Distribución de Aedes albopictus en Ibagué: potencial riesgo de brotes de arbovirosis

Introducción. Aedes albopictus es un vector de arbovirus como dengue, Zika, chikungunya y fiebre amarilla. Los primeros reportes en el continente americano datan de 1985 y dada su capacidad de adaptación ecológica y fisiológica, se ha distribuido rápidamente en el territorio colombiano desde su primer reporte en 1998. Objetivo. Determinar la distribución de A. albopictus en las comunas de Ibagué, Colombia. Materiales y métodos. Los muestreos se realizaron entre mayo y noviembre de 2022 en zonas con abundante vegetación de las 13 comunas de Ibagué. Se emplearon aspiradores y redes entomológicas. Los mosquitos fueron transportados al Laboratorio de Investigaciones en Parasitología Tropical de la Universidad del Tolima para su determinación taxonómica. Resultados. Se identificaron 708 ejemplares de A. albopictus, distribuidos en las comunas de Ibagué. La mayor abundancia del vector se presentó en las comunas 10, 11, 7, 8, 2 y 9. Las comunas 3, 4, 5, 6, 12 y 13 presentaron abundancias relativas cercanas al 3 %, y la comuna 1 tuvo una abundancia del 2 %. Conclusiones. Aedes albopictus está distribuido en todas las comunas de Ibagué, probablemente su dispersión se ha visto favorecida por las condiciones ambientales y sociales de esta región. Se recomienda hacer seguimiento anual a las poblaciones de este vector y realizar una caracterización molecular de los arbovirus encontrados. Además, el conocer la distribución de este mosquito en la ciudad permitirá focalizar las estrategias de control entomológico y prevenir futuros brotes de arbovirosis.

Introduction. Aedes albopictus is a vector for arboviruses, such as dengue, Zika, chikungunya, and yellow fever. The first A. albopictus reports on the American continent date back to 1985. It has spread rapidly throughout Colombia since its first report in 1998 due to its ecological and physiological adaptation capability. Objective. To determine A. albopictus distribution in the 13 communes of Ibagué, Colombia. Materials and methods. Samples were collected between May and November 2022 in the 13 communes of Ibagué. Vacuum sampling and sweep-netting entomological nets were used in areas with abundant vegetation. The mosquitoes were transported to the Laboratorio de Investigaciones en Parasitología Tropical at the Universidad del Tolima for taxonomic determination. Results. We identified 708 A. albopictus specimens distributed throughout Ibague's 13 communes. The highest vector abundance occurred in communes 10, 11, 7, 8, 2, and 9; communes 3, 4, 5, 6, 12, and 13 had a relative abundance of around 3%, while commune 1 had 2% of relative abundance. Conclusions. Aedes albopictus is distributed throughout all the communes of Ibague. Its dispersion has probably been favored by this region's environmental and social conditions. We recommend annual monitoring of these vectors populations and molecular characterization of the found arboviruses. Ascertaining this mosquito's distribution throughout the city will enable focusing entomological control strategies and preventing future arbovirus outbreaks.

Aedes albopictus distribution in Ibagué, Colombia: Potential risk of arboviral outbreaks / Distribución de Aedes albopictus en Ibagué: potencial riesgo de brotes de arbovirosis

Introduction: Aedes albopictus is a vector for arboviruses, such as dengue, Zika, chikungunya, and yellow fever. The first A. albopictus reports on the American continent date back to 1985. It has spread rapidly throughout Colombia since its first report in 1998 due to its ecological and physiological adaptation capability. Objective: To determine A. albopictus distribution in the 13 communes of Ibagué, Colombia. Materials and methods: Samples were collected between May and November 2022 in the 13 communes of Ibagué. Vacuum sampling and sweep-netting entomological nets were used in areas with abundant vegetation. The mosquitoes were transported to the Laboratorio de Investigaciones en Parasitología Tropical at the Universidad del Tolima for taxonomic determination. Results: We identified 708 A. albopictus specimens distributed throughout Ibague's 13 communes. The highest vector abundance occurred in communes 10, 11, 7, 8, 2, and 9; communes 3, 4, 5, 6, 12, and 13 had a relative abundance of around 3%, while commune 1 had 2% of relative abundance. Conclusions: Aedes albopictus is distributed throughout all the communes of Ibague. Its dispersion has probably been favored by this region's environmental and social conditions. We recommend annual monitoring of these vectors populations and molecular characterization of the found arboviruses. Ascertaining this mosquito's distribution throughout the city will enable focusing entomological control strategies and preventing future arbovirus outbreaks.

