Infectious Hematopoietic Necrosis Virus 11290 Infectious hematopoietic necrosis IHNV causes the disease known as infectious hematopoietic necrosis in salmonid fish like trout and salmon (Wiki: Infectious Hematopoietic Necrosis Virus). Infectious hematopoietic necrosis virus (IHNV), is a negative-sense single-stranded RNA virus that is a member of the Rhabdoviridae family, and from the genus Novirhabdovirus. It causes the disease known as infectious hematopoietic necrosis in salmonid fish like trout and salmon. IHNV is commonly found in the Pacific Coast of Canada and the USA, and has also been found in Europe and Japan. The first reported epidemics of IHNV occurred in the United States at the Washington and the Oregon fish hatcheries during the 1950s. IHNV is transmitted following shedding of the virus in the feces, urine, sexual fluids, and external mucus and by direct contact or close contact with surrounding water. The virus gains entry into fish at the base of the fins. Clinical signs of infection with IHNV include abdominal distension, bulging of the eyes, skin darkening, anemia and fading of the gills. Infected fish commonly hemorrhage in several areas; the mouth and behind the head, the pectoral fins, muscles near the anus, and (in fry) the yolk sac. Diseased fish weaken eventually floating “belly-up” on the surface of the water. Necrosis is common in the kidney and spleen, and sometimes in the liver (Wiki: Infectious Hematopoietic Necrosis Virus). 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 Apex-IHN Apex-IHN Novartis (Aqua Health) VO_0000941 DNA vaccine Licensed Intramuscular injection (i.m.) DNA vaccine Intramuscular injection (i.m.) IHNV DNA vaccine pCDNA-G encoding G VO_0004420 DNA vaccine Research pCDNA3.1 [Ref2338:Corbeil et al., 1999] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction Vector pCDNA3.1 expressed the glycoprotein (G) (Corbeil et al., 1999). The G protein expressed by the DNA vaccine construct did confer significant protection in fry up to 80 days post-immunization and induced protective neutralizing antibodies (Corbeil et al., 1999). VO_0000286 Cumulative percent mortality in fish immunized with pCDNA-G ranged from 0 to 2% whereas mortality in fish immunized with DNA vaccines that contained other proteins or were left unhandled ranged from 36 to 76%. Significant protection was elicited up to 80 days post-immunization, and the vaccine also induced protective neutralizing antibodies (Corbeil et al., 1999). IHNV DNA vaccine pCMV4-G VO_0004321 DNA vaccine Research pCMV4 [Ref2156:Anderson et al., 1996] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction Vector pCMV4 expressed the infectious hematopoietic necrosis virus (IHNV) glycoprotein (Anderson et al., 1996). Fish injected with the glycoprotein-encoding plasmid pCMV4-G, either alone or in combination with the nucleoprotein-encoding plasmid pCMV4-N, generated glycoprotein-specific and virus-neutralizing antibody responses (Anderson et al., 1996). VO_0000286 The vaccinated fish were protected from subsequent IHNV challenge (Anderson et al., 1996). IHNV DNA vaccine pCMV4-N encoding the nucleoprotein VO_0004322 DNA vaccine Research pCMV4 [Ref2156:Anderson et al., 1996] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction Vector pCMV4 expressed the infectious hematopoietic necrosis virus (IHNV) nucleoprotein (Anderson et al., 1996). VO_0000286 Fish