Ehrlichia and Borrelia spp. Infection in German Shepherd Dogs in Korea

독일 셰퍼드 개에서 Ehrlichia와 Borrelia spp.의 감염

  • Hwang, Cheol-Yong (Department of Veterinary Internal Medicine, Research Institute and BK21 Program for Veterinary Science and College of Veterinary Medicine) ;
  • Seo, Sang-Ho (Department of Veterinary Internal Medicine, Research Institute and BK21 Program for Veterinary Science and College of Veterinary Medicine) ;
  • Kang, Jun-Gu (Department of Veterinary Internal Medicine, Research Institute and BK21 Program for Veterinary Science and College of Veterinary Medicine) ;
  • Youn, Hwa-Young (Department of Veterinary Internal Medicine, Research Institute and BK21 Program for Veterinary Science and College of Veterinary Medicine) ;
  • Chae, Joon-Seok (Department of Veterinary Internal Medicine, Research Institute and BK21 Program for Veterinary Science and College of Veterinary Medicine)
  • Accepted : 2011.04.16
  • Published : 2011.04.30

Abstract

The presence of the tick-borne pathogens Ehrlichia and Borrelia in German Shepherd dogs in Korea was determined by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). A total of 291 dogs were randomly selected from five Korean provinces from October 2005 through September 2006. The seroprevalence of antibodies to canine Ehrlichia and Borrelia agents detected by ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ Test, IDEXX Laboratories) was 7.56% (22 dogs) and 1.72% (5 dogs) respectively, throughout the country. Positive antibodies against both pathogens were detected in two dogs (0.69%). The provincial distribution of seroprevalence against Ehrlichia was 1.28% (1 of 78) in Gyeonggi-do, 12.64% (11 of 87) in Gangwon-do, 9.76% (4 of 41) in Chungcheong-do, 8.93% (5 of 56) in Gyeongsang-do, and 3.45% (1 of 29) in Jeolla-do. According to PCR analysis, Ehrlichia chaffeensis target DNA was amplified in 3.09% (9 of 291 dogs) of blood samples, 2.41% (7 of 291) from Gangwon-do and 0.69% (2 of 291) from Chungcheong-do. The oligonucleotide sequences (SNU-EC3 and SNU-EC5) from the PCR fragment examined in Korea were closely related to E. chaffeensis isolated from the tick Haemaphysalis longicornis, in China and the state of Arkansas in the US. Based on these results, the presence of E. chaffeensis infection was identified in German Shepherds being bred in Korea. These results bring to light the importance of paying close attention to tick-borne infections such as Lyme disease during clinical diagnosis. This infectious disease should be included as a differential diagnosis for patients who participate in outdoor activity from spring to fall or who have thrombocytopenia or leucopenia.

국내 독일 셰퍼드로부터 에를리히아증과 라임병에 대한 항체가 조사를 위하여 효소면역측정법을 시용하였으며, 중합효소연쇄반응을 이용하여 진드기매개성 병원체를 조사하였다. 2005년 10월부터 2006년 9월까지 국내 5개 지방으로부터 291마리 독일 셰퍼드를 선정하였다. 개 에를리히아 항체 조사를 위한 ELISA (Snap$^{(R)}$ 3Dx$^{(R)}$ test kit, IDEXX) 검사결과 전국적으로 7.56%(22마리)의 에를리히아 양성 항체 보균개와 1.72%(5마리)의 보렐리아 양성 항체 보균개로 조사되었다. 이 중 2마리에서는 두 가지 병원체 모두에 대한 양성 항체가 검출되었다. 에를리히아에 대한 양성항체를 지방별로 보면 경기도는 1.28%(78마리 중 1마리), 강원도는 12.64%(87마리 중 11마리), 충청도는 9.76%(41마리 중 4마리), 경상도는 8.93%(56마리 중 5마리), 전라도는 3.45%(56마리 중 5마리)로 조사되었다. Ehrlichia chaffeensis 유전자를 검출한 결과 전국 평균이 3.09%(291 마리 중 9마리), 강원도는 2.41%(291 마리 중 7마리) 그리고 충청도는 0.69%(291마리중 2마리)로 조사되었다. 중합효소연쇄반응으로 증폭된 유전자(SNU-EC3과 SNU-EC5)에 대한 유전자염기서열분석결과 한국(Haemaphysalis longicornis 진드기)과 중국 그리고 Arkansas 분리주인 E. chaffeensis와 매우 유사하였다. 이 연구의 결과 국내에서 사육되는 독일셰퍼드에서 에를리히아증의 병원체와 라임병에 대한 항체가 검출된 것으로 볼 때 임상 진단에 있어서 진드기매개전염병에 대한 중요성을 인식해야 할 것으로 판단된다. 따라서 혈소판감소증이나 백혈구감소증이 있는 경우 그리고 봄부터 가을까지 야외활동을 한 경력이 있는 환자에 대한 감별진단 항목에 반드시 포함시켜야 할 것을 제안한다.

