Canine Rabies Elimination from the United States: an Epidemiological and Virological Perspective

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

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

4. Rabies in Wild and Domestic Animals in the United States, 1950-2005 History of rabies control and its elimination from the United States Last dog rabies enzootics in the United States

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

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

7. General and objective 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.

8. Results

9. 2587txcy94 2193txhm79 D1 1979-2003 835txhm76 2160txdg85 92 l * 2361txdg94 834txhm79 247coahcy01 75 4506tampct00 1989-2000 4508tampct02 * 91 l 3288michdg00 D2 848ormichihm89 3147qrodg95 l 1978-1995 3151mxuk * 555dfdg78 D3 92 3127tlxgt91 3124puedg91 1993-1996 3126puepg93 2184camorhm93 3138michdk91 D4 4823slpdg96 l 3267mxdg01 4511mxdg02 85 1999-2003 3260dfdg00 l 3325hgodg03 87 D5 3269zacbv00 3284mxdg99 * 4514mxdg03 4516mxdg02 D6 3134dfpg91 1990-1998 1278dfdg91 * l 3121dfhm91 60 3244mxdg98 1282dfdg91 1994-1999 3142dfferr90 3278grodg99 4822puedg96 3143puebv94 l 3299dfdg99 D7 3281puedg00 61 4513puedg02 1999-2006 4519puedg02 l 3303tlxdg02 3279puedg03 63 D8 3275gtoct01 3302tlxdg01 4491mxdg02 3313jaluk03 559dfdg78 1992-2006 l 4815slpdg99 50 3125oaxdg92 84 l 59 3251grodg00 D9 3285grodg01 3287grodg03 D10 D 1995-2006 l 4476yucdg95 4480yucdg01 97 4481yucpg02 87 1995-2006 4485yucdg03 55 674cagroct87 4486chisdg03 66 l 1303orchihdg91 D11 947azsonhm81 l * 3123michbv91 73 85 3122michct90 953cachihhm79 1961-1995 2830txcy95 l 3144chihbv94 2520cacy90 63 726cahm61 3128dgodg91 671sondg88 672sondg88 65 1274sondg88 673sondg88 Searching the origin of re emerging enzootic dog rabies in the US: the domestic dog/coyote case, part I ? Potentially, more cases during 2006 West US-Mx border Dog-coyote D11 East Texas-Mex Border Dog-coyote D1 * ? Yucatan Dog D10 Estado de Mexico Dog D5 and D6 Puebla-Tlaxcala Dog D3, D7, D8 * * * * Michoacan Dog D2, D4 Guerrero-Oaxaca Dog D9 Chiapas Dog *Extinct dog rabies focus

