Virome analyses of Amblyomma cajennense and Rhipicephalus microplus ticks collected in Colombia.

Tick-borne viruses (TBV) have gained public health relevance in recent years due to the recognition of human-associated fatal cases and the increase in tick-borne disease and transmission. However, many tick species have not been studied for their potential to transmit pathogenic viruses, especially those found in Latin America. To gain better understanding of the tick virome, we conducted targeted amplification using broadly-reactive consensus-degenerate pan-viral targeting viruses from the genera Flavivirus, Bandavirus, Uukuvirus, and Orthonairovirus genus. Additionally, we conducted unbiased metagenomic analyses to investigate the presence of viral RNA sequences in Amblyomma cajennense, A. patinoi and Rhipicephalus microplus ticks collected from a horse slaughter plant in Medellín, Colombia. While no viral products were detected by PCR, results of the metagenomic analyses revealed the presence of viral genomes belonging to the genera Phlebovirus, Bandavirus, and Uukuvirus, including Lihan Tick Virus (LTV), which was previously reported in Rhipicephalus microplus from Colombia. Overall, the results emphasized the enormous utility of the next-generation sequencing in identifying virus genetic diversity presents in ticks and other species of vectors and reservoirs.

Leptospira Infection in Rural Areas of Urabá Region, Colombia: A Prospective Study.

The objective of this study was to analyze the eco-epidemiological aspects of Leptospira seroprevalence and seroincidence and its associated factors in two municipalities of northwest Colombia. A prospective study was performed in rural areas of Urabá, Antioquia, Colombia. The study enrolled 597 people between November 2015 and January 2016, of which 274 were followed up 1 year later. Serologic testing was performed by a microscopic agglutination. The outcomes were seroprevalent and seroincident cases, and the primary exposure was an outdoor occupation. A binary and mixed-effect multinomial logistic regression model was used to estimate factors associated with seroprevalent or seroincident cases of Leptospira infection. The overall Leptospira seroprevalence was 27.81% (95% confidence interval: 23.62-32.49), and the overall cumulative seroincidence for Leptospira was 14.60% (95% confidence interval: 10.33-20.23). Multivariable analysis showed that factors associated with L. interrogans serogroups seropositivity were outdoor occupation, male gender, older age, the presence of dirt floor in the household, and the presence of piglets and opossums. It also showed that factors associated with other Leptospira species serogroups were the presence of pit latrines and of turkeys. In addition, the multivariable model of seroincident cases of L. interrogans serogroups evidenced outdoor occupations, the presence of rats, and corn cultivation as risk factors. Likewise, the multivariable model for seroincident cases of other Leptospira species showed that the presence of hunting canines and cassava cultivation were risk factors. We found specific factors associated with the transmission of Leptospira serogroups contribute to the understanding of the epidemiology of Leptospira infection in rural areas of Urabá, Colombia.

Presence of two species of the Amblyomma cajennense complex (Acari: Ixodidae) and probable zones of sympatry in northwestern Colombia.

The A. cajennense tick complex has been thought to be the main vector of Rickettsia rickettsii in Central and South America. Studies in Colombia have determined the presence of species A. patinoi and A. mixtum of the A. cajennense complex, but it is unknown which species of this complex exist in northwestern Colombia. Our aim was to identify the species of the A. cajennense complex that are present in northwestern Colombia. We sampled ticks of A. cajennense sensu lato infesting equids. Females identified according to the morphology of their genital pore were selected for genetic confirmation. Specimens from each locality were selected to perform molecular and genetic analysis. Specimens were analyzed from five departments (Antioquia, Bolívar, Córdoba, Magdalena, and Sucre). Morphologically 65 specimens were identified as A. patinoi and 5 as A. mixtum. Molecular analysis allowed to confirm the morphological identification of 27 specimens. In this study A. patinoi was widely distributed in the departments of Antioquia, Bolívar, and Córdoba with allopatric and sympatric distribution in some places. These two species in the region could have unexpected effects on the epidemiology of rickettsiosis.

Epidemiology of Rickettsial Infection in the Municipality of Uramita, Colombia.

