Koi herpesvirus infection in experimentally infected common carp Cyprinus carpio (Linnaeus, 1758) and three potential carrier fish species Carassius carassius (Linnaeus, 1758); Rutilus rutilus (Linnaeus, 1758); and Tinca tinca (Linnaeus, 1758) by quantitative real-time PCR and in-situ hybridization

Only single cells in the carrier fish species Carassius carassius (Linnaeus, 1758) for koi herpesvirus (KHV) are infected in contrast to large numbers in the susceptible species common carp Cyprinus carpio (Linnaeus 1758). Several species of the family Cyprinidae have been described as virus carrier species, showing no clinical signs of a KHV disease but able to transmit the virus to other susceptible fish. In this study, 72 common carp Cyprinus carpio (Linnaeus, 1758), 36 tench Tinca tinca (Linnaeus, 1758), 36 crucian carp Carassius carassius (Linnaeus, 1758) and 36 common roach Rutilus rutilus (Linnaeus, 1758) were experimentally infected with KHV (isolate Israel) by immersion and kept at 20 degrees C. The fish were euthanized at 12 timepoints over a period of 90days and virus DNA was quantified in tissues by a real-time TaqMan PCR. Whereas KHV-DNA was found in Cyprinus carpio for up to 90days, the virus DNA was detectable only in single individuals of Rutilus rutilus, Tinca tinca and Carassius carassius for up to 25days after experimental virus exposure. Tissue samples of Cyprinus carpio and Carassius carassius were screened by in-situ hybridization. Positive signals were found in various organs of the common carp tested crucian carp. In the latter species a much smaller number of virus-positive stained cells was detected compared to the infected carp.