• 1.

    Thompson CN, et al., 2015. Undifferentiated febrile illness in Kathmandu, Nepal. Am J Trop Med Hyg 92: 875878.

  • 2.

    Dirección de Vigilancia y Análisis del Riesgo en Salud Pública Instituto Nacional deSalud (INS), 2015. Boletín Epidemiológico Semanal. Semana Epidemiológica Número 52 de 2015 (11 dic. al 02 ene.). Available at: http://www.ins.gov.co/boletin-epidemiologico/Boletn%20Epidemiolgico/2015%20Boletin%20epidemiologico%20Semana%2052.pdf. Accessed January 15, 2016.

    • Search Google Scholar
    • Export Citation
  • 3.

    Patino L, Afanador A, Paul JH, 1937. A spotted fever in Tobia, Colombia. Am J Trop Med 17: 639653.

  • 4.

    Patiño-Camargo L, 1941. Nuevas observaciones sobre un tercer foco de fiebre petequial (maculosa) en el hemisferio americano. Bol Of Sanit Panam (Engl) 20: 11121124.

    • Search Google Scholar
    • Export Citation
  • 5.

    Hidalgo M, et al., 2007. Rocky Mountain spotted fever, Colombia. Emerg Infect Dis 13: 10581060.

  • 6.

    Hidalgo M, Sánchez R, Orejuela L, Hernández J, Walker DH, Valbuena G, 2007. Prevalence of antibodies against spotted fever group rickettsiae in a rural area of Colombia. Am J Trop Med Hyg 77: 378380.

    • Search Google Scholar
    • Export Citation
  • 7.

    Hidalgo M, Vesga JF, Lizarazo D, Valbuena G, 2009. A survey of antibodies against Rickettsia rickettsii and Ehrlichia chafeensis in domestic animals from a rural area of Colombia. Am J Trop Med Hyg 80: 10291030.

    • Search Google Scholar
    • Export Citation
  • 8.

    Campos SDE, da Cunha NC, Almosny NRP, 2016. Brazilian spotted fever with an approach in veterinary medicine and One Health perspective. Vet Med Int 2016: 17.

    • Search Google Scholar
    • Export Citation
  • 9.

    Eremeeva ME, Dasch GA, 2015. Challenges posed by tick-borne rickettsiae: eco-epidemiology and public health implications. Front Public Health 3: 55.

    • Search Google Scholar
    • Export Citation
  • 10.

    Faccini-Martínez ÁA, Costa FB, Hayama-Ueno TE, Ramírez-Hernández A, Cortés-Vecino JA, Labruna M, 2015. Rickettsia rickettsii in Amblyomma patinoi ticks, Colombia. Emerg Infect Dis 21: 537539.

    • Search Google Scholar
    • Export Citation
  • 11.

    Faccini-Martínez ÁA, Gárcia-Álvarez L, Hidalgo M, Oteo JA, 2014. Syndromic classification of rickettsioses: an approach for clinical practice. Int J Infect Dis 28: 126139.

    • Search Google Scholar
    • Export Citation
  • 12.

    Hidalgo M, et al., 2013. Flea-borne rickettsioses in the North of Caldas Province, Colombia. Vector Borne Zoonotic Dis 13: 289294.

  • 13.

    Barros-Battesti D, Arzua M, Bechara G, 2006. Carrapatos de Importância Médico-Veterinária da Região Neotropical: Um Guia Ilustrado para Identificação de Espécies. São Paulo, Brazil: International Consortium on Ticks and Tick-borne Diseases, 223.

    • Search Google Scholar
    • Export Citation
  • 14.

    Martins TF, Onofrio VC, Barros-Battesti DM, Labruna M, 2010. Nymphs of the genus Amblyomma (Acari: Ixodidae) of Brazil: descriptions, redescriptions, and identification key. Ticks Tick Borne Dis 1: 7599.

    • Search Google Scholar
    • Export Citation
  • 15.

    Ramírez-Hernández A, et al., 2013. Molecular detection of Rickettsia felis in different flea species from Caldas, Colombia. Am J Trop Med Hyg 89: 453459.

    • Search Google Scholar
    • Export Citation
  • 16.

    Black WC, Piesman J, 1994. Phylogeny of hard- and soft-tick taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences. Proc Natl Acad Sci USA 91: 1003410038.

    • Search Google Scholar
    • Export Citation
  • 17.

    Roux V, Fournier PE, Raoult D, 1996. Differentiation of spotted fever group rickettsiae by sequencing and analysis of restriction fragment length polymorphism of PCR-amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol 34: 20582065.

    • Search Google Scholar
    • Export Citation
  • 18.

    Regnery RL, Spruill CL, Plikaytis BD, 1991. Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes. J Bacteriol 173: 15761589.

    • Search Google Scholar
    • Export Citation
  • 19.

    Choi YJ, Jang WJ, Ryu JS, Lee SH, Park KH, Paik HS, Koh YS, Choi MS, Kim IS, 2005. Spotted fever group and typhus group rickettsioses in humans, South Korea. Emerg Infect Dis 11: 237244.

    • Search Google Scholar
    • Export Citation
  • 20.

    Bello S, Rodríguez M, Paredes A, Mendivelso F, Walteros D, Rodríguez F, Realpe ME, 2013. Comportamiento de la vigilancia epidemiológica de leptospirosis humana en Colombia, 2007–2011. Biomedica 33: 153160.

    • Search Google Scholar
    • Export Citation
  • 21.

    Bharti AR, et al., 2003. Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3: 757771.

  • 22.

    Arroyave E, Londoño AF, Quintero JC, Agudelo-Flórez P, Arboleda M, Díaz FJ, Rodas JD, 2013. Etiología y caracterización epidemiológica del síndrome febril no palúdico en tres municipios del Urabá antioqueño, Colombia. Biomedica 33: 99107.

    • Search Google Scholar
    • Export Citation
  • 23.

    Acosta J, et al., 2006. Brote de rickettsiosis en Necoclí, Antioquia, febrero-marzo de 2006. Informe Quincenal Epidemiológico Nacional 11: 177192.

    • Search Google Scholar
    • Export Citation
  • 24.

    Pacheco-García OE, Giraldo MR, Hidalgo M, Galeano A, Echeverri I, Echavarría-Rodríguez L, Parra E, Rey G, 2008. Estudio de brote febril hemorrágico en el corregimiento de Alto de Mulatos: Distrito Especial Portuario de Turbo, Antioquia, enero de 2008. Informe Quincenal Epidemiológico Nacional 13: 145160.

    • Search Google Scholar
    • Export Citation
  • 25.

    Delisle J, Mendell NL, Stull-Lane A, Bloch KC, Bouyer DH, Moncayo AC, 2016. Human infections by multiple spotted fever group rickettsiae in Tennessee. Am J Trop Med Hyg 94: 12121217.

    • Search Google Scholar
    • Export Citation
  • 26.

    Mattar S, Parra M, 2006. Detection of antibodies to Anaplasma, Bartonella, and Coxiella in rural inhabitants of the Caribbean area of Colombia. Rev MVZ Cordoba 11: 781789.

    • Search Google Scholar
    • Export Citation
  • 27.

    Rar V, Golovljova I, 2011. Anaplasma, Ehrlichia, and “Candidatus Neorhrlichia” bacteria: pathogenicity, biodiversity, and molecular genetic characteristics, a review. Infect Genet Evol 11: 18421861.

