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Publication Date: 2013-06-21
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Rojas Triviño, A. ., Rueda Hurtado, A. ., Díaz Molano, D. M. ., Mesa Cobo, N. C. ., Benavides Montaño, J. A. ., Imbachi López, K. ., Álvarez Ríos, L. ., & López Bermúdez, R. . (2013). Identification of Ehrlichia canis (Donatien & Lestoquard) Moshkovski using nested-PCR. Revista Veterinaria Y Zootecnia (On Line), 7(1), 37–48. Retrieved from https://revistasojs.ucaldas.edu.co/index.php/vetzootec/article/view/4403
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Abstract
ABSTRACT: Canine monocytic ehrlichiosis is a multisystem disease in which the symptoms are not very clear, and is caused by the rickettsia Ehrlichia canis, which is transmitted by the Rhipicephalus sanguineus tick. The presence of Ehrlichia canis in dogs found in animal home refuges located in the Department of Valle del Cauca, in the municipalities of Cali, Palmira, Buga, Ginebra, Caicedonia and Cartago was determined. E. canis molecular identification was carried out through nested-PCR from DNA extracted from blood samples of animals with known tick contact, and which was amplified using ECC-ECB choke pairs during the first reaction and ECAN5 and HE3 chokes for the second PCR reaction; these two last choke pairs are specific for the E. canis species. Samples were also randomly analyzed to determine the presence of E. canis through serologic test. Ehrlichia canis was found in all areas sampled in the Department of Valle del Cauca, registering the highest prevalence in the municipalities of Palmira and Cartago, with 92.8 and 90%, respectively. The medium prevalence was observed in Santiago de Cali with 68.75% and low prevalence places such as the municipalities of Caicedonia (10%), Ginebra (20%) and Buga (30%). This research represents the first report of canine monocytic ehrlichiosis diagnosed through nested-PCR in Valle del Cauca, Colombia.
References
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Beaufils, J.P. Ehrlichiosis: Clinical aspects in dogs and cats. International Forum on Ticks and Tick-Borne Disease, Compendium on Continuing Education for the Practicing Veterinarian, v.19, p.57-61, 1997.
Benavides, J.A.; Ramírez, G. Ehrlichiosis canina. Revista Colombiana de Ciencias Pecuarias, v.16, n.3, p.268-274, 2003.
Breitschwerdt, E.B. Ehrlichiosis: One or many diseases? A supplement to Compendium on Continuing Education for the Practicing Veterinarian, v.19, p.62-63, 1997.
Centre for Food Security and Public Health –CFSPH–. Ehrlichiosis and Anaplasmosis: Zoonotic Species. Iowa State University, 2013. Disponible en: http://www.cfsph.iastate.edu/Factsheets/pdfs/ehrlichiosis.pdf Accesado en: 01/08/2013.
Dawson, J.E.; Anderson, B.; Fishbein, D. et al. Isolation and characterization of an Ehrlichia sp. from a patient diagnosed with human ehrlichiosis. Journal of Clinical Microbiology, v.29, n.12, p.2741-2745, 1991.
Dawson, J.E.; Stallknecht, D.E.; Howerth, E.W. et al. Susceptibility of white-tailed deer (Odocoileus virginianus) to infection with Ehrlichia chaffeensis, the etiologic agent of human ehrlichiosis. Journal of Clinical Microbiology, v.32, n.11, p.2725-2728, 1994.
Dumler, J.S.; Barbet, A.F.; Bekker, C.P. et al. Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and ‘HGE agent’ as subjective synonyms of Ehrlichia phagocytophila. International Journal of Systematic and Evolutionary, v.51, p.2145- 2165, 2001.
Eddlestone, S.M.; Diniz, P.P.; Neer, T.M. et al. Doxycycline clearance of experimentally induced chronic Ehrlichia canis infection in dogs. Journal of Veterinary Internal Medicine, v.21, n.6, p.1237-1242, 2007.