This study aimed to analyze epidemiological indicators related to seroprevalent and seroincident cases of spotted fever group rickettsiae (SFGR) and to identify housing conditions related to tick infestation. A prospective study (2016-2018) was conducted to examine rickettsial seropositivity in humans, domestic animals, and wild mammals in the municipality of Uramita, Antioquia, Colombia, where a previous Rickettsia rickettsii outbreak was reported from 2014 to 2015. The seroprevalence and its associated factors were estimated at baseline, and the seroincidence and its risk factors for humans were estimated 20 months later. A cross-sectional analysis was performed to evaluate the housing conditions linked to tick infestation. The SFGR seroprevalence was 26.70% (95% confidence interval [CI], 20.79-31.37), and the factors associated with SFGR seropositivity were male sex (adjusted prevalence ratio [PRa], 1.67; 95% CI, 1.19-2.32), age (evaluated in 5-year increments) (PRa, 1.05; 95% CI, 1.01-1.09), and household proximity (PRascattered vs. very near=3.87; 95% CI, 1.12-8.66). The overall seroincidence was 7.40% (95% CI, 4.71-11.06), and the factors associated with SFGR seroincident cases were the presence of wild animals (adjusted relative risk [RRa], 2.46; 95% CI, 1.06-4.72) and the presence of trees in the peri-domiciliary area (RRa, 0.47; 95% CI, 0.23-0.94). The prevalence of house tick infestation was 27.81% (95% CI, 21.93-34.80), and the factors associated with infestation were dirt floors (PRa, 5.88; 95% CI, 2.28-10.31), fiber cement roofs (PRa, 1.76; 95% CI, 1.07-2.83), and the presence of canines in peri-domiciliary or intra-domiciliary areas (PRa, 5.05; 95% CI, 3.31-7.19). Seropositivity rates for canines and opossums were 35.62% (26/73) and 100% (6/6), respectively. Identification of these factors will help to implement efficient surveillance programs in Colombia.

Human Rickettsiosis Caused by Rickettsia parkeri Strain Atlantic Rainforest, Urabá, Colombia.

We describe the clinical, serologic, and molecular findings of a new human rickettsiosis in Colombia. Antibodies against Rickettsia spp. were detected. PCR showed amplification of genes for R. parkeri strain Atlantic Rainforest. This new rickettsiosis of minor virulence could explain some of the undifferentiated acute febrile diseases in Colombia.

Whole-Genome Sequence of Rickettsia parkeri Strain Atlantic Rainforest, Isolated from a Colombian Tick.

Rickettsia parkeri is classified as a member of the alphaproteobacterial microorganisms, genus Rickettsia Here, we report the complete genome sequence of Rickettsia parkeri strain Atlantic Rainforest, which was isolated from an Amblyomma ovale tick collected in the municipality of Necoclí, Colombia.

Infection by Rickettsia felis in Ctenocephalides felis felis Fleas from North of Colombia.

BACKGROUND: Rickettsia felis is an emergent Rickettsial agent whose main vector is Ctenocephalides felis, but ticks, mites and lice are also infected. We aimed to search for molecular evidence of Rickettsia spp. in fleas collected from dogs and wild rodents (Heteromys anomalous) from three villages of Córdoba and Antioquia provinces (Northern of Colombia), where outbreaks of rickettsioses have occurred, and discuss the possible role of fleas on endemic/enzootic regions for rickettsia. METHODS: During 2010 and 2012, 649 Ctenocephalides felis felis and 24 Pulex irritans fleas were removed from dogs and wild rodents (Heteromys anomalous), respectively, in 3 locations from Córdoba and Antioquia provinces (Colombia). These fleas were tested into pools for Rickettsial infection by PCR, targeting gltA, ompB, and ompA Rickettsial genes. RESULTS: Almost 20% (30/153) of C. felis felis pools contained Rickettsial DNA. The fragments of ompB gene showed high identity values between sequences from Necocli and Los Cordobas with R. felis strain from Senegal (100% and 99.7% respectively) and all were highly related by phylogenetic analyses. Rickettsial DNA in pools of P. irritans was not detected. CONCLUSION: Our findings highlighted the endemicity of the infection by R. felis in fleas from northern of Colombia and showed the likely importance of dogs as hosts of C. felis felis fleas and their potential role as reservoirs of R. felis.

A Cluster of Cases of Rocky Mountain Spotted Fever in an Area Of Colombia Not Known to be Endemic for This Disease.