    • Search Google Scholar
    • Export Citation
  • 28.

    Guglielmone AA, et al., 2006. Ticks (Ixodidae) on humans in South America. Exp Appl Acarol 40: 83100.

  • 29.

    Santos HA, et al., 2013. Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil. Parasit Vectors 6: 348.

    • Search Google Scholar
    • Export Citation
  • 30.

    Estrada-Peña A, Guglielmone AA, Mangold AJ, 2004. The distribution and ecological ‘preferences’ of the tick Amblyomma cajennense (Acari: Ixodidae), an ectoparasite of humans and other mammals. Ann Trop Med Parasitol 98: 283292.

    • Search Google Scholar
    • Export Citation
  • 31.

    Wells EA, D'Alessandro A, Morales GA, Angel D, 1981. Mammalian wildlife diseases as hazards to man and livestock in an area of the Llanos Orientales of Colombia. J Wildl Dis 17: 153162.

    • Search Google Scholar
    • Export Citation
  • 32.

    Paternina LE, Díaz-Olmos Y, Paternina-Gómez M, Bejarano EE, 2009. Canis familiaris, Un nuevo hospedero de Ornithodoros (A.) puertoricensis FOX, 1947 (Acari: Ixodida) en Colombia. Acta Biolo Colomb 14: 153160.

    • Search Google Scholar
    • Export Citation
  • 33.

    Londoño AF, Díaz FJ, Valbuena G, Gazi M, Labruna M, Hidalgo M, Mattar S, Contreras V, Rodas JD, 2014. Infection of Amblyomma ovale by Rickettsia sp. strain Atlantic rainforest, Colombia. Ticks Tick Borne Dis 5: 672675.

    • Search Google Scholar
    • Export Citation
  • 34.

    Miranda J, Portillo A, Oteo JA, Mattar S, 2012. Rickettsia sp. strain Colombianensi (Rickettsiales: Rickettsiaceae): a new proposed Rickettsia detected in Amblyomma dissimile (Acari: Ixodidae) from iguanas and free-living larvae ticks from vegetation. J Med Entomol 49: 960965.

    • Search Google Scholar
    • Export Citation
  • 35.

    Miranda J, Mattar S, 2014. Molecular detection of Rickettsia bellii and Rickettsia sp. strain Colombianensi in ticks from Cordoba, Colombia. Ticks Tick Borne Dis 5: 208212.

    • Search Google Scholar
    • Export Citation
  • 36.

    Faccini-Martínez ÁA, Ramírez-Hernández A, Forero-Becerra E, Cortés-Vecino JA, Escandón P, Rodas JD, Palomar AM, Portillo A, Oteo JA, Hidalgo M, 2016. Molecular evidence of different Rickettsia species in Villeta, Colombia. Vector Borne Zoonotic Dis 16: 8587.

    • Search Google Scholar
    • Export Citation
  • 37.

    Ramírez-Hernández A, Escandón P, Cortés-Vecino JA, Rodas JD, Hidalgo M, 2013. Detección molecular de Rickettsia spp. (Da Rocha-Lima, 1916) en garrapatas recolectadas en tres regiones de Colombia. Acta Med Costarric 55: 8384.

    • Search Google Scholar
    • Export Citation
  • 38.

    Bermúdez SE, Eremeeva ME, Karpathy SE, Samudio F, Zambrano ML, Zaldivar Y, Motta JA, Dasch GA, 2009. Detection and identification of rickettsial agents in ticks from domestic mammals in eastern Panama. J Med Entomol 46: 856861.

    • Search Google Scholar
    • Export Citation
  • 39.

    Merhej V, Angelakis E, Socolovschi C, Raoult D, 2014. Genotyping, evolution and epidemiological findings of Rickettsia species. Infect Genet Evol 25: 122137.

    • Search Google Scholar
    • Export Citation
  • 40.

    Monje LD, Costa FB, Colombo VC, Labruna MB, Antoniazzi LR, Gamietea I, Nava S, Beldomenico PM, 2016. Dynamics of exposure to Rickettsia parkeri in cattle in the Paraná River Delta, Argentina. J Med Entomol 53: 660665.

    • Search Google Scholar
    • Export Citation
  • 41.

    Pesquera C, Portillo A, Palomar AM, Oteo JA, 2015. Investigation of tick-borne bacteria (Rickettsia spp., Anaplasma spp., Ehrlichia spp. and Borrelia spp.) in ticks collected from Andean tapirs, cattle and vegetation from a protected area in Ecuador. Parasit Vectors 8: 46.

    • Search Google Scholar
    • Export Citation
  • 42.

    Blanton LS, Mendell NL, Walker DH, Bouyer DH, 2014. Rickettsia amblyommii” induces cross protection against lethal Rocky Mountain spotted fever in a guinea pig model. Vector Borne Zoonotic Dis 14: 557562.

    • Search Google Scholar
    • Export Citation
  • 43.

    Rivas JJ, Moreira-Soto A, Alvarado G, Taylor L, Calderón-Arguedas O, Hun L, Corrales-Aguilar E, Morales JA, Troyo A, 2015. Pathogenic potential of a Costa Rican strain of ‘Candidatus Rickettsia amblyommii’ in guinea pigs (Cavia porcellus) and protective immunity against Rickettsia rickettsii .Ticks Tick Borne Dis 6: 805811.

    • Search Google Scholar
    • Export Citation
  • 44.

    Walker DH, 2016. Changing dynamics of human-rickettsial interactions. Am J Trop Med Hyg 94: 34.

  • 45.

    Ogrzewalska M, Saraiva DG, Moraes-Filho J, Martins TF, Costa FB, Pinter A, Labruna MB, 2012. Epidemiology of Brazilian spotted fever in the Atlantic Forest, state of São Paulo, Brazil. Parasitology 139: 12831300.

    • Search Google Scholar
    • Export Citation
  • 46.

    Wood CL, Lafferty KD, 2013. Biodiversity and disease: a synthesis of ecological perspectives on Lyme disease transmission. Trends Ecol Evol 28: 239247.

    • Search Google Scholar
    • Export Citation
 
 
 

 

 

 

 

 

 

Epidemiology of Spotted Fever Group Rickettsioses and Acute Undifferentiated Febrile Illness in Villeta, Colombia

View More View Less
  • 1 Grupo de Enfermedades Infecciosas, Departamento de Microbiología, Pontificia Universidad Javeriana, Bogotá, Colombia;
  • | 2 Programa de Pós-Graduação em Doenças Infecciosas, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES, Brazil;
  • | 3 Grupo Parasitología Veterinaria, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, Bogotá, Colombia;
  • | 4 Departamento de Medicina Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brazil;
  • | 5 Departamento de Medicina Veterinária, Universidade Federal de Viçosa, Viçosa, MG, Brazil;
  • | 6 Grupo Salud Pública Veterinaria, Departamento Salud Animal, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, Bogotá, Colombia;
  • | 7 Departamento de Biología, Pontificia Universidad Javeriana, Bogotá, Colombia;
  • | 8 Departamento de Enfermedades Infecciosas, Centro de Rickettsiosis y Enfermedades Transmitidas por Artrópodos Vectores, Hospital San Pedro–CIBIR, Logroño, Spain