Introducción: Aedes albopictus es un vector de arbovirus como dengue, Zika, chikungunya y fiebre amarilla. Los primeros reportes en el continente americano datan de 1985 y dada su capacidad de adaptación ecológica y fisiológica, se ha distribuido rápidamente en el territorio colombiano desde su primer reporte en 1998. OBJETIVO: Determinar la distribución de A. albopictus en las comunas de Ibagué, Colombia. Materiales y métodos: Los muestreos se realizaron entre  mayo y noviembre de 2022 en zonas con abundante  vegetación de las 13 comunas de Ibagué. Se emplearon aspiradores y redes entomológicas. Los mosquitos fueron transportados al Laboratorio de Investigaciones en  Parasitología Tropical de la Universidad del Tolima para su determinación taxonómica. RESULTADOS: Se identificaron 708 ejemplares de A.  lbopictus, distribuidos en las comunas de Ibagué. La mayor abundancia del vector se presentó en las comunas 10, 11, 7, 8, 2 y 9. Las comunas 3, 4, 5, 6, 12 y 13 presentaron abundancias relativas cercanas al 3 %, y la comuna 1 tuvo una abundancia del 2 %. CONCLUSIONES: Aedes albopictus está distribuido en todas las comunas de Ibagué, probablemente su dispersión se ha visto favorecida por las condiciones ambientales y sociales de esta región. Se recomienda hacer seguimiento anual a las poblaciones de este vector y realizar una caracterización molecular de los arbovirus encontrados. Además, el conocer la distribución de este mosquito en la ciudad permitirá focalizar las estrategias de control entomológico y prevenir futuros brotes de arbovirosis.

Helicobacter pylori virulence factors: relationship between genetic variability and phylogeographic origin.

BACKGROUND: Helicobacter pylori is a pathogenic bacteria that colonize the gastrointestinal tract from human stomachs and causes diseases including gastritis, peptic ulcers, gastric lymphoma (MALT), and gastric cancer, with a higher prevalence in developing countries. Its high genetic diversity among strains is caused by a high mutation rate, observing virulence factors (VFs) variations in different geographic lineages. This study aimed to postulate the genetic variability associated with virulence factors present in the Helicobacter pylori strains, to identify the relationship of these genes with their phylogeographic origin. METHODS: The complete genomes of 135 strains available in NCBI, from different population origins, were analyzed using bioinformatics tools, identifying a high rate; as well as reorganization events in 87 virulence factor genes, divided into seven functional groups, to determine changes in position, number of copies, nucleotide identity and size, contrasting them with their geographical lineage and pathogenic phenotype. RESULTS: Bioinformatics analyses show a high rate of gene annotation errors in VF. Analysis of genetic variability of VFs shown that there is not a direct relationship between the reorganization and geographic lineage. However, regarding the pathogenic phenotype demonstrated in the analysis of many copies, size, and similarity when dividing the strains that possess and not the cag pathogenicity island (cagPAI), having a higher risk of developing gastritis and peptic ulcer was evidenced. Our data has shown that the analysis of the overall genetic variability of all VFs present in each strain of H. pylori is key information in understanding its pathogenic behavior.

Mitochondrial genomics of human pathogenic parasite Leishmania (Viannia) panamensis.

BACKGROUND: The human parasite Leishmania (V.) panamensis is one of the pathogenic species responsible for cutaneous leishmaniasis in Central and South America. Despite its importance in molecular parasitology, its mitochondrial genome, divided into minicircles and maxicircles, haven't been described so far. METHODS: Using NGS-based sequencing (454 and ILLUMINA), and combining de novo genome assembly and mapping strategies, we report the maxicircle kDNA annotated genome of L. (V.) panamensis, the first reference of this molecule for the subgenus Viannia. A comparative genomics approach is performed against other Leishmania and Trypanosoma species. RESULTS: The results show synteny of mitochondrial genes of L. (V.) panamensis with other kinetoplastids. It was also possible to identify nucleotide variants within the coding regions of the maxicircle, shared among some of them and others specific to each strain. Furthermore, we compared the minicircles kDNA sequences of two strains and the results show that the conserved and divergent regions of the minicircles exhibit strain-specific associations.

Genomic Analysis of Colombian Leishmania panamensis strains with different level of virulence.

