No abstract

Hybrids of koi, Cyprinus carpio x crucian carp, Carassius carassius and koi x goldfish, Carassius auratus, proved to be susceptible to koi herpesvirus (KHV, syn. CyHV-3) and developed KHV disease (KHVD). While hybrids of koi x goldfish were partly resistant to mortality following infection by immersion, most koi x crucian carp hybrids died after bath infection. KHV DNA was detected in dead fish but also in all surviving animals by different polymerase chain reactions (PCRs). According to these results, hybrid crossbreeding does not seem to prevent severe losses associated with KHV in terms of inducing KHVD. The present study showed severe losses after a waterborne KHV infection of between 35% and 100% in koi x goldfish and koi x crucian carp hybrids as well as in SPF carp.

Background. Sturgeons have long been extinct in Polish inland waters. A substantial effort has recently been put into their restitution, covering the drainage areas of two major Polish rivers, the Oder and the Vistula. The stocked fishes are clinically healthy, but very little is known about their potential to transmit viral diseases including koi herpes virus (KHV) to healthy fishes of other species, which may pose a threat to the disease-free zones. This study was intended to determine if sturgeons could be asymptomatic carriers of KHV. Materials and Methods. A total of 29 sturgeons (two species; length 8-37 cm) originating from fish farms in northern Poland with a known KHV history in common carp or koi in the area were examined: 15 Russian sturgeons, Acipenser gueldenstaedtii, with clinical signs of a disease and 14 asymptomatic Atlantic sturgeons, A. oxyrinchus. The former were sent to the laboratory alive while the latter were sent fixed in ethanol. As it is required for detection of a latent KHV infection in acipenserids, two independent procedures were applied. The preliminary results were obtained using PCR. Those findings were subsequently confirmation by nested PCR. The latter procedure consists of sequence analysis of PCR products and direct detection of KHV infected cells in tissue materials by in-situ hybridization on nucleic acid level or indirect immunofluorescence on KHV protein level. Results. KHV genome parts were found in nine Russian sturgeons and four Atlantic sturgeons. Comparison of PCR results obtained from three primer pairs used for KHV diagnostic in sturgeon showed that those designed by Bercovier et al. were most sensitive and robust for this purpose. In order to confirm the presence of viral particles the most useful method was in-situ hybridization (ISH), allowing the detection of KHV in gill samples obtained from live sturgeons.Conclusion. This preliminary study shows that sturgeons can be carriers of KHV. Therefore a viral diagnostics is highly recommended not only for sturgeons obtained from the environment but also for fertilized eggs, fry, and fish intended for re-stocking measurements of inland waters.

Worldwide koi herpesvirus (KHV) causes high mortalities in Cyprinus carpio L. aquaculture. So far, it is unknown how the different variants of KHV have developed and how they spread in the fish, but also in the environmental water bodies. Therefore, a phylogenetic method based on variable number of tandem repeats (VNTR) was improved to gain deeper insights into the phylogeny of KHV and its possible worldwide distribution. Moreover, a VNTR-3 qPCR was designed which allows fast virus typing. This study presents a useful method for molecular tracing of diverse KHV types, variants, and lineages.

Koi herpesvirus (KHV) causes a highly infectious disease afflicting common carp and koi, Cyprinus carpio L. Various molecular and antibody-based detection methods have been used to elucidate the rapid attachment and dissemination of the virus throughout carp tissues, facilitating ongoing development of effective diagnostic approaches. In situ hybridization (ISH) was used here to determine the target tissues of KHV during very early infection, after infecting carp with a highly virulent KHV isolate. Analysis of paraffin-embedded tissues (i.e. gills, skin, spleen, kidney, gut, liver and brain) during the first 8 h and following 10 days post-infection (hpi; dpi) revealed positive signals in skin mucus, gills and gut sections after only 1 hpi. Respiratory epithelial cells were positive as early as 2 hpi. Viral DNA was also detected within blood vessels of various tissues early in the infection. Notable increases in signal abundance were observed in the gills and kidney between 5 and 10 dpi, and viral DNA was detected in all tissues except brain. This study suggests that the gills and gut play an important role in the early pathogenesis of this Alloherpesvirus, in addition to skin, and demonstrates ISH as a useful diagnostic tool for confirmation of acutely infected carp.