Etiology of acute undifferentiated febrile syndrome (AUFS) is often unknown, leading to inaccurate diagnosis and treatment. Villeta town has been identified as an endemic area for spotted fever group (SFG) rickettsioses but little is known about possible amplifier hosts and other Rickettsia species different from Rickettsia rickettsii. Besides, few studies have approached other AUFS etiologies in the region. We investigated the role of dengue, leptospirosis, rickettsioses, human anaplasmosis, and Q fever as possible causes of AUFS in patients from Villeta. Sera specimens and ticks from animals as well as ticks from vegetation were studied for the presence of different Rickettsia spp. Among 104 sera from patients with AUFS, 16.4%, 24.0%, and 2.9% patients seroconverted to dengue, Leptospira, and SFG Rickettsia, respectively, with a case of probable coinfection or cross-reaction with Anaplasma phagocytophilum. None of the samples were reactive for Coxiella burnetii. Sera samples from 74 horses, 118 dogs, and 62 bovines were collected and showed 33.8%, 14.4%, and 50.0% of seroprevalence for SFG Rickettsia, respectively. A total of 1,287 ixodid ticks were collected from animals/vegetation and processed in pools for polymerase chain reaction. Among them, 1.7% was positive for Rickettsia genes, and Rickettsia amblyommii, R. rickettsii, and Rickettsia spp. were found. These results confirm the circulation of dengue, different SFG Rickettsia species and the relevance of other etiologies like leptospirosis and human anaplasmosis. Further studies must identify different epidemiological variables to establish proper surveillance and control programs.

INTRODUCTION

Acute undifferentiated febrile syndrome (AUFS) is defined as fever without a focus of infection on initial physical examination or in basic laboratory tests.1 Etiologic nature of AUFS has been a challenge everywhere. Although AUFS is common in tropical regions, the specific etiology is often unknown, making accurate diagnosis, treatment, and surveillance very difficult. Villeta town in Colombia has been considered an endemic area for dengue, spotted fever group (SFG) rickettsioses, and recently, for chikungunya and Zika fever.2,3 Since the first descriptions in 1937, high fatality rates of Rocky Mountain spotted fever (RMSF) have been reported in this region. In the beginning of the 21st century, fatal cases of RMSF and significant seroprevalences against SFG Rickettsia spp. in humans and domestic animals have been recorded.37 The relationship and connection between vertebrate species, including humans, through the human–animal–ecosystem interface, could have an impact on human health and should be considered.8 Nowadays, rickettsioses have generated a serious concern in public health because Rickettsia spp. are expanding their geographical distribution, hosts, and vectors leading to zoonotic processes that might affect humans.9 Recent studies have confirmed the presence of Rickettsia rickettsii in Amblyomma patinoi ticks collected from cattle in Naranjal village, Villeta.10 Despite this new evidence, since 2004, there are no new reported cases of SFG rickettsioses in this region. Several authors have confirmed the important role of domestic animals, mainly horses and dogs, as sentinels for monitoring the circulation of rickettsiae in urban areas.8 A previous study in Villeta analyzed seroprevalence against Rickettsia spp. in dogs and horses as a first approach to the dynamics of the infection in domestic animals from this geographic area, showing values of 18.2% and 16.3%, respectively.7 On the other hand, the role of the small wild mammals as amplifiers of rickettsioses has not been studied in the area. Besides, the circulating Rickettsia species in ticks and its possible relationship with the recent disease epidemiology for this area is unknown. In addition, no studies have evaluated the etiology of AUFS, except dengue and recent epidemiological surveys of chikungunya and Zika.2

The aim of this study was to approach the probable etiologies of AUFS. Since dengue can be misdiagnosed and/or overlapped with rickettsial syndromes,11 we investigated (among others) the role of rickettsioses as possible causes of AUFS in patients with presumptive diagnostic of dengue in Villeta, including clinical characteristics, demographics, and epidemiology from a One Health perspective. In addition, sera specimens and ticks from animals as well as ticks from vegetation were studied for the presence of different Rickettsia spp.

MATERIALS AND METHODS

Study area.

The study area was Villeta, Cundinamarca, Colombia (5°0′53″N, 74°28′29″W), located at 842 m above sea level and with an annual mean temperature of 26°C, a relative humidity varying between 80% and 97%, and a total area of 140 km2. According to the 2014 data from the National Department of Statistics (DANE), the estimated population is 25,061 inhabitants, distributed in 22 villages. Agriculture and ecotourism are the predominant economic activities (www.villeta-cundinamarca.gov.co).

Patients.

Patients included in the study were those who attended Salazar Hospital (Villeta) with presumptive diagnosis of dengue from October 2011 to March 2013. Sera specimens from acute and convalescent phase (> 15 days and < 2 months from the onset of symptoms) were obtained. Only those patients who had paired samples were included in the study. All of them accepted the informed consent that was approved by the ethical committee from Pontificia Universidad Javeriana.

Serological assays in humans.

Sera samples were centrifuged and stored at −20°C for subsequent analyses. The following diagnostic tests were performed: 1) capture enzyme-linked immunosorbent assay (Panbio Diagnostic®, San Diego, CA) for the detection of IgM antibodies against dengue virus in the acute illness phase; 2) microagglutination test (MAT) for five Leptospira pathogenic serovars (Icterohaemorrhagiae, Hardjo, Pomona, Grippotyphosa, and Canicola) for the detection of IgG antibodies with a cutoff value of 1:25; 3) indirect immunofluorescence assay (IFA) for the detection of IgG antibodies against R. rickettsii (strain Taiaçu) and Rickettsia amblyommii (strain Ac37) with a cutoff value of 1:6412; 4) IFA using the commercial kit “Rickettsia IFA IgG” (Focus Diagnostics, Cypress, CA) for the detection of IgG antibodies of Rickettsia typhi, with a cutoff value of 1:64; 5) IFA using the commercial kit “Anaplasma phagocytophilum IFA IgG” (Focus Diagnostics) for the detection of IgG antibodies against A. phagocytophilum, with a cutoff value of 1:64; 6) IFA using the commercial kit “Q Fever IgG” (Focus Diagnostics) for the detection of IgG antibodies against Coxiella burnetii (phase I and II antigens), with a cutoff value of 1:64. Throughout the text, for IFA and MAT, seroconversion was defined as a difference of 4-fold titers between acute and convalescent phases or when a negative sample turned into positive.

Serum samples from domestic mammals.

From November to December 2011, domestic animals (dogs, horses, and cattle) were sampled for sera. Sera samples were analyzed by in-house IFA identifying IgG antibodies against R. rickettsii (strain Taiaçu). Samples with IgG titers ≥ 64 were considered positive for Rickettsia.7 Reactive sera (R. rickettsii antigen) obtained from a dog and a horse in the same town in 20087 and a reactive serum for SFG Rickettsia spp. (Rickettsia africae antigen) obtained from cattle from Caribbean Islands, were used as positive controls.

Ticks from domestic, wild mammals, and vegetation.

From November to December 2011, ticks were removed with tweezers from the above-mentioned domestic animals and during July 2012, from wild mammals captured (by Sherman and Tomahawk traps) in 17 villages, previously selected for high seroprevalence against SFG Rickettsia in domestic animals and humans.6,7 Ticks from vegetation were also captured by flagging and dragging methods. Collected specimens were kept in 70% ethanol and further classified by taxonomic keys.13,14

Molecular detection of Rickettsia species in ticks collected from Villeta.