Keywords

References

  1. Adelson ME, Rao RY, Tilton RC, Cabets K, Eskow E, Fein, L, Occi JL, Mordechai E. Prevalence of Borrelia burgdorferi, Batonella spp., Babesia microti, and Anaplasma phagocytophila in Ixodes scapularis ticks collected in northern New Jersey. J Clin Microbiol 2004; 42: 2799-2801. https://doi.org/10.1128/JCM.42.6.2799-2801.2004
  2. Anderson BE, Greene CE, Jones DC. Dawson JE. Ehrlichia ewingii sp. nov., the etiologic agent of canine granulocytic ehrlichiosis. lnt J Syst Bacteriol 1992a; 42: 299-302. https://doi.org/10.1099/00207713-42-2-299
  3. Anderson BE, Sims KG, Olson JG. Childs JE, Piesman JF, Happ CM, Maupin GO, Johnson BJ. Amblyomma americanum: a potential vector of human ehrlichiosis. Am J Trop Med Hyg 1993; 49: 239-244.
  4. Anderson BE, Summer JW, Dawson JE, Tzianabos T, Greene CR, Olson JG, Fishbein DB, Olsen-Rasmussen M, Holloway BP, George EH. Detection of the etiologic agent of human ehrlichiosis by polymerase chain reaction. J Clin Microbiol 1992b; 30: 775-780.
  5. Anziani OS, Ewing SA, Barker RW. Experimental transmission of a granulocytic form of the tribe Ehrlichieae by Dermacentor variabilis and Amblyomma americanum to dogs. Am J Vet Res 1990; 51: 929-931.
  6. Baumgartner BU, Rollinghoff M, Bogdan C. Prevalence of Borrelia burgdorferi and granulocytic and monocytic ehrlichiae in Ixodes ricinus ticks from southern Germany. J Clin Microbiol 1999; 37: 3448-3451.
  7. Cao WC, Gao YM, Zhang PH, Zhang XT, Dai QH, Dumler JS, LQ, Yang H. ldentification of Ehrlichia chaffeensis by nested PCR in ticks from southern China. J Clin Microbiol 2000; 38: 2778-2780.
  8. Castro MB, Nicholson WL, Kramer VL, Childs JE. Persistent infection in Neotoma fuscipes (Muridae: Sigmodontinae) with Ehrlichia phagocytophila sensu lato. Am J Trop Med Hyg 2001; 65: 261-267.
  9. Chae JS, Kim CM, Kim EH, Hur EJ, Klein TA, Kang TK, Lee HC, Song JW. Molecular epidemiological study for tick-borne disease (Ehrlichia and Anaplasma spp.) surveillance at selected U.S. military training sites/installations in Korea. Ann N Y Acad Sci 2003; 990: 118-125. https://doi.org/10.1111/j.1749-6632.2003.tb07349.x
  10. Chen SM, Dumler JS, Bakken JS. ldentification of the granulocytrotropic Ehrlichia species as the etiologic agent of human disease. J Clin Microbiol 1994; 32: 589-595.
  11. Dawson JE, Anderson BE, Fishbein DB, Sanchez JL, Goldsmith CS, Wilson KH, Duntley CW. Isolation and characterization of an Ehrlichia sp. from a patient diagnosed with human ehrlichiosis. J Clin Microbiol 1991; 29: 2741-2745.
  12. Dawson .JE, Biggie KL, Warner CK, Cookson K, Jenkins S, Levine JF, Olson JG. Polymerse chain reaction evidence of Ehrlichia chaffeensis, and etiologic agent of human ehrlichiosis, in dogs from southeast Virginia. Am Jet Res 1996; 57: 1175-1179.
  13. Dawson JE, Ewing SA. Susceptibility of dogs to infection with Ehrlichia chaffeensis, causative agent of human ehrlichiosis. Am J Vet Res 1992; 53 : 1322-1327.
  14. Donatein A, Lestoquard F. State of the present knowledge concerning rickettsiosis of animals. Arch Inst Pasteur Alger 1937; 15: 142-187.
  15. Dumler JS. Molecular methods for ehrlichiosis and Lyme disease. Clin Lab Med 2003; 23: 867-884. https://doi.org/10.1016/S0272-2712(03)00080-5
