1. Canine Rabies Elimination from the United States: an Epidemiological and Virological Perspective Canine Rabies Elimination from the United States: an Epidemiological and Virological Perspective Andrés Velasco Villa, Serena Reeder, Jesse Blanton,Lillian A Orciari and Charles E. Rupprecht Rabies Unit Poxvirus and Rabies Branch Division of Viral and Rickettsial Diseases Coordinating Center of Infectious Diseases Centers for Disease Control and Prevention The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the funding agencies. Use of trade names and commercial sources are for identification only and do not imply endorsement by the US Department of Health and Human services.

2. Rabies is an acute viral encephalomyelitis caused by the members of the Lyssavirus genus and Rhabdoviridae family Disease definition Rabies causes more than 55, 000 human deaths, every year, throughout the world.

3. Rabies epizootiology RABIES RESERVOIRS URBAN RABIES WILD RABIES RABIES ASSOCIATED WITH TERRESTRIAL CARNIVORES SUSCEPTIBLE SPECIES (DEAD ENDS) Cats VECTORS Ferrets Perros Dogs Vampire bat Big brown bat Silver hair bat Red bat Free tail bat Mapaches Raccoons Gray foxes Stripped skunk Spotted skunk Arctic foxesCoyotes Mongooses RABIES ASSOCIATED WITH BATS Cows Goats and Sheep Humans and horses Pigs Beavers and woodchucks Raccoon dogs ENZOOTIC RABIES

4. Rabies status CityYear*Controlled EpizooticMemphis, TN19481958 EnzooticChicago, IL19541958 EnzooticPontiac, MI19541958 EnzooticSt Louis, MO19541958 & EnzooticLos Angeles, CA1922-19571975 & EnzooticDallas, TX19541976 & EnzooticPhoenix, AZ19501976 & EnzooticHouston/Harris, TX19541977 Last dog rabies enzootics in the United States Rabies in Wild and Domestic Animals in the United States, 1950-2005 History of rabies control and its elimination from the United States

5. 1988 Major rabies enzootics in terrestrial carnivores and the last rabies outbreak that involved both dogs and coyotes in the United States FOX

6. Major Terrestrial RABV Biotypes: Geographic distribution in the US and Mexico. Major Terrestrial RABV Biotypes: Geographic distribution in the US and Mexico. Northcentral Skunk Southcentral Skunk Dog/ Coyote Raccoon CA Skunk Baja CA Skunk Red Fox Yucatan Skunk Yucatan Dog Chiapas Dog Guerrero-Oaxaca Dog Puebla -Tiaxcala Dog Estado de Mexico Dog Michoacan Dog US- MX Dog AZ Gray Fox Sinaloa- Durango Skunk North Central Mexico Skunk Puerto Rico Mongoose TX Gray Fox

7. To characterized, at genetic level, all RABV variants and lineages historically associated with enzootic rabies in dogs in the US in order to understand and follow up its elimination, evolution and re-emergence in wildlife canines. General and objective

8. Results

9. East Texas-Mex Border Dog-coyote D1 Yucatan Dog D10 Puebla-Tlaxcala Dog D3, D7, D8 Guerrero-Oaxaca Dog D9 Michoacan Dog D2, D4 Estado de Mexico Dog D5 and D6 Chiapas Dog West US-Mx border Dog-coyote D11 * Extinct dog rabies focus * * * * * Searching the origin of re emerging enzootic dog rabies in the US: the domestic dog/coyote case, part I ? Potentially, more cases during 2006 ?