receiving pCMV4-N and pCMV4-G were protected from subsequent IHNV challenge (Anderson et al., 1996). IHNV DNA vaccine pIHN-G encoding glycoproteins VO_0004495 DNA vaccine Research pcDNA3.1(+) vector [Ref2434:Lorenzen et al., 2002] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction The pcDNA3.1(+) vector expressed viral glycoproteins (G proteins) (Lorenzen et al., 2002). VO_0000286 Challenge experiments revealed that immunity established shortly after vaccination was cross-protective between IHNV and VHSV whereas no increased survival was found upon challenge with bacterial pathogens. Within two months after vaccination, the cross-protection disappeared while the specific immunity to homologous virus remained high (Lorenzen et al., 2002). IHNV DNA vaccine pIHNw-G VO_0011398 DNA vaccine Research pCDNA 3.1 vector (Invitrogen) Intramuscular injection (i.m.) Intramuscular injection (i.m.) Infectious Hematopoietic Necrosis Virus glycoprotein antigen DNA vaccine construction The open reading frame of the G gene of IHNV WRAC strain was amplifed by reverse transcriptase-polymerase chain reaction using sequence-specic primers and cloned into the pT7-blue vector (Novagen, Madison, WI). The gene was then subcloned downstream of the immediate-early enhancer-promoter sequences of human cytomegalovirus (CMV) of the pCDNA 3.1 vector (Invitrogen). The resulting plasmid was designated pIHNw-G (previously pCDNA-G) (Corbeil et al., 2000). Fish from cohort 1 were used for viral challenge experiments, serological analyses, histology, and passive transfer experiments 1 and 2. For these studies two groups of 500 juvenile rainbow trout from cohort 1 (mean weight 2.5 g) were anaesthetized by immersion in 100 μg/ml tricaine methane sulfonate (MS-222; Argent Chemical Laboratories, Redmond, WA) and vaccinated by i.m. injection with 0.1 μg of either pIHNw-G or pLuc in a total volume of 50 μl phosphate buffered saline (PBS). A group of 200 fish was left unhandled, and all fish were maintained at 12 °C for the entire study period (Kurath et al., 2006). The DNA vaccine pIHNw-G encodes the glycoprotein of the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV). Vaccine performance in rainbow trout was measured 3, 6, 13, 24, and 25 months after vaccination. At three months all fish vaccinated with 0.1 microg pIHNw-G had detectable neutralizing antibody (NAb) and they were completely protected from lethal IHNV challenge with a relative percent survival (RPS) of 100% compared to control fish (Kurath et al., 2006). . At 25 months post-vaccination a challenge experiment was done with three subgroups of 18 fish per treatment. For each experiment duplicate groups of pIHNw-G vaccinated fish were i.p. injected with IHNV as described above, and the third group was injected with 200 μl MEM without virus. Similarly, two groups of pLuc vaccinated fish were challenged with IHNV, and the third group was mock-challenged (Kurath et al., 2006). Infectious Hematopoietic Necrosis Virus NV mutant vaccine VO_0002979 Live, attenuated vaccine Research Intraperitoneal injection (i.p.) Intraperitoneal injection (i.p.) Gene mutation This NV mutant is from Infectious hematopoietic necrosis virus (Thoulouze et al., 2004). A NV mutant is attenuated in rainbow trout (Thoulouze et al., 2004). An NV mutant induces significant protection in rainbow trout from challenge with wild type IHNV (Thoulouze et al., 2004). G