of classical swine fever virus.an experimental infection with classical swine fever (csf) virus in e2 sub-unit marker vaccine vaccinated gilts was conducted in order to evaluate the effect of vaccination on virus transmission and course of the disease. therefore, clinical signs as well as horizontal and vertical virus transmission were monitored in two inoculated, non-vaccinated and 10 vaccinated conventional gilts, housed in individual sow boxes. within 10 days post-inoculation, all vaccinated gilts became infected. dependin ...200111567750
molecular epidemiology of classical swine fever in italy.to gain an insight into the molecular epidemiology of classical swine fever (csf) in italy, virus isolates originating from outbreaks that occurred between 1985 and 2000 in wild boar or in domestic pigs in mainland italy and in sardinia were analysed by genetic typing. for this, a fragment (190 nucleotides) of the e2 glycoprotein gene was sequenced and phylogenetic analyses were performed, including older italian isolates and isolates from recent outbreaks in europe for comparison. the results s ...200111574170
morphological and immunohistochemical changes in splenic macrophages of pigs infected with classical swine fever.classical swine fever (csf) was induced in 20 pigs by inoculation with a virulent strain of csf virus to determine sequential changes (2, 4, 7, 10 and 14 days post-inoculation) in the number and morphology of splenic macrophages (red pulp and lymphoid marginal zone) and thus to assess the role of these cells in the pathogenesis of the disease. the first splenic cells to be infected with csf virus were macrophages in the marginal zone followed by other macrophage populations. the initial phase of ...200111578124
evaluation of diagnostic tests for the detection of classical swine fever in the field without a gold standard.knowledge of the sensitivity of diagnostic tests for infectious diseases under field conditions can be used to design a surveillance program that increases the effectiveness of the control policy. in this study, the sensitivity of tests for the detection of classical swine fever (csf) virus (csfv) under field conditions was estimated without knowledge of the true disease status of the animals tested. during the csf epidemic of 1997-1998 in the netherlands, tonsil samples from pigs of csf suspect ...200111580058
african swine fever virus infection of porcine aortic endothelial cells leads to inhibition of inflammatory responses, activation of the thrombotic state, and apoptosis.african swine fever (asf) is an asymptomatic infection of warthogs and bushpigs, which has become an emergent disease of domestic pigs, characterized by hemorrhage, lymphopenia, and disseminated intravascular coagulation. it is caused by a large icosohedral double-stranded dna virus, african swine fever virus (asfv), with infection of macrophages well characterized in vitro and in vivo. this study shows that virulent isolates of asfv also infect primary cultures of porcine aortic endothelial cel ...200111581405
erns protein of pestiviruses. 200111586914
factors associated with the introduction of classical swine fever virus into pig herds in the central area of the 1997/98 epidemic in the netherlands.a matched case-control study of 135 infected and 99 uninfected pig herds from the central area of the 1997 to 1998 epidemic of classical swine fever (csf) in the netherlands was undertaken to identify factors associated with the introduction of the virus. the herds were matched on the basis of herd type and the shortest geographical distance between pairs of herds. data on management, hygienic measures, experiences during the depopulation of an infected nearest neighbour, and the frequency of co ...200111601514
the effect of vaccination with the pav-250 strain classical swine fever (csf) virus on the airborne transmission of csf virus.the airborne transmission of classical swine fever (csf) virus to susceptible pigs, as well as the effect of vaccination with the csf virus pav-250 strain was investigated on this mode of transmission. experiment i: four pigs were inoculated with the ald csfv strain (10(4.3) 50% tcid) by the intramuscular route, and at the onset of fever, they were introduced into an enclosed chamber. at the end of the experiment surviving pigs were sedated, anesthetized and euthanatized. experiment ii: four pig ...200111642288