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

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

12. 1700Mxdfdg99 1766Mxdg02 575Mxtlxdg00 409 01550F 73 D8 Dog Puebla Tlaxcala 1999 2002 457Mxpuedg01 84 2749Mxpuepg00 177 02 2609Mxgtoct01 1769Mxpuedg02 67Mxtlxdg02 1096-00 440Mxgrodg99 D7 Dog Puebla Tlaxcala 1994 2001 657Mxdgosk01 75 1390Mxmxdg99 3143Mxpuebv94 99 D6 Estado de Mexico 1991 1998 3134Mxdfpg91 3121Mxdfhm91 3142Mxdffer90 D4 Michoacan 1991 1993 3138Mxmichdk91 1407Mxmxdg00 95 1634-99 83 D5 Estado de Mexico 1991 2002 1587Mxdfdg00 100 1027 02 2509Mxmxdg02 2411-99 202Mxgrodg00 D9 Guerrero Oaxaca 1992 2003 99 250Mxgrodg01 Mexican dog lineages 251Mxgrodg01 175Mxyucdg98 10Mxyucdg95 89 110Mxyucdg97 100 D10 Yucatan 1995 2002 151Mxyuc98 428Mxyucct03 182Mxyucdg98 412Mxyucdg02 71 Chiapas and Central America 2001 2003 60Honddg01 3066Mxchisdg02 100 97 Michoacan 1990 1991 3122Mxmichct90 95 3123Mxmichbv91 D11 Sonora Dog West US Mexico border 1961 1991 3128Mxdgodg91 99 3144Mxchihbv94 Cosmopolitan dog variant 3124Mxpuedg91 D3 Dog Puebla-Tlaxcala 1978 1995 D2Michoacan 1989 2000 100 3126Mxpuepg93 3127Mxtlxgt91 90 3147Mxqrodg95 3151Puedg95 490Mxmichdg00 77 247Mxcoahcy01 99 3306Mxct02 B14Ustxcyken92 D1 Coyote/Dog East US Mexico border 1976 2002 H2USTXWillcy98 98 E15USTXzavdg95 F2USTXlasallecy96 Rabies associated with terrestrial carnivores E14USTXhgocy95 G1USTXWebbcy97 B15UScyduv92 100 3907Mxsonbct W2 Arizona gray fox variant 3140Mxsonbct94 3148Mxchihbct94 100 Flury LEP and HEP FluryLEP HEP-flury W3 Texas gray fox variant 65 D9USTxdglub94 76 YC63China CVS11 99 91 PM1503 Vaccine strains related with CVS MRVChinahenan 100 RAVNUCM61047 75 CVSFR AV01 78 RdfxEurope 98 RacdgfxEastEu European and Middle east red fox StepfxRdefxFareast RedfxMiddleast African 1 dog Africa1b 89 72 Africa1a RCHLAB009663 Vaccine strains red fox related 86 NiCEAB128149 100 NishigaraAB044824 84 South American Dog variant BrdgV2 100 SADB19M31046 SRV9AF499686 SADB19 and Puerto Rico 2840PuertoRico 68 PrdgV1 88 100 2839PuertoRico Baja California Sur, skunk 1629Mxbcssk01 3908Mxslpsk 100 100 OrgTIndAY956319 ArctfxPakistan Arctic fox and arctic like variant U11734ontario24 100 U03769arctica1 77 100 AY352499ArctAK 83 ArctfxSibCan 100 African mongoose 76 1500AFS Viverrid 100 Viver African AF467949 West African dog, Africa 2 8660GUIAFR2 100 100 88636HAVAFR2 Asian dog AsiandgInd-SriL AsiandgThai-Viet 71 100 2311Mxzacsk01 73 85 2818Mxslpsk02 North American Skunk and raccoon variants 1516-01 RaccoonCAnUs 100 SHBRVAY705373 LC8 57 100 3901Mxmichbv03 3902Mxmichbv03 98 100 TB1 100 AF394876 TB Rabies associated with bats 1475 00 North American RV bat variants 2153Salvhm02 3888Mxbvtamp03 99 460 91 3891verbv03 99 3858Mxhgoov03 3861Mxhgobv04 3878Mxslpbv04 AY573943flyfxGT7 Outgroup, other Lyssavirus species U22845GT5 U22846GT6 99 0.05