  16. Ewing SA, Dawson JE, Mathew JS. Attempted transmission of human granulocytotropic Ehrlichia (HGE) by Amblyomma americanum and Amblyomma maculatum. Vet Parasitol 1997; 70: 183-190. https://doi.org/10.1016/S0304-4017(96)01157-0
  17. Ewing SA, Roberson WR, Buckner RG, Hayat CS. A new strain of Ehrlichia canis. J Am Vet Med Assoc 1971; 159: 1771-1774.
  18. Foley .JE, Foley P, Brown RN, Lane RS, Dumlers JS, Madigan JE. Ecology of Anaplasma phagocytophilum and Borrelia burgdorferi in the western United States. J Vector Ecol 2004; 29: 41-50.
  19. Furuya Y, Katayama T, Yoshida Y, Kaiho I. Specific amplification of Rickettsia japonica DNA from clinical specimens by PCR. J Clin Microbiol 1995; 33: 487-489.
  20. Green CE. Ehrlichiosis, neorickettsiosis, anaplasmosis, and Wolbachia infection. In: Infectious diseases of the dog and cat, 3rd Ed. Saunders, St. Louis, 2006: 203-231.
  21. Greig B, Frigo E, Nelson PM. Seroprevalence of a newly emerging zoonotic granulocytic ehrlichial disease in Minnesota and Wisconsin dogs. J Am Coll Vet Intern Med 1996; 10: 180
  22. Groves MG, Dennis GL, Amyx HL, HuxsolI. DL. Transmission of Ehrlichia canis to dogs by ticks (Rhipicephalus sanguineous). Am J Vet Res 1975; 36: 937-940.
  23. Heo EJ, Park JH, Koo JR, Park MS, Park MY, Dumler JS, Chae JS. Serologic and molecular detection of Ehrlichia chaffeensis and Anaplasma phagcytophila (Human granulocytic ehrlichiosis agent) in Korean patients. J Clin Microbiol 2002; 40: 3082-3085. https://doi.org/10.1128/JCM.40.8.3082-3085.2002
  24. lnokuma H, Beppu T, Okuda M, Shimada Y, Sakata Y. Detection of ehrlichial DNA in Haemaphysalis ticks recovered from dogs in Japan that is closely related to a novel Ehrlichia sp. found in cattle ticks from Tibet, Thailand, and Africa. J Clin Microbiol 2004; 42: 1353-1355. https://doi.org/10.1128/JCM.42.3.1353-1355.2004
  25. Johnson EM, Ewing SA, Barker RW. Experimental transmission of Ehrlichia canis (Rickettsiales: Ehrlichieae) by Dermacentor variabilis (Acari: lxodidae). Vet Parasitol 1998; 74: 277-288. https://doi.org/10.1016/S0304-4017(97)00073-3
  26. Kim CM, Kim MS, Park MS, Park JH, Chae JS. Identification of Ehrlichia chaffeensis, Anaplasma phagocytophilum, and A, bovis in Haemaphysalis longicornis and Ixodes persulcatus ticks from Korea. Vector Borne Zoonotic Dis 2003; 3: 17-26. https://doi.org/10.1089/153036603765627424
  27. Kim CM, Kim JY, Yi YH, Lee MJ, Cho MR, Shah DH, Klein TA, Kim HC, Song JW, Chong ST, O'Guinn ML, Lee JS, Lee IY, Park JH, Chae JS. Detection of Bartonella species from ticks, mites and small mammals in Korea. J Vet Sci 2005; 6: 327-334.
  28. Kim CM, Yi YH, Yu DH, Lee MJ, Cho MR, Desai AR, Shringi S, Klein TA, Kim HC, Song JW, Baek LJ, Chong ST, O'guinn ML, Lee JS, Lee IY, Park JH, Foley J, Chae JS. Tick-borne rickettsial pathogens in ticks and small mammals in Korea. Appl Environ Microbiol 2006; 72: 5766-5776. https://doi.org/10.1128/AEM.00431-06
  29. Kim TH, Choi EH, Lee MG, Ahn SK. Serologically diagnosed Lyme disease manifesting erythema migrans in Korea. J Korean Med Sci 1999; 14: 85-88.
  30. Kordick SK, Breitschwerdt EB, Hegarty BC, Southwick KL, Colitz CM, Hancock SI, Bradley JM, Rumbough R, Mcpherson JT, MacCormack JN. Coinfection with multiple tick-borne pathogens in a Walker Hound kennel in North Carolina. J Clin Microbiol 1999; 37: 2631-2638.