phylogenetic comparison of classical swine fever virus in china.an n-terminal fragment of the e2 gene of classical swine fever (csf) virus encoding major immunogenic sites was amplified by rt-pcr directly from 110 clinical specimens representing 109 epizootic sites during the last decade in china. phylogenetic relationships between these viruses as well as 20 reference strains were determined by comparison of their nucleotide sequences. a phylogenetic tree showed that 103 of the 110 field viruses (93.6%) were clustered within group 2 and subdivided into thre ...200111682122
[sero-monitoring of notifiable diseases in wild boar in the netherlands 1999-2001].within the framework of a sero-monitoring system, in operation since 1996. blood samples from wild boar shot during the hunting seasons 1999-2000 and 2000-2001 in the netherlands were screened for the presence of antibodies against classical swine fever virus (csfv), swine vesicular disease virus (svdv), and anjeszky's disease virus (adv). the results indicate that csfv, svdv, and adv are uncommon in the wild boar population in the netherlands. because of the recent foot-and-mouth disease (fmd) ...200111780256
a 250-nucleotide ua-rich element in the 3' untranslated region of xenopus laevis vg1 mrna represses translation both in vivo and in vitro.xenopus laevis vgl mrna undergoes both localization and translational control during oogenesis. vg1 protein does not appear until late stage iv, after localization is complete. to determine whether vg1 translation is regulated by cytoplasmic polyadenylation, the race-pat method was used. vg1 mrna has a constant poly(a) tail throughout oogenesis, precluding a role for cytoplasmic polyadenylation. to identify cis-acting elements involved in vg1 translational control, the vg1 3' utr was inserted do ...200111780632
pathogenesis of african swine fever virus in ornithodoros ticks.african swine fever virus (asfv) is the only known dna arbovirus and the sole member of the family asfarviridae. it causes a lethal, hemorrhagic disease in domestic pigs. asfv is enzootic in sub-saharan africa and is maintained in a sylvatic cycle by infecting both wild members of the suidae (e.g. warthogs) and the argasid tick ornithodoros porcinus porcinus. the pathogenesis of asfv in o. porcinus porcinus ticks is characterized by a low infectious dose, lifelong infection, efficient transmissi ...200111831434
sequences and replication of genomes of the archaeal rudiviruses sirv1 and sirv2: relationships to the archaeal lipothrixvirus sifv and some eukaryal viruses.the double-stranded dna genomes of the viruses sirv1 and sirv2, which infect the extremely thermophilic archaeon sulfolobus and belong to the family rudiviridae, were sequenced. they are linear, covalently closed at the ends, and 32,312 and 35,502 bp long, respectively, with an a+t content of 75%. the genomes of sirv1 and sirv2 carry inverted terminal repeats of 2029 and 1628 bp, respectively, which contain multiple direct repeats. sirv1 and sirv2 genomes contain 45 and 54 orfs, respectively, of ...200111878892
genetic typing of ruminant pestivirus strains from northern ireland and the republic of ireland.a study was performed to investigate the genotypes and sub-groups of pestiviruses present in ruminants in ireland. these comprised one ovine and eighteen bovine pestiviruses from northern ireland and six bovine pestiviruses from the republic of ireland. a 288 base pair (bp) portion of the 5'-non coding region (5'-ncr) from each of 25 pestiviruses collected over a period of 31 years was amplified by reverse-transcription-polymerase chain reaction (rt-pcr) and the product directly sequenced. from ...200111883891
solution structure of a viral dna repair polymerase.dna polymerase x (pol x) from the african swine fever virus (asfv) specifically binds intermediates in the single-nucleotide base-excision repair process, an activity indicative of repair function. in addition, pol x catalyzes dna polymerization with low nucleotide-insertion fidelity. the structural mechanisms by which dna polymerases confer high or low fidelity in dna polymerization remain to be elucidated. the three-dimensional structure of pol x has been determined. unlike other dna polymeras ...200111685238
solution structure of a viral dna polymerase x and evidence for a mutagenic function.the african swine fever virus dna polymerase x (asfv pol x or pol x), the smallest known nucleotide polymerase, has recently been reported to be an extremely low fidelity polymerase that may be involved in strategic mutagenesis of the viral genome. here we report the solution structure of pol x. the structure, unique within the realm of nucleotide polymerases, consists of only palm and fingers subdomains. despite the absence of a thumb subdomain, which is important for dna binding in other polym ...200111685239
