Molecular Characterization of Tandem Repeat Protein 36 Gene of Ehrlichia canis Detected in Naturally Infected Dogs from Peru

Joseph Geiger Graduate College of Biomedical Sciences, Western University of Health Sciences, Pomona, California;

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Bridget A. Morton College of Veterinary Medicine, Western University of Health Sciences, Pomona, California;
Affiliated Veterinary Specialists, Orange Park, Florida;

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Elton Jose Rosas Vasconcelos College of Veterinary Medicine, Western University of Health Sciences, Pomona, California;

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Maryam Tngrian College of Agriculture, California State Polytechnic University, Pomona, Pomona, California;

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Malika Kachani College of Veterinary Medicine, Western University of Health Sciences, Pomona, California;

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Eduardo A. Barrón Laboratorio de Medicina Veterinaria Preventiva, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima, Peru;

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Cesar M. Gavidia Laboratorio de Medicina Veterinaria Preventiva, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima, Peru;

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Robert H. Gilman Department of International Health, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland;
Laboratorio de Investigación de Enfermedades Infecciosas, Departamento de Microbiología, Universidad Peruana Cayetano Heredia, Lima, Peru;
Asociación Benéfica Proyectos en Informática, Salud, Medicina y Agricultura, Lima, Peru;

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Noelia P. Angulo Laboratorio de Medicina Veterinaria Preventiva, Facultad de Medicina Veterinaria, Universidad Nacional Mayor de San Marcos, Lima, Peru;

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Richard Lerner Pan American Zoonotic Research and Prevention (PAZ), Framingham, Massachusetts;

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Tamerin Scott Los Angeles County Department of Public Health, Los Angeles, California

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N. Hannah Mirrashed College of Veterinary Medicine, Western University of Health Sciences, Pomona, California;

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Brian Oakley College of Veterinary Medicine, Western University of Health Sciences, Pomona, California;

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Pedro Paulo V. P. Diniz College of Veterinary Medicine, Western University of Health Sciences, Pomona, California;

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Ehrlichia spp. are emerging infectious pathogens, especially in the Americas. Although Ehrlichia canis is primarily a parasite of dogs, polymerase chain reaction-confirmed human infections have been reported from Mexico, Venezuela, and Costa Rica. This study reports the presence of E. canis DNA in 13.7% of 205 dogs from urban areas in Peru and of those, five were analyzed for phylogenetic variation using the Tandem Repeat Protein 36 (TRP36) gene. The use of the TRP36 gene for such analysis was validated against 16S rRNA and heat shock protein genes using Shannon’s entropy bioinformatic approach. When compared with other E. canis strains previously reported, three unique and novel E. canis strains were detected. In addition, the TRP36 amino acid tandem repeat sequences of the Peruvian strains share close similarity to an E. canis strain detected from four human blood bank samples in Costa Rica. This study reports for the first time domestic dogs infected with E. canis strains closely related to a zoonotic strain, which may be of public health concern as dogs can be chronically infected with this pathogen.

Author Notes

Address correspondence to Pedro Paulo V. P. Diniz, College of Veterinary Medicine, Western University of Health Sciences, 309 E. Second St., Pomona, CA 91766-1854. E-mail: pdiniz@westernu.edu

Authors’ addresses: Joseph Geiger, Graduate College of Biomedical Sciences, Western University of Health Sciences, Pomona, CA, E-mail: joseph.geiger@westernu.edu. Bridget A. Morton, College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA, and Affiliated Veterinary Specialists, Orange Park, FL, E-mail: bmorton@westernu.edu. Elton Jose Rosas Vasconcelos, Malika Kachani, N. Hannah Mirrashed, Brian Oakley, and Pedro Paulo V. P. Diniz, College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA, E-mails: evasconcelos@westernu.edu, mkachani@westernu.edu, hmirrashed@westernu.edu, boakley@westernu.edu, and pdiniz@westernu.edu. Maryam Tngrian, College of Agriculture, California State Polytechnic University, Pomona, Pomona, CA, E-mail: mtngrian@cpp.edu. Eduardo A. Barrón, College of Veterinary Medicine, National Mayor of San Marcos, Lima, Peru, E-mail: edubarron1@hotmail.com. Cesar M. Gavidia, College of Veterinary Medicine, National University of San Marcos, Lima, Peru, E-mail: cgavidiac@unmsm.edu.pe. Robert H. Gilman, Department of International Health, John Hopkins Bloomberg School of Public Health, Baltimore, MD, E-mail: gilmanbob@gmail.com. Noelia P. Angulo, Laboratorio de Investigación en Enfermedades Infecciosas, Universidad Peruana Cayetano Heredia, Lima, Peru, E-mail: shaki2700@yahoo.es. Richard Lerner, Pan American Zoonotic Research and Prevention, Framingham, MA, E-mail: riccardolerner@gmail.com. Tamerin Scott, Los Angeles County Department of Public Health, Los Angeles, CA, E-mail: tscott@ph.lacounty.gov.