Ticks were grouped by species, stage, locality, and host as main criteria. Females were processed individually and males, nymphs, and larvae in pools of 2–4, 7–12, and 10–20 individuals, respectively. They were processed for DNA extraction using a modified protocol of the commercial kit DNeasy Blood and Tissue (QIAGEN Inc., Valencia, CA) with the addition of guanidine thiocyanate (DNAzol; Invitrogen, Life Technologies Corp., Grand Island, NY).15

DNA extracts were used for molecular detection of Rickettsia spp. by conventional and nested polymerase chain reaction (PCR) assays, ensuring 150 ng/μL of DNA per reaction. An initial PCR targeting 16S rRNA mitochondrial tick gene (as internal control) was performed.16 Positive pools were further tested for rickettsial infection using ompA (seminested assay with primers Rr190.70p-Rr190.701n/Rr190.602n)17,18 and ompB (nested assay with primers rompB OF-rompB OR/rompB SFG IF-rompB SFG IR)19 as PCR target genes. DNA of Rickettsia slovaca strain S14ab (from the Center of Rickettsiosis and Arthropod-borne Diseases, CIBIR, La Rioja-Spain) and water were used as positive and negative controls, respectively. All PCR products with expected sizes (532 and 420 base pairs for partial ompA and ompB genes, respectively) were sequenced, and nucleotide sequences were compared with GenBank data by BLAST analysis. Minimum infection rates for positive pools were calculated as the percentage of the ratio between the number of pools positive for SFG Rickettsia, and the total number of individual ticks included in the specific sample.

Data analysis.

Official Colombian report documents for dengue from patients included in the study were evaluated. Gender, age, village, symptoms, laboratory analysis (hematocrit, platelets, and leukocytes), were analyzed.

RESULTS

AUFS cases from Salazar Hospital.

A total of 104 paired samples (31%) were obtained from 335 patients who attended Salazar Hospital with presumptive diagnoses of dengue during the inclusion period. Seventeen of 104 patients (16.4%) presented only IgM positive for dengue in the acute illness phase. Twenty-five (24%) presented unique seroconversion to one or more pathogenic serovars of Leptospira. Individually, the Icterohaemorrhagiae serovar was the most representative one (15 patients). Three patients (2.9%) showed unique seroconversion to SFG Rickettsia (one patient for R. rickettsii [< 1:64/1:128] and two for R. amblyommii [1:64/1:256 and < 1:64/1:128]).

We detected probable coinfection or cross-reaction in 17/104 (16.4%) for dengue/leptospirosis, 6/104 (5.7%) leptospirosis/SFG rickettsioses, 5/104 (4.8%) leptospirosis/SFG rickettsioses/dengue, and 1/104 (0.9%) for SFG rickettsioses/dengue. Likewise, a probable case of human anaplasmosis/dengue coinfection was detected in one sample (0.9%). Curiously, 70/104 (67.3%) and 7/104 patients (6.7%) presented seropositivity to SFG Rickettsia (R. rickettsii and/or R. amblyommii) and A. phagocytophilum, respectively, in at least one of the two sera specimens (in acute or convalescent phase), without evidence of seroconversion in paired samples.

None of the paired samples were reactive for C. burnetii (phase I and II antigens) or for R. typhi. Finally, for 29/104 samples (27.8%), all the tests were negative. No patients developed clinical complications or death.

A total of 88 Official Colombian report documents for dengue were evaluated from the 104 patients with paired serum samples. For each etiology, we analyzed all patients that had shown unique probable diagnosis for dengue, leptospirosis, and rickettsioses (N = 45) and sociodemographic features like gender, age, and area of origin (Table 1). The most common symptoms for the three etiologies were fever, myalgia, headache, arthralgia, and retro-orbital pain. Specifically, two-thirds of patients with SFG rickettsioses (66.67%) showed rash (Table 2). Patients with unique seroconversion for SFG Rickettsia presented the following clinical signs: fever, myalgia, nonpurulent conjunctival injection, vomit, and headache, for the R. rickettsii-seropositive case; and fever, myalgia, arthralgia, retro-orbital pain, abdominal pain, rash, and headache, for both R. amblyommii-seropositive cases.

Table 1

Sociodemographic features of patients with acute febrile syndrome by etiology attended in Villeta town (October 2011–March 2013)

VariablesDengue n (%)Leptospirosis n (%)SFG rickettsioses n (%)
Gender
 Male9 (53)11 (44)2 (66.7)
 Female8 (47)14 (56)1 (33.3)
Age (years)
 < 106 (35.3)6 (24)1 (33.3)
 10–207 (41.2)5 (20)
 21–301 (5.9)5 (20)1 (33.3)
 31–403 (17.6)4 (16)
 41–503 (12)
 51–601 (4)
 > 601 (4)1 (33.3)
Area
 Urban11 (64.7)14 (56)2 (66.7)
 Rural4 (23.5)2 (8)1 (33.3)
 Undetermined2 (11.8)9 (36)
No. of patients17 (100)25 (100)3 (100)

SFG = spotted fever group.

Table 2

More frequent symptoms presented in dengue fever, leptospirosis, and SFG rickettsioses from patients attended in Villeta town (October 2011–March 2013)

SymptomsDengue fever (%)Leptospirosis (%)SFG rickettsioses (%)
Fever16/17 (94.12)25/25 (100)3/3 (100)
Myalgia13/17 (76.5)18/25 (72)3/3 (100)
Headache11/17 (64.7)15/25 (60)3/3 (100)
Arthralgia10/17 (58.8)15/25 (60)1/3 (33.3)
Retro-orbital pain9/17 (52.9)13/25 (52)1/3 (33.3)
Rash2/17 (11.8)7/25 (28)2/3 (66.7)
Vomit4/17 (23.5)8/25 (32)1/3 (33.3)
Abdominal pain3/17 (17.6)8/25 (32)1/3 (33.3)
Red eye illness3/17 (17.6)4/25 (16)1/3 (33.3)
Diarrhea3/17 (17.6)4/25 (16)ND
Jaundice2/17 (11.8)NDND
Tachycardia1/17 (5.8)3/25 (12)1/3 (33.3)
Hypotension1/17 (5.8)NDND

ND = no data; SFG = spotted fever group.

According to the laboratory results, the initial average hematocrit was 37.2% ± 12.0 for dengue, 39.7% ± 10.2 for leptospirosis, and 31% ± 16.5 for SFG rickettsioses (normal range: 39–50); the average leukocyte counts were 3,282.3 ± 714.3, 4,420 ± 2,626.3 and 3,300 ± 1,473.0 cells/mm3 (normal range: 4,600–10,600) and platelet counts were 134,352 ± 26,542, 124,300 ± 34,563.6, and 154,666 ± 17,214 cells/mm3 (normal range: 160,000–380,000), respectively.

Seroprevalence against SFG Rickettsia in domestic mammals.

Sera samples were collected from 74 horses, 118 dogs, and 62 cows. Among the sera samples from domestic animals, 25/74 from horses (33.8%), 17/118 from dogs (14.4%), and 31/62 from cattle (50%) showed titers equal to or higher than 64, and three of them reached titers of 8,192 (Table 3).

