Diagnosis of canine monocytotropic ehrlichiosis: an update review
Abstract
Canine monocytic ehrlichiosis (CME) is caused by the infection with a rickettsial organism, Ehrlichia canis. This organism is replicated only in the cytoplasm of blood monocytes and tissue macrophages. CME has been reported throughout the world but at higher frequencies in tropical and subtropical regions. CME is often under-recognized since the disease is difficult to diagnose due to variable clinical signs, its different stages, and frequent co—infection with other tick-borne pathogens or other infectious agents. Clinical manifestations and the traditional diagnostic techniques such as hematology and demonstration of typical cytoplasmic E. canis-morulae in monocytes in blood smears have been used for the diagnosis of CME; however, the PCR—based diagnostic tests and serology methods based on specific antigens, are the most sensitive and specific for this disease. This article reviews the current literature covering the diagnosis of infection by E. canis in dogs.
Keywords
ehrlichia canis, dogs diseases, thrombocytopenia, molecular diagnostic techniques, Inmuneadsortion Enzimatic test.
References
- Garrity GM, Bell JA, Lilburn TG. Taxonomic outline of the prokaryotes Bergey´s manual of systematic bacteriology. Second edition, New York, Springer, 2004.
- Harrus S, Warner T. Diagnosis of canine monocytotropic ehrlichiosis (Ehrlichia canis): An overview. Vet J 2011; 187, 292-296.
- Harrus S, Waner T, Bark H, Jongejan F, Corneliensse AW. Recent advances in determining the pathogenesis of canine monocytic ehrlichiosis. J Clin Microbiol 1997; 37: 2745-2749.
- Harrus S. Perspectives on the pathogenesis and treatmentof canine monocytic ehrlichiosis (Ehrlichia canis). Vet J 2015; 204(3): 239-240.
- Sainz Á, Roura X, Miró G, Estrada-Peña A, Kohn B, Harrus S et al. Guideline for veterinary practitioners on canine ehrlichiosis and anaplasmosis in Europe. Parasit Vectors 2015; 8: 75.
- René-Martellet M, Lebert I, Chêne J, Massot R, Leon M, Leal A. et al. Diagnosis and incidence risk of clinical canine monocytic ehrlichiosis under field conditions in Southern Europe. Parasit Vectors 2015; 8: 3.
- Rodríguez-Vivas RI, Albornoz REF, Bolio GME. Ehrlichia canis in dogs in Yucatan, México: seroprevalence, prevalence of infection and associated factors. Vet Parasitol 2005; 127(1): 75-79.
- Maazi N; Malmasi A, Shayan P, Nassiri SM, Salehi TZ, Fard MS. Molecular and serological detection of Ehrlichia canis in naturally exposed dogs in Iran: an analysis on associated risk factors. Rev Bras Parasitol, 2014; 23(1): 16-22.
- Allison RW, Little SE. Diagnosis of rickettsial diseases in dogs and cats. Vet Clin Pathol 2013; 42(2): 127-144.
- Mylonakis ME, Koutinas AF, Breitschwerdt EB, Hegarty BC, Billinis CD, Leontides LS et al. Chronic canine ehrlichiosis (Ehrlichia canis): a retrospective study of 19 natural cases. J Am Anim Hosp Assoc, 2004; 40: 174-184.
- Mavromatis K, Doyle CK, Lykidis A, Ivanova N, Francino MP, Chain P et al. The genome of the obligately intracellular bacterium Ehrlichia canis reveals themes of complex membrane structure and immune evasion strategies. J Bacteriol, 2006; 188: 4015-4023.
- Singu V, Peddireddi L, Sirigireddy KR, Cheng C, Munderloh U, Ganta RR. Unique macrophage and tick cell-specific protein expression from the p28/p30-outer membrane protein multigene locus in Ehrlichia chaffeensis and Ehrlichia canis. Cell Microbiol, 2006; 8: 1475-1487.
- McBride JW, Walker DH. Molecular and cellular pathobiology of Ehrlichia infection: Targets for new therapeutics and immunomodulation strategies. Exp Rev Mol Med, 2011; 13: 1-19.
- Alves RN, Levenhagen MA, Levenhagen MM, Rieck SE, Labruna MB, Beletti ME. The spreading process of Ehrlichia canis in macrophages is dependent on actin cytoskeleton, calcium and iron influx and lysosomal evasion. Vet Microbiol, 2014; 168: 442-446.
- Harrus S. Perspectives on the pathogenesis and treatment of canine monocytic ehrlichiosis (Ehrlichia canis). Vet J, 2015; 204(3): 239-240.