These authors contributed equally to this work.

  • 1.

    Dantas-Torres F, Chomel BB, Otranto D, 2012. Ticks and tick-borne diseases: a one health perspective. Trends Parasitol 28: 437446.

  • 2.

    Eremeeva ME et al. 2007. Bacteremia, fever, and splenomegaly caused by a newly recognized Bartonella species. N Engl J Med 356: 23812387.

  • 3.

    Arraga-Alvarado CM, Qurollo BA, Parra OC, Berrueta MA, Hegarty BC, Breitschwerdt EB, 2014. Molecular evidence of Anaplasma platys infection in two women from Venezuela. Am J Trop Med Hyg 91: 11611165.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 4.

    Allison RW, Little SE, 2013. Diagnosis of rickettsial diseases in dogs and cats. Vet Clin Pathol 42: 127144.

  • 5.

    de Castro MB, Machado RZ, de Aquino LP, Alessi AC, Costa MT, 2004. Experimental acute canine monocytic ehrlichiosis: clinicopathological and immunopathological findings. Vet Parasitol 119: 7386.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6.

    Vinasco J, Li O, Alvarado A, Diaz D, Hoyos L, Tabachi L, Sirigireddy K, Ferguson C, Moro MH, 2007. Molecular evidence of a new strain of Ehrlichia canis from South America. J Clin Microbiol 45: 27162719.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7.

    Gray J, Dantas-Torres F, Estrada-Pena A, Levin M, 2013. Systematics and ecology of the brown dog tick, Rhipicephalus sanguineus. Ticks Tick Borne Dis 4: 171180.

  • 8.

    Uspensky I, Ioffe-Uspensky I, 2002. The dog factor in brown dog tick Rhipicephalus sanguineus (Acari: Ixodidae) infestations in and near human dwellings. Int J Med Microbiol 291 (Suppl 33): 156163.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Eiras DF, Craviotto MB, Vezzani D, Eyal O, Baneth G, 2013. First description of natural Ehrlichia canis and Anaplasma platys infections in dogs from Argentina. Comp Immunol Microbiol Infect Dis 36: 169173.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 10.

    Vieira RF et al. 2011. Ehrlichiosis in Brazil. Rev Bras Parasitol Vet 20: 112.

  • 11.

    McCown ME, Monterroso VH, Cardona W, 2014. Surveillance for Ehrlichia canis, Anaplasma phagocytophilum, Borrelia burgdorferi, and Dirofilaria immitis in dogs from three cities in Colombia. J Spec Oper Med 14: 8690.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Diniz PP et al. 2013. Infection of domestic dogs in Peru by zoonotic Bartonella species: a cross-sectional prevalence study of 219 asymptomatic dogs. PLoS Negl Trop Dis 7: e2393.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Huerto-Medina E, Damaso-Mata B, 2015. Factors associated with Ehrlichia canis infection in dogs infested with ticks from Huanuco, Peru. Rev Peru Med Exp Salud Publica 32: 756760.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Diniz PPVP, Schwartz DS, De Morais HSA, Breitschwerdt EB, 2007. Surveillance for zoonotic vector-borne infections using sick dogs from southeastern Brazil. Vector Borne Zoonotic Dis 7: 689697.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15.

    Perez M, Rikihisa Y, Wen B, 1996. Ehrlichia canis-like agent isolated from a man in Venezuela: antigenic and genetic characterization. J Clin Microbiol 34: 21332139.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Unver A, Perez M, Orellana N, Huang H, Rikihisa Y, 2001. Molecular and antigenic comparison of Ehrlichia canis isolates from dogs, ticks, and a human in Venezuela. J Clin Microbiol 39: 27882793.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17.

    Moro PL, Shah J, Li O, Gilman RH, Harris N, Moro MH, 2009. Short report: serologic evidence of human ehrlichiosis in Peru. Am J Trop Med Hyg 80: 242244.

  • 18.

    Aguiar DM et al. 2013. Genetic diversity of Ehrlichia canis in Brazil. Vet Microbiol 164: 315321.

  • 19.

    Harrus S, Waner T, 2011. Diagnosis of canine monocytotropic ehrlichiosis (Ehrlichia canis): an overview. Vet J 187: 292296.

  • 20.

    Aguiar DM, Zhang X, Braga IA, Taques II, McBride JW, 2016. Detection of genotype-specific Ehrlichia canis exposure in Brazilian dogs by TRP36 peptide ELISA. Ticks Tick Borne Dis 7: 142145.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21.

    Aguiar DM et al. 2014. A novel Ehrlichia genotype strain distinguished by the TRP36 gene naturally infects cattle in Brazil and causes clinical manifestations associated with ehrlichiosis. Ticks Tick Borne Dis 5: 537544.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22.

