Hirame rhabdovirus 38142 Hirame rhabdovirus (HIRRV) is one of the four recognized species within the Novirhabdovirus genus, represented by the type species Infectious Haematopoietic Necrosis (IHNV). HIRRV was first isolated during an outbreak on cultured flounder (Paralichthys olivaceus) and ayu (Plecoglossus altivelis) in Japan. It was also found on other marine fish in Asia, such as stone flounder (Kareius bicoloratus) in China. Furthermore, it was shown to be pathogenic for a range of salmonids species, including rainbow trout, experimentally challenged in freshwater. The major clinical signs of HIRRV infection were congestion of the gonads, focal hemorrhages of the skeletal muscle and fins and ascitic fluid collection (Borzym et al., 2012). Baboon Papio cynocephalus 9556 Bank vole Clethrionomys glareolus 447135 Bear Ursus americanus 9643 Birds Passeroidea 175121 Brown Trout Salmo trutta 8032 Buffalo Bison bison 9901 Carnivores Vulpes 9625 Cat Felis catus 9685 Catfishes Siluriformes 7995 Cattle Bos taurus 9913 Chicken Gallus gallus 9031 Chimpanzee Pan troglodytes 9598 chinchillas Chinchillidae 10150 Copper Pheasant Syrmaticus soemmerringii 9067 Deer Cervus elaphus 9860 Deer mouse Peromyscus maniculatus 10042 Dog Canis familiaris 9615 Ducks Anas 8835 Ferret Mustela putorius furo 9669 Fish Hyperotreti 117565 Gerbil Gerbillina 10045 Goat Capra hircus 9925 Gray wolf Canis lupus 9612 Guinea pig Cavia porcellus 10141 Hamster Mesocricetus auratus 10036 Horse Equus caballus 9796 Human Homo sapiens 9606 Macaque Macaca fascicularis 9541 Mongolian Gerbil Meriones unguiculatus 10047 Monkey Platyrrhini 9479 Mouse Mus musculus 10090 None None Parrot Psittacidae 9224 Pig Sus scrofa 9823 Rabbit Oryctolagus cuniculus 9986 Rainbow trout Oncorhynchus mykiss 8022 Rat Rattus 10114 Raven Corvus corax 56781 sei whale Balaenoptera borealis 9768 Sheep Ovis aries 9940 Squirrel Spermophilus richardsonii 37591 Tree shrew Tupaiidae 9393 Trouts, salmons & chars Salmoninae 504568 Turkey Meleagris gallopavo 9103 Vole Microtus ochrogaster 79684 Water buffalo Bubalus bubalis 391902 H. rhabdovirus DNA Vaccine encoding G VO_0004583 Live, attenuated vaccine Licensed Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction Both cellular and humoral immune responses are involved in the protection of fish immunized with DNA vaccine. Induction of specific antibody response has been observed from rainbow trout immunized with rhabdovirus G protein constructs. However, despite high levels of protection, neutralizing antibodies could not be detected in all vaccinated fish at the time of challenge, or survivors of the challenge (Seo et al., 2006). Five groups of forty flounder fry with a mean weight of 3 g were prepared for injection with pcDNA-G, pcDNA-N, a pcDNA-G and pcDNA-N mixture, pcDNA3.1(+) vector, or PBS (Seo et al., 2006). VO_0003057 Groups injected with the pcDNA-G showed only 5% cumulative mortality. Control groups injected with plasmid only or PBS only showed at least 95% mortality. These results demonstrated the protective effect of this DNA vaccine against HIRRV infection (Seo et al., 2006). At 21 days post vaccination, flounder fry injected with different DNA vaccines were challenged by immersion in water containing 1.2 × 104 TCID ml−1 HIRRV (Seo et al., 2006). H. rhabdovirus DNA Vaccine encoding N and G VO_0004567 DNA