Rocky Mountain spotted fever (RMSF) has been reported in Colombia since 1937. Most recent outbreaks were in 2006-2008, followed by the absence of recognized cases. This report describes new clinical cases of RMSF and epidemiologic investigations. Medical records were reviewed, and fieldwork, serological and molecular diagnostic testing, and bacterial isolation were performed. Fever, hypotension, abdominal discomfort, and rash accompanied by thrombocytopenia and leukopenia were the most characteristic manifestations. Two convalescent sera from the index case and sera from two dogs revealed high anti-spotted fever group Rickettsia antibody titers. Rickettsia rickettsii was isolated from case 4. Cases 2 and 3 were identified by epidemiological connection with the index case. Thus, a new cluster of cases of RMSF was identified in Antioquia, Colombia, with the occurrence of fatal cases, which indicates the active circulation of the bacteria and a potential risk for the population.

Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks / Diseño y validación analítica de una PCR dúplex para la detección de Ehrlichia y Rickettsia en garrapatas / Projeto e validação analítica de uma PCR duplex para detecção de Ehrlichia e Rickettsia em carrapatos

Abstract Background: Ehrlichia and Rickettsia are two major rickettsial genera transmitted by ticks that affect a number of wild and domestic animal species and human populations around the world. Objective: To design and validate a duplex PCR for Ehrlichia and Rickettsia in ticks. Methods: Assay validation included testing for sensitivity, specificity, reproducibility, and robustness of the PCR. The groEL and 23sr RNA genes were used for Ehrlichia and Rickettsia, respectively. Results: The limit of detection was one hundred gene copies per 50 μL of reaction for Ehrlichia spp, and one gene copy of Rickettsia per 50 μL of reaction. In general, the primers of the test only amplified in silico those bacterial agents for which they were originally designed, with the exception of the primers for Rickettsia that also amplified Methylocystis sp. The test was reproducible (intermediate precision) 96.7% of the times for both agents. The test was robust enough to tolerate concentration changes of all reagents with the exception of Taq DNA polymerase. Conclusions: The validation results indicated that this PCR is useful for detection in both bacterial genera and it is a good candidate for diagnostic validation.

Resumen Antecedentes: Ehrlichia spp. y Rickettsia spp. son dos de los principales géneros rickettsiales transmitidos por garrapatas que afectan a animales silvestres, domésticos y humanos alrededor del mundo. Objetivo: Diseñar y validar una prueba PCR dúplex para Ehrlichia y Rickettsia en garrapatas. Métodos: La validación de la prueba incluyó ensayos de sensibilidad, especificidad, reproducibilidad y robustez. En la PCR se usó groEL y ARNr 23S como genes blanco para Ehrlichia y Rickettsia, respectivamente. Resultados: El límite de detección fue de 100 copias del gen por 50 μL de reacción para Ehrlichia spp y una copia del gen de Rickettsia por 50 μL de reacción. En general, los cebadores de la prueba solo amplificaron in silico los agentes bacterianos para los cuales fueron originalmente diseñados, con la excepción de los cebadores de Rickettsia que también amplificaron Methylocystis sp. La prueba fue reproducible (precisión intermedia) en un 96.7% de las veces para ambos agentes. La prueba fue suficientemente robusta como para tolerar cambios de concentración de los diferentes reactivos, con excepción de la Taq DNA polimerasa. Conclusión: Los resultados de validación indican que la PCR es útil para detectar ambos géneros bacterianos y podria usarse para validación diagnostica.

Resumo Antecedentes: Ehrlichia e Rickettsia são dois dos principais gêneros de rickettsias transmitidos por carrapatos que infectam tanto animais selvagens quanto animais domésticos e até homens em todo o mundo. Objetivo: O objetivo principal foi elaborar e validar uma PCR duplex para Ehrlichia e Rickettsia em carrapatos. Métodos: A validação incluiu testes de sensibilidade, especificidade, reprodução e robustez. Para o PCR, utilizamos os genes groEl e 23Sr-RNA para Ehrlichia e Rickettsia, respectivamente. Resultados: O limite de detecção foi de 100 cópias de genes por 50 ml de reação para Erliquia spp e uma cópia de gene de Rickettsia por 50 ml de reação. Em geral, os iniciadores dos testes amplificaram em modelos computacionais os agentes bacterianos para os quais eles foram projetados, exceto os primers de Rickettsia que também amplificou Methylocystis sp. Os testes foram reproduzíveis (precisão intermediária) 96,7% para ambos os agentes e foram também robustos para tolerar mudanças de concentração em todos os reagentes, exceto o reagente Taq DNA polymerase. Conclusões: Os resultados da validação indicaram que o PCR é útil para detecção em ambos os gêneros bacterianos, portanto, um bom exame para validação diagnóstica.