epitope mapping and affinity purification of monospecific antibodies by escherichia coli cell surface display of gene-derived random peptide libraries.we report a method for the precise mapping of linear epitopes by presenting a peptide library on the surface of escherichia coli cells. a random library of gene fragments derived from the classical swine fever virus (csfv) envelope protein e(rns) was generated by dnase i cleavage and cloned into a specially designed bacterial surface display vector. a carboxyterminally truncated intimin, an adhesin from enteropathogenic e. coli, serves as a carrier protein to present foreign peptides on the surf ...200111687250
the trans golgi network is lost from cells infected with african swine fever virus.the cellular secretory pathway is important during the assembly and envelopment of viruses and also controls the transport of host proteins, such as cytokines and major histocompatibility proteins, that function during the elimination of viruses by the immune system. african swine fever virus (asfv) encodes at least 26 proteins with stretches of hydrophobic amino acids suggesting entry into the secretory pathway (r. j. yanez, j. m. rodriguez, m. l. nogal, l. yuste, c. enriquez, j. f. rodriguez, ...200111689656
antigenic and immunogenic properties of a chimera of two immunodominant african swine fever virus proteins.a chimera of the two immunodominant african swine fever (asf) virus proteins p54 and p30 was constructed by insertion of the gene cp204l into a not i restriction site of e183l gene. the resulting chimeric protein p54/30, expressed by a recombinant baculovirus in insect cells and in trichoplusia ni larvae, retained antigenic determinants present in both proteins and reacted in western blot with a collection of sera from inapparent asf virus carrier pigs. remarkably, pigs immunized with the chimer ...200111699955
oral and sub-cutaneous vaccination of commercial pigs with a recombinant porcine adenovirus expressing the classical swine fever virus gp55 gene.a recombinant porcine adenovirus expressing the classical swine fever virus (csfv) gp55/e2 gene was administered to commercially available pigs via oral or subcutaneous routes and their susceptibility to oral and subcutaneous challenge with csfv was determined. 100% of animals vaccinated and challenged subcutaneously were protected. in the groups of pigs vaccinated either orally or subcutaneously and then challenged orally, 60% of animals were protected. before challenge, neutralising antibodies ...200111699963
adsorption of colostral antibodies against classical swine fever, persistence of maternal antibodies, and effect on response to vaccination in baby pigs.to determine kinetics of antibody absorption, persistence of antibody concentrations, and influence of titers on vaccination of baby pigs with a vaccine against classical swine fever (csf).200111703028
an experimental infection with classical swine fever virus in pregnant sows: transmission of the virus, course of the disease, antibody response and effect on gestation.an experimental infection with classical swine fever (csf) virus in 12 conventional gilts, housed in a sow-box housing system, was conducted in order to evaluate horizontal transmission, clinical, virological and serological response, and the effect on gestation. two of the 12 gilts, of which 10 were pregnant, were experimentally inoculated. they became viraemic for the first time 6 days post-inoculation (dpi). the contact gilts became viraemic between 18 and 21 days post inoculation. on the bas ...200111708677
are low-density granulocytes the major target cells of classical swine fever virus in the peripheral blood?the target cells of classical swine fever (csf) virus in the peripheral blood of pigs infected with recent field isolates from germany were studied. eight weaned pigs were inoculated oronasally with the csf virus field isolate visbek/han 95 and three weaners were inoculated with the isolate losten/freese 98. all pigs showed severe clinical signs typical of csf and died or had to be euthanized between 9 and 24 days post-infection (dpi). the first cells in the peripheral blood which became infecte ...200111708678
the ns5a protein of bovine viral diarrhoea virus interacts with the alpha subunit of translation elongation factor-1.a cellular protein that interacts with the ns5a polypeptide of bovine viral diarrhoea virus (bvdv) was identified in a yeast two-hybrid screen. the ns5a interactor was identified as the alpha subunit of bovine translation elongation factor 1a (eef1a). cell-free binding studies were performed with chimeric ns5a fused to glutathione s-transferase (gst-ns5a) expressed in bacteria. gst-ns5a bound specifically to both in vitro-translated and mammalian cell-expressed eef1a. moreover, purified eef1a bo ...200111714969