vaccine Research pcDNA-G and pcDNA-N [Ref2693:Seo et al., 2006] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction DNA vaccine construction Both cellular and humoral immune responses are involved in the protection of fish immunized with DNA vaccine. Induction of specific antibody response has been observed from rainbow trout immunized with rhabdovirus G protein constructs. However, despite high levels of protection, neutralizing antibodies could not be detected in all vaccinated fish at the time of challenge, or survivors of the challenge (Seo et al., 2006). Five groups of forty flounder fry with a mean weight of 3 g were prepared for injection with pcDNA-G, pcDNA-N, a pcDNA-G and pcDNA-N mixture, pcDNA3.1(+) vector, or PBS (Seo et al., 2006). VO_0003057 Groups injected with the mixed pcDNA-N+pcDNA-G vaccine showed only 2.5% cumulative mortality. Control groups injected with plasmid only or PBS only showed at least 95% mortality. These results demonstrated the protective effect of this DNA vaccine against HIRRV infection (Seo et al., 2006). At 21 days post vaccination, flounder fry injected with different DNA vaccines were challenged by immersion in water containing 1.2 × 10^4 TCID ml−1 HIRRV (Seo et al., 2006). H. rhabdovirus DNA vaccine pHRV-G encoding HIRRV and VHSV G proteins VO_0004419 DNA vaccine Research pCI-neo [Ref2337:Yasuike et al., 2007] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction A type I IFN system was stimulated by vaccination with pHRV-G in the Japanese flounder, which contributes not only to innate immunity but also to acquired immunity. In addition, type I IFN plays a role in Th1 polarization of the immune response induced by DNA vaccination (Yasuike et al., 2007). Fourteen days after the HIRRV challenge, cumulative mortalities of the fish injected with pHRV-G was 13%, as compared to the cumulative mortalities of the control group which was at 98%. This confirmed previous protection studies which demonstrated the efficacy of this vaccine (Yasuike et al., 2007). G glycoprotein Hirame novirhabdovirus 2559534 34610118 HIRRVgp4 AF104985 NP_919033 1980915 2923 4449 ? glycoprotein 6.67 53029.85 508 >NC_005093.1:2923-4449 Hirame rhabdovirus, complete genome CATGGACCCTCGAATAATGTATTACACTGTTCTCCTCACGACAGCTGCCAGAGTCTATGGTCAAACCATC AAGCCTGGAGTTGACAGCGTATCCGATCAACCCACCTGGGCGAATCCCCTGTTTACATACCCTGTGGACT GTCCTGCGGCTAAGCTATCCAAGGTGAGTCCCTCACAGTTGAGGTGCCCACGCATATTTGATGATGAAAA CCGAGGACTCGTTGCCTACCCTGCTGTCATCAGATCCCTATCGGTAGGGAACAATCTTGGGGACATTCAT ACTCAGGGGGAATACGTCCACAAGGTCCTTTACCGAACCACATGTTCAACAGGGTTCTTTGGGGGCCAAA CCATAGAGAAGGCCCTAGTAGAGATGAAACTGGCCCCCAGGGAGGTGGGAGTGTATGACACCACGACAGC CTCAGCACTGTACTTCCCAGCTCCAAGGTGCCAATGGTACACGGACAATGTACATAACGACCTGACATTC TACTATACGACCGCCAAGAGTGTCCTTAGAGATCCTTACACCCTGGGATTCCTTGATTCTGATTTTATTG AAGGGAAGTGCTCCAAGTCGCCGTGTCAGACCCATTGGTCAAATGTGGTGTGGAAGGGAGACTCGGGAGT TGCCGCCTGTGACACGGGGTCTGAGATAAAGGGTCACATATTTGTGGACAAGACCTCCCACCATGTCGTC AAGGCAACGAGCTATGGACACCACCCATGGGGACTTCATCGAGCTTGCATGATTACCTTCTGTGGAAAAC CATGGATCAGGACAGACCTAGGGGACCTCATCGCAATCGAATACAATGGCGGGGCGACCCTTCTCGCATT CCCTGCTTGCAAGGACACCACGGTTGGAATGAGAGGGAGTCTTGACGACTTTGCCTACTTGGATGATCTG GTGAAGTCATCTGAAAGTCGGGAGGAATGTCTAGAGGCACATGCGGAGATCATCGCTACCAATAGTGTGA CTCCTTACCTGCTATCCAAATTCAGATCTCCCCACCCAGGAATAAACGATGTCTACGCCATGCATGACGG GTCCATATACCACGGAAAGTGCATGACCGTTGCCATTGATGAGGTTTCCAAAGATAGACGAACTTATCGT