a-dV. Code assigned:

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1 This form should be used for all taxonomic proposals. Please complete all those modules that are applicable (and then delete the unwanted sections). For guidance, see the notes written in blue and the separate document Help with completing a taxonomic proposal Please try to keep related proposals within a single document; you can copy the modules to create more than one genus within a new family, for example. MODULE 1: TITLE, AUTHORS, etc Code assigned: a-dV (to be completed by ICTV officers) Short title: 1. Create a new species (Cadicivirus A) in a new genus (Dicipivirus) within the Picornaviridae. (e.g. 6 new species in the genus Zetavirus) Modules attached (modules 1 and 9 are required) Author(s) with address(es) of the proposer: Nick Knowles (nick.knowles@iah.ac.uk) on behalf of the Picornaviridae Study Group. List the ICTV study group(s) that have seen this proposal: A list of study groups and contacts is provided at If in doubt, contact the appropriate subcommittee chair (fungal, invertebrate, plant, prokaryote or vertebrate viruses) Picornaviridae Study Group ICTV-EC or Study Group comments and response of the proposer: Date first submitted to ICTV: 01/07/2012 Date of this revision (if different to above): Page 1 of 7

2 MODULE 2: NEW SPECIES creating and naming one or more new species. If more than one, they should be a group of related species belonging to the same genus. All new species must be placed in a higher taxon. This is usually a genus although it is also permissible for species to be unassigned within a subfamily or family. Wherever possible, provide sequence accession number(s) for one isolate of each new species proposed. Code aV (assigned by ICTV officers) To create 1 new species within: Genus: Dicipivirus (new) Subfamily: - Family: Picornaviridae Order: Picornavirales And name the new species: Cadicivirus A Fill in all that apply. If the higher taxon has yet to be created (in a later module, below) write (new) after its proposed name. If no genus is specified, enter unassigned in the genus box. GenBank sequence accession number(s) of reference isolate: JN8102, JN8103 & JN8104 Reasons to justify the creation and assignment of the new species: Explain how the proposed species differ(s) from all existing species. o If species demarcation criteria (see module 3) have previously been defined for the genus, explain how the new species meet these criteria. o If criteria for demarcating species need to be defined (because there will now be more than one species in the genus), please state the proposed criteria. Further material in support of this proposal may be presented in the Appendix, Module 9 A novel small RNA virus has recently been found in dogs (Woo et al., 2012). The authors named the virus canine picodicistrovirus (CPDV). The sequences of the most highly conserved genome regions (P1, 2C, 3C, 3D) are most closely related to the picornaviruses. The virus has a similar genome organization to picornaviruses, except that the P1 and P2/P3 regions are encoded in two separate ORFs separated by an intergenic region (of 588 nt) containing a second internal ribosome entry site (IRES). Woo et al. (2012) have demonstrated that both IRESs are functional using a dual-luciferase activity assay. The virus sequences were detected in 47 faecal and urine samples from dogs over a 22 month period in Hong Kong. Four samples, positive for CPDV, were cultured in MDCK (Madin-Darby canine kidney), DH82 (canine macrophagemonocyte), RD (human rhadomyosarcoma), Vero E6 (monkey kidney), and HEL (human embryonic lung fibroblast) cells, but no growth was detected in any of the cell cultures. Likewise no virus replication was detected when inoculated, by a variety of routes, into in suckling mice. Quantitative RT-PCR showed that numbers of viral RNAs ranged from 7.55 x 10 6 to 1.26 x 10 9 copies/ml of urine and 1.82 x 10 6 to 4.97 x copies/ml of faecal sample suggesting the likely replication of this virus in dogs. However, the use of nucleotide composition analysis (NCA) to infer hosts (Kapoor et al., 2010) suggests a possible invertebrate origin for the virus (Fig. 1). Despite have two ORFs, phylogenetically (using distance or maximum likelihood algorithms) the virus falls within the Picornaviridae when homologous conserved proteins are compared; P1 capsid (Fig. 2), 2C (Woo et al., 2012), 3C pro (data not shown) and 3D pol (Fig. 3; Woo et al., 2012). Page 2 of 7

3 The genome organization of the proposed species Cadicivirus A is: VPg+5'UTR IRES [1A-1B-1C-1D]IGR IRES [2A-2B-2C/3A-3B VPg -3C pro -3D pol ]3'UTR-poly(A) [ ] open reading frame /, primary cleavages -, secondary/tertiary cleavages IGR, intergenic region IRES, internal ribosome entry site The name of the species is derived from canine dicistronic virus (canine only indicating that this was the species in which this virus was first recognized and not implying that dogs are the actual host). Page 3 of 7