Gutiérrez, C.N.; Martínez, M.; Sánchez, E. et al. Cultivation and molecular identification of Ehrlichia canis and Ehrlichia chaffeensis form a naturally con-infected dog in Venezuela. Veterinary Clinical Pathology, v.37, n.3, p.258-265, 2008.
Harrus, S.; Waner, T. Diagnosis of canine monocytotropic ehrlichiosis (Ehrlichia canis): an overview. Veterinary Journal, v.187, n.3, p.292- 296, 2011.
Hegarty, B.C.; De Paiva Diniz, P.P.; Bradley, J.M. et al. Clinical relevance of annual screening using a commercial enzyme-linked immunosorbent assay (SNAP 3Dx) for canine ehrlichiosis. Journal of the American Animal Hospital Association, v.45, n.3, p.118-124, 2009.
Hidalgo, M.; Vesga, J.F.; Lizarazo, D. et al. Short Report: A Survey of Antibodies against Rickettsia rickettsii and Ehrlichia chafeensis in Domestic Animals from a Rural Area of Colombia. The American Journal of Tropical Medicine and Hygiene, v.80, n.6, p.1029-1030, 2009.
Kelly, P.J.; Xu, C.; Lucas. H.; Loftis, A. et al. Ehrlichiosis, Babesiosis, Anaplasmosis and Hepatozoonosis in dogs from St. Kitts, West Indies. PLoS One, v.8, n.1, p.1-10, 2013.
Labruna, M.B.; Machado, R.Z. Agentes transmitidos por carrapatos na Regiao Neotropical. En: Barros-Batesti, D.M.; Arzua. M.; Bechara, G.H. (Eds.). Carrapatos de importância médico-veterinária da Região Neotropical: Um guia ilustrado para identificação de espécies. São Paulo: Vox/ICTTD/Butantan, 2006. p.155-162.
Lakshmanan, B.; John, L.; Gomathinayagam, S. et al. Molecular detection of Ehrlichia canis from blood of naturally infected dogs in India. Veterinarski arhiv, v.77, n.4, p.307-312, 2007.
Lanza-Perea, M.; Kumthekar-Sachin, M.; Sabarinath, A. et al. Doxycycline treatment of asymptomatic dogs seropositive for Ehrlichia canis. West Indian Veterinary Journal, v.9, n.2, p.11-13, 2009.
López, G. Bioecología y distribución de garrapatas en Colombia. En: Control de garrapatas. Medellín: Instituto Colombiano Agropecuario, Compendio No. 39, 1980. p.33-43.
McClure, J.C.; Crothers, M.L.; Schaefer, J.J. et al. Efficacy of a doxycycline treatment regimen initiated during three different phases of experimental ehrlichiosis. Antimicrobial Agents and Chemotherapy, v.54, n.12, p.5012-5020, 2010.
Mylonakis, M.E.; Koutinas, A.F.; Billinis, C. et al. Evaluation of cytology in the diagnosis of acute canine monocytic ehrlichiosis (Ehrlichia canis): a comparison between five methods. Veterinary Microbiology, v.91, n.2-3, p.197-204, 2003.
Murphy, L.S.; Ewing, L; Whitworth, J. et al. A molecular and serologic survey of Ehrlichia canis, E. chaffensis and E. ewingii in dogs and ticks from Oklahoma. Veterinary Parasitology, v.79, n.4, p.325-339, 1998.
Nakaghi, A.C.; Zacarias-Machado, R.; Tinucci Costa, M. et al. Canine ehrlichiosis: clinical, hematological, serological and molecular aspects. Ciência Rural, Santa Maria, v.38, n.3, p.766-770, 2008.
National Committee for Clinical Laboratory Standards –NCCLS–. Procedure for Determining Packed Cell Volume by the Microhematocrit Method; Approved. 3.ed. Standard. 2000. 7p.