cloning and sequencing of full-length cdna of classical swine fever virus lpc strain.the cdnas of classical swine fever virus (lpc vaccine strain) were cloned by transcriptase-polymerase chain reaction, and their nucleotide sequences were determined. in this work, we obtained the sequence information of the 786 bases of the 5'-terminal region, 6049 bases of the middle region, and 1648 bases of the 3'-terminal region. taking our previous results and present data together, the entire genomic sequence of lpc strain was completed (12344 nucleotides in length). the genome of lpc has ...200111724273
1h, 15n, and 13c resonance assignments for a 20 kda dna polymerase from african swine fever virus. 200111727982
mechanism of interferon resistance in hepatitis c. 200111728538
the correlation of virus-specific interferon-gamma production and protection against classical swine fever virus infection.the level of antigen-specific interferon-gamma (ifn-gamma) production can be used as an indicator of cellular immunity. in this study, we investigated the role of cellular immune response in protection against classical swine fever virus (csfv). pigs were vaccinated once with csfv vaccine and challenged 6 days post-vaccination (dpv). vaccinated animals had significantly higher csfv-specific ifn-gamma secreting cells than the unvaccinated pigs (p<0.05) at the time of challenge and were protected ...200111730928
a serodiagnostic elisa using recombinant antigen of swine transmissible gastroenteritis virus nucleoprotein.a serodiagnostic elisa utilizing the recombinant nucleoprotein (rn protein) of transmissible gastroenteritis virus (tgev) was developed, and evaluated by examining a panel of 141 virus neutralization (vn) positive and 101 negative sera. the rn protein-based elisa (rnelisa) appeared to be highly sensitive and specific (98.6% and 98.0%, respectively) when it was compared to the vn test. the result was similar to that of an elisa based on purified viral antigens with showing good correlation (r=0.8 ...200111767065
identification and characterization of a spliced c-type lectin-like gene encoded by rat cytomegalovirus.the english isolate of rat cytomegalovirus (rcmv) encodes a 20-kda protein with a c-type lectin-like domain that is expressed in the delayed-early and late phases of the viral replication cycle. genomic sequence analysis of the restriction fragment kpnr of rcmv revealed significant homology to several c-type lectin-containing molecules implicated in natural killer (nk) and t-cell interactions, as well as genes from four poxviruses and african swine fever virus. the gene is spliced into five exon ...200111134273
african swine fever virus protease, a new viral member of the sumo-1-specific protease family.african swine fever virus (asfv) is a complex dna virus that employs polyprotein processing at gly-gly-xaa sites as a strategy to produce several major core components of the viral particle. the virus gene s273r encodes a 31-kda protein that contains a "core domain" with the conserved catalytic residues characteristic of sumo-1-specific proteases and the adenovirus protease. using a cos cell expression system, it was found that protein ps273r is capable of cleaving the viral polyproteins pp62 an ...200111031264
african swine fever virus iap homologue inhibits caspase activation and promotes cell survival in mammalian cells.african swine fever virus (asfv) a224l is a member of the inhibitor of apoptosis protein (iap) family. we have investigated the antiapoptotic function of the viral iap both in stably transfected cells and in asfv-infected cells. a224l was able to substantially inhibit caspase activity and cell death induced by treatment with tumor necrosis factor alpha and cycloheximide or staurosporine when overexpressed in vero cells by gene transfection. we have also observed that asfv infection induces caspa ...200111222676
secretory pathway limits the enhanced expression of classical swine fever virus e2 glycoprotein in insect cells.the 3' untranslated region (utr) is an important element that determines the level of recombinant protein expression via baculovirus vectors. previous work using chloramphenicol acetyl transferase as reporter has shown that p10-promoter based baculovirus vectors with the authentic p10 3' utr resulted in higher expression levels than vectors carrying an sv40 early terminator, as part of a lacz selection cassette. to examine whether a similar increase in expression levels could be obtained for bac ...200111223142
