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1.
Vet Microbiol ; 166(1-2): 195-9, 2013 Sep 27.
Article in English | MEDLINE | ID: mdl-23838147

ABSTRACT

Bovine viral diarrhea virus (BVDV) belongs together with Classical swine fever virus (CSFV) and Border disease virus (BDV) to the genus Pestivirus in the Flaviviridae family. BVDV has been subdivided into two different species, BVDV1 and BVDV2 based on phylogenetic analysis. Subsequent characterization of both strains revealed major antigenic differences. Because the envelope glycoprotein E2 is the most immunodominant protein for all pestiviruses, the present study focused on epitope mapping by constructing chimeric BVDV type 1 and 2 E2 genes in expression plasmids. These plasmids with chimeric E2-genes were transfected in SK6 cells and transient expression was studied by immunostaining with a panel of MAbs specific for E2 of BVDV1 or BVDV2, resulting in the localization of type-specific antigenic domains at similar regions. These results indicate that E2 glycoproteins of both BVDV types exhibit a comparable antigenic structure, but with type specific epitopes. In addition, the antigenic resemblance with envelope glycoprotein E2 of Classical swine fever virus is discussed.


Subject(s)
Diarrhea Virus 1, Bovine Viral/immunology , Diarrhea Virus 2, Bovine Viral/immunology , Epitopes/immunology , Viral Envelope Proteins/immunology , Amino Acid Sequence , Animals , Cattle , Conserved Sequence , Diarrhea Virus 1, Bovine Viral/chemistry , Diarrhea Virus 1, Bovine Viral/genetics , Diarrhea Virus 2, Bovine Viral/chemistry , Diarrhea Virus 2, Bovine Viral/genetics , Epitope Mapping , Epitopes/chemistry , Epitopes/genetics , Molecular Sequence Data , Phylogeny , Sequence Alignment , Viral Envelope Proteins/chemistry , Viral Envelope Proteins/genetics
2.
Vet Microbiol ; 136(3-4): 240-5, 2009 May 12.
Article in English | MEDLINE | ID: mdl-19128896

ABSTRACT

Swine can be infected with classical swine fever virus (CSFV), as well as ruminant pestiviruses: bovine viral diarrhoea virus (BVDV), and Border disease virus (BDV). Cross-reactions between pestiviruses occur, both regarding protective immunity and in diagnostic tests. The presence of BVDV and BDV in a swine population may thus affect the transmission of CSFV, but also the diagnosis of a CSFV infection. In this study, the seroprevalence against BVDV and BDV in two categories of swine, sows and finishing pigs, in the Netherlands was determined. Furthermore, several risk factors, associated with the presence of swine and ruminants on the same farm or in the immediate surroundings, were evaluated. In sows, the seroprevalence against BVDV was 2.5% on the animal level, and 11.0% on herd level. In finishing pigs these prevalences were 0.42% and 3.2%, respectively. Antibodies against BDV were found in three sows only. Risk factors, associated with a BVDV-seropositive status in breeding pigs, were the presence of cattle on the same premises and a high density of sheep and/or goats herds in a radius of 3km. While BVDV and BDV hardly pose any threat to the swine population themselves, knowledge, and therefore regular monitoring, on the presence of these viruses in the swine population is important with respect to CSF eradication. It will allow for a better interpretation of diagnostic test results, both in terms of possible false positives and false negatives, but may also bring about additional measures or surveillance protocols in times of CSF outbreaks to avoid surprises caused by cross-reactivity with ruminant pestiviruses.


Subject(s)
Border Disease/epidemiology , Border disease virus/isolation & purification , Bovine Virus Diarrhea-Mucosal Disease/epidemiology , Classical Swine Fever/epidemiology , Classical Swine Fever/virology , Diarrhea Viruses, Bovine Viral/isolation & purification , Animals , Antibodies, Viral/blood , Border Disease/immunology , Border Disease/virology , Bovine Virus Diarrhea-Mucosal Disease/immunology , Bovine Virus Diarrhea-Mucosal Disease/virology , Cattle , Classical Swine Fever/immunology , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Netherlands/epidemiology , Risk Factors , Seroepidemiologic Studies , Swine
3.
Appl Environ Microbiol ; 64(11): 4546-54, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9797321

ABSTRACT

During the past few years, Ralstonia (Pseudomonas) solanacearum race 3, biovar 2, was repeatedly found in potatoes in Western Europe. To detect this bacterium in potato tissue samples, we developed a method based on fluorescent in situ hybridization (FISH). The nearly complete genes encoding 23S rRNA of five R. solanacearum strains and one Ralstonia pickettii strain were PCR amplified, sequenced, and analyzed by sequence alignment. This resulted in the construction of an unrooted tree and supported previous conclusions based on 16S rRNA sequence comparison in which R. solanacearum strains are subdivided into two clusters. Based on the alignments, two specific probes, RSOLA and RSOLB, were designed for R. solanacearum and the closely related Ralstonia syzygii and blood disease bacterium. The specificity of the probes was demonstrated by dot blot hybridization with RNA extracted from 88 bacterial strains. Probe RSOLB was successfully applied in FISH detection with pure cultures and potato tissue samples, showing a strong fluorescent signal. Unexpectedly, probe RSOLA gave a less intense signal with target cells. Potato samples are currently screened by indirect immunofluorescence (IIF). By simultaneously applying IIF and the developed specific FISH, two independent targets for identification of R. solanacearum are combined, resulting in a rapid (1-day), accurate identification of the undesired pathogen. The significance of the method was validated by detecting the pathogen in soil and water samples and root tissue of the weed host Solanum dulcamara (bittersweet) in contaminated areas.


Subject(s)
Nucleic Acid Conformation , Pseudomonas/isolation & purification , RNA, Ribosomal, 23S/chemistry , RNA, Ribosomal, 23S/genetics , Solanum tuberosum/microbiology , Base Sequence , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Phylogeny , Plant Diseases , Polymerase Chain Reaction/methods , Pseudomonas/genetics , RNA, Bacterial/chemistry , RNA, Bacterial/genetics , Sequence Alignment , Sequence Homology, Nucleic Acid
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