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R.T. Trout 1 , C.D. Steelman 1 , A.L. Szalanski 1 , K. Kvamme 2 , and P.C. Williamson 3

Genetic & Spatial Identification of Borrelia & Rickettsia Pathogens from Arkansas Ticks & Mammals. R.T. Trout 1 , C.D. Steelman 1 , A.L. Szalanski 1 , K. Kvamme 2 , and P.C. Williamson 3 1 University of Arkansas Entomology • Fayetteville, Arkansas 72701

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R.T. Trout 1 , C.D. Steelman 1 , A.L. Szalanski 1 , K. Kvamme 2 , and P.C. Williamson 3

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  1. Genetic & Spatial Identification of Borrelia & Rickettsia Pathogensfrom Arkansas Ticks & Mammals R.T. Trout1, C.D. Steelman1, A.L. Szalanski1, K. Kvamme2, and P.C. Williamson3 1University of Arkansas Entomology • Fayetteville, Arkansas 72701 2University of Arkansas Anthropology • Fayetteville, Arkansas 72701 3University of North Texas Health Science Center • Ft. Worth, Texas 76107-2699

  2. Larvae Nymph Adult Tick - Host Associations

  3. Ixodes scapularis Amblyomma americanum Borrelia Pathogens B. burgdorferi B. lonestari • Several Species • Variable Symptoms • Infection variable • Cases • Misdiagnosed • Under reported

  4. Dermacentor variabilis Ixodes scapularis Amblyomma americanum Rickettsia Pathogens • Spotted fevers • Several Species • Variable Symptoms • Fever, edema, spreading rash • Infection variable • Cases

  5. Arkansas Tick-borne Diseases • Changing State • Fragmented habitat • top-10 growing metropolitan areas (Anonymous 2001) • 1996: military cases • R. rickettsii • R. typhus • B. burgdorferi (Warner et al. 1996)

  6. Borrelia in Arkansas 26 cases of LD in Arkansas, since 2002

  7. Rickettsia in Arkansas 231 cases of RMSF in 45 Ctys., since 2002

  8. Objectives • Genetically identify bacteria in Arkansas by examining ticks & hosts using molecular techniques • Borreliapathogens • Rickettsiapathogens • Identify potential risk areas using geographic information systems

  9. Outline • General Methods for the Project • Results • Tick Collections • Borrelia from Ticks & Hosts • Rickettsia from Ticks • Co-infection • Summary & Future Direction

  10. Methods: Tick Collections Lyme disease risk increases in areas with Ab positive canines & ticks on white-tailed deer (Daniels et al. 1993)

  11. Methods: Extraction & PCR Tick DNA- Ethanol Vials: Qiagen DNEasy Protocol 4 Reactions 1 Tick DNA (16S mtDNA) 1 Borrelia (flaB) 2 Rickettsia (gltA & rompB) Host DNA- Whatman FTA Cards: Protocol

  12. Methods: PCR • Tick DNA Reaction • 16S mt DNA ribosomal RNA (Black & Piesman 1994) • Borrelia genus specific reaction • flaB gene (Barbour et al. 1996) • Rickettsia genus specific reactions • gltA (Kollars & Kengluecha 2001) • rompB (Regnery et al. 1991)

  13. Methods: Sample Verification 16S tick mtDNA gltA rompB flaB Sequenced at UNT Health Science Center, Ft. Worth TX Map Incidence GenBank

  14. Methods: Host Distribution 247 Clinics 35 Participated (14%) Moran’s I: Deer- 0.46 (1.19) Pet- 0.01 (0.47)

  15. Methods: GIS Mapping IDW2 Mask Buffer Results!

  16. Results: Tick Collections Canines • 1498 ticks from 173 canines • 8 ± 4.0 ticks/host (range 1-100) Felines • 84 ticks from 16 felines • 5 ± 2.4 ticks/host (range 1-17) Deer • 1581 ticks from 250 deer • 6 ± 0.3 ticks/host (range 0-32)