enzyme-linked immunosorbent assay using a virus type-specific peptide based on a subdomain of envelope protein e(rns) for serologic diagnosis of pestivirus infections in swine.peptides deduced from the c-terminal end (residues 191 to 227) of pestivirus envelope protein e(rns) were used to develop enzyme-linked immunosorbent assays (elisas) to measure specifically antibodies against different types of pestiviruses. the choice of the peptide was based on the modular structure of the e(rns) protein, and the peptide was selected for its probable independent folding and good exposure, which would make it a good candidate for an antigenic peptide to be used in a diagnostic ...200111230402
african swine fever virus multigene family 360 and 530 genes are novel macrophage host range determinants.pathogenic african swine fever virus (asfv) isolates primarily target cells of the mononuclear-phagocytic system in infected swine and replicate efficiently in primary macrophage cell cultures in vitro. asfvs can, however, be adapted to grow in monkey cell lines. characterization of two cell culture-adapted viruses, ms16 and ba71v, revealed that neither virus replicated in macrophage cell cultures. cell viability experiments and ultrastructural analysis showed that infection with these viruses r ...200111238833
evaluation of genetic vaccine against classical swine fever.classical swine fever is important diseases affecting pigs. it results in great losses in their population and in limitations in the commercial international trade of pigs. the aim of the study was the preparation of the genetic vaccine against csf and the estimation of its safety, protection value and immunogenicity. clinical observations, body temperature and the immune response (haematological and facs analyses) were monitored. pigs vaccinated with the dna vaccine were protected from the chal ...200111257381
evolution of hepatitis c virus genome in chronically infected patients receiving ribavirin monotherapy.recent results of clinical trials suggest that combination of interferon and ribavirin exhibits an enhanced antiviral effect in the treatment of chronic hepatitis c. to investigate the effect of ribavirin on hepatitis c virus (hcv) infection, we analysed the evolution of the genetic heterogeneity of hcv in relation to the anti-hcv humoral response in patients treated by ribavirin alone. the study population included 35 patients with liver biopsy proven chronic hepatitis c infected with hcv genot ...200111264732
immunogenicity of the e1e2 proteins of hepatitis c virus expressed by recombinant adenoviruses.the e1 and e2 proteins of hepatitis c virus (hcv) are believed to be the viral envelope glycoproteins that are major candidate antigens for hcv vaccine development. we reported previously that the replication-competent recombinant adenovirus encoding core-e1-e2 genes of hcv (ad/hcv) produces serologically reactive e1 and e2 proteins forming a heterodimer in substantial amounts. here, we examined immunogenicity of the e1e2 proteins copurified from hela cells infected with ad/hcv virus in mice. fu ...200111282207
a prime-boost vaccination strategy using naked dna followed by recombinant porcine adenovirus protects pigs from classical swine fever.weaned pigs (6-week-old) and 7-day-old pre-weaned piglets were vaccinated with naked plasmid dna expressing the gp55/e2 gene from classical swine fever virus (csfv). both groups of pigs were then given a booster dose of recombinant porcine adenovirus expressing the gp55 gene (rpav-gp55). following challenge with csfv, 100% of weaned pigs and 75% pre-weaned piglets were protected from disease. weaned pigs given a single dose of rpav-gp55 were also protected, but showed a slight increase in temper ...200111295331
vitamin b12 and hepatitis c: molecular biology and human pathology.cobalamins are stored in high concentrations in the human liver and thus are available to participate in the regulation of hepatotropic virus functions. we show that cyanocobalamin (vitamin b12) inhibited the hcv internal ribosome entry site (ires)-dependent translation of a reporter gene in vitro in a dose-dependent manner without significantly affecting the cap-dependent mechanism. vitamin b12 failed to inhibit translation by ires elements from encephalomyocarditis virus (emcv) or classical sw ...200111296247
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