glycoprotein Infectious hematopoietic necrosis virus VO_0011109 1489848 9628086 IHNVgp4 KP216199 NP_042679 11290 2998 4524 ? glycoprotein 6.5 54073.98 508 >NC_001652.1:2998-4524 Infectious hematopoietic necrosis virus, complete genome AATGGACACCATGATCACCACTCCGCTCATTCTCATTCTGATCACCTGCGGAGCAAACAGCCAAACCGTC AAACCCGACACCGCAAGCGAATCAGACCAACCCACCTGGTCAAACCCGCTCTTCACCTATCCCGAGGGAT GCACTCTGGACAAGCTCTCTAAGGTCAATGCCTCTCAACTGAGATGCCCAAGGATCTTCGACGATGAGAA CAGGGGGCTAATTGCCTATCCCACATCCATCCGGTCCCTGTCAGTCGGAAACGACCTCGGGGACATTCAC ACCCAAGGGAACCACATCCACAAAGTCCTGTACCGCACAATCTGCTCAACAGGGTTCTTCGGGGGTCAGA CGATAGAGAAGGCGCTTGTAAAAATGAAACTCTCTACGAAAGAAGCAGGGGCGTATGACACCACAACCGC AGCCGCTCTGTACTTCCCAGCTCCCCGATGCCAATGGTACACCGACAACGTACAAAACGATCTCATCTTC TACTACACAACACAAAAGAGTGTTCTTAGAGATCCCTACACCAGAGACTTTCTGGACTCAGATTTTATTG GAGGAAAATGTACCAAATCACCCTGCCAGACTCATTGGTCCAACGTAGTTTGGATGGGTGATGCAGGGAT ACCAGCCTGTGATTCCAGCCAAGAGATAAAAGCTCACCTCTTTGTTGATAAAATCTCCAATCGAGTCGTG AAGGCAACGAGCTACGGACACCACCCCTGGGGACTGCATCAGGCCTGTATGATTGAATTCTGTGGGCAAC AGTGGATACGGACAGATCTCGGTGACCTAATATCTGTCGTATACAATTCTGGATCAGAAATCCTCTCGTT CCCGAAGTGTGAAGACAAGACCGTGGGAATGAGGGGAAACTTGGATGACTTTGCCTATCTAGACGATCTG GTGAAGGCCTCTGAGAGCAGAGAGGAATGTCTTGAGGCGCACGCCGAGATAATATCAACAAACAGTGTGA CTCCATACCTCCTATCCAAATTCCGATCTCCACATCCCGGAATAAATGACGTCTACGCTATGCACAAAGG CTCCATCTATCACGGGATGTGCATGACGGTCGCTGTGGACGAGGTATCCAAGGACAGGACAACGTACAGG GCCCATCGCGCTACCAGCTTCACGAAATGGGAACGACCCTTTGGGGATGAGTGGGAGGGCTTTCACGGAT TGCACGGAAACAACACCACCATTATTCCAGACCTGGAGAAATACGTCGCCCAGTACAAGATGAGCATGAT GGAACCGATGAGCATCAAATCCGTACCCCATCCAAGCATCCTGGCCCTCTACAATGAGACAGACGTGTCA GGGATCTCCATCAGGAAACTGGACTCATTCGACCTTCAATCACTCCACTGGAGTTTCTGGCCCACAATCT CCGCACTGGGTGGAATTCCCTTTGTTCTCCTCCTTGCTGTTGCCGCGTGCTGCTGCTGGTCAGGGAGACC TCCCACTCCCTCCGTGCCGCAGAGTATCCCCATGTATCACCTGGCAAACCGGTCCTA >NP_042679.1 glycoprotein [Infectious hematopoietic necrosis virus] MDTMITTPLILILITCGANSQTVKPDTASESDQPTWSNPLFTYPEGCTLDKLSKVNASQLRCPRIFDDEN RGLIAYPTSIRSLSVGNDLGDIHTQGNHIHKVLYRTICSTGFFGGQTIEKALVKMKLSTKEAGAYDTTTA AALYFPAPRCQWYTDNVQNDLIFYYTTQKSVLRDPYTRDFLDSDFIGGKCTKSPCQTHWSNVVWMGDAGI PACDSSQEIKAHLFVDKISNRVVKATSYGHHPWGLHQACMIEFCGQQWIRTDLGDLISVVYNSGSEILSF PKCEDKTVGMRGNLDDFAYLDDLVKASESREECLEAHAEIISTNSVTPYLLSKFRSPHPGINDVYAMHKG SIYHGMCMTVAVDEVSKDRTTYRAHRATSFTKWERPFGDEWEGFHGLHGNNTTIIPDLEKYVAQYKMSMM EPMSIKSVPHPSILALYNETDVSGISIRKLDSFDLQSLHWSFWPTISALGGIPFVLLLAVAACCCWSGRP PTPSVPQSIPMYHLANRS Protective antigen The DNA vaccine pIHNw-G encodes the glycoprotein of the fish rhabdovirus infectious hematopoietic necrosis virus (IHNV). Vaccine performance in rainbow trout was measured 3, 6, 13, 24, and 25 months after vaccination. At three months all fish vaccinated with 0.1 microg pIHNw-G had detectable neutralizing antibody (NAb) and they were completely protected from lethal IHNV challenge with a relative percent survival (RPS) of 100% compared to control fish [Ref1353:Kurath et al., 2006]. N gene Infectious hematopoietic necrosis virus 1489845 9628083 IHNVgp1 L40883 NP_042676 11290 174 1349 ? nucleocapsid protein 4.56 39826.36 391 >gi|9628082:174-1349 Infectious hematopoietic necrosis virus, complete genome GATGACAAGCGCACTCAGAGAGACGTTCACTGGACTCAGAGACATCAAGGGGGGAGTCCTCGAGGATCCA GAGACGGAGTATCGTCCCAGTACGATAACCCTCCCTCTATTTTTCTCCAAGACAGACTTAGACCTAGAGA TGATCAAGCGGGCGGTGAGTCAAGTCGGAGGAGAGGGAACGAGAAAGGCATTGAGCCTCCTGTGCGCGTT CGTCATTGCAGAGACGGTCCCATCGGAGGCAGGTACGGTCGCCGAACTTCTGGAAGCCCTGGGTTTCGTG