- Cardenas AM, Doyle CK, Zhang X, Nethery K, Corstvet RE, Walker et al. Enzyme-linked immunosorbent assay with conserved immunoreactive glycoproteins gp36 and gp19 has enhanced sensitivity and provides species-specific immunodiagnosis of Ehrlichia canis infection. Clin Vaccine Immunol, 2007; 14: 123-128.
- Skotarczak B. 2003. Canine Ehrlichiosis. Ann Agric Environ Med, 2003; 10: 137-141.
- Bulla C, Takahira RK, Araújo JP, Trinca LA, Lopes RS, Wiedmeyer CE. The relationship between the degree of thrombocytopenia and infection with Ehrlichia canis in an endemic area. Vet Res, 2004; 35: 141-146.
- Komnenou AA, Mylonakis ME, Kouti V, Tendoma L, Leontides L, Skountzou E et al. Ocular manifestations of natural canine monocytic ehrlichiosis (Ehrlichia canis): a retrospective study of 90 cases. Vet Ophthalmol, 2007; 10: 137-142.
- Mark Neer T. Ehrlichiosis. En: Greene, Enfermedades infecciosas en perros y gatos. Capítulo 28. Segunda Edición, Ed. McGraw-Hill Interamericana. Philadelphia, USA 2000; pp. 153-161.
- De Castro MB, Machado RZ, de Aquino LP, Alessi AC, Costa MT. Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings. Vet Parasitol, 2004; 119: 73-86.
- Shimada T, Ishida Y, Shimizu M, Nomura M, Kawato K, Iguchi K, Jinbo T. Monitoring C-reactive protein in beagle dogs experimentally inoculated with Ehrlichia canis. Vet Res Commun, 2002; 26: 171-177.
- Gyles CL, Prescott JF, Songer JO, Thoen CO. Pathogenesis of bacterial infections in animals. Chapter 31. Third Edition. Iowa, USA 2004; pp. 429-431.
- Brandão LP, Hasegawa MY., Hagiwara MK., Kohayagawa A. Platelet aggregation studies in acute experimental canine ehrlichiosis. Vet Clin Pathol, 2006; 35(1): 78-81.
- Waner T, Harrus S. Canine monocytic ehrlichiosis from pathology to clinical manifestations. Isr J Vet Med, 2013; 68(1): 12-18.
- Davoust B, Parzy D, Demoncheaux JP, Tine R, Diarra M, Marié JL et al. Usefulness of a rapid immuno-migration test for the detection of canine monocytic ehrlichiosis in Africa. Comp Immunol Microbiol Infect Dis, 2014; 37(1): 31-37.
- Waner T, Harrus S, BarkH, Bogin E, Avidar Y, Keysary A. Characterization of the subclinical phase of canine ehrlichiosis in experimentally infected beagle dogs. Vet Parasitol, 1997; 69: 307-317.
- Bowman DD. Georgis’ Parasitology for veterinarians. Ninth Edition. Chapter 5. Saunders Elsevier NY USA; 2009. pp. 244-246.
- McBride JW, Corstvet RE, Gaunt SD, Boudreaux C, Guedry T, Walker DH. Kinetics of antibody response to Ehrlichia canis immunoreactive proteins. Infect Immun, 2003; 71: 2516-2524.
- Waner T, Harrus S, Jongejan F, Bark H, Keysary A, Cornelissen AW. Significance of serological testing for ehrlichial diseases in dogs with special emphasis on the diagnostic of canine monocytic ehrlichiosis caused by Ehrlichia canis. Vet Parasitol, 2001; 95: 1-15.
- Harrus S, Alleman AR, Bark H, Mahan SM, Waner T. Comparison of three antibody test for the diagnosis of canine infection with Ehrlichia canis. Vet Microbiol, 2002; 8: 361-368.
- Rikihisa Y. Cross-reacting antigens between Neorickettsia helminthoeca and Ehrlichia species, shown by immunofluorescence and Western immunoblotting. J Clin Microbiol, 1991; 29: 2024-2029.
- Krawczak Fda S, Reis IA, Silveira JA, Avelar DM, Marcelino AP, Werneck GL et al. Leishmania, Babesia and Ehrlichia in urban pet dogs: co-infection or cross-reaction in serological methods? Rev Soc Bras Med Trop, 2015; 48(1): 64-68.
- Waner T, Strenger C, Keysary A, Harrus S. Kinetics of serologic crossreactions between Ehrlichia canis and the Ehrlichia phagocytophila genogroups in experimental E. canis infection in dogs. Vet Immunol Immunop, 1998; 66: 237-243.