13. 1700Mxdfdg99 1766Mxdg02 575Mxtlxdg00 409 01550F 73 D8 Dog Puebla Tlaxcala 1999 2002 457Mxpuedg01 84 2749Mxpuepg00 177 02 2609Mxgtoct01 1769Mxpuedg02 67Mxtlxdg02 1096-00 440Mxgrodg99 D7 Dog Puebla Tlaxcala 1994 2001 1700Mxdfdg99 1766Mxdg02 657Mxdgosk01 75 575Mxtlxdg00 409 01550F 73 1390Mxmxdg99 457Mxpuedg01 84 2749Mxpuepg00 3143Mxpuebv94 177 02 99 2609Mxgtoct01 D6 Estado de Mexico 1991 1998 3134Mxdfpg91 1769Mxpuedg02 67Mxtlxdg02 3121Mxdfhm91 1096-00 440Mxgrodg99 3142Mxdffer90 657Mxdgosk01 75 D4 Michoacan 1991 1993 1390Mxmxdg99 3138Mxmichdk91 3143Mxpuebv94 99 3134Mxdfpg91 1407Mxmxdg00 3121Mxdfhm91 95 3142Mxdffer90 1634-99 83 D5 Estado de Mexico 1991 2002 3138Mxmichdk91 1407Mxmxdg00 1587Mxdfdg00 100 95 1634-99 83 1027 02 1587Mxdfdg00 100 1027 02 2509Mxmxdg02 2509Mxmxdg02 2411-99 2411-99 202Mxgrodg00 99 250Mxgrodg01 202Mxgrodg00 D9 Guerrero Oaxaca 1992 2003 251Mxgrodg01 99 175Mxyucdg98 250Mxgrodg01 10Mxyucdg95 89 Mexican dog lineages 110Mxyucdg97 100 251Mxgrodg01 151Mxyuc98 428Mxyucct03 175Mxyucdg98 182Mxyucdg98 412Mxyucdg02 71 10Mxyucdg95 60Honddg01 89 3066Mxchisdg02 100 110Mxyucdg97 97 3122Mxmichct90 100 95 3123Mxmichbv91 D10 Yucatan 1995 2002 151Mxyuc98 3128Mxdgodg91 99 3144Mxchihbv94 428Mxyucct03 3124Mxpuedg91 100 3126Mxpuepg93 182Mxyucdg98 3127Mxtlxgt91 90 3147Mxqrodg95 412Mxyucdg02 71 3151Puedg95 490Mxmichdg00 Chiapas and Central America 2001 2003 60Honddg01 77 247Mxcoahcy01 99 3306Mxct02 3066Mxchisdg02 100 B14Ustxcyken92 97 Michoacan 1990 1991 H2USTXWillcy98 98 3122Mxmichct90 E15USTXzavdg95 95 F2USTXlasallecy96 3123Mxmichbv91 E14USTXhgocy95 D11 Sonora Dog West US Mexico border 1961 1991 G1USTXWebbcy97 3128Mxdgodg91 99 B15UScyduv92 100 3907Mxsonbct 3144Mxchihbv94 3140Mxsonbct94 Cosmopolitan dog variant 3124Mxpuedg91 3148Mxchihbct94 100 FluryLEP D3 Dog Puebla-Tlaxcala 1978 1995 D2Michoacan 1989 2000 100 3126Mxpuepg93 HEP-flury 65 D9USTxdglub94 3127Mxtlxgt91 76 YC63China 90 CVS11 99 91 3147Mxqrodg95 PM1503 MRVChinahenan 100 3151Puedg95 RAVNUCM61047 75 CVSFR 490Mxmichdg00 AV01 78 RdfxEurope 77 247Mxcoahcy01 98 RacdgfxEastEu StepfxRdefxFareast 99 3306Mxct02 RedfxMiddleast Africa1b 89 72 B14Ustxcyken92 Africa1a RCHLAB009663 D1 Coyote/Dog East US Mexico border 1976 2002 86 H2USTXWillcy98 98 NiCEAB128149 100 NishigaraAB044824 84 E15USTXzavdg95 BrdgV2 100 SADB19M31046 F2USTXlasallecy96 SRV9AF499686 2840PuertoRico E14USTXhgocy95 68 PrdgV1 88 100 2839PuertoRico G1USTXWebbcy97 1629Mxbcssk01 3908Mxslpsk 100 B15UScyduv92 100 OrgTIndAY956319 ArctfxPakistan 100 3907Mxsonbct U11734ontario24 W2 Arizona gray fox variant 100 U03769arctica1 3140Mxsonbct94 77 100 AY352499ArctAK 83 ArctfxSibCan 3148Mxchihbct94 100 100 76 1500AFS Viverrid 100 Viver African AF467949 Flury LEP and HEP FluryLEP 8660GUIAFR2 100 100 88636HAVAFR2 HEP-flury W3 Texas gray fox variant AsiandgInd-SriL 65 AsiandgThai-Viet D9USTxdglub94 71 100 2311Mxzacsk01 73 76 YC63China 85 2818Mxslpsk02 1516-01 RaccoonCAnUs CVS11 99 100 91 SHBRVAY705373 PM1503 LC8 57 100 Vaccine strains related with CVS 3901Mxmichbv03 MRVChinahenan 3902Mxmichbv03 98 100 100 TB1 RAVNUCM61047 100 AF394876 TB Rabies associated with bats 1475 00 75 CVSFR 2153Salvhm02 3888Mxbvtamp03 99 AV01 460 78 91 3891verbv03 RdfxEurope 99 3858Mxhgoov03 3861Mxhgobv04 98 RacdgfxEastEu European and Middle east red fox 3878Mxslpbv04 AY573943flyfxGT7 StepfxRdefxFareast U22845GT5 U22846GT6 99 RedfxMiddleast African 1 dog 0.05 Africa1b 89 72 Africa1a RCHLAB009663 Vaccine strains red fox related 86 NiCEAB128149 100 NishigaraAB044824 84 South American Dog variant BrdgV2 100 SADB19M31046 SRV9AF499686 SADB19 and Puerto Rico 2840PuertoRico PrdgV1 88 100 2839PuertoRico Baja California Sur, skunk 1629Mxbcssk01 3908Mxslpsk

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

15. 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. 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.

16. Acknowledgements 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.