Generation of a potential koi herpesvirus live vaccine by simultaneous deletion of the viral thymidine kinase and dUTPase genes.

Koi herpesvirus (KHV, Cyprinidherpesvirus 3) causes a fatal disease of koi and common carp. To obtain safe and efficacious live vaccines, we generated deletion mutants of KHV lacking the nonessential genes encoding two enzymes of nucleotide metabolism, thymidine kinase (TK, ORF55) and deoxyuridine-triphosphatase (DUT, ORF123). Since single-deletion mutants based on a KHV isolate from Israel (KHV-I) only exhibited partial attenuation (Fuchs W, Fichtner D, Bergmann SM, Mettenleiter TC.

Identification of structural proteins of koi herpesvirus.

As a prerequisite for development of improved vaccines and diagnostic tools for control of the fish pathogen koi herpesvirus, or cyprinid herpesvirus 3 (CyHV-3), we have started to identify putative viral envelope and capsid proteins. The complete or partial CyHV-3 open reading frames ORF25, ORF65, ORF92, ORF99, ORF136, ORF138, ORF146, ORF148, and ORF149 were expressed as bacterial fusion proteins, which were then used for preparation of monospecific rabbit antisera. All of the sera that were obtained detected their target proteins in cells transfected with the corresponding eukaryotic expression plasmids.

Virus isolate from carp: genetic characterization reveals a novel picornavirus with two aphthovirus 2A-like sequences.

Picornaviruses have been isolated from a variety of hosts, mainly mammals and birds. Here, we describe the sequence analysis of carp picornavirus 1 (CPV-1) F37/06 that was isolated from an organ pool (heart, brain, liver) of a common carp (Cyprinus carpio). This carp perished after an accidental discharge of liquid manure into a fish pond and presented without obvious clinical symptoms.

Characterization of a novel picornavirus isolate from a diseased European eel (Anguilla anguilla).

A novel picornavirus was isolated from specimens of a diseased European eel (Anguilla anguilla). This virus induced a cytopathic effect in eel embryonic kidney cells and high mortality in a controlled transmission study using elvers. Eel picornavirus has a genome of 7,496 nucleotides that encodes a polyprotein of 2,259 amino acids.

Development of a Bovine leukemia virus polymerase gene-based real-time polymerase chain reaction and comparison with an envelope gene-based assay.

Bovine leukemia virus (BLV) causes a persistent infection with provirus formation in B-lymphocytes. A real-time polymerase chain reaction (PCR) based on the conserved BLV polymerase (BLV pol) gene sequences was developed. Dually labeled probes were used to permit detection by the 5' exonuclease assay.

Generation and characterization of koi herpesvirus recombinants lacking viral enzymes of nucleotide metabolism.

Koi herpesvirus (KHV) causes a fatal disease in koi and common carp, but no reliable and genetically characterized vaccines are available up to now. Therefore, we generated KHV recombinants possessing deletions within the viral ribonucleotide reductase (RNR), thymidine kinase (TK), dUTPase, or TK and dUTPase genes, and their corresponding rescuants. All KHV mutants were replication competent in cultured cells.

Evolution of infectious hematopoietic necrosis virus (IHNV), a fish rhabdovirus, in Europe over 20 years: implications for control.

The fish pathogenic rhabdovirus infectious hematopoietic necrosis virus (IHNV) causes substantial losses in European aquaculture. IHNV was first detected in Europe in 1987 and has since undergone considerable spread. Phylogenetic analyses of the full G-gene sequences of 73 isolates obtained from 4 countries in Europe (France, n = 18; Italy, 9; Switzerland, 4; Germany, 42) enable determination of the evolution of the virus in Europe since the first detection, and identification of characteristic changes within the G-genes of European strains.

Investigation on the diagnostic sensitivity of molecular tools used for detection of koi herpesvirus.

Previous and new PCRs for KHV detection were compared by estimation of their sensitivity in recognizing KHV DNA in plasmids, cell culture extracted KHV DNA and total DNA obtained from field tissue samples. A modified real-time PCR (Gilad et al., 2004), combined with an internal control system (IC2, Hoffmann et al.

Identification of envelope protein pORF81 of koi herpesvirus.

Koi herpesvirus (KHV), an emerging pathogen causing mass mortality in koi and common carp, possesses the largest known herpesvirus genome of 295 kbp predicted to encode 156 different proteins. However, none of them has been identified or functionally characterized up to now. In this study, a rabbit antiserum was prepared against a bacterial fusion protein that permitted detection of the predicted type III membrane protein encoded by ORF81 of KHV.

Development of an oral vaccine for immunisation of rainbow trout (Oncorhynchus mykiss) against viral haemorrhagic septicaemia.

In the European Union Viral Haemorrhagic Septicaemia (VHS) eradication is still based on stamping out. Due to the lack of effective low cost vaccines immune prophylaxis is currently not used to combat VHS. This paper describes a new oral delivery method for immunisation of trout with attenuated virus.

A process for making cutaneous radiation applicators based on digital data.

An optical modeling process for facial regions and other body surfaces has been developed. The body part in question is digitized using optical 3-D metrology to obtain a comprehensive dataset. The data is then prepared for further use by smoothing the point clusters.

Optical modeling of extraoral defects.

In order to reduce the stress caused to patients by conventional methods of modeling using computed tomography (CT) or magnetic resonance imaging (MRI), an optical modeling process has been developed for extraoral defects and body areas. The selected body part is digitized using optical 3-coordinate measuring technology, providing an extensive data record. This is adapted for further use by equalizing the point clouds to obtain a Computer Aided Design (CAD) model, which is converted to a physical model by means of a stereolithographic process.

Age- and weight-dependent susceptibility of rainbow trout Oncorhynchus mykiss to isolates of infectious haematopoietic necrosis virus (IHNV) of varying virulence.

The virulence of 5 European and 1 North American isolate of infectious haematopoietic necrosis virus (IHNV) was compared by infecting female sibling rainbow trout ('Isle of Man' strain) of different weights and ages (2, 20 and 50 g). The fish were exposed to 10(4) TCID50 IHNV per ml of water by immersion, and the mortality was recorded for 28 d. Two new IHNV isolates from Germany were included in the investigation.

Identification and ultrastructural characterization of a novel virus from fish.

During routine investigations on fish, a virus (isolate DF 24/00) with novel morphological features and hitherto undescribed morphogenesis was isolated from a white bream (Blicca bjoerkna L.; Teleostei, order Cypriniformes). Cell-free virions consist of a rod-shaped nucleocapsid (120-150x19-22 nm) similar to that seen in baculoviruses.

Expression of VP5 of infectious pancreatic necrosis virus strain VR299 is initiated at the second in-frame start codon.

Infectious pancreatic necrosis virus (IPNV), a member of the BIRNAVIRIDAE: with two double-stranded RNA genome segments, encodes five proteins designated VP1 to VP5. To study the function of the 17 kDa nonstructural protein VP5 during virus replication several mutated IPNV genome segments A were constructed and included in a reverse genetics system for IPNV to obtain recombinant virus. Mutations between nt 68 and 85 or nt 94 and 103 in the noncoding region failed to yield viable virus.