First isolation of Rickettsia amblyommatis from Amblyomma mixtum in Colombia.

BACKGROUND: Rickettsiae are obligate intracellular Gram-negative bacteria that are the causative agent of rickettsioses and are spread to vertebrate hosts by arthropods. There are no previous reports of isolation of Rickettsia amblyommatis for Colombia. METHODS: A convenience sampling was executed in three departments in Colombia for direct collection of adult ticks on domestic animals or over vegetation. Ticks were screened for the presence of Rickettsia spp. by real-time polymerase chain reaction (qPCR) amplifying the citrate synthase gene (gltA), and the positive sample was processed for isolation and further molecular characterization by conventional PCR. The absolute and relative frequencies were calculated for several tick species variables. All products from conventional PCR were further purified and sequenced by the Sanger technique. Representative sequences of 18 Rickettsia species were downloaded from GenBank. Consensus phylogenetic trees were constructed for the gltA, ompB, ompA, and htrA genes with 1000 replicates, calculating bootstrap values through the maximum likelihood method and the generalized time reversible substitution model in the MEGA 7.0 software program. RESULTS: One female Amblyomma mixtum collected on vegetation was amplified by qPCR (gltA), indicating a frequency of 1.6% (1/61) for Rickettsia spp. INFECTION: Sequence analysis of a rickettsial isolate from this tick in BLASTn showed 100% identity with gltA (340 base pairs [bp]), 99.87% for ompB (782 bp), 98.99% for htrA (497 bp), and 100% for ompA (488 bp) to R. amblyommatis. Concatenated phylogenetic analysis confirmed these findings indicating that the isolate is grouped with other sequences of Amblyomma cajennense complex from Panama and Brazil within the R. amblyommatis clade. CONCLUSIONS: This paper describes the isolation and early molecular identification of a R. amblyommatis strain from A. mixtum in 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.

Seroepidemiology of Sarcocystis neurona and Neospora spp. in horses, donkeys, and mules from Colombia.

Sarcocystis neurona and Neospora spp. are related protozoa that can cause equine protozoal myeloencephalitis (EPM). The present study aimed to determine the frequency of antibodies to these parasites in 649 equids (351 horses, 267 donkeys, and 31 mules) from six departments in the North and Northwest of Colombia. For this purpose, the indirect fluorescent antibody test (IFAT) was used for detecting antibodies against S. neurona and Neospora spp. with a cut-off point of 1:20 and 1:50, respectively. A binomial logistic regression model was selected to predict variables associated with exposure. The frequency of anti-S. neurona antibodies was 14.24% (95% CI: 10.84-18.44) for horses, 2.99% (95% CI: 1.39-6.04) for donkeys, and 16.13% (95% CI: 6.09-34.47) for mules. The risk for S. neurona infection was significantly lower in donkeys (OR: 0.18 [0.08-0.38]; p<0.001) than horses and mules, and higher in animals with a poor body condition (OR: 2.82 [1.45-6.05]; p<0.05). Additionally, older animals (>12y) had a higher risk of seropositivity (OR: 5.26 [1.88-19.1]; p<0.05), as well as animals that inhabit climatic conditions associated with tropical very dry forest (OR: 1.85 [1.01-3.51]; p<0.05). Córdoba and Antioquia departments presented the highest seropositivity to S. neurona with 13.01 and 8.3%, respectively. The frequency of anti-Neospora spp. antibodies was 1.42% (95% CI: 0.52-3.48) for horses, 1.12% (95% CI:0.29-3.52) for donkeys and 0% (95%, CI: 0-0) for mules. Atlántico was the state with the highest seropositivity to Neospora spp. (10%). No risks associated with Neospora spp. infection were found. These findings allow us to conclude that equids from these regions of Colombia are exposed to S. neurona, but antibodies to Neospora spp. are uncommon. Further studies are necessary to explore the presence of these two agents in other areas of the country. In addition, we need to prove the importance of the above-mentioned risk factors over the susceptibility of horses to these protozoal agents and the epidemiological impact of these underdiagnosed coccidia.

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.

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.

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.

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.