Table 3

Seropositive dogs, horses, and cattle from Villeta town against Rickettsia rickettsii antigen by IFA (IgG titer ≥ 64); November–December 2011

VillageHorsesDogsCattle
No. positive /no. testedMaximum titerNo. positive /no. testedMaximum titerNo. positive /no. testedMaximum titer
Alto de Paja1/2640/5
Alto de Torres0/30/5
Bagazal0/10/92/364
Balsal1/31280/2
Chapaima3/8644/54,0961/464
Chorrillo1/4641/8640/5
Cune0/51/58,1923/464
El Puente1/45120/6
Ilo Grande1/6640/6
La Bolsa3/4640/20/1
La Esmeralda0/50/2
La Mazata0/11/364
Maní1/11280/37/864
Mave1/5643/8640/1
Naranjal2/41281/35126/1264
Payandé2/2640/4
Potrero Grande1/4642/264
Quebrada Honda0/4
Río Dulce1/3641/164
Salitre Blanco0/32/118,1922/964
Salitre Negro2/71280/10/2
San Isidro1/41280/27/8128
Urban area5/78,1922/1464
Total (%)25/74 (33.8)17/118 (14.4)31/62 (50.0)

IFA = immunofluorescence assay.

Molecular detection of Rickettsia species in ticks.

A total of 516 ticks were collected from domestic animals and were identified as follows: Dermacentor nitens and Amblyomma cajennense sensu lato (s.l.) from horses; Rhipicephalus sanguineus s.l., A. cajennense s.l., and Amblyomma ovale from dogs; and Rhipicephalus microplus and A. cajennense s.l. from cattle (Table 4). Besides, 13 wild mammals (five Didelphis marsupialis, three Marmosa robinsoni, three Mus musculus, one Rattus rattus, and one Sigmodon hirsutus) were captured and tick samples were only collected from D. marsupialis and classified as Ixodes luciae and Ixodes spp. (Table 4). A total of 744 ticks were obtained from vegetation and identified as R. microplus, Amblyomma sp., A. cajennense s.l., and Dermacentor sp. (Table 4).

Table 4

Number and species of ticks collected from animals and vegetation in Villeta town (November–December 2011 and July 2012)

No. of individualsRickettsia infection*
Species of ticksSourceMFNLTotal
Amblyomma cajennense s.l.Horses35267169
Dogs33
Cattle1819138Positive
Vegetation22158162Positive
Amblyomma ovaleDogs11
Amblyomma sp.Vegetation222222Positive
Dermacentor nitensHorses611205010241Positive
Dermacentor sp.Vegetation6464
Ixodes luciaeDidelphis marsupialis141125
Ixodes sp.D. marsupialis22
Rhipicephalus microplusCattle931242Positive
Vegetation296296Positive
Rhipicephalus sanguineus s.l.Dogs66479122

F = female adults; L = larvae; M = male adults; N = nymphs.

Positive for Rickettsia ompA or ompB genes.

As it is showed in Table 5, ticks were grouped into 446 pools. Among them, 358 pools were positive for the 16S rRNA PCR and further screened for Rickettsia spp. (ompA and ompB genes). Rickettsia spp. was found in 6/358 pools (1.7%). Rickettsia amblyommii (GenBank accession no. KJ433807) was detected in larvae of R. microplus (pool M841). In addition, R. rickettsii (GenBank accession nos. KJ433802, KJ433806, and KJ433805, respectively) was found in male adults of D. nitens (pool M181), in nymphs of A. cajennense s.l. (pool M827), and in larvae of Amblyomma sp. (pool M822). Moreover, Rickettsia spp. were detected in male adults of A. cajennense s.l. (pool M235) and a female specimen of R. microplus (pool M196), showing 94.9% identity with Rickettsia conorii (GenBank accession no. KJ433804) and 99.7% identity with Rickettsia monacensis (GenBank accession no. KJ433803), respectively, as highest identities with validly published Rickettsia species (Table 5).

Table 5

Tick pools evaluated for the presence of Rickettsia spp. in Villeta town (November–December 2011, and July 2012)

VillageTick species (source)Positive pools*/total of poolsTotal of ticks (% MIR)BLAST analysis
ChapaimaAmblyomma cajennense s.l. (DA)0/5
Dermacentor nitens (DA)0/81
Rhipicephalus sanguineus s.l. (DA)0/3
Rhipicephalus microplus (DA)0/1
CuneA. cajennense s.l. (DA)0/7
D. nitens (DA)0/11
R. sanguineus s.l. (DA)0/1
R. microplus (DA)0/22
ManiR. microplus (V)1/26229 (0.4)Rickettsia amblyommii
NaranjalA. cajennense s.l. (DA)1/4343 (2.3)Rickettsia sp.
A. cajennense s.l. (V)1/3251 (2.0)R. rickettsii§
Amblyomma ovale (DA)0/1
Amblyomma sp. (V)1/23104 (1.0)R. rickettsii
Dermacentor sp. (V)0/8
Ixodes luciae (WA)0/16
Ixodes sp. (WA)0/2
R. sanguineus (DA)0/1
Urban areaA. cajennense s.l. (DA)0/12
D. nitens (DA)0/58
R. sanguineus s.l. (DA)0/69
Salitre BlancoA. cajennense s.l. (DA)0/5
D. nitens (DA)1/410 (10)R. rickettsii
R. microplus (DA)1/1516 (6.3)Rickettsia sp.**

DA = domestic animals; MIR = minimum infection rate (no. of positive pools/total no. of individual ticks) × 100; V = Vegetation; WA = wild animals.

Positive pools for ompA or ompB.

Pool “M841”: 15 larvae from vegetation positive for ompA gene.

Pool “M235”: four males from cattle positive for ompA gene (94.9% identity with Rickettsia conorii).

Pool “M827”: 10 nymphs from vegetation positive for ompA gene.

Pool “M822”: 15 larvae from vegetation positive for ompA gene.

Pool “M181”: three males from a horse positive for ompB gene.

Pool “M196”: one female from a cow positive for ompB gene (99.7% identity with Rickettsia monacensis).

DISCUSSION

The probable etiology of AUFS for patients who attended Salazar Hospital from October 2011 to March 2013 was mainly leptospirosis, with 24% of seroconversion, especially against the Icterohaemorrhagiae serovar (60%). In a national study that showed the epidemiology of human leptospirosis from 2007 to 2011, the Icterohaemorrhagiae serovar represented 7.56% of the circulation.20 Our study evidences the circulation of Leptospira serovars and indicates that leptospirosis, a common worldwide zoonotic disease with a predominant presence in tropical regions,21 is one of the main causes of AUFS in Villeta. Our results of possible etiologies of AUFS are comparable with those from a recent study in the region of Urabá (Antioquia),22 where R. rickettsii cases were confirmed.23,24 In Antioquia, dengue was probably the main cause of AUFS (37.3%) with higher seroprevalence than the one found herein (16.4%). In addition, percentages of Rickettsia infection (2.7%) and coinfection of SFG rickettsiosis/dengue (0.5%) were comparable to those found in our study (2.9% and 0.9%, respectively).22

Herein, the continuous circulation of SFG Rickettsia spp. in humans and in domestic animals (cattle, horses, and dogs) from Villeta has been confirmed, showing 67.3%, 50.0%, 33.8%, and 14.4% of seroprevalence, respectively. These data suggest that pathogenic and nonpathogenic SFG Rickettsia species may cause human diseases or asymptomatic infections in the area. Also, the finding of three patients with unique SFG Rickettsia seroconversion is noteworthy; those with R. amblyommii seroconversion presented clinical signs previously reported from probable cases of R. amblyommii rickettsiosis (i.e., fever, headache, myalgia, and rash)25; unfortunately it was not possible to confirm (with molecular or Western Immunoblotting tests) the specific SFG Rickettsia species involved in these patients.

