DAO 92:21-29 (2010)  -  doi:10.3354/dao02251

First isolation and genotyping of viruses from recent outbreaks of viral haemorrhagic septicaemia (VHS) in Slovenia

Ivan Toplak1,*, Peter Hostnik1, Danijela Rihtarič1, Niels Jørgen Olesen3, Helle Frank Skall3, Vlasta Jenčič2

1Virology Unit, Institute of Microbiology and Parasitology, and 2Institute for Health Care and Breeding of Wild Animals Fishes and Bees, Veterinary Faculty, University of Ljubljana, Gerbičeva 60, 1115 Ljubljana, Slovenia
3Fish Diseases Section , Technical University of Denmark, National Veterinary Institute, Hangøvej 2, 8200 Århus N, Denmark

ABSTRACT: In November and December 2007, the virus causing viral haemorrhagic septicaemia (VHS) was detected in rainbow trout Oncorhynchus mykiss from 2 fish farms in Slovenia. During 2008 and 2009 the infection spread only among rainbow trout farms and 4 new outbreaks were confirmed. High mortality and clinical signs of VHS were observed among the diseased fish. VHSV was confirmed by virus isolation, immunoperoxidase test, reverse transcriptase polymerase chain reaction (RT-PCR) and phylogenetic analysis. Based on 1 complete (1524 nucleotides [nt]) and 9 partial (600 nt) glycoprotein gene nucleotide sequences, 9 VHSV isolates from the 6 VHS outbreaks were genetically closely related (99 to 100% identity), and were classified into the Subgroup I-a of Genotype I, most closely related to the German isolates Dstg21-07, Dstg36-06, and Dstg54-1-07 (99 to 100% identity). Phylogenetic analysis and epidemiological investigations confirmed that the VHS virus had been (re)introduced with imported live fish, and that subsequent outbreaks were linked to the initial infection. Our study shows that direct nucleotide sequencing of RT-PCR products, amplified from the tissue of VHSV-infected fish, represents a reliable tool for fast routine genotyping in diagnostic laboratories. This is the first report of a natural epidemic associated with VHSV infection in Slovenia since the eradication of the disease in 1977.


KEY WORDS: Viral haemoragic septicaemia virus · VHSV · Rainbow trout · Epidemiology · Slovenia


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Cite this article as: Toplak I, Hostnik P, Rihtarič D, Olesen NJ, Skall HF, Jenčič V (2010) First isolation and genotyping of viruses from recent outbreaks of viral haemorrhagic septicaemia (VHS) in Slovenia. Dis Aquat Org 92:21-29

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