The establishment of Leishmania infection in mammalian hosts and the subsequent manifestation of clinical symptoms require internalization into macrophages, immune evasion and parasite survival and replication. Although many of the genes involved in these processes have been described, the genetic and genomic variability associated to differences in virulence is largely unknown. Here we present the genomic variation of four Leishmania (Viannia) panamensis strains exhibiting different levels of virulence in BALB/c mice and its application to predict novel genes related to virulence. De novo DNA sequencing and assembly of the most virulent strain allowed comparative genomics analysis with sequenced L. (Viannia) panamensis and L. (Viannia) braziliensis strains, and showed important variations at intra and interspecific levels. Moreover, the mutation detection and a CNV search revealed both base and structural genomic variation within the species. Interestingly, we found differences in the copy number and protein diversity of some genes previously related to virulence. Several machine-learning approaches were applied to combine previous knowledge with features derived from genomic variation and predict a curated set of 66 novel genes related to virulence. These genes can be prioritized for validation experiments and could potentially become promising drug and immune targets for the development of novel prophylactic and therapeutic interventions.

Curvas de fusión de regiones genómicas específicas: una herramienta prometedora para el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia / High-resolution melting analysis based on specific genomic regions: A promising tool for the diagnosis and typing of species causing cutaneous leishmaniasis in Colombia

Resumen Introducción. La leishmaniasis cutánea es una enfermedad causada por parásitos del género Leishmania que tiene gran incidencia en Colombia. El diagnóstico y la identificación de la especie infecciosa son factores críticos en el momento de escoger e iniciar el tratamiento. Actualmente, los métodos de diagnóstico y tipificación requieren procedimientos complejos, por lo que es necesario validar nuevos marcadores moleculares y métodos que simplifiquen el proceso. Objetivo. Desarrollar una herramienta basada en la reacción en cadena de la polimerasa (PCR) con curvas de fusión (High Resolution Melting; PCR-HRM) para el diagnóstico y tipificación de las tres especies de Leishmania de importancia epidemiológica en casos de leishmaniasis cutánea en Colombia. Materiales y métodos. Los genomas de Leishmania panamensis, L. braziliensis y L. guyanensis se compararon mediante métodos bioinformáticos. Las regiones específicas de especie identificadas se validaron mediante PCR. Para los marcadores seleccionados se diseñó una PCR-HRM y se estimaron algunos parámetros de validez y seguridad usando aislamientos de pacientes colombianos caracterizados previamente mediante PCR y análisis de polimorfismos en la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism - RFLP; PCR-RFLP) del gen hsp70. Resultados. El análisis genómico comparativo mostró 24 regiones específicas de especie. Sin embargo, la validación mediante PCR solo identificó un marcador específico para cada especie de Leishmania. Los otros marcadores mostraron amplificación cruzada. El límite de detección para los tres marcadores seleccionados fue de un parásito, mientras que la sensibilidad, la especificidad, el valor predictivo positivo y el negativo fueron de 91,4, 100, 100 y 75 %, respectivamente. Conclusiones. Las tres regiones seleccionadas pueden emplearse como marcadores moleculares en el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

Abstract Introduction: Cutaneous leishmaniasis, caused by parasites of the genus Leishmania, is a disease with high incidence in Colombia. The diagnosis and identification of the infectious species are critical factors when selecting and initiating treatment. Currently, the methods for diagnosing and typing cutaneous leishmaniasis require complicated procedures and there is a need for the validation of new molecular markers and methods to simplify the process. Objective: To develop a tool based in PCR melting curves (PCR-HRM) for the diagnosis and typing of the three Leishmania species of epidemiological importance for cutaneous leishmaniasis in Colombia. Materials and methods: The genomes of Leishmania panamensis, L. braziliensis and L. guyanensis were compared with bioinformatic methods. The species-specific regions were then validated using PCR. For the selected markers, a PCR-HRM was designed, and validity and security parameters were estimated using isolates from Colombian patients previously characterized by PCR-RFLP of the hsp70 gene. Results: The comparative genomic analysis yielded 24 species-specific regions. However, the PCR validation identified only one marker that was specific to each Leishmania species. The other markers showed cross amplification. The detection limit for the three selected markers was one parasite. The sensitivity, specificity, predictive positive and negative values were 91.4%, 100%, 100% and 75%, respectively. Conclusions: The three selected regions can be used as molecular markers in the diagnosis and typing of the causative species of cutaneous leishmaniasis in Colombia.