In addition, this study represents the second report of probable exposition to A. phagocytophilum in Colombia,26 based on the probable case of infection defined by seroconversion and the seroprevalence found in our study (6.7%), which is lower than that reported in Cordoba.26 Although A. phagocytophilum is typically transmitted by Ixodes ticks,27 which did not represent a huge concern in Latin America,28 this bacterium has been recently detected in A. cajennense s.l.29 This tick species is present in our area but the finding of A. phagocytophilum in Amblyomma ticks does not confirm their role as vectors. Besides, to confirm the circulating Anaplasma species and its vector, further studies must attempt to isolate it from vertebrate and/or invertebrate hosts.

Herein, A. cajennense s.l. was parasitizing the three species of domestic mammals sampled (horses, dogs, and cattle) with the highest levels of infestation in cattle (47.5%). The multihost feeding habits and the anthropophilic behavior of this tick28,30 represent a high risk of exposition to SFG Rickettsia for the human population of Villeta, even more considering that this species has been incriminated as vector of R. rickettsii in the region.4,10 Furthermore, the tick species A. ovale and I. luciae have been recorded for the first time in this geographical area of Colombia. Previous studies documented the presence of I. luciae parasitizing D. marsupialis in the Orinoquia region,31 and A. ovale parasitizing dogs in Sucre and Urabá (Antioquia).32,33

Molecular evidence of SFG Rickettsia spp. in ticks from animals has been previously reported in Colombia.3336 In this study, R. amblyommii was found in R. microplus from vegetation and R. rickettsii in D. nitens from a horse and also in A. cajennense s.l. and Amblyomma sp. from vegetation. In the same region, A. cajennense s.l. harboring R. amblyommii and D. nitens infected with R. rickettsii as well as the isolation of R. rickettsii (strain Villeta) from A. patinoi have been previously published.10,36,37 The detection of R. microplus infected with R. amblyommii has been previously reported in Panama.38 The cofeeding phenomenon could explain the association of R. rickettsii with D. nitens since this tick species and A. cajennense s.l. are common parasites of horses in Latin America.13,39

Furthermore, our study shows the first detection of SFG Rickettsia spp. in cattle from this country, and it is the second report from South America.40 Specifically, A. cajennense s.l. and R. microplus removed from cows were found to be infected with Rickettsia spp. that showed the highest identities with R. conorii (94.9%) and R. monacensis (99.7%), respectively. New rickettsial genotypes related to R. monacensis had been found in other Colombian areas34 or in neighboring countries such as Ecuador.41 Further studies are necessary for a better characterization of these bacteria.

Despite the high lethality caused by R. rickettsii 80 years ago,3 the occurrence of human fatal cases of RMSF in 2003 and 2004,5 the high rates of seroprevalence for SFG Rickettsia spp. in humans and domestic mammals,57 and the recent isolation of R. rickettsii from A. patinoi ticks,10 there are no new reports of human cases caused by R. rickettsii in this endemic region of Villeta (Cundinamarca). This situation could be explained by misdiagnosis of this illness due to lack of clinical suspicions and of compulsory notification in Colombia, by possible cross-immune protection caused by less pathogenic rickettsiae,4244 and by ecological transformations which modulate the epidemiological pattern of the disease.45 To better understand the latter scenario, further research must identify the vertebrate hosts which sustain the natural cycle of these pathogens (mainly as amplifier hosts) and comprehend the related epidemiological determinants and the effects of environmental changes on the tick–pathogen–vertebrate interface as has been studied for similar diseases.46

The results presented herein confirm the relevance of rickettsioses as a differential diagnosis in patients with AUFS and the importance of the clinical suspicion and laboratory confirmation of different etiologies with an early treatment. Nonetheless, other diseases (e.g., leptospirosis, anaplasmosis, and dengue), their interrelationships, and other epidemiological variables should be further studied in the region.

Acknowledgments:

We acknowledge all personnel from Hospital Salazar, UMATA, and local farms from Villeta for their invaluable help during all phases of the study. Additionally, we thank Patrick Kelly from Ross University School of Veterinary Medicine (Basseterre, St. Kitts) for kindly providing bovine sera for IFAT, Patricia Hernández (Universidad de La Salle) for kindly providing MAT-Leptospira diagnosis, and Marcelo Labruna from University of São Paulo for kindly providing antigens of R. rickettsii and R. amblyommii for IFAT. We also thank the Pontificia Universidad Javeriana (PUJ) research project: “Caracterización de factores climáticos y ecológicos de una especie de garrapata y su relación con la epidemiologia de las rickettsiosis en un área endémica” (ID Cod ppta 4344).

REFERENCES

  • 1.

    Thompson CN, et al., 2015. Undifferentiated febrile illness in Kathmandu, Nepal. Am J Trop Med Hyg 92: 875878.

  • 2.

    Dirección de Vigilancia y Análisis del Riesgo en Salud Pública Instituto Nacional deSalud (INS), 2015. Boletín Epidemiológico Semanal. Semana Epidemiológica Número 52 de 2015 (11 dic. al 02 ene.). Available at: http://www.ins.gov.co/boletin-epidemiologico/Boletn%20Epidemiolgico/2015%20Boletin%20epidemiologico%20Semana%2052.pdf. Accessed January 15, 2016.

    • Search Google Scholar
    • Export Citation
  • 3.

    Patino L, Afanador A, Paul JH, 1937. A spotted fever in Tobia, Colombia. Am J Trop Med 17: 639653.

  • 4.

    Patiño-Camargo L, 1941. Nuevas observaciones sobre un tercer foco de fiebre petequial (maculosa) en el hemisferio americano. Bol Of Sanit Panam (Engl) 20: 11121124.

    • Search Google Scholar
    • Export Citation
  • 5.

    Hidalgo M, et al., 2007. Rocky Mountain spotted fever, Colombia. Emerg Infect Dis 13: 10581060.

  • 6.

    Hidalgo M, Sánchez R, Orejuela L, Hernández J, Walker DH, Valbuena G, 2007. Prevalence of antibodies against spotted fever group rickettsiae in a rural area of Colombia. Am J Trop Med Hyg 77: 378380.

    • Search Google Scholar
    • Export Citation
  • 7.

    Hidalgo M, Vesga JF, Lizarazo D, Valbuena G, 2009. A survey of antibodies against Rickettsia rickettsii and Ehrlichia chafeensis in domestic animals from a rural area of Colombia. Am J Trop Med Hyg 80: 10291030.

    • Search Google Scholar
    • Export Citation
  • 8.

    Campos SDE, da Cunha NC, Almosny NRP, 2016. Brazilian spotted fever with an approach in veterinary medicine and One Health perspective. Vet Med Int 2016: 17.