CTGGAGTCTTTGGACACTGGGGCACCACCAGACGCTACCTTCGCAGATCCCAACAACAAGCTTGCAGAAA CGATCGTAAAGGAAAATGTCCTTGAGGTTGTGACCGGCCTCCTCTTCACCTGCGCCCTACTGACTAAGTA TGATGTGGACAAGATGGCCACATACTGCCAAAACAAGCTCGAGCGTCTTGCAACCAGCCAAGGGATTGGC GAGTTGGTTAACTTCAACGCCAACAGGGGAGTCCTGGCCAAGATCGGGGCGGTGCTTAGACCCGGACAGA AGCTCACCAAGGCTATCTATGGGATCATTCTCATCAACCTGTCCGACCCAGCCATCGCTGCCAGAGCCAA GGCACTGTGCGCCATGAGACTGAGCGGGACAGGAATGACAATGGTGGGGCTGTTCAACCAAGCCGCAAAG AACCTGGGCGCCCTTCCAGCCGACCTTTTAGAGGATCTGTGCATGAAGTCAGTGGTGGAGTCCGCCAGAC GCATTGTCAGACTGATGAGGATCGTAGCAGAGGCCCCAGGGGTAGCAGCAAAGTACGGTGTCATGATGAG CAGGATGCTCGGGGAGGGGTACTTCAAGGCCTACGGGATCAACGAGAACGCCAGGATCACCTGCATTCTC ATGAACATCAACGATAGGTATGACGATGGGACCTCGAGAGGACTGACAGGGATAAAGGTCTCCGACCCTT TCAGGAAGCTGGCGAGGGAGATCGCTCGTCTCCTTGTCCTCAAGTACGATGGCGATGGCTCAACCGGAGA GGGGGCGTCAGAGCTGATCCGCCGGGCGGAGATGGCATCTCGGGGACCAGACATGGGTGAGGAGGAGGAG GAGGACGAGGAGGACGACGACTCCAGTGAGCCAGGAGACTCCGACTCATTCCACTG >gi|9628083|ref|NP_042676.1| nucleocapsid protein [Infectious hematopoietic necrosis virus] MTSALRETFTGLRDIKGGVLEDPETEYRPSTITLPLFFSKTDLDLEMIKRAVSQVGGEGTRKALSLLCAF VIAETVPSEAGTVAELLEALGFVLESLDTGAPPDATFADPNNKLAETIVKENVLEVVTGLLFTCALLTKY DVDKMATYCQNKLERLATSQGIGELVNFNANRGVLAKIGAVLRPGQKLTKAIYGIILINLSDPAIAARAK ALCAMRLSGTGMTMVGLFNQAAKNLGALPADLLEDLCMKSVVESARRIVRLMRIVAEAPGVAAKYGVMMS RMLGEGYFKAYGINENARITCILMNINDRYDDGTSRGLTGIKVSDPFRKLAREIARLLVLKYDGDGSTGE GASELIRRAEMASRGPDMGEEEEEDEEDDDSSEPGDSDSFH Other NV Infectious hematopoietic necrosis virus 1489849 9628087 IHNVgp5 L40883 NP_042680 11290 4594 4929 ? non-virion protein 6.96 12583.43 111 >gi|9628082:4594-4929 Infectious hematopoietic necrosis virus, complete genome AATGGACCACCGCGACATAAACACGAACATGGAGGCACTCAGAGAAGCTCTGCGATACAAGAACGAGGTG GCCGGACACGGCTTCTCTTTTGATGACGGAGACCTGGTATGGCGTGAAGAGGACGACGCAACGTGGAGGA GGCTTTGCGATGTCGTCAACGCACTGATTTCCTCCAAGAGGATGCAGCGAGTATTGTACATGGACCTCAG CATCACCAAGGGCGAGGGGCATCTACTTCTTGTGGATCTCCAGGGGACCAAGAACCGCCTGTACAAAGAA CCCCGATTCAGGAGACATCTGATCCTGATTGAAGACTTTCTTGCTTATCCCAGATA >gi|9628087|ref|NP_042680.1| non-virion protein [Infectious hematopoietic necrosis virus] MDHRDINTNMEALREALRYKNEVAGHGFSFDDGDLVWREEDDATWRRLCDVVNALISSKRMQRVLYMDLS ITKGEGHLLLVDLQGTKNRLYKEPRFRRHLILIEDFLAYPR Virmugen A mutation in the non-virion protein of IHNV causes attenuated growth in rainbow trout and is able to provide significant protection from subsequent challenge with wild type IHNV [Ref2056:Thoulouze et al., 2004]. Anderson et al., 1996 journal Anderson ED, Mourich DV, Fahrenkrug SC, LaPatra S, Shepherd J, Leong JA 1996 5 2 114-122 Molecular marine biology and biotechnology Corbeil et al., 1999 journal Corbeil S, Lapatra SE, Anderson ED, Jones J, Vincent B, Hsu YL, Kurath G 1999 39 1 29-36 Diseases of aquatic organisms Corbeil et al., 2000 journal Corbeil S, LaPatra SE, Anderson ED, Kurath G 2000 18 25 2817-2824 Vaccine Einer-Jensen et al., 2009 journal Einer-Jensen K, Delgado L, Lorenzen E, Bovo G, Evensen Ø, Lapatra S, Lorenzen N 2009 27 8 1248-1253 Vaccine Kurath et al., 2006 journal Kurath G, Garver KA, Corbeil S, Elliott DG, Anderson ED, LaPatra SE 2006 24 3 345-354 Vaccine Lorenzen et al., 2002 journal Lorenzen N, Lorenzen E, Einer-Jensen K, LaPatra SE 2002 26 2 173-179 Developmental and comparative immunology Thoulouze et al., 2004 journal Thoulouze MI, Bouguyon E, Carpentier C, Brémont M 2004 78 8 4098-4107 Journal of virology Wiki: Infectious Hematopoietic Necrosis Virus website http://en.wikipedia.org/wiki/Infectious_hematopoietic_necrosis_virus