Modelos potenciales de distribución geográfica y climática del complejo Amblyomma cajennense (Acari: Ixodidae), potencial vector de Rickettsia rickettsii en Colombia / Potential models of the geographic and climatic distribution of the Amblyomma cajennense complex (Acari: Ixodidae), a potential vector of Rickettsia rickettsii in Colombia

Resumen Introducción. Las especies del complejo Amblyomma cajennense, potenciales vectores implicados en la transmisión de Rickettsia rickettsii en Centroamérica y Suramérica, presentan una amplia distribución geográfica en el neotrópico. En Colombia, todavía es incipiente el conocimiento sobre las especies de este complejo, su distribución y su ecología. Objetivo. Explorar la ecología del complejo A. cajennense en Colombia a partir del estudio de su selección de hábitat y la generación de modelos potenciales de distribución geográfica y climática. Materiales y métodos. Se utilizaron registros únicos de la presencia de estas garrapatas en el estudio de sus patrones de selección del hábitat, mediante el análisis factorial del nicho ecológico (Ecological Niche Factor Analysis, ENFA), y se construyeron modelos de distribución potencial con las metodologías MaxEnt y GARP. Resultados. El complejo A. cajennense se encontró en las cinco regiones naturales de Colombia, con 64,71 % de los registros procedentes de los departamentos de Antioquia, Cundinamarca y Huila. La selección del hábitat evidenció que estas garrapatas escogen preferentemente las áreas geográficas (ENFA: 96,03 %) según sus condiciones de índice de estabilidad térmica y de precipitación en el trimestre más seco del año. Los modelos de distribución potencial indicaron la idoneidad del hábitat en 31 a 51 % del área del país dependiendo del algoritmo empleado. Conclusiones. En Colombia, el complejo A. cajennense presenta una distribución muy influenciada por el comportamiento en la selección del hábitat climático, característica que facilitó la creación de modelos de distribución. En futuros estudios de campo, se podrían validar los modelos presentados; se requieren, además, otros estudios genéticos para la identificación de especies del complejo y el manejo de sus potenciales zonas de hibridación en el país.

Abstract Introduction: The species of the Amblyoma cajennense complex, potential vectors involved in the transmission of Rickettsia rickettsii in Central and South America, have a wide geographical distribution in the Neotropics. In Colombia, the knowledge about these species, their distribution, and many aspects of their ecology is still limited. Objective: To explore the ecology of A. cajennense complex in Colombia based on the study of its habitat selection, as well as the generation of potential geographic and climatic distribution models. Materials and methods: We used unique records of these ticks to study the selection patterns of the climatic habitat using the Ecological Niche Factor Analysis (ENFA), and we built distribution models using the MaxEnt and GARP algorithms. Results: The record of distributions showed that A. cajennense complex is present in all five natural regions of Colombia with 64.71% of the records found in the departments of Antioquia, Cundinamarca, and Huila. The analysis of their habitat selection showed that these ticks preferably select geographical areas (ENFA: 96.03%) according to the isothermality conditions and the precipitation of the driest quarter of the year. The potential distribution models indicated habitat suitability in 31 to 51% of the area of the country depending on the algorithm. Conclusions: The distribution of A. cajennense complex in Colombia is highly influenced by a climatebased habitat selection, a characteristic that favored the creation of distribution models. Field studies will allow the validation of the potential distribution models and further genetic studies will be required in the country to identify species within the complex and study their potential hybridization areas.

Potential models of the geographic and climatic distribution of the Amblyomma cajennense complex (Acari: Ixodidae), a potential vector of Rickettsia rickettsii in Colombia / Modelos potenciales de distribución geográfica y climática del complejo Amblyomma cajennense (Acari: Ixodidae), potencial vector de Rickettsia rickettsii en Colombia