Osorno-Mesa, E. Las garrapatas de la República de Colombia. Revista del Instituto Nacional de Salud Biomédica, v.26, n.3, p.317-36, 2006.
Paraná da Silva, B.M.; Leal, D.C.; Portela Medina Barboza, D.C. et al. Prevalence of ehrlichial infection among dogs and ticks in Northeastern Brazil. Revista Brasileira de Parasitologia Veterinária, v.19, n.2, p.89-93, 2010.
Parrado, M.; Vargas, F.; Hernández, G. et al. Asociación de los resultados de una prueba serológica (ELISA) y frotis sanguíneo en caninos con sintomatología compatible de Ehrlichiosis. Orinoquía, v.7, n.1-2, p.6-11, 2003.
Rar, V.; Golovljova, I. Anaplasma, Ehrlichia, and “Candidatus Neoehrlichia” bacteria: pathogenicity, biodiversity, and molecular genetic characteristics, a review. Infection, Genetics and Evolution, v.11, n.8, p.1842-1861, 2011.
Ríos, R.; Franco, S.; Mattar, S. et al. Seroprevalencia de Leptospira sp., Rickettsia sp. y Ehrlichia sp. en trabajadores rurales del departamento de Sucre, Colombia. Asociación Colombiana de Infectología, v.12, n.2, p.319-324, 2008.
Sambrook, J., Fritsch, E.F.; Maniatis, T. Molecular Cloning: A Laboratory manual. 2.ed. N.Y.: Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, 1989. 1659p.
Silva-Molano, R.F.; Sánchez-Ucrós, N.; Loaiza-Echeverri, A.M. Remove from marked records recurrence report of Ehrlichia canis in dog blood samples in Cali, Colombia. Veterinaria y Zootecnia, v.2, n.1, p.27-31, 2008.
Vargas-Hernández, G.; André, M.R.; Faria, J.L. et al. Molecular and serological detection of Ehrlichia canis and Babesia vogeli in dogs in Colombia. Veterinary Parasitology, v.186, p.254-260, 2012.
Voltzit, O.V. A review of Neotropical Amblyomma species (Acari: Ixodidae). Acarina, v.15, n.1, p.3-134, 2007.
Wallerstein, R.O. Laboratory evaluation of anemia. Western Journal of Medicine, v.146, n.4, p.443-451, 1987.
Wen, B.; Rikihisa, Y.; Mott, J.M. et al. Comparison of nested PCR with immunofluorescent-antibody assay for detection of Ehrlichia canis infection in dogs treated with doxycycline. Journal of Clinical Microbiology, v.35, n.7, p.1852-1855, 1997.
Woody, B.J.; Hoskins, J.D. Ehrlichial disease of dogs. The Veterinary Clinics of North America, Small Animal Practice, v.21, n.1, p.75-98, 1991.
Barros-Battesti, D.M.; Arzua, M.; Bechara, G.H. Carrapatos de importância médico-veterinária da Região Neotropical: Um guia ilustrado para identificação de espécies. 1.ed. São Paulo, Brasil: Editorial Vox, 2006. 223p.
Beaufils, J.P. Ehrlichiosis: Clinical aspects in dogs and cats. International Forum on Ticks and Tick-Borne Disease, Compendium on Continuing Education for the Practicing Veterinarian, v.19, p.57-61, 1997.
Benavides, J.A.; Ramírez, G. Ehrlichiosis canina. Revista Colombiana de Ciencias Pecuarias, v.16, n.3, p.268-274, 2003.
Breitschwerdt, E.B. Ehrlichiosis: One or many diseases? A supplement to Compendium on Continuing Education for the Practicing Veterinarian, v.19, p.62-63, 1997.
Centre for Food Security and Public Health –CFSPH–. Ehrlichiosis and Anaplasmosis: Zoonotic Species. Iowa State University, 2013. Disponible en: http://www.cfsph.iastate.edu/Factsheets/pdfs/ehrlichiosis.pdf Accesado en: 01/08/2013.
