Comparative diagnostic methods for canine ehrlichiosis

The efficacy of blood and buffy coat smear examination, commercially available dot-enzyme-linked immunosorbent assay (ELISA) kit, and nested polymerase chain reaction (PCR) was evaluated in the diagnosis of canine ehrlichiosis in 40 dogs exhibiting symptoms suggestive of the disease. The intracytoplasmic rickettsial organisms were seen in the smears of only 1 dog (2.5%) with an extremely low level of parasitemia. Nested PCR with 16S rRNA revealed characteristic 378 bp bands indicating positive reaction in 8 (20.0%) dogs and sera samples of 21 (52.5%) dogs revealed anti-Ehrlichia antibodies. The dog that revealed organisms in the blood also showed positive results with the other 2 methods. Similarly all the sera samples of dogs with nPCR positive results were also positive for anti-Ehrlichia antibodies. The 8 dogs that exhibited positive results by nPCR and dot-ELISA were studied for clinical, hematological, and biochemical characterization of the disease. Fever, depression, anorexia, anemia, splenomegaly, and bleeding tendency were the common symptoms associated with the disease. Thrombocytopenia was the most consistent hematological finding, followed by anemia. Biochemical analysis revealed hyperglobulinemia with a reverse albumin:globulin ratio in 3 cases.

Comparative diagnostic methods for canine ehrlichiosis

The efficacy of blood and buffy coat smear examination, commercially available dot-enzyme-linked immunosorbent assay (ELISA) kit, and nested polymerase chain reaction (PCR) was evaluated in the diagnosis of canine ehrlichiosis in 40 dogs exhibiting symptoms suggestive of the disease. The intracytoplasmic rickettsial organisms were seen in the smears of only 1 dog (2.5%) with an extremely low level of parasitemia. Nested PCR with 16S rRNA revealed characteristic 378 bp bands indicating positive reaction in 8 (20.0%) dogs and sera samples of 21 (52.5%) dogs revealed anti-Ehrlichia antibodies. The dog that revealed organisms in the blood also showed positive results with the other 2 methods. Similarly all the sera samples of dogs with nPCR positive results were also positive for anti-Ehrlichia antibodies. The 8 dogs that exhibited positive results by nPCR and dot-ELISA were studied for clinical, hematological, and biochemical characterization of the disease. Fever, depression, anorexia, anemia, splenomegaly, and bleeding tendency were the common symptoms associated with the disease. Thrombocytopenia was the most consistent hematological finding, followed by anemia. Biochemical analysis revealed hyperglobulinemia with a reverse albumin:globulin ratio in 3 cases.