Curvas de fusión de regiones genómicas específicas: una herramienta prometedora para el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

Introducción. La leishmaniasis cutánea es una enfermedad causada por parásitos del género Leishmania que tiene gran incidencia en Colombia. El diagnóstico y la identificación de la especie infecciosa son factores críticos en el momento de escoger e iniciar el tratamiento. Actualmente, los métodos de diagnóstico y tipificación requieren procedimientos complejos, por lo que es necesario validar nuevos marcadores moleculares y métodos que simplifiquen el proceso.Objetivo. Desarrollar una herramienta basada en la reacción en cadena de la polimerasa (PCR) con curvas de fusión (High Resolution Melting; PCR-HRM) para el diagnóstico y tipificación de las tres especies de Leishmania de importancia epidemiológica en casos de leishmaniasis cutánea en Colombia.Materiales y métodos. Los genomas de Leishmania panamensis, L. braziliensis y L. guyanensis se compararon mediante métodos bioinformáticos. Las regiones específicas de especie identificadas se validaron mediante PCR. Para los marcadores seleccionados se diseñó una PCR-HRM y se estimaron algunos parámetros de validez y seguridad usando aislamientos de pacientes colombianos caracterizados previamente mediante PCR y análisis de polimorfismos en la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism - RFLP; PCR-RFLP) del gen hsp70.Resultados. El análisis genómico comparativo mostró 24 regiones específicas de especie. Sin embargo, la validación mediante PCR solo identificó un marcador específico para cada especie de Leishmania. Los otros marcadores mostraron amplificación cruzada. El límite de detección para los tres marcadores seleccionados fue de un parásito, mientras que la sensibilidad, la especificidad, el valor predictivo positivo y el negativo fueron de 91,4, 100, 100 y 75 %, respectivamente.Conclusiones. Las tres regiones seleccionadas pueden emplearse como marcadores moleculares en el diagnóstico y tipificación de las especies causantes de la leishmaniasis cutánea en Colombia.

[Giardia duodenalis genotypes found in the Instituto Colombiano de Bienestar Familiar day care centers and dogs in Ibagué, Colombia]. / Genotipos de Giardia duodenalis en muestras de niños de las guarderías del Instituto Colombiano de Bienestar Familiar y de perros en Ibagué, Colombia.

INTRODUCTION: Eight Giardia duodenalis genotypes (A-H) have been described to date. Genotypes A and B have been isolated from humans and a wide range of mammals; however, genotypes C-H have shown greater host specificity. OBJECTIVE: Identifying G. duodenalis genotypes from cysts in faeces obtained from children attending the Instituto Colombiano de Bienestar Familiar (ICBF) day care centres and from dogs in Ibagué by PCR-RFLP targeting both the b -giardin and glutamate dehydrogenase genes. MATERIALS AND METHODS: Cysts from G. duodenalis positive samples were concentrated, DNA was extracted and the b -giardin and glutamate dehydrogenase genes were analysed by PCR-RFLP. The MHOM/CO/04/G40 strain was used as positive control (this was obtained from the Grupo de Parasitología at the Instituto Nacional de Salud ). RESULTS: Of the total human samples, 11/23 (48%) were genotyped as A and 12/23 (52%) as B; PCR-RFLP revealed that four canine samples were genotypes C and D, these being host-specific. CONCLUSIONS: Only genotypes associated with human infection (AII, BIII and BIV) were found in the children and host-specific genotypes were observed in canines (C and D). No interaction could be established between animal and human transmission cycles due to the small canine sample size and as the former did not come into contact with children attending ICBF day-care centres.

Genotipos de Giardia duodenalis en muestras de niños de las guarderías del Instituto Colombiano de Bienestar Familiar y de perros en Ibagué, Colombia / Giardia duodenalis genotypes found in the Instituto Colombiano de Bienestar Familiar day care centers and dogs in Ibagué, Colombia