- Gusa AA, Buller RS, Storch GA, Huycke MM, Machado LJ, Slater LN et al. Identification of a p28 gene in Ehrlichia ewingii: evaluation of gene for use as a target for a species-specific PCR diagnostic assay. J Clin Microbiol, 2001; 39: 3871-3876.
- O’Connor TP, Saucier JM, Daniluk D, Stillman BA, Krah R, Rikihisa Y et al. Evaluation of peptide- and recombinant protein-based assays for detection of anti-Ehrlichia ewingii antibodies in experimentally and naturally infected dogs. Am J Vet Res, 2010; 71: 1195-1200.
- Sumner JW, Childs JE, Paddock CD. Molecular cloning and characterization of the Ehrlichia chaffeensis variable-length PCR target: an antigen-expressing gene that exhibits interstrain variation. J Clin Microbiol, 1999; 37: 1447-1453.
- Beall MJ, Alleman RA, Breitschwerdt EB, Cohn LA, Couto CG, Dryden MW et al. Seroprevalence of Ehrlichia canis, Ehrlichia chaffeensis and Ehrlichia ewingii in dogs in North America. Parasit Vectors, 2012; 5: 29.
- Qurollo BA, Chandrashekar R, Hegarty BC, Beall MJ, Stillman B, Liu J et al. A serological survey of tick-borne pathogens in dogs in North America and the Caribbean as assessed by Anaplasma phagocytophilum, A. platys, Ehrlichia canis, E. chaffeensis, E. ewingii, and Borrelia burgdorferi species-specific peptides. Infect Ecol Epidemiol, 2014; 4: 24699.
- Starkey LA, Barrett AW, Chandrashekar R, Stillman BA, Tyrrell P, Thatcher B et al. Development of antibodies to and PCR detection of Ehrlichia spp. in dogs following natural tick exposure. Vet Microbiol, 2014; 173: 379-384.
- Mylonakis ME, Koutinas AF, Billinis C, Leontides LS, Kontos V, Papadopoulos O et al. Evaluation of cytology in the diagnosis of acute canine monocytic ehrlichiosis (Ehrlichia canis): a comparison between five methods. Vet Microbiol, 2003; 91(2-3): 197-204.
- Mylonakis ME, Borjesson DL, Leontides L, Siarkou VI, Theodorou K, Koutinas AF. Cytologic patterns of lymphadenopathy in canine monocytic ehrlichiosis. Vet Clin Pathol, 2011; 40(1): 78-83.
- Alves RN, Rieck SE, Ueira-Vieira C, Labruna MB, Beletti ME. Isolation, in vitro propagation, genetic analysis, and immunogenic characterization of an Ehrlichia canis strain from southeastern Brazil. J Vet Scien, 2014a; 15(2): 241- 248.
- Tesouro DMA, Sainz R E. Etiología y transmisión. En: Ehrlichiosis canis et felis. Aula Veterinaria. 2001. Madrid, España. 51: 1.
- Aguiar DM, Hagiwara MK, Labruna MB. In vitro isolation and molecular characterization of an Ehrlichia canis strain from São Paulo, Brazil. Braz J Microbiol, 2008; 39: 489-493.
- Dawson JE, Candal FJ, George VG, Ades EW. Human endothelial cells as an alternative to DH82 cells for isolation of Ehrlichia chaffeensis, E. canis, and Rickettsia rickettsii. Pathobiol, 1993; 61: 293-296.
- Rivas LV, Morales AD, Saenz M, Bonilla JL. Hallazgo de Ehrlichiosis canina causada por E. canis en una Comunidad del municipio de León, Nicaragua. RedVet, 2010; 11(3): 1-10.
- 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 Chemoth, 2004; 48: 4488-4490.
- Harrus S, Waner T, Bark H, Jongejan F, Cornelissen AW. Recent advances in determining the pathogenesis of canine monocytic ehrlichiosis. J Clin Microbiol, 1999; 37(9): 2745-2749.
- Gal A, Loeb E, Yisaschar-Mekuzas Y, Baneth G. Detection of Ehrlichia canis by PCR in different tissues obtained during necropsy from dogs surveyed for naturally occurring canine monocytic ehrlichiosis. Vet J, 2008; 175(2): 212-217.
- Baneth G, Harrus S, Ohnona FS, Schlesinger Y. Longitudinal quantification of Ehrlichia canis in experimental infection with comparison to natural infection. Vet Microbiol, 2009; 136: 321-325.