Dawson, J.E.; Anderson, B.; Fishbein, D. et al. Isolation and characterization of an Ehrlichia sp. from a patient diagnosed with human ehrlichiosis. Journal of Clinical Microbiology, v.29, n.12, p.2741-2745, 1991.
Dawson, J.E.; Stallknecht, D.E.; Howerth, E.W. et al. Susceptibility of white-tailed deer (Odocoileus virginianus) to infection with Ehrlichia chaffeensis, the etiologic agent of human ehrlichiosis. Journal of Clinical Microbiology, v.32, n.11, p.2725-2728, 1994.
Dumler, J.S.; Barbet, A.F.; Bekker, C.P. et al. Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and ‘HGE agent’ as subjective synonyms of Ehrlichia phagocytophila. International Journal of Systematic and Evolutionary, v.51, p.2145- 2165, 2001.
Eddlestone, S.M.; Diniz, P.P.; Neer, T.M. et al. Doxycycline clearance of experimentally induced chronic Ehrlichia canis infection in dogs. Journal of Veterinary Internal Medicine, v.21, n.6, p.1237-1242, 2007.
Gutiérrez, C.N.; Martínez, M.; Sánchez, E. et al. Cultivation and molecular identification of Ehrlichia canis and Ehrlichia chaffeensis form a naturally con-infected dog in Venezuela. Veterinary Clinical Pathology, v.37, n.3, p.258-265, 2008.
Harrus, S.; Waner, T. Diagnosis of canine monocytotropic ehrlichiosis (Ehrlichia canis): an overview. Veterinary Journal, v.187, n.3, p.292- 296, 2011.
Hegarty, B.C.; De Paiva Diniz, P.P.; Bradley, J.M. et al. Clinical relevance of annual screening using a commercial enzyme-linked immunosorbent assay (SNAP 3Dx) for canine ehrlichiosis. Journal of the American Animal Hospital Association, v.45, n.3, p.118-124, 2009.
Hidalgo, M.; Vesga, J.F.; Lizarazo, D. et al. Short Report: A Survey of Antibodies against Rickettsia rickettsii and Ehrlichia chafeensis in Domestic Animals from a Rural Area of Colombia. The American Journal of Tropical Medicine and Hygiene, v.80, n.6, p.1029-1030, 2009.
Kelly, P.J.; Xu, C.; Lucas. H.; Loftis, A. et al. Ehrlichiosis, Babesiosis, Anaplasmosis and Hepatozoonosis in dogs from St. Kitts, West Indies. PLoS One, v.8, n.1, p.1-10, 2013.
Labruna, M.B.; Machado, R.Z. Agentes transmitidos por carrapatos na Regiao Neotropical. En: Barros-Batesti, D.M.; Arzua. M.; Bechara, G.H. (Eds.). Carrapatos de importância médico-veterinária da Região Neotropical: Um guia ilustrado para identificação de espécies. São Paulo: Vox/ICTTD/Butantan, 2006. p.155-162.
Lakshmanan, B.; John, L.; Gomathinayagam, S. et al. Molecular detection of Ehrlichia canis from blood of naturally infected dogs in India. Veterinarski arhiv, v.77, n.4, p.307-312, 2007.
Lanza-Perea, M.; Kumthekar-Sachin, M.; Sabarinath, A. et al. Doxycycline treatment of asymptomatic dogs seropositive for Ehrlichia canis. West Indian Veterinary Journal, v.9, n.2, p.11-13, 2009.
López, G. Bioecología y distribución de garrapatas en Colombia. En: Control de garrapatas. Medellín: Instituto Colombiano Agropecuario, Compendio No. 39, 1980. p.33-43.
McClure, J.C.; Crothers, M.L.; Schaefer, J.J. et al. Efficacy of a doxycycline treatment regimen initiated during three different phases of experimental ehrlichiosis. Antimicrobial Agents and Chemotherapy, v.54, n.12, p.5012-5020, 2010.