10. Searching the origin of re emerging enzootic dog rabies in the US: the domestic dog/coyote case, part II

11. D5 E19 D13 E20 E17 A14 A15 G5 F6 B12 C8 D6 E14 C7 G3 H2 A16 C5 C12 D8 B19 H5 B13 D15 C3 C14 A8 B14 D13 A6 A9 C13 G4 B10 B2 E12 I6 H4 C9 A1 F17 C1 B17 I4 A4 H3 B20 B16 I5 F5 B5 D2 G1 A10 C21 B4 E11 E3 A13 A3 D1 H1 C19 B7 C2 F1 E8 I3 D4 F12 A12 I2 A18 C17 A5 F11 D14 A17 B1 D3 I1 B3 D16 G6 A2 E1 D17 B15 B11 B18 C11 C6 D7 F13 E10 G2 E7 E9 E2 F10 E5 D10 F14 E4 E16 F15 F3 F2 F4 E6 E15 D2 Michoacan Dog D3 Puebla Tlaxcala Dog D9 D6DFEdoMexDg D8PueTlxDg D7PueTlxDg D5EdoMexDg D4MichMorDg D10YucDg 3853 El Salvador, Central America Hm 2003 4486chisdg02 D11 West US-Mexico border dog foci 1961-1995 “Sonora Dog variant” C16 C20 W6UScaliforniaSK7494 W7Yucunidentifiedreservoir W9MxBCSSk9002 W4732californiaSK W5USNorthcentralSk8498 WestCAnSK W2UsMxGrayfox7902 3886Sonsk2003 W1SinDgoSk01 E13 D12 C15 C10 W3 US Texas gray fox 83-97 D9 A11 E18 D11 B6 C18 C4 SD53304 SD55304 SD54304 11104304 F016304 SNP11104304 W11 US Southcentral Skunk 84-02 B9 W12CanadaUSRaccoon W12 W13MxNorthcentralSk LCNA LCMx LCCi LBNA LMx LCi LANNA HTCi HTAg HTMx Mevcaluc9295 MCUS MCAzCAUSA8693 PipistrellusUSA EF1aCOUSA8502 EF1 71USA72EF1b AF394869ap93ca AF394868ap91ca MCiAg TdbrCidmtc EF2AZ EF2US TBCiAg Tadarida brasiliensis mexicana North America B8 Dr7 Dr6 Dr1 Dr2 Dr3 DrSA Dr4 Dr5 Duvenhage2 EBL2 99 92 99 68 70 75 99 89 88 77 97 96 61 65 60 89 99 87 98 66 99 96 94 88 67 95 70 84 67 71 65 63 64 62 89 0.05 W11 US Southcentral Skunk 84-99 Tadarida brasiliensis mexicana North America D11 West Texas US Mexico border Dog “Sonora Dog” W3 US Texas gray fox 83-97 Coyote/Dog East US- Mexico border 1976-2002 Consensus amino acid change with respect Dog variants. Amino acid Serine change to Threonine in position 426. D2, D3 Puebla-Tlaxcala 1978-1995 Michoacan 1989-2000 Northcentral Skunk Southcentral Skunk Dog/Coyote Raccoon CA Skunk Red Fox Yucatan Dog Chiapas Dog Guerrero- Oaxaca Dog Puebla -Tlaxcala Dog Estado de Mexico Dog Michoacan Dog US- MX Dog AZ Gray Fox TX Gray Fox