4 MODULE 3: NEW GENUS creating a new genus Ideally, a genus should be placed within a higher taxon. Code bV (assigned by ICTV officers) To create a new genus within: Subfamily: - Family: Picornaviridae Order: Picornavirales Fill in all that apply. If the higher taxon has yet to be created (in a later module, below) write (new) after its proposed name. If no family is specified, enter unassigned in the family box naming a new genus Code cV (assigned by ICTV officers) To name the new genus: Dicipivirus Assigning the type species and other species to a new genus Code dV (assigned by ICTV officers) To designate the following as the type species of the new genus Cadicivirus A Every genus must have a type species. This should be a well characterized species although not necessarily the first to be discovered The new genus will also contain any other new species created and assigned to it (Module 2) and any that are being moved from elsewhere (Module 7b). Please enter here the TOTAL number of species (including the type species) that the genus will contain: 1 Reasons to justify the creation of a new genus: Additional material in support of this proposal may be presented in the Appendix, Module 9 The canine picodicistrovirus is clearly phylogenetically distinct from other presently-classified picornaviruses and has a novel genome organization (two ORFs under the translational control of two IRESs). Origin of the new genus name: The genus name Dicipivirus is derived from dicistronic picornavirus. Reasons to justify the choice of type species: There is only a single known species in the proposed genus. Species demarcation criteria in the new genus: If there will be more than one species in the new genus, list the criteria being used for species demarcation and explain how the proposed members meet these criteria. There is only a single known species in the proposed genus. Page 4 of 7

5 MODULE 9: APPENDIX: supporting material additional material in support of this proposal References: Kapoor, A., Simmonds, P,, Lipkin, W.I., Zaidi, S. and Delwart, E. (2010). Use of nucleotide composition analysis to infer hosts for three novel picorna-like viruses. J Virol. 84: Woo, P.C., Lau, S.K., Choi, G.K., Huang, Y., Teng, J.L., Tsoi, H.W., Tse, H., Yeung, M.L., Chan, K.H., Jin, D.Y. and Yuen, K.Y. (2012). Natural occurrence and characterization of two internal ribosome entry site elements in a novel virus, canine picodicistrovirus, in the picornavirus-like superfamily. J. Virol. 86: Epub 2011 Dec 28. Annex: Include as much information as necessary to support the proposal, including diagrams comparing the old and new taxonomic orders. The use of Figures and Tables is strongly recommended but direct pasting of content from publications will require permission from the copyright holder together with appropriate acknowledgement as this proposal will be placed on a public web site. For phylogenetic analysis, try to provide a tree where branch length is related to genetic distance. Fig. 1. Nucleotide composition analysis (NCA) to infer hosts (Kapoor et al., 2010). Page 5 of 7