Mylonakis, M.E.; Koutinas, A.F.; Billinis, C. et al. Evaluation of cytology in the diagnosis of acute canine monocytic ehrlichiosis (Ehrlichia canis): a comparison between five methods. Veterinary Microbiology, v.91, n.2-3, p.197-204, 2003.
Murphy, L.S.; Ewing, L; Whitworth, J. et al. A molecular and serologic survey of Ehrlichia canis, E. chaffensis and E. ewingii in dogs and ticks from Oklahoma. Veterinary Parasitology, v.79, n.4, p.325-339, 1998.
Nakaghi, A.C.; Zacarias-Machado, R.; Tinucci Costa, M. et al. Canine ehrlichiosis: clinical, hematological, serological and molecular aspects. Ciência Rural, Santa Maria, v.38, n.3, p.766-770, 2008.
National Committee for Clinical Laboratory Standards –NCCLS–. Procedure for Determining Packed Cell Volume by the Microhematocrit Method; Approved. 3.ed. Standard. 2000. 7p.
Osorno-Mesa, E. Las garrapatas de la República de Colombia. Revista del Instituto Nacional de Salud Biomédica, v.26, n.3, p.317-36, 2006.
Paraná da Silva, B.M.; Leal, D.C.; Portela Medina Barboza, D.C. et al. Prevalence of ehrlichial infection among dogs and ticks in Northeastern Brazil. Revista Brasileira de Parasitologia Veterinária, v.19, n.2, p.89-93, 2010.
Parrado, M.; Vargas, F.; Hernández, G. et al. Asociación de los resultados de una prueba serológica (ELISA) y frotis sanguíneo en caninos con sintomatología compatible de Ehrlichiosis. Orinoquía, v.7, n.1-2, p.6-11, 2003.
Rar, V.; Golovljova, I. Anaplasma, Ehrlichia, and “Candidatus Neoehrlichia” bacteria: pathogenicity, biodiversity, and molecular genetic characteristics, a review. Infection, Genetics and Evolution, v.11, n.8, p.1842-1861, 2011.
Ríos, R.; Franco, S.; Mattar, S. et al. Seroprevalencia de Leptospira sp., Rickettsia sp. y Ehrlichia sp. en trabajadores rurales del departamento de Sucre, Colombia. Asociación Colombiana de Infectología, v.12, n.2, p.319-324, 2008.
Sambrook, J., Fritsch, E.F.; Maniatis, T. Molecular Cloning: A Laboratory manual. 2.ed. N.Y.: Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, 1989. 1659p.
Silva-Molano, R.F.; Sánchez-Ucrós, N.; Loaiza-Echeverri, A.M. Remove from marked records recurrence report of Ehrlichia canis in dog blood samples in Cali, Colombia. Veterinaria y Zootecnia, v.2, n.1, p.27-31, 2008.
Vargas-Hernández, G.; André, M.R.; Faria, J.L. et al. Molecular and serological detection of Ehrlichia canis and Babesia vogeli in dogs in Colombia. Veterinary Parasitology, v.186, p.254-260, 2012.
Voltzit, O.V. A review of Neotropical Amblyomma species (Acari: Ixodidae). Acarina, v.15, n.1, p.3-134, 2007.
Wallerstein, R.O. Laboratory evaluation of anemia. Western Journal of Medicine, v.146, n.4, p.443-451, 1987.
Wen, B.; Rikihisa, Y.; Mott, J.M. et al. Comparison of nested PCR with immunofluorescent-antibody assay for detection of Ehrlichia canis infection in dogs treated with doxycycline. Journal of Clinical Microbiology, v.35, n.7, p.1852-1855, 1997.
Woody, B.J.; Hoskins, J.D. Ehrlichial disease of dogs. The Veterinary Clinics of North America, Small Animal Practice, v.21, n.1, p.75-98, 1991.
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