  31. Lee MG, Chung KY, Choi YS, Cho SN. Lyme diseases. Korean J Dermatol 1993; 31: 601-605
  32. Lee SH, Kim BJ, Kim JH, Park KH, Yeo SJ, Kim SJ, Kook YH. Characterization of Borrelia burgdorferi strains isolated from Korea by 16S rDNA sequence analysis and PCR-RFLP analysis of rrf (5S)-rrl (23S) intergenic spacer amplicons. Int J Syst Evol Microbiol 2000; 50 pt 2: 857-863. https://doi.org/10.1099/00207713-50-2-857
  33. Lissman BA, Bosler EM, Camay H, Ormiston BG, Benach JL. Spirochete associated arthritis (Lyme disease) in a dog. J Am Vet Med Assoc 1984; 185: 219-220.
  34. Maeda K, Markowitz N, Hawley RC, Ristic M, Cox D, McDade JE. Human infection with Ehrlichia canis, a leukocytic rickettsia. N Engl J Med 1987; 316: 853-856. https://doi.org/10.1056/NEJM198704023161406
  35. Murphy GL, Ewing SA, Whitworth LC, Fox JC, Kocan AA. A molecular and serologic survey of Ehrlichia canis, E. chaffeensis, and E. ewingii in dogs and ticks from Oklahoma. Vet Parasitol 1998; 79: 325-339 https://doi.org/10.1016/S0304-4017(98)00179-4
  36. Park JH, Heo EJ, Choi KS, Dumler JS, Chae JS. Detection of antibodies to Anaplasma phagocytophilum and Ehrlichia chaffeensis antigens in sera of Korean patients by western immunoblotting and indirect immunofluorescence assays. Clin Diagn Lab Immunol 2003; 10: 1059-1064
  37. Park KH, Chang WH, Schwan TG Identification and characterlzation of Lyme disease spirochetes, Borrelia burgdorferi sensu lato, isolated in Korea. J Clin Microbiol 1993; 31 1831-1837.
  38. Park KH, Lee SH, Won WJ, Jang WJ, Chang WH. Isolation of Borrelia burgdorferi, the causative agent of Lyme disease, from Ixodes ticks in Korea. J Korean Soc Microbiol 1992; 27: 307-312.
  39. Regnery RL, Spruill CL, Plikaytis BD. Genotypic identification of rickettsiae and estimation of intraspecies sequences divergence for portions of two rickettsial genes. J Bacteriol 1991; 173: 1576-1589.
  40. Rikihisa Y. Cross - reacting antigens between Neorickettsia helminthoeca and Ehrlichia species, shown by immunofluorescence and Western immunoblotting. J Clin Microbiol 1991; 29: 2024-2029.
  41. Ristic M, Dawson JE, Holland CJ. Susceptibility of dogs to infection with E. risticii, the causative agent of equine monocytic ehrlichiosis (Potomac horse fever). Am J Vet Res 1988; 49: 1497-1500
  42. Rodgers SJ, Morton RJ, Baldwin CA. A serological survey of Ehrlichia canis, Ehrlichia equi, Rickettsia rickettsii and Borrelia burgdorferi in dogs in Oklahoma. J Vet Diagn Invest 1989; 1: 154-159. https://doi.org/10.1177/104063878900100212
  43. Sachar DS. Ehrlichia chaffeensis infection in an active duty soldier stationed in Korea. Medical Surveillance Monthly Report 2000; 6(10): 9-11
  44. Schouls LM, Van De Pol I. Rijpkema SGT, Schot CS. Detection and identification of Ehrlichia, Borrelia burgdorferi sensu lato, and Bartonella species in Dutch Ixodes ricinus ticks. J Clin Microbiol 1999; 37: 2215-2222.
  45. Stockham SL, Schmidt DA, Curtis KS. Evaluation of granulocytic ehrlichiosis in dogs of Missouri, including serologic status to Ehrlichia canis, Ehrlichia equi, and Borrelia burgdorferi. Am J Vet Res 1992; 53: 63-68.
  46. Wang G, van Dam AP, Schwartz I. Molecular typing of Borrelia burgdorferi sensu lato: taxonomic, epidemiological, and clinical implications. Clin Microbiol Rev 1990; 12: 633-653.