Introducción. Se han descrito ocho genotipos de Giardia duodenalis, del A al H. Los genotipos A y B se han aislado de humanos y de una gran variedad de mamíferos; sin embargo, los genotipos del C al H han mostrado mayor especificidad de huésped. Objetivo. Identificar los genotipos de G. duodenalis a partir de quistes obtenidos en heces de niños de las guarderías del Instituto Colombiano de Bienestar Familiar (ICBF) y de perros en Ibagué, mediante PCR-RFLP de los genes de la beta giardina y la glutamato deshidrogenasa. Materiales y métodos. Los quistes de las muestras positivas para G. duodenalis fueron sometidos a concentración; se extrajo su ADN y se efectuó el análisis de PCR-RFLP de los genes de la beta giardina y de la glutamato deshidrogenasa. Como control positivo se utilizó la cepa MHOM/CO/04/G40 procedente del Grupo de Parasitología del Instituto Nacional de Salud. Resultados. De las muestras tomadas de niños, 11/23 (48 %) correspondieron al genotipo A y, 12/23 (52 %), al genotipo B. Cuatro muestras de perros presentaron los genotipos C y D, específicos de este huésped. Conclusiones. En los niños solamente se encontraron los genotipos asociados a infecciones humanas (AII, BIII y BIV) y en los perros, los genotipos específicos para este huésped (C y D). Debido al reducido tamaño de las muestras analizadas provenientes de perros, y dado que estos no estuvieron en contacto con los niños de las guarderías del ICBF, no fue posible determinar una interacción entre el ciclo de transmisión de los humanos y el de los animales.

Introduction: Eight Giardia duodenalis genotypes (A-H) have been described to date. Genotypes A and B have been isolated from humans and a wide range of mammals; however, genotypes C-H have shown greater host specificity. Objective: Identifying G. duodenalis genotypes from cysts in faeces obtained from children attending the Instituto Colombiano de Bienestar Familiar (ICBF) day care centres and from dogs in Ibagué by PCR-RFLP targeting both the b -giardin and glutamate dehydrogenase genes. Materials and methods: Cysts from G. duodenalis positive samples were concentrated, DNA was extracted and the b -giardin and glutamate dehydrogenase genes were analysed by PCR-RFLP. The MHOM/CO/04/G40 strain was used as positive control (this was obtained from the Grupo de Parasitología at the Instituto Nacional de Salud ). Results: Of the total human samples, 11/23 (48%) were genotyped as A and 12/23 (52%) as B; PCR-RFLP revealed that four canine samples were genotypes C and D, these being host-specific. Conclusions: Only genotypes associated with human infection (AII, BIII and BIV) were found in the children and host-specific genotypes were observed in canines (C and D). No interaction could be established between animal and human transmission cycles due to the small canine sample size and as the former did not come into contact with children attending ICBF day-care centres.

The identification of two Trypanosoma cruzi I genotypes from domestic and sylvatic transmission cycles in Colombia based on a single polymerase chain reaction amplification of the spliced-leader intergenic region.

A single polymerase chain reaction (PCR) reaction targeting the spliced-leader intergenic region of Trypanosoma cruzi I was standardised by amplifying a 231 bp fragment in domestic (TcIDOM) strains or clones and 450 and 550 bp fragments in sylvatic strains or clones. This reaction was validated using 44 blind coded samples and 184 non-coded T. cruzi I clones isolated from sylvatic triatomines and the correspondence between the amplified fragments and their domestic or sylvatic origin was determined. Six of the nine strains isolated from acute cases suspected of oral infection had the sylvatic T. cruzi I profile. These results confirmed that the sylvatic T. cruzi I genotype is linked to cases of oral Chagas disease in Colombia. We therefore propose the use of this novel PCR reaction in strains or clones previously characterised as T. cruzi I to distinguish TcIDOMfrom sylvatic genotypes in studies of transmission dynamics, including the verification of population selection within hosts or detection of the frequency of mixed infections by both T. cruzi I genotypes in Colombia.

Sequence analysis of the spliced-leader intergenic region (SL-IR) and random amplified polymorphic DNA (RAPD) of Trypanosoma rangeli strains isolated from Rhodnius ecuadoriensis, R. colombiensis, R. pallescens and R. prolixus suggests a degree of co-evolution between parasites and vectors.

Spliced leader intergenic region (SL-IR) sequences from 23 Trypanosoma rangeli strains isolated from the salivary glands of Rhodnius colombiensis, R. ecuadoriensis, R. pallescens and R. prolixus and two human strains revealed the existence of 4 genotypes with CA, GT, TA, ATT and GTAT microsatellite repeats and the presence of insertions/deletions (INDEL) and single nucleotide polymorphism (SNP) characterizing each genotype. The strains isolated from the same vector species or the same Rhodnius evolutionary line presented the same genotypes, even in cases where strains had been isolated from vectors captured in geographically distant regions. The dendrogram constructed from the SL-IR sequences separated all of them into two main groups, one with the genotypes isolated from R. prolixus and the other group containing three well defined sub-groups with the genotypes isolated from R. pallescens, R. colombiensis and R