GCCCACCAGACCTCAGCTTTCGTAGCATGGGGGCATCCCTTCGGTGATGAATGGGGCGGGTTTCACGGTC TTCATGGCAACGATACTCCTGTGATTCCGGATCTTGAGAAATACGTCGCTCAGTACAAGGTGAGCATGAT GGACAAGATGGACATTCGCCCCGTTCCGCATCCTAGTGTCCAGATACTCTACAACGACACAGACACCGCA GACATCACAATCAGGAAGATAGACTCGTTTGATCTGCAATCACTCAACTGGAGCTTCTGGCCATCATTGT CAGCACTGGGAGGGGTTCCAATACTCCTCGCCCTCGTATTCTTTCTGTACTGCTGCATGAACAGAAGACC CTCCATGCCTGCAGCACCCCAAGAGATCCCCATGTACCACCTCGCCAGTCGAGGGTA >NP_919033.1 glycoprotein [Hirame novirhabdovirus] MDPRIMYYTVLLTTAARVYGQTIKPGVDSVSDQPTWANPLFTYPVDCPAAKLSKVSPSQLRCPRIFDDEN RGLVAYPAVIRSLSVGNNLGDIHTQGEYVHKVLYRTTCSTGFFGGQTIEKALVEMKLAPREVGVYDTTTA SALYFPAPRCQWYTDNVHNDLTFYYTTAKSVLRDPYTLGFLDSDFIEGKCSKSPCQTHWSNVVWKGDSGV AACDTGSEIKGHIFVDKTSHHVVKATSYGHHPWGLHRACMITFCGKPWIRTDLGDLIAIEYNGGATLLAF PACKDTTVGMRGSLDDFAYLDDLVKSSESREECLEAHAEIIATNSVTPYLLSKFRSPHPGINDVYAMHDG SIYHGKCMTVAIDEVSKDRRTYRAHQTSAFVAWGHPFGDEWGGFHGLHGNDTPVIPDLEKYVAQYKVSMM DKMDIRPVPHPSVQILYNDTDTADITIRKIDSFDLQSLNWSFWPSLSALGGVPILLALVFFLYCCMNRRP SMPAAPQEIPMYHLASRG Protective antigen N nucleocapsid protein Hirame novirhabdovirus 2559538 34610115 HIRRVgp1 AF104985 NP_919030 1980915 166 1344 ? nucleocapsid protein 4.48 39984.35 392 >NC_005093.1:166-1344 Hirame rhabdovirus, complete genome TATGGCGAACCTTAAGGAAGAATTTGCAGGACTCCGAGGAGTGAAGGGAGGAGCCCTCGAAGACTCTGGC ACGGAGTATGATCCCACCAAGATCAACCTGACGCTCTACGGGACAGACAAGCTCTATACCCTGGCGATCA TCAAGAGGGCAGTCAGTCAGGTCGGAGGGTCCCAAACCAACAAAGCACTAGGAATCCTTTGTGCTTTTGT CACGTCAGAGAACAACCCTGACATGACAGATGCAGCAGTCAAGCTCCTGGTGGACATGAGGTTCAAGGTG GACGTGGTGCCCGTGGACGACAGGCTCGGTGACAATCTGGATGACCCCAACTCAAAGCTGGCGGAGGTCC TGACGGAAGAGAACATGGTGGACCTGGTGAAAGGCCTTCTGTTCACTTGTGCCCTGATGGTCAAGTACGA TGTGGACAAGATGGCCACATACTGCCAGCAGAAGCTGGAGCGACTGGCCAACAGTCAGGGACTCAACGAG CTGACCCTGATAAGCACGAGTAGGGCAGTTCTGGCTCGGATTGGGGCAGCGGTAAGACCGGGGCAGAAGT TAACCAAGGCCATCTACGGGATCATTCTTATCAACCTGATGGATCCCGCCACTGCTGCCAGGGCAAAGGC CCTTTGTGCCATGAGGCTGAGCGGAACCGGAATGACCATGGTGGGTCTATTCAATCAGGCCTCTAAGAAC CTTGGAGCTCCACCTGCAGATCTGCTGGAAGATCTCTGCATGAAGTCCATCATCGACTCTGCCAGGAGGA TTGTCAAGCTGATGAGGATCGTTGCAGACGTCGAGGACATGACCGCAAAGTATGCCATCATGATGAGCAG GATGCTTGGGGATGGGTACTTCAAGTCCTACGGGATCAATGAGAACTCACGGATCACCTGTATCTTGATG AACATCAACGAGCAGTACGATGAGGGGACAACCGGAGGACTGGCAGGAGTCAGGGTTTCACCCCCCTTCC GAAAGCTGGCAACCGAGATTGCTCGCCTTCTGGTCAAAAAGTATGACGGGAACGGATCTGCCGGGCCAGG TGCCTCAGATCTTGTCCGCCAGGCCGAGCAGGCAGCGCAAGAGACCGAGGGCGAGGATGATAAGTACGAT GAGGAGGGCGAGGAAGACGGAGGGGAGGGAGAGGGGGGAGAGGACGAAGAGTACTACTG >NP_919030.1 nucleocapsid protein [Hirame novirhabdovirus] MANLKEEFAGLRGVKGGALEDSGTEYDPTKINLTLYGTDKLYTLAIIKRAVSQVGGSQTNKALGILCAFV TSENNPDMTDAAVKLLVDMRFKVDVVPVDDRLGDNLDDPNSKLAEVLTEENMVDLVKGLLFTCALMVKYD VDKMATYCQQKLERLANSQGLNELTLISTSRAVLARIGAAVRPGQKLTKAIYGIILINLMDPATAARAKA LCAMRLSGTGMTMVGLFNQASKNLGAPPADLLEDLCMKSIIDSARRIVKLMRIVADVEDMTAKYAIMMSR MLGDGYFKSYGINENSRITCILMNINEQYDEGTTGGLAGVRVSPPFRKLATEIARLLVKKYDGNGSAGPG ASDLVRQAEQAAQETEGEDDKYDEEGEEDGGEGEGGEDEEYY Protective antigen Borzym et al., 2012 website Ewa Borzym, Marek Matras, Joanna Maj, Agnieszka Sandomierska, Niels Jørgen Olesen, Mette Eliassen, Marine Baud, Chiraz Talbi, Laurent Bigarré 2012 http://orbit.dtu.dk/fedora/objects/orbit:112698/datastreams/file_9708708/content Seo et al., 2006 journal Seo JY, Kim KH, Kim SG, Oh MJ, Nam SW, Kim YT, Choi TJ 2006 24 7 1009-1015 Vaccine Yasuike et al., 2007 journal Yasuike M, Kondo H, Hirono I, Aoki T 2007 23 3 531-541 Fish & shellfish immunology