    • Search Google Scholar
    • Export Citation
  • 9.

    Eremeeva ME, Dasch GA, 2015. Challenges posed by tick-borne rickettsiae: eco-epidemiology and public health implications. Front Public Health 3: 55.

    • Search Google Scholar
    • Export Citation
  • 10.

    Faccini-Martínez ÁA, Costa FB, Hayama-Ueno TE, Ramírez-Hernández A, Cortés-Vecino JA, Labruna M, 2015. Rickettsia rickettsii in Amblyomma patinoi ticks, Colombia. Emerg Infect Dis 21: 537539.

    • Search Google Scholar
    • Export Citation
  • 11.

    Faccini-Martínez ÁA, Gárcia-Álvarez L, Hidalgo M, Oteo JA, 2014. Syndromic classification of rickettsioses: an approach for clinical practice. Int J Infect Dis 28: 126139.

    • Search Google Scholar
    • Export Citation
  • 12.

    Hidalgo M, et al., 2013. Flea-borne rickettsioses in the North of Caldas Province, Colombia. Vector Borne Zoonotic Dis 13: 289294.

  • 13.

    Barros-Battesti D, Arzua M, Bechara G, 2006. Carrapatos de Importância Médico-Veterinária da Região Neotropical: Um Guia Ilustrado para Identificação de Espécies. São Paulo, Brazil: International Consortium on Ticks and Tick-borne Diseases, 223.

    • Search Google Scholar
    • Export Citation
  • 14.

    Martins TF, Onofrio VC, Barros-Battesti DM, Labruna M, 2010. Nymphs of the genus Amblyomma (Acari: Ixodidae) of Brazil: descriptions, redescriptions, and identification key. Ticks Tick Borne Dis 1: 7599.

    • Search Google Scholar
    • Export Citation
  • 15.

    Ramírez-Hernández A, et al., 2013. Molecular detection of Rickettsia felis in different flea species from Caldas, Colombia. Am J Trop Med Hyg 89: 453459.

    • Search Google Scholar
    • Export Citation
  • 16.

    Black WC, Piesman J, 1994. Phylogeny of hard- and soft-tick taxa (Acari: Ixodida) based on mitochondrial 16S rDNA sequences. Proc Natl Acad Sci USA 91: 1003410038.

    • Search Google Scholar
    • Export Citation
  • 17.

    Roux V, Fournier PE, Raoult D, 1996. Differentiation of spotted fever group rickettsiae by sequencing and analysis of restriction fragment length polymorphism of PCR-amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol 34: 20582065.

    • Search Google Scholar
    • Export Citation
  • 18.

    Regnery RL, Spruill CL, Plikaytis BD, 1991. Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes. J Bacteriol 173: 15761589.

    • Search Google Scholar
    • Export Citation
  • 19.

    Choi YJ, Jang WJ, Ryu JS, Lee SH, Park KH, Paik HS, Koh YS, Choi MS, Kim IS, 2005. Spotted fever group and typhus group rickettsioses in humans, South Korea. Emerg Infect Dis 11: 237244.

    • Search Google Scholar
    • Export Citation
  • 20.

    Bello S, Rodríguez M, Paredes A, Mendivelso F, Walteros D, Rodríguez F, Realpe ME, 2013. Comportamiento de la vigilancia epidemiológica de leptospirosis humana en Colombia, 2007–2011. Biomedica 33: 153160.

    • Search Google Scholar
    • Export Citation
  • 21.

    Bharti AR, et al., 2003. Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3: 757771.

  • 22.

    Arroyave E, Londoño AF, Quintero JC, Agudelo-Flórez P, Arboleda M, Díaz FJ, Rodas JD, 2013. Etiología y caracterización epidemiológica del síndrome febril no palúdico en tres municipios del Urabá antioqueño, Colombia. Biomedica 33: 99107.

    • Search Google Scholar
    • Export Citation
  • 23.

    Acosta J, et al., 2006. Brote de rickettsiosis en Necoclí, Antioquia, febrero-marzo de 2006. Informe Quincenal Epidemiológico Nacional 11: 177192.

    • Search Google Scholar
    • Export Citation
  • 24.

    Pacheco-García OE, Giraldo MR, Hidalgo M, Galeano A, Echeverri I, Echavarría-Rodríguez L, Parra E, Rey G, 2008. Estudio de brote febril hemorrágico en el corregimiento de Alto de Mulatos: Distrito Especial Portuario de Turbo, Antioquia, enero de 2008. Informe Quincenal Epidemiológico Nacional 13: 145160.

    • Search Google Scholar
    • Export Citation
  • 25.

    Delisle J, Mendell NL, Stull-Lane A, Bloch KC, Bouyer DH, Moncayo AC, 2016. Human infections by multiple spotted fever group rickettsiae in Tennessee. Am J Trop Med Hyg 94: 12121217.

    • Search Google Scholar
    • Export Citation
  • 26.

    Mattar S, Parra M, 2006. Detection of antibodies to Anaplasma, Bartonella, and Coxiella in rural inhabitants of the Caribbean area of Colombia. Rev MVZ Cordoba 11: 781789.

    • Search Google Scholar
    • Export Citation
  • 27.

    Rar V, Golovljova I, 2011. Anaplasma, Ehrlichia, and “Candidatus Neorhrlichia” bacteria: pathogenicity, biodiversity, and molecular genetic characteristics, a review. Infect Genet Evol 11: 18421861.

    • Search Google Scholar
    • Export Citation
  • 28.

    Guglielmone AA, et al., 2006. Ticks (Ixodidae) on humans in South America. Exp Appl Acarol 40: 83100.

  • 29.

    Santos HA, et al., 2013. Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil. Parasit Vectors 6: 348.

    • Search Google Scholar
    • Export Citation
  • 30.

    Estrada-Peña A, Guglielmone AA, Mangold AJ, 2004. The distribution and ecological ‘preferences’ of the tick Amblyomma cajennense (Acari: Ixodidae), an ectoparasite of humans and other mammals. Ann Trop Med Parasitol 98: 283292.

    • Search Google Scholar
    • Export Citation
  • 31.

    Wells EA, D'Alessandro A, Morales GA, Angel D, 1981. Mammalian wildlife diseases as hazards to man and livestock in an area of the Llanos Orientales of Colombia. J Wildl Dis 17: 153162.

    • Search Google Scholar
    • Export Citation
  • 32.

    Paternina LE, Díaz-Olmos Y, Paternina-Gómez M, Bejarano EE, 2009. Canis familiaris, Un nuevo hospedero de Ornithodoros (A.) puertoricensis FOX, 1947 (Acari: Ixodida) en Colombia. Acta Biolo Colomb 14: 153160.

    • Search Google Scholar
    • Export Citation
  • 33.

    Londoño AF, Díaz FJ, Valbuena G, Gazi M, Labruna M, Hidalgo M, Mattar S, Contreras V, Rodas JD, 2014. Infection of Amblyomma ovale by Rickettsia sp. strain Atlantic rainforest, Colombia. Ticks Tick Borne Dis 5: 672675.

    • Search Google Scholar
    • Export Citation
  • 34.