  17. Results: Tick Collections

  18. Results: Tick Collections All ticks from pets were tested but Only 56% of the ticks collected from deer were tested

  19. Ixodes scapularis Amblyomma americanum Results: Borrelia

  20. Results: Borrelia in Ticks Ticks from Canines 23% 24% 24% 36% 10% 12% 24% Ticks from Felines 20% 34% 27% 0% 34% 0% 0% Ticks from Deer 37% 37% 38% 33% 43% 57% 42%

  21. Results: Borrelia in Ticks Felines Canines Deer Total

  22. Results: Borrelia species in ticks 613 ticks positive 3 B. burgdorferi 5 B. lonestari strains 22 SSc26 38 M2002-V1 21 Tx/06 44 Nc/Md 1 AA115 *487 ticks not sequenced

  23. Results: Borrelia spp. in Hosts 25% 15% 4% Host Animal

  24. Results: Borrelia in Hosts N = 21 N = 14 N = 3 N = 1

  25. Results: Borrelia Incidence Ticks from Pets (n = 355 / 1585 ticks) Ticks from Deer n = 259 / 695 ticks)

  26. Results: Borrelia Incidence Blood from Pets (n = 9/174 hosts) Blood from Deer (n = 55/220 hosts)

  27. Results: Borrelia Incidence IDW2 Ticks from Pets (n = 355 / 1585 ticks) Ticks from Deer n = 259 / 695 ticks)

  28. Results: Borrelia Incidence IDW2 Blood from Pets (n = 9/174 hosts) Blood from Deer (n = 55/220 hosts)

  29. Results: Borrelia Incidence IDW2 Map Algebra: (4 maps) * State Mask

  30. Results: Borrelia Risk Areas 2007 Cases (n=8) Benton, Carroll, Crawford, Saline, Washington Co.

  31. Results: Borrelia Risk Areas

  32. Results: Borrelia Risk Areas

  33. Dermacentor variabilis Ixodes scapularis Amblyomma americanum Results: Rickettsia

  34. Results: Rickettsia in Ticks Ticks from Canines 26% 37% 9% 32 3% 40% 40% Ticks from Felines 31% 40% 14% 10% 39% 0% 0% Ticks from Deer 38% 37% 56% 37% 0% 11% 33%

  35. Results: Rickettsia in Ticks

  36. Results: Rickettsia Incidence Ticks from Deer (n = 120/315 ticks) Ticks from Pets (n = 425/1580 ticks)

  37. Results: Rickettsia Incidence Ticks from Deer (n = 120/315 ticks) Ticks from Pets (n = 425/1580 ticks)

  38. Results: Rickettsia Incidence 2007 Cases (n=121): Benton, Cleburne, Crawford, Faulkner, Hot Spring, Pope, Pulaski, Van Buren, White (>4 cases)

  39. Results: Rickettsia Risk Areas

  40. Results: Rickettsia Risk Areas

  41. Results: Co-infection

  42. Results: Co-infection Borrelia Risk Map Rickettsia Risk Map Map Algebra: Add the 2 Maps

  43. Results: Co-infection

  44. Results: Co-infection

  45. Summary Borrelia Rickettsia

  46. Summary Borrelia All Tick Species Positive R. amblyommii, R. montanensis, R. parkeri, R. rickettsii Rickettsia • All Tick Species Positive • All Hosts Positive • Borrelia burgdorferi & Borrelia lonestari

  47. Future Direction Determine Population Structure of Aa & Is Conduct Field Work in Designated Risk Areas Identify the Pathogens in Humans Determine Symptoms & Pathogenesis Determine Vector Competency… Identify Environmental Variables with Risk Areas

  48. Acknowledgements • Arkansas Veterinary Medical Association • University of Arkansas • Dr. D. Steelman • Dr. A. Szalanski • Dr. K. Loftin • Dr. K. Kvamme • University of North Texas • Phillip Williamson • Peggy Billingsly • Janel • Veterinary Entomology Laboratory • J. Shen • Insect Genetics Laboratory • J. McKern • M. Tolivar • Friends and Family 

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