- Aguirre E, Sainz A, Dunner S, Amusategui I, López L, Rodríguez- Franco F et al. First isolation and molecular characterization of Ehrlichia canis in Spain. Vet Parasitol, 2004; 125: 365-372.
- Kocan A, Crowder Levesque G, Whitworth LC, Murphy GL, Ewing SA, Barker RW. Naturally ocurring Ehrlichia chaffeensis infection in coyotes from Oklahoma. Emerg Infect Dis, 2000; 6: 477-480.
- Gutiérrez GCN, Martínez AMC, Triana-Alonso FJ. Identificación microscópica y molecular de ehrlichias en perros del estado Aragua-Venezuela. Salus, 2008; 12: 197-204.
- NakaghiI ACH, Machado RZ, Costa MT, André MR, Baldani CD. Canine ehrlichiosis: clinical, hematological, serological and molecular aspects. Cienc Rural, 2008; 38(3):766-770.
- Rojas-Triviño A, Rueda-Hurtado A, Díaz-Molano DM, Mesa-Cobo NC, Benavides-Montaño JA, Imbachi-López K et al. Identificación de Ehrlichia canis (Donatien & Lestoquard) Moshkovski mediante PCR anidada. Vet Zootec, 2013; 7: 37-48.
- Harrus S, Waner T, Aizenberg I, Foley JE, Poland AM, Bark H. Amplification of ehrlichial DNA from dogs 34 months after infection with Ehrlichia canis. J Clin Microbiol, 1998; 36(1): 73-76.
- Pat-Nah H, Rodríguez-Vivas RI, Bolio-Gonzalez ME, Villegas-Perez SL, Reyes-Novelo N. Molecular Diagnosis of Ehrlichia canis in dogs and ticks Rhipicephalus sanguineus (Acari: Ixodidae) in Yucatan, Mexico. J Med Entomol, 2015; 52(1): 101-104.
- Peleg O, Baneth G, Eyal O, Inbar J, Harrus S. Use of chimeric DNA–RNA primers in quantitative PCR for detection of Ehrlichia canis and Babesia canis. Appl Environ Microbiol, 2009; 75: 6393-6398.
- Peleg O, Baneth G, Eyal O, Inbar J, Harrus S. Multiplex real-time qPCR for the detection of Ehrlichia canis and Babesia canis vogeli. Vet Parasitol, 2010; 173: 292-299.
- Hsieh YC, Lee CC, Tsang CL, Chung YT. Detection and characterization of four novel genotypes of Ehrlichia canis from dogs. Vet Microbiol, 2010; 146: 70-75.
- Kamani J, Lee CC, Haruna AM, Chung PJ, Weka PR, Chung YT. First detection and molecular characterization of Ehrlichia canis from dogs in Nigeria. Res Vet Sci, 2013; 94: 27-32.
- Zweygarth E, Cabezas-Cruz A, Josemans AI, Oosthuizen MC, Matjila PT, Lis K et al. In vitro culture and structural differences in the major immunoreactive protein gp36 of geographically distant Ehrlichia canis isolates. Ticks Tick Borne Dis, 2013; 5(4): 423-431.
- Faria JL, Dagnone AS, Munhoz TD, Joao CF, Pereira WA, Machado RZ et al. Ehrlichia canis morulae and DNA detection in whole blood and spleen aspiration samples. Rev Bras Parasitol Vet, 2010; 19: 98-102.
- Muangchuen A, Chaumpluk P, Suriyasomboon A, Ekgasit S. Colorimetric detection of Ehrlichia canis via nucleic acid hybridization in gold nano-colloids. Sensors, 2014; 14: 14472-14487.
- Jiménez-Coello M, Pérez-Osorio C, Vado-Solís I, Rodríguez-Buenfil JC, Ortega-Pacheco A. Serological survey of Ehrlichia canis in stray dogs from Yucatán, México, using two different diagnostic tests. VBZD, 2009; 9(2): 209-211.
- Munderloh UG, Liu Y, Wang M, Chen C, Kurtti TJ. Establishment, maintenance and description of cell lines from the tick Ixodes scapularis. J Parasitol, 1994; 80: 533-543.
- De Farias RTE, Araújo AHK, da Silva KF, Lira SV, Fernandes VI, Labruna MB et al. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil. Braz J Vet Parasitol, 2015; 24(1): 52-58.
- Wei L, Kelly P, Ackerson K, El-Mahallawy HS, Kaltenboeck B, Wang C. Molecular detection of Dirofilaria immitis, Hepatozoon canis, Babesia spp., Anaplasma platys and Ehrlichia canis in dogs on Costa Rica. Acta Parasitologica, 2014; 60(1): 21-25.