    Miranda J, Portillo A, Oteo JA, Mattar S, 2012. Rickettsia sp. strain Colombianensi (Rickettsiales: Rickettsiaceae): a new proposed Rickettsia detected in Amblyomma dissimile (Acari: Ixodidae) from iguanas and free-living larvae ticks from vegetation. J Med Entomol 49: 960965.

    • Search Google Scholar
    • Export Citation
  • 35.

    Miranda J, Mattar S, 2014. Molecular detection of Rickettsia bellii and Rickettsia sp. strain Colombianensi in ticks from Cordoba, Colombia. Ticks Tick Borne Dis 5: 208212.

    • Search Google Scholar
    • Export Citation
  • 36.

    Faccini-Martínez ÁA, Ramírez-Hernández A, Forero-Becerra E, Cortés-Vecino JA, Escandón P, Rodas JD, Palomar AM, Portillo A, Oteo JA, Hidalgo M, 2016. Molecular evidence of different Rickettsia species in Villeta, Colombia. Vector Borne Zoonotic Dis 16: 8587.

    • Search Google Scholar
    • Export Citation
  • 37.

    Ramírez-Hernández A, Escandón P, Cortés-Vecino JA, Rodas JD, Hidalgo M, 2013. Detección molecular de Rickettsia spp. (Da Rocha-Lima, 1916) en garrapatas recolectadas en tres regiones de Colombia. Acta Med Costarric 55: 8384.

    • Search Google Scholar
    • Export Citation
  • 38.

    Bermúdez SE, Eremeeva ME, Karpathy SE, Samudio F, Zambrano ML, Zaldivar Y, Motta JA, Dasch GA, 2009. Detection and identification of rickettsial agents in ticks from domestic mammals in eastern Panama. J Med Entomol 46: 856861.

    • Search Google Scholar
    • Export Citation
  • 39.

    Merhej V, Angelakis E, Socolovschi C, Raoult D, 2014. Genotyping, evolution and epidemiological findings of Rickettsia species. Infect Genet Evol 25: 122137.

    • Search Google Scholar
    • Export Citation
  • 40.

    Monje LD, Costa FB, Colombo VC, Labruna MB, Antoniazzi LR, Gamietea I, Nava S, Beldomenico PM, 2016. Dynamics of exposure to Rickettsia parkeri in cattle in the Paraná River Delta, Argentina. J Med Entomol 53: 660665.

    • Search Google Scholar
    • Export Citation
  • 41.

    Pesquera C, Portillo A, Palomar AM, Oteo JA, 2015. Investigation of tick-borne bacteria (Rickettsia spp., Anaplasma spp., Ehrlichia spp. and Borrelia spp.) in ticks collected from Andean tapirs, cattle and vegetation from a protected area in Ecuador. Parasit Vectors 8: 46.

    • Search Google Scholar
    • Export Citation
  • 42.

    Blanton LS, Mendell NL, Walker DH, Bouyer DH, 2014. Rickettsia amblyommii” induces cross protection against lethal Rocky Mountain spotted fever in a guinea pig model. Vector Borne Zoonotic Dis 14: 557562.

    • Search Google Scholar
    • Export Citation
  • 43.

    Rivas JJ, Moreira-Soto A, Alvarado G, Taylor L, Calderón-Arguedas O, Hun L, Corrales-Aguilar E, Morales JA, Troyo A, 2015. Pathogenic potential of a Costa Rican strain of ‘Candidatus Rickettsia amblyommii’ in guinea pigs (Cavia porcellus) and protective immunity against Rickettsia rickettsii .Ticks Tick Borne Dis 6: 805811.

    • Search Google Scholar
    • Export Citation
  • 44.

    Walker DH, 2016. Changing dynamics of human-rickettsial interactions. Am J Trop Med Hyg 94: 34.

  • 45.

    Ogrzewalska M, Saraiva DG, Moraes-Filho J, Martins TF, Costa FB, Pinter A, Labruna MB, 2012. Epidemiology of Brazilian spotted fever in the Atlantic Forest, state of São Paulo, Brazil. Parasitology 139: 12831300.

    • Search Google Scholar
    • Export Citation
  • 46.

    Wood CL, Lafferty KD, 2013. Biodiversity and disease: a synthesis of ecological perspectives on Lyme disease transmission. Trends Ecol Evol 28: 239247.

    • Search Google Scholar
    • Export Citation

Author Notes

Address correspondence to Marylin Hidalgo, Departamento de Microbiología, Facultad de Ciencias, Pontificia Universidad Javeriana, Cra 7a No 43-82, Bogotá, Colombia. E-mail: hidalgo.m@javeriana.edu.co

Financial support: The study was supported by Departamento Administrativo de Ciencia, Tecnología e Innovación from Colombia (COLCIENCIAS, Code: 120351929098). Regulatory permits for this work were Permit in scientific research in biodiversity No. 005 of June 19, 2012, given by Corporación Autónoma Regional (CAR) de Cundinamarca and contract for access to genetic resources for scientific research without commercial interest No. 85 of 2013, given by Ministerio de Ambiente y Desarrollo Sostenible from Colombia.

Authors' addresses: Álvaro A. Faccini-Martínez, Programa de Pós-Graduação em Doenças Infecciosas, Centro de Ciências da Saúde, Universidade Federal do Espírito Santo, Vitória, ES, Brazil, E-mail: afaccini@gmail.com. Alejandro Ramírez-Hernández, Departamento de Medicina Veterinária Preventiva e Saúde Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brazil, E-mail: aramirezhe@unal.edu.co. Christian Barreto, Diego Millán, Elkin Valbuena, Andrea C. Sánchez-Alfonso, and Marylin Hidalgo, Grupo de Enfermedades Infecciosas, Departamento de Microbiología, Pontificia Universidad Javeriana, Bogotá, Colombia, E-mails: davidbarreto_02@hotmail.com, dmillan19@hotmail.com, notengokorreo8@hotmail.com, asanchez-a@javeriana.edu.co, and hidalgo.m@javeriana.edu.co. Elkin Forero-Becerra, Departamento de Medicina Veterinária, Universidade Federal de Viçosa, Viçosa, MG, Brazil, E-mail: egforerob@unal.edu.co. Wilson O. Imbacuán-Pantoja and Jesús A. Cortés-Vecino, Grupo Parasitología Veterinaria, Universidad Nacional de Colombia, Ciudad Universitaria, Bogotá, Colombia, E-mails: woimbacuanp@unal.edu.co and jacortesv@unal.edu.co. Luis J. Polo-Terán and Néstor Yaya-Lancheros, Grupo Salud Pública Veterinaria, Departamento Salud Animal, Facultad de Medicina Veterinaria y de Zootecnia, Universidad Nacional de Colombia, Bogotá, Colombia, E-mails: luijpolot@unal.edu.co and nyayal@unal.edu.co. Jorge Jácome, Departamento de Biología, Pontificia Universidad Javeriana, Bogotá, Colombia, E-mail: jacomej@javeriana.edu.co. Ana M. Palomar, Sonia Santibáñez, Aránzazu Portillo, and José A. Oteo, Infectious Diseases Department, Center of Rickettsioses and Arthropod-borne Diseases, Hospital San Pedro–CIBIR, Logroño, Spain, E-mails: ampalomar@riojasalud.es, ssantibanez@riojasalud.es aportillo@riojasalud.es, and jaoteo@riojasalud.es.

Save