Introduction: The species of the Amblyoma cajennense complex, potential vectors involved in the transmission of Rickettsia rickettsii in Central and South America, have a wide geographical distribution in the Neotropics. In Colombia, the knowledge about these species, their distribution, and many aspects of their ecology is still limited. Objective: To explore the ecology of A. cajennense complex in Colombia based on the study of its habitat selection, as well as the generation of potential geographic and climatic distribution models. Materials and methods: We used unique records of these ticks to study the selection patterns of the climatic habitat using the Ecological Niche Factor Analysis (ENFA), and we built distribution models using the MaxEnt and GARP algorithms. Results: The record of distributions showed that A. cajennense complex is present in all five natural regions of Colombia with 64.71% of the records found in the departments of Antioquia, Cundinamarca, and Huila. The analysis of their habitat selection showed that these ticks preferably select geographical areas (ENFA: 96.03%) according to the isothermality conditions and the precipitation of the driest quarter of the year. The potential distribution models indicated habitat suitability in 31 to 51% of the area of the country depending on the algorithm. Conclusions: The distribution of A. cajennense complex in Colombia is highly influenced by a climatebased habitat selection, a characteristic that favored the creation of distribution models. Field studies will allow the validation of the potential distribution models and further genetic studies will be required in the country to identify species within the complex and study their potential hybridization areas.

Introducción. Las especies del complejo Amblyomma cajennense, potenciales vectores implicados en la transmisión de Rickettsia rickettsii en Centroamérica y Suramérica, presentan una amplia distribución geográfica en el neotrópico. En Colombia, todavía es incipiente el conocimiento sobre las especies de este complejo, su distribución y su ecología.Objetivo. Explorar la ecología del complejo A. cajennense en Colombia a partir del estudio de su selección de hábitat y la generación de modelos potenciales de distribución geográfica y climática.Materiales y métodos. Se utilizaron registros únicos de la presencia de estas garrapatas en el estudio de sus patrones de selección del hábitat, mediante el análisis factorial del nicho ecológico (Ecological Niche Factor Analysis, ENFA), y se construyeron modelos de distribución potencial con las metodologías MaxEnt y GARP.Resultados. El complejo A. cajennense se encontró en las cinco regiones naturales de Colombia, con 64,71 % de los registros procedentes de los departamentos de Antioquia, Cundinamarca y Huila. La selección del hábitat evidenció que estas garrapatas escogen preferentemente las áreas geográficas (ENFA: 96,03 %) según sus condiciones de índice de estabilidad térmica y de precipitación en el trimestre más seco del año. Los modelos de distribución potencial indicaron la idoneidad del hábitat en 31 a 51 % del área del país dependiendo del algoritmo empleado.Conclusiones. En Colombia, el complejo A. cajennense presenta una distribución muy influenciada por el comportamiento en la selección del hábitat climático, característica que facilitó la creación de modelos de distribución. En futuros estudios de campo, se podrían validar los modelos presentados; se requieren, además, otros estudios genéticos para la identificación de especies del complejo y el manejo de sus potenciales zonas de hibridación en el país.

Wild and domestic animals likely involved in rickettsial endemic zones of Northwestern Colombia.

Between 2006 and 2008, three outbreaks of human rickettsiosis occurred in Northwestern Colombia (municipalities of Necoclí, Los Córdobas and Turbo), with case fatality rates between 27% and 54%. The aim of this study was to determine previous exposure of wild and domestic animals to spotted fever group (SFG) rickettsiae through serological tests, to detect rickettsial evidence in their ectoparasites, and to analyze their possible role in the epidemiology of rickettsial diseases in this zone of the country. A cross-sectional association study was performed from 2010 to 2011. Blood and ectoparasite samples were collected from domestic animals and small mammals. A statistically significant association (p<0.05) between seropositive animals and the study zones was observed. A total of 2937 ticks, 672 fleas and 74 lice were collected and tested in pools by PCR. The minimum infection rate (MIR) of the positive pools was 5% in ticks, 4% in fleas, and 0% in lice. Phylogenetic analyses showed circulation of three 4.Rickettsia species: R. felis in fleas, and R. bellii and Rickettsia sp. strain Atlantic rainforest, both in Amblyomma ovale ticks. In conclusion, this study demonstrated the occurrence of SFG rickettsiae in domestic, synanthropic and wild animals, and suggests the use of equines and canines as good sentinels of infection, in the study zone. We speculate that a transmission cycle exist involving rodents in the areas where these outbreaks have occurred. Tomes' spiny rats (Proechimys semispinosus) and common opossums (Didelphis marsupialis) could be good candidates as amplifier hosts for SFG rickettsiae in enzootic/endemic zones.

[First evidence of lymphocytic choriomeningitis virus (Arenavirus) infection in Mus musculus rodents captured in the urban area of the municipality of Sincelejo, Sucre, Colombia].