    Bouza-Mora L, Dolz G, Solorzano-Morales A, Romero-Zuniga JJ, Salazar-Sanchez L, Labruna MB, Aguiar DM, 2017. Novel genotype of Ehrlichia canis detected in samples of human blood bank donors in Costa Rica. Ticks Tick Borne Dis 8: 3640.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23.

    Diniz PP et al. 2010. High prevalence of tick-borne pathogens in dogs from an Indian reservation in northeastern Arizona. Vector Borne Zoonotic Dis 10: 117123.

  • 24.

    INEI, 2007. Instituto Nacional de Estadística e Informática, Censo Nationales 2007. Lima, Peru: Presidencia del Consejo de Ministros.

    • PubMed
    • Export Citation
  • 25.

    Barber RM et al. 2010. Evaluation of brain tissue or cerebrospinal fluid with broadly reactive polymerase chain reaction for Ehrlichia, Anaplasma, spotted fever group Rickettsia, Bartonella, and Borrelia species in canine neurological diseases (109 cases). J Vet Intern Med 24: 372378.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26.

    Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ, 1990. Basic local alignment search tool. J Mol Biol 215: 403410.

  • 27.

    Kimura M, 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol 16: 111120.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 28.

    Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S, 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 27312739.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 29.

    Edgar RC, 2004. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32: 17921797.

  • 30.

    Waterhouse AM, Procter JB, Martin DM, Clamp M, Barton GJ, 2009. Jalview version 2—a multiple sequence alignment editor and analysis workbench. Bioinformatics 25: 11891191.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31.

    Shenkin PS, Erman B, Mastrandrea LD, 1991. Information-theoretical entropy as a measure of sequence variability. Proteins 11: 297313.

  • 32.

    Jari Oksanen et al. 2017. Ordination Methods, Diversity Analysis and Other Functions for Community and Vegetation Ecologists. Available at: https://cran.r-project.org/web/packages/vegan/index.html. Accessed June 1, 2017.

    • PubMed
    • Export Citation
  • 33.

    Wickham H, 2016. Use R! ggplot2: Elegant Graphics for Data Analysis. New York, NY: Springer.

    • PubMed
    • Export Citation
  • 34.

    R Core Team, 2017. R: A language and Environment for Statistical Computing. Available at: http://www.R-project.org. Accessed June 1, 2017.

    • PubMed
    • Export Citation
  • 35.

    Perez M, Bodor M, Zhang C, Xiong Q, Rikihisa Y, 2006. Human infection with Ehrlichia canis accompanied by clinical signs in Venezuela. Ann N Y Acad Sci 1078: 110117.

  • 36.

    Bronson E, Emmons LH, Murray S, Dubovi EJ, Deem SL, 2008. Serosurvey of pathogens in domestic dogs on the border of Noel Kempff Mercado National Park, Bolivia. J Zoo Wildl Med 39: 2836.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 37.

    Lopez J, Abarca K, Mundaca MI, Caballero C, Valiente-Echeverria F, 2012. Molecular identification of Ehrlichia canis in a dog from Arica, Chile. Rev Chilena Infectol 29: 527530.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 38.

    McCown ME, Opel T, Grzeszak B, 2013. Vector-borne disease surveillance in Puerto Rico: pathogen prevalence rates in canines? Implications for public health and the U.S. Military? Applying the one health concept. J Spec Oper Med 13: 5963.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 39.

    Barbet AF, Bekker CP, Dasch GA, Palmer GH, Ray SC, Rikihisa Y, Rurangirwa FR, 2001. 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. Int J Syst Evol Microbiol 51: 21452165.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 40.

    Walker DH, Dumler JS, 1996. Emergence of the ehrlichioses as human health problems. Emerg Infect Dis 2: 1829.

  • 41.

    Sumner JW, Nicholson WL, Massung RF, 1997. PCR amplification and comparison of nucleotide sequences from the groESL heat shock operon of Ehrlichia species. J Clin Microbiol 35: 20872092.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 42.

    Hsieh YC, Lee CC, Tsang CL, Chung YT, 2010. Detection and characterization of four novel genotypes of Ehrlichia canis from dogs. Vet Microbiol 146: 7075.

  • 43.

    McBride JW, Doyle CK, Zhang X, Cardenas AM, Popov VL, Nethery KA, Woods ME, 2007. Identification of a glycosylated Ehrlichia canis 19-kilodalton major immunoreactive protein with a species-specific serine-rich glycopeptide epitope. Infect Immun 75: 7482.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 44.

    Cárdenas AM, Doyle CK, Zhang X, Nethery K, Corstvet RE, Walker DH, McBride JW, 2007. 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 14: 123128.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 45.

    Yu X-j, McBride JW, Walker DH, 2007. Restriction and expansion of Ehrlichia strain diversity. Vet Parasitol 143: 337346.

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