Sano N, Moriwake M, Sano T, 1993. Whom should I contact if I suspect my fish have KHV or if I want more information? 5C). The principal histopathological changes were observed in gill filaments. Thus, all but one of the junctions in set II that were supported by >10 reads were detected by RT-PCR. 2B). In one of the more elegant detection methods for CyHV-3 by Soliman & El-Matbouli (2005), the product of LAMP-PCR is visualized by mixing with SYBR-Green I to confirm positive results for CyHV-3 ().
Statistical analysis was performed using the statistical software R (3.0.2) (27). The similarity of CyHV-3 TK to the homologous pox virus enzymes explains why we and others failed to inhibit CyHV-3 propagation with Acyclovir and Gancyclovir . The terminal direct repeats are shaded gray. It consisted of a tube in the shape of a “U” made of Plexiglas pipes (5-cm diameter). Results of the PCR detection of linear CyHV-3 DNA. Journal of Aquatic Animal Health. Understanding the ecology of CyHV-3 in natural environments may be useful in preventing the spread of CyHV-3.
The location of the structural proteins is indicated. Specificity and sensitivity are the most important parameters to evaluate an assay. The optimal conditions of LAMP reaction for the detection were determined at 64°C for 60min with 6mMMgCl2. Forty microlitres of labelled RNA was added to 100 μL of hybridization buffer (final concentration of 2.5x SSPE, 20% Formamide, 0.01% Tween-20, 0.01 mg/mL Acetylated BSA, 6.5% w/v Dextran Sulphate and 2% SDS), this was then heated to 65°C for 5 min, kept on ice for 5 min and hybridized to the array at 42°C for 8 h in a Tecan HS 400 Pro Hybridization Station. In addition, the conserved genes include one encoding a large membrane glycoprotein (ORF99) and five encoding proteins with unknown functions (ORF47, ORF61, ORF80, ORF90, and ORF56). Transparent 12-well plates (Nunc, Denmark) were used to culture KF-1 cells and CCB cells seeded onto sterile 1.6 mm2 glass cover slips (Fisher Scientific, UK) for 24 h at 20 °C. BMC Microbiol.
DNA was extracted from 100–150 mg of individual gill and brain tissues using a Gentra Puregene Tissue Kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. If initial inoculation failed to produce KHV CPE the first time, total cell lysate was reinoculated onto CCB cells seeded in 12-well plates and incubated at 22°C. To establish the detection limit of the LAMP assay, serial dilutions of each plasmid DNA which had been quantified by measuring the optical density at 260 nm were tested. Other external signs of infection may include sunken eyes (enophthalmos) and pale swollen gills (Figure 2, Haenen et al. Additionally an inflammation of the anus was detected in many cases. The best way to remove a viral infection is for the immune system to try to seek and destroy virions (virus particles) as fast as the hijacked cell is producing them. The present study was devoted to CyHV-3 ORF134 encoding an IL-10 homologue.
Despite this low diversity, molecular markers enabling discrimination among 9 genotypes (7 from Europe and 2 from Asia) have been identified (12). However, a minor product corresponding to the unspliced transcript of ORF134 was also observed (see the faint 624 bp band in Figure 2 ). Common carp is a host for two highly contagious viruses: spring viraemia of carp virus (Rhabdovirus carpio, SVCV) and the Cyprinid herpesvirus 3 (CyHV-3), which belong to Rhabdoviridae and Alloherpesviridae families, respectively. View Full Text PDF Listings View primary source full text article PDFs. View Full Text PDF Listings View primary source full text article PDFs. Assigned abbreviations ( ) are also listed. Proteins were extracted by different methods and identified by mass spectrometry.
Op basis van deze argumenten adviseert de COGEM MuHV-1 als strikt dierpathogeen virus in te delen in pathogeniteitsklasse 2. The recovery of seeded CyHV-3 based on the cation-coated filter method varied by more than 3 orders of magnitude among the water samples. Affected individuals develop most prominent lesions in gills, skin and kidney, in tissues which are involved in the osmotic regulation of freshwater teleosts. Comparisons generated from the aligned nucleic and deduced amino acid sequences revealed significant similarities between KHV, CyHV-1, and CyHV-2 as well as lower but still significant similarities between the three cyprinid herpesviruses and the channel catfish virus. The basic algorithm is simple and robust; it can be implemented in a number of ways and applied in a variety of contexts including straightforward DNA and protein sequence database searches, motif searches, gene identification searches, and in the analysis of multiple regions of similarity in long DNA sequences. In the present study, we investigated the portal of entry of KHV in carp by using bioluminescence imaging. Cyprinid herpesvirus-3 (CyHV-3, koi herpesvirus, KHV) is the causative agent of an economically important disease in carp.
In the early summer of 2014, mass mortality of sichel (Pelecus cultratus) was observed in Lake Balaton, Hungary.