INTRODUCTION: The lymphocytic choriomeningitis virus is an Old World arenavirus that infects Mus musculus, and can cause congenital hydrocephalus, chorioretinitis and multisystemic failure in transplant human recipients. Although the disease has not been clinically diagnosed in Colombia yet, there have been reports of infection with the Pichindé virus in rodents from Cauca and Valle del Cauca departments, and with the Guanarito virus in rodents from Córdoba department. OBJECTIVE: To identify the lymphocytic choriomeningitis virus from Mus musculus captured in the municipality of Sincelejo. MATERIALS AND METHODS: We evaluated 80 samples of plasma by ELISA using antigen from lymphocytic choriomeningitis virus. Additionally, a nested RT-PCR was performed to seropositive and seronegative samples for the S-segment. RESULTS: We found a 10% seroprevalence (8/80) and the viral genome was detected in 16 brain samples; the alignment (BLAST) and the phylogenetic analysis (MrBayes, version 3.2.2) confirmed the presence of the lymphocytic choriomeningitis virus. CONCLUSION: The results indicated that human infection with the lymphocytic choriomeningitis virus in humans could occur in the urban area of Sincelejo, although no cases have been reported so far.

Primera evidencia de infección por el virus de la coriomeningitis linfocítica (arenavirus) en roedores Mus musculus capturados en la zona urbana del municipio de Sincelejo, Sucre, Colombia / First evidence of lymphocytic choriomeningitis virus (Arenavirus) infection in Mus musculus rodents captured in the urban area of the municipality of Sincelejo, Sucre, Colombia

Resumen Introducción. El virus de la coriomeningitis linfocítica es un arenavirus del Viejo Mundo que se hospeda en el ratón casero (Mus musculus), y puede causar infecciones congénitas, hidrocefalia, coriorretinitis y falla orgánica múltiple en pacientes receptores de trasplantes. En Colombia aún no se ha reportado la enfermedad mediante diagnóstico clínico, pero en estudios serológicos se ha detectado la infección por el virus Pichindé en roedores en los departamentos del Cauca y Valle del Cauca, y por el virus Guanarito, en roedores en Córdoba. Objetivo. Detectar el virus de la coriomeningitis linfocítica en M. musculus en el municipio de Sincelejo. Materiales y métodos. Se evaluaron 80 muestras de plasma mediante la prueba ELISA usando antígeno del virus de la coriomeningitis linfocítica. Además, se empleó la reacción en cadena de la polimerasa con transcriptasa inversa (RT-PCR) anidada en muestras de animales seropositivos y seronegativos para la detección del segmento S. Resultados. Se encontró una seroprevalencia de 10% (8/80) y se detectó el genoma viral en 16 muestras de cerebro; el alineamiento (en la Basic Local Alignment Search Tool, BLAST) y el análisis filogenético (mediante el programa MrBayes, versión 3.2.2) confirmaron que correspondía al virus de la coriomeningitis linfocítica. Conclusión. Los resultados indicaron que la infección por el virus de la coriomeningitis linfocítica en humanos podría ocurrir en el área urbana de Sincelejo, aunque hasta la fecha no se hayan reportado casos.

Abstract Introduction: The lymphocytic choriomeningitis virus is an Old World arenavirus that infects Mus musculus, and can cause congenital hydrocephalus, chorioretinitis and multisystemic failure in transplant human recipients. Although the disease has not been clinically diagnosed in Colombia yet, there have been reports of infection with the Pichindé virus in rodents from Cauca and Valle del Cauca departments, and with the Guanarito virus in rodents from Córdoba department. Objective: To identify the lymphocytic choriomeningitis virus from Mus musculus captured in the municipality of Sincelejo. Materials and methods: We evaluated 80 samples of plasma by ELISA using antigen from lymphocytic choriomeningitis virus. Additionally, a nested RT-PCR was performed to seropositive and seronegative samples for the S-segment. Results: We found a 10% seroprevalence (8/80) and the viral genome was detected in 16 brain samples; the alignment (BLAST) and the phylogenetic analysis (MrBayes, version 3.2.2) confirmed the presence of the lymphocytic choriomeningitis virus. Conclusion: The results indicated that human infection with the lymphocytic choriomeningitis virus in humans could occur in the urban area of Sincelejo, although no cases have been reported so far.

Human prevalence of the spotted fever group (SFG) rickettsiae in endemic zones of Northwestern Colombia.