6 Bat-PV-1/LMH22A (HQ595341) Bat-PV-1/NC16A (HQ595340) Bat-PV-2/MH9F (HQ595342) Bat-PV-2/SK17F (HQ595343) CanPV-1 325F (JN831356) Ia io PV-1 China/2010 (JQ814852) Bat-PV-3/TLC21F (HQ595345) Bat-PV-3/TLC5F (HQ595344) Feline PV 127F (JN572116) Feline PV 661F (JN572118) Feline PV 073F (JN572115) Feline PV 356F (JN572117) Feline PV 1021F (JN572119) Avian sapelovirus/asv-1/tw90a (AY563023) Quail PV1 HUN/2010 (JN674502) Calif sea lion (sapelo)virus/cslsv-1/1162 (JN420368) Porcine sapelovirus/psv-1/v13 (AF406813) Simian sapelovirus/ssv-1/sv (AY064708) Human rhinovirus A/HRV-A1/2060 (FJ445111) Human rhinovirus C/HRV-C1/NAT001 (EF077279) Human rhinovirus B/HRV-B3/FEB (DQ473485) Human enterovirus B/CV-B3/Nancy (M88483) Human enterovirus C/PV-1/Mahoney (J02281) Simian enterovirus A/SEV-A1/SV UWB (AF326759) Human enterovirus D/EV-D70/J670/71 (D00820) Bovine enterovirus/bev-1/vg(5)27 (D00214) Bovine enterovirus/bev-2/bev-261 (DQ0770) Porcine enterovirus B/PEV-9/UKG/410/73 (AF363453) Human enterovirus A/CV-A2/Fleetwood (AY421760) Enterovirus J/SV6/1631 (AF326766) Enterovirus J/EV-J103/USA/GA-POo-1 (FJ007373) Enterovirus J/EV-J108/N125 (AF414372) Pigeon-PV-B/PPV-B/03/641 (FR727144) Bovine hungarovirus (JQ941880) Ovine hungarovirus (HM153767) Porcine teschovirus/ptv-1/talfan (AF231769) Bovine rhinitis A virus/brav-1/sd1 Bovine rhinitis A virus/brav-2/h-1 Bovine rhinitis B virus/brbv-1/ec11 (EU236594) Foot-and-mouth diease virus/fmdv-o/ov1 (AY593829) Equine rhinitis A virus/erav-1/perv (DQ272578) Equine rhinitis B virus/erbv-1/p1436/71 (X96871) Tortoise-PV/TPV-1/TGT1A/96 Mosavirus/MoSV-1/M-7 (JF973687) Seneca Valley virus/svv-1/svv-001 (DQ641257) Encephalomyocarditis virus/emcv-1/r (M81861) Theilovirus/TMEV/GDVII (M20562) Miniopterus schreibersii PV 1 China/2010 (JQ814851) Cosavirus D/HCoSV-D1/5004 (FJ438908) Cosavirus B/HCoSV-B1/2263 (FJ438907) Cosavirus E/HCoSV-E1/Australia/81 (FJ555055) Cosavirus F/PK5006 (JN867758) Cosavirus A/HCoSV-A1/0553 (FJ438902) Cosavirus A/HCoSV-A2/6344 (FJ438903) Cadicivirus A (236)(JN8103) Cadicivirus A (209)(JN8102) Cadicivirus A (244U)(JN8104) Turkey hepatitis virus/thv-1/ (HQ189775) Turkey hepatitis virus/thv-1/23d (HM7511) Turdivirus-2/TV-2/ (GU182409) Turdivirus-2/TV-2/10717 (GU182408) Turdivirus-3/TV-3/00742 (GU182411) Turdivirus-3/TV-3/10878 (GU182410) Turdivirus-1/TV-1/00356 (GU182406) Turdivirus-1/TV-1/00805 (GU182407) Salivirus A/SaV-1/ (GQ184145) Salivirus A/SaV-1/NG-J1 (GQ179640) Canine kobuvirus/cakv-1/an211d (JN387133) Murine kobuvirus/mkv-1/m-5 (JF755427) Aichi virus/aiv-1/a846/88 (AB040749) Porcine kobuvirus/pkv-1/s-1-hun (EU787450) Bovine kobuvirus/bkv-1/u-1 (AB084788) Sheep kobuvirus/skv-1/sheep/tb3/hun/2009 (GU245693) Hepatitis A virus/hav-1/hm-175 (M14707) Avian encephalomyelitis virus/aev-1/calnek (AJ225173) Bluegill-PV/BGPV-1/ Carp-PV/CPV-1/F37/06 Aquamavirus A/SePV-1/HO (EU142040) Eel-PV/EPV-1/F15-05 Duck hepatitis A virus/dhav-1/r85952 (DQ226541) Rhinolophus affinis PV 1 China/2010 (JQ814853) Swine pasivirus 1 Human Parechovirus/HPeV-1/Harris (L02971) Ljungan virus/lv-1/ (AF3270) Enterovirus Teschovirus Aphthovirus Erbovirus Senecavirus Cardiovirus Cosavirus Dicipivirus Megrivirus Salivirus Kobuvirus Hepatovirus Tremovirus Aquamavirus Avihepatovirus Parechovirus 0.2 Fig. 2. Neighbor-joining tree comparing the P1 capsid of canine picodicistrovirus and other picornaviruses. Page 6 of 7