12. 1700Mxdfdg99 1766Mxdg02 575Mxtlxdg00 409 01550F 457Mxpuedg01 2749Mxpuepg00 177 02 2609Mxgtoct01 1769Mxpuedg02 67Mxtlxdg02 1096-00 440Mxgrodg99 657Mxdgosk01 1390Mxmxdg99 3143Mxpuebv94 3134Mxdfpg91 3121Mxdfhm91 3142Mxdffer90 3138Mxmichdk91 1407Mxmxdg00 1634-99 1587Mxdfdg00 1027 02 2509Mxmxdg02 2411-99 202Mxgrodg00 250Mxgrodg01 251Mxgrodg01 175Mxyucdg98 10Mxyucdg95 110Mxyucdg97 151Mxyuc98 428Mxyucct03 182Mxyucdg98 412Mxyucdg02 60Honddg01 3066Mxchisdg02 3122Mxmichct90 3123Mxmichbv91 3128Mxdgodg91 3144Mxchihbv94 3124Mxpuedg91 3126Mxpuepg93 3127Mxtlxgt91 3147Mxqrodg95 3151Puedg95 490Mxmichdg00 247Mxcoahcy01 3306Mxct02 B14Ustxcyken92 H2USTXWillcy98 E15USTXzavdg95 F2USTXlasallecy96 E14USTXhgocy95 G1USTXWebbcy97 B15UScyduv92 3907Mxsonbct 3140Mxsonbct94 3148Mxchihbct94 FluryLEP HEP-flury D9USTxdglub94 YC63China CVS11 PM1503 MRVChinahenan RAVNUCM61047 CVSFR AV01 RdfxEurope RacdgfxEastEu StepfxRdefxFareast RedfxMiddleast Africa1b Africa1a RCHLAB009663 NiCEAB128149 NishigaraAB044824 BrdgV2 SADB19M31046 SRV9AF499686 2840PuertoRico PrdgV1 2839PuertoRico 1629Mxbcssk01 3908Mxslpsk OrgTIndAY956319 ArctfxPakistan U11734ontario24 U03769arctica1 AY352499ArctAK ArctfxSibCan 1500AFS Viverrid Viver African AF467949 8660GUIAFR2 88636HAVAFR2 AsiandgInd-SriL AsiandgThai-Viet 2311Mxzacsk01 2818Mxslpsk02 1516-01 RaccoonCAnUs SHBRVAY705373 LC8 3901Mxmichbv03 3902Mxmichbv03 TB1 AF394876 TB 1475 00 2153Salvhm02 3888Mxbvtamp03 460 3891verbv03 3858Mxhgoov03 3861Mxhgobv04 3878Mxslpbv04 AY573943flyfxGT7 U22845GT5 U22846GT6 99 100 99 91 100 99 100 73 83 100 98 85 100 57 100 71 76 75 76 91 100 78 100 77 98 86 68 99 89 88 84 72 65 77 100 98 100 90 100 99 71 95 97 99 89 83 100 95 75 99 73 84 0.05 D1 Coyote/Dog East US Mexico border 1976 2002 D3 Dog Puebla-Tlaxcala 1978 1995 D2 Michoacan 1989 2000 W2 Arizona gray fox variant W3 Texas gray fox variant Vaccine strains related with CVS Flury LEP and HEP D11 Sonora Dog West US Mexico border 1961 1991 Michoacan 1990 1991 Chiapas and Central America 2001 2003 D10 Yucatan 1995 2002 D9 Guerrero Oaxaca 1992 2003 D5 Estado de Mexico 1991 2002 D4 Michoacan 1991 1993 D6 Estado de Mexico 1991 1998 D7 Dog Puebla Tlaxcala 1994 2001 D8 Dog Puebla Tlaxcala 1999 2002 Mexican dog lineages European and Middle east red fox African 1 dog SADB19 and Puerto Rico South American Dog variant Baja California Sur, skunk Arctic fox and arctic like variant African mongoose West African dog, Africa 2 Asian dog Vaccine strains red fox related North American Skunk and raccoon variants North American RV bat variants Outgroup, other Lyssavirus species Cosmopolitan dog variant Rabies associated with terrestrial carnivores Rabies associated with bats

14. Dog Rabies and Human Migrations Northcentral Skunk Southcentral Skunk Dog/ Coyote Raccoon CA Skunk Baja CA Skunk Red Fox Yucatan Dog Chiapas Dog Guerrero-Oaxaca Dog Puebla -Tiaxcala Dog Estado de Mexico Dog Michoacan Dog US- MX Dog AZ Gray Fox Puerto Rico Mongoose TX Gray Fox SAD B19 Puerto Rico Monngoose conection

15. We were able to detect historic movement patterns of major dog rabies enzootics which in turn are consistent with the demographics and immigration patterns of the Mexican and Central American populations described by the Sociologist Jorge Durand 2005. We also showed potential historic disseminations of enzootic dog rabies into wild life populations such as those illustrated by the phylogenetic analysis in gray foxes and coyotes in Texas. This findings stress out the importance of the historic follow up of rabies epizootics in order to assess future burdens for animal and public health. Final Remarks Our study showed that rabies virus associated with enzootics in dog populations has evolved over time, in discrete geographic lineages which may be translocated by human related activities.

16. To: Texas Department of Health, Austin, TX L. Zuckero P. Hunt S. Oertli L. Robinson National Zoonoses Program from the Ministry of Health, Mexico. Dr Fernando Vargas Pino and Rabies Laboratory at InDRE, from the Ministry of Health, Mexico. Acknowledgements