In February 2006, an outbreak of human rickettsiosis occurred in the municipality of Necoclí Colombia, with 35% of lethality. This episode was, followed by two more, one in the municipality of Los Cordobas in 2007 with a 54% of lethality and the other one in the municipality of Turbo in 2008 with 27% of lethality. The aim of this study was to perform serological tests in healthy persons to determine the seroprevalence of antibodies against spotted fever group (SFG) rickettsiae and develop a survey to study some infection risk-related factors. A cross-sectional study was performed in 2011 and 2012. A blood sample and survey of associated factors was performed in healthy persons. A prevalence of 32%-41% was found in healthy people. From the multivariate analysis, we found that people living more than 16 years in these sites had a 79% higher risk of being seropositive and a 46% higher risk when they reported having birds in their houses if the variable of having a horse was included in the model. In conclusion, this study shows endemicity of at least one spotted fever group Rickettsia in the study zone.

[Serological evidence of Leptospira spp., dengue, hantavirus and arenavirus infection in Embera-Katío indigenous population, Colombia]. / Evidencia serológica retrospectiva de infecciones por Leptospira spp., dengue, hantavirus y arenavirus en indígenas Emberá-Katío, Colombia.

We have developed a study in the Embera-Katío indigenous community and we found that the seropositivity for IgG antibodies to Leptospira spp. was 18.1%; to dengue was 61.1%; to arenavirus was 3.1% and to hantavirus was 1.5%. We conclude that this indigenous community live in conditions that promote the reemerging and emerging, infections.

Evidencia serológica retrospectiva de infecciones por Leptospira spp, dengue, hantavirus y arenavirus en indígenas Emberá-Katío, Colombia / Serological evidence of Leptospira spp, dengue, hantavirus and arenavirus infection in Embera-Katío indigenous population, Colombia

We have developed a study in the Embera-Katío indigenous community and we found that the seropositivity for IgG antibodies to Leptospira spp. was 18.1%; to dengue was 61.1%; to arenavirus was 3.1% and to hantavirus was 1.5%. We conclude that this indigenous community live in conditions that promote the reemerging and emerging, infections.

Genetic Characterization of Northwestern Colombian Chikungunya Virus Strains from the 2014-2015 Epidemic.

Chikungunya fever, an acute and often chronic arthralgic disease caused by the mosquito-borne alphavirus, chikungunya virus (CHIKV), spread into the Americas in late 2013. Since then it has caused epidemics in nearly all New World countries, the second largest being Colombia with over 450,000 suspected cases beginning in September, 2014, and focused in Bolivar Department in the north. We examined 32 human sera from suspected cases, including diverse age groups and both genders, and sequenced the CHIKV envelope glycoprotein genes, known determinants of vector host range. As expected for Asian lineage CHIKV strains, these isolates lacked known Aedes albopictus-adaptive mutations. All the Colombian strains were closely related to those from the Virgin Islands, Saint Lucia, Mexico, Puerto Rico, and Brazil, consistent with a single, point-source introduction from the southeast Asia/Pacific region. Two substitutions in the E2 and E1 envelope glycoprotein genes were found in the Colombian strains, especially E1-K211E involving a residue shown previously to affect epistatically the penetrance of the E1-A226V A. albopictus-adaptive substitution. We also identified two amino acid substitutions unique to all American CHIKV sequences: E2-V368A and 6K-L20M. Only one codon, 6K-47, had a high nonsynonymous substitution rate suggesting positive selection.

Molecular Evidence of Different Rickettsia Species in Villeta, Colombia.

The aim of this work was to detect and identify Rickettsia species in ticks collected in rural areas of Villeta, Colombia. Tick specimens were collected from domestic animals and walls of houses in five rural villages of Villeta town and from humans in Naranjal village (same town). Moreover, a flea collected from the same area was also processed. DNA was extracted and tested by conventional, semi-nested, and nested PCR reactions targeting rickettsial genes. In the ticks collected from humans from Naranjal village, a nymph of Amblyomma cajennense sensu lato was amplified using primers for ompA and sequenced (100% identity with "Candidatus Rickettsia amblyommii"). Last, three amplicons from the Ctenocephalides felis flea, corresponding to gltA, ompB, and 16S rRNA genes, showed high identity with R. felis (98.5%, 97.3%, and 99.2%, respectively) and "Candidatus Rickettsia asemboensis" (99.7% and 100%, respectively). To our knowledge, these results correspond to the first molecular detection in Colombia of "Candidatus Rickettsia amblyommii" and "Ca. Rickettsia asemboensis" in fleas.