7 Canine kobuvirus/cakv-1/an211d (JN387133) Murine kobuvirus/mkv-1/m-5 (JF755427) Aichi virus/aiv-1/a846/88 (AB040749) Porcine kobuvirus/pkv-1/s-1-hun (EU787450) Bovine kobuvirus/bkv-1/u-1 (AB084788) Sheep kobuvirus/skv-1/sheep/tb3/hun/2009 (GU245693) Gallivirus/GV-1/chicken/CHK1/USA/2010 (JF424824) Turdivirus-1/TV-1/00356 (GU182406) Turdivirus-1/TV-1/00805 (GU182407) Salivirus A/SaV-1/ (GQ184145) Salivirus A/SaV-1/NG-J1 (GQ179640) Turdivirus-2/TV-2/ (GU182409) Turdivirus-2/TV-2/10717 (GU182408) Turdivirus-3/TV-3/00742 (GU182411) Turdivirus-3/TV-3/10878 (GU182410) Turkey hepatitis virus/thv-1/ (HQ189775) Turkey hepatitis virus/thv-1/23d (HM7511) Rosavirus/RoSV/M-7 (JF973686) Cadicivirus A (236)(JN8103) Cadicivirus A (209)(JN8102) Cadicivirus A (244U)(JN8104) Tortoise-PV/TPV-1/TGT1A/96 Mosavirus/MoSV-1/M-7 (JF973687) Bovine hungarovirus (JQ941880) Ovine hungarovirus (HM153767) Porcine teschovirus/ptv-1/talfan (AF231769) Equine rhinitis B virus/erbv-1/p1436/71 (X96871) Bovine rhinitis A virus/brav-1/sd1 Bovine rhinitis A virus/brav-2/h-1 Bovine rhinitis B virus/brbv-1/ec11 (EU236594) Foot-and-mouth diease virus/fmdv-o/ov1 (AY593829) Equine rhinitis A virus/erav-1/perv (DQ272578) Encephalomyocarditis virus/emcv-1/r (M81861) Theilovirus/TMEV/GDVII (M20562) Seneca Valley virus/svv-1/svv-001 (DQ641257) Miniopterus schreibersii PV 1 China/2010 (JQ814851) Cosavirus D/HCoSV-D1/5004 (FJ438908) Cosavirus E/HCoSV-E1/Australia/81 (FJ555055) Cosavirus B/HCoSV-B1/2263 (FJ438907) Cosavirus A/HCoSV-A1/0553 (FJ438902) Cosavirus A/HCoSV-A2/6344 (FJ438903) Pigeon-PV-A/PPV-A/3/603-7 (FR727145) Pigeon-PV-B/PPV-B/03/641 (FR727144) Quail PV1 HUN/2010 (JN674502) Enterovirus J/EV-J103/USA/GA-POo-1 (FJ007373) Enterovirus J/SV6/1631 (AF326766) Enterovirus J/EV-J108/N125 (AF414372) Human enterovirus B/CV-B3/Nancy (M88483) Human enterovirus C/PV-1/Mahoney (J02281) Human enterovirus D/EV-D70/J670/71 (D00820) Porcine enterovirus B/PEV-9/UKG/410/73 (AF363453) Bovine enterovirus/bev-1/vg(5)27 (D00214) Bovine enterovirus/bev-2/bev-261 (DQ0770) Human enterovirus A/CV-A2/Fleetwood (AY421760) Human rhinovirus B/HRV-B3/FEB (DQ473485) Simian enterovirus A/SEV-A1/SV UWB (AF326759) Human rhinovirus A/HRV-A1/2060 (FJ445111) Human rhinovirus C/HRV-C1/NAT001 (EF077279) Avian sapelovirus/asv-1/tw90a (AY563023) Porcine sapelovirus/psv-1/v13 (AF406813) Simian sapelovirus/ssv-1/sv (AY064708) Feline PV 356F (JN572117) Feline PV 1021F (JN572119) Feline PV 073F (JN572115) Feline PV 127F (JN572116) Feline PV 661F (JN572118) CanPV-1 325F (JN831356) Bat-PV-3/TLC21F (HQ595345) Bat-PV-3/TLC5F (HQ595344) Ia io PV-1 China/2010 (JQ814852) Bat-PV-1/LMH22A (HQ595341) Bat-PV-1/NC16A (HQ595340) Bat-PV-2/MH9F (HQ595342) Bat-PV-2/SK17F (HQ595343) Hepatitis A virus/hav-1/hm-175 (M14707) Avian encephalomyelitis virus/aev-1/calnek (AJ225173) Aquamavirus A/SePV-1/HO (EU142040) Swine pasivirus 1 Duck hepatitis A virus/dhav-1/r85952 (DQ226541) Eel-PV/EPV-1/F15-05 Human Parechovirus/HPeV-1/Harris (L02971) Ljungan virus/lv-1/ (AF3270) Bluegill-PV/BGPV-1/ Carp-PV/CPV-1/F37/06 Kobuvirus Salivirus Megrivirus Dicipivirus Teschovirus Erbovirus Aphthovirus Cardiovirus Senecavirus Cosavirus Enterovirus Hepatovirus Tremovirus Aquamavirus Avihepatovirus Parechovirus 0.1 Fig. 3. Neighbor-joining tree comparing the 3D pol capsid of canine picodicistrovirus and other picornaviruses. Page 7 of 7