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  • values (Avg.) 6–18 12–30 60–77 3–10 2–10 2–5 Waner, T., Harrus, S.: Canine monocytic ehrlichiosis, in recent advances in canine infectious disease. In: Carichael, L.E., Ed. International Veterinary Information Service (cited from www. Ivis.org). 2000.
  • Varshney, J.P., Kumar, A., Hoque, M.: Ehrlichia platys and Babesia canis infection in a dog: a clinical report. J. Vet. Parasitol. 2004; 18: 35–37.
  • Banerjee, P.S., Mylonakis, M.E., Garg, R., Vatsya, S., Yadav, C.L.: Concurrent hepatozoonosis and granulocytic ehrlichiosis in a dog. J. Vet. Parasitol., 2008; 22: 9–11.
  • Harrus, S., Waner, T., Keysary, I., Aroch, I., Voet, H., Bark, H.: Investigation of splenic functions in canine monocytic ehrlichiosis. Vet. Immunol. Immunopathol., 1998; 62: 15–27.
  • Waner, T., Keysary, A., Bark, H., Sharabani, E., Harrus, S.: Canine monocytic ehrlichiosis – an overview. Israel J. Vet. Med., 1999; 54: 103–107.
  • Iqbal, Z., Chaichanasiriwithaya, W., Rikihisa, Y.: Comparison of PCR with other tests for early diagnosis of canine ehrlichiosis. J. Clinic. Microbiol., 1994; 32: 1658–1662.
  • Harrus, S., Kenny, M., Miara, L., Aizenberg, I., Waner, T., Shaw, S.: Comparison of simultaneous splenic sample PCR with blood sample PCR for diagnosis and treatment of experimental Ehrlichia canis infection. Antimicrob. Agents Chemother., 2004; 48: 4488–4490.
  • Rathore, V.S., Sengar, Y.S.: Diagnostic Parasitology. 1st edn., Pointer Publishers, Jaipur, India. 2005; 72–82.
  • Stich, R.W., Rikihisa, Y., Ewing, S.A., Needham, G.R., Grover, D.L., Jittapalapong, S.: Detection of Ehrlichia canis in canine carrier blood and in individual experimentally infected ticks with a p30-based PCR assay. J. Clinic. Microbiol., 2002; 40: 540–546.
  • Lakshmanan, B.: Diagnosis of canine ehrlichiosis using polymerase chain reaction. MVSc Thesis. The Tamil Nadu Veterinary and Animal Sciences University, Chennai, India. 200 Benjamin, M.M.: Outline of Veterinary Clinical Pathology. 3rd edn., Kalyani Publisher, New Delhi. 1985; 49–98.
  • Bool, P.H., Sutmoller, P.: Ehrlichia canis infection in dogs on Aruba (Netherlands Antilles). J. Am. Vet. Med. Assoc., 1957; 130: 418–421.
  • Philip, C.B.: Ehrlichia In: Breed, R.S., Murphy, E.G.D., Smith, N.R., Eds. Bergey’s Manual of Determinative Bacteriology. 7th edn., Williams and Wilkins, Baltimore, Maryland. 1957; 948–950.
  • McDade, J.E.: Ehrlichiosis – a disease of animals and humans. J. Infect. Dis., 1990; 161: 609–617.
  • Woody, B.J., Hoskins, J.D.: Ehrlichial disease of dogs. Vet. Clinic. N. Am. Small Anim. Pract., 1991; 21: 75–98.
  • Juyal, P.D., Kalra, I.S., Singhla, L.D.: Prevalence of haemoprotozoan in the domestic animals in Punjab. Sixth National Congress of Veterinary Parasitology, Abst. 1994; 4.
  • Thriunavukkarasu, P.S., Dhanapalan, P., Gnanaprakasam, V.: Incidence of canine ehrlichiosis in Madras city. Cherion., 1993; 22: 222–224.
  • Nakaghi, A.C.H., Machado, R.Z., Costa, M.T., André, M.R., Baldani, C.D.: Canine ehrlichiosis: clinical, hematological, serological and molecular aspects. Ciência Rural, 2008; 38: 766–770.
  • Katyal, D.: Studies on the prevalence of canine blood parasitic infections, with special reference to diagnosis and chemotherapy of Ehrlichia canis. MVSc Thesis. Konkan Krishi Vidyapeeth, Dapoli, India. 2000.
  • Mallapur, S.S.: Studies on ehrlichiosis in dogs of Mumbai. MVSc Thesis. The Maharashtra Animal and Fishery Sciences University, Nagpur. India. 2002.
  • Samaradhni, D., Maske, D.K., Kolte, S.W., Shinde, P.N.: Ehrlichiosis in dogs in Nagpur. J. Vet. Parasitol., 2003; 17: 165–166.
  • Huxsoll, D.L., Hildebrandt, P.K., Nims, R.M., Amyx, H.L., Ferguson, J.A.: Epizootiology of tropical canine pancytopenia. J. Wildlife Dis., 1970; 6: 220–225.
  • Thriunavukkarasu, P.S., Nambi, A.P., Rajan, T.S.S., Gnanaprakasam, V.: Clinical and haematological findings in canine ehrlichiosis in Madras City. Ind. Vet. J., 1994; 71: 825– 8
  • Ashuma, P.K., Juyal, P.D.: Canine ehrlichiosis: an emerging silent tick-borne disease of dogs. Intas. Polivet., 2005; 6: 346– 3
  • McBride, J.W., Corstvet, R.E., Breitschwerdt, E.B., Walker, D.H.: Immunodiagnosis of Ehrlichia canis infection with recombinant proteins. J. Clinic. Microbiol., 2001; 39: 315–322.
  • Waner, T., Harrus, S., Jongejan, F., Keysary, A., Cornelissen, A.W.: Significance of serological testing for ehrlichial diseases in dogs with special emphasis on the diagnosis of canine monocytic ehrlichiosis caused by Ehrlichia canis. Vet. Parasitol., 2001; 95: 1–15.
  • McBride, J.W., Corstvet, R.E., Gaunt, S.D., Boudreaux, C., Guedry, T., Walker, D.H.: Kinetics of antibody response to Ehrlichia canis immunoreactive proteins. Infection Immun., 2003; 71: 2516–2524.
  • McBride, J. W., Corstvet, R.E., Gaunt, S.D., Chinsangaram, J., Akita, G.Y., Osburn, B.I.: PCR detection of acute Ehrlichia canis infection in dogs. J. Vet. Diagnos. Investigat., 1996; 8: 441–447.
  • Inokuma, H., Raoult, D., Brouqui, P.: Detection of Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) in Okinawa Island, Japan. J. Clinic. Microbiol., 2000; 38: 4219– 42
  • Unver, A., Perez, M., Orellana, N., Huang, H., Rikihisa, Y.: Molecular and antigenic comparison of Ehrlichia canis isolates from dogs, ticks, and a human in Venezuela. J. Clinic. Microbiol. 2001; 39: 2788–2793.
  • Smith, R.D., Ristic, M., Huxsoll, D.L., Baylor, R.A.: Platelet kinetics in canine ehrlichiosis: evidence for increased platelet destruction as the cause of thrombocytopenia. Infection Immun., 1975; 11: 1216–1221.
  • Waner, T.: Hematopathological changes in dogs infected with Ehrlichia canis. Israel Vet. Med. J., 2008; 63: 1–8.
Turkish Journal of Veterinary and Animal Sciences-Cover
  • ISSN: 1300-0128
  • Yayın Aralığı: Yılda 6 Sayı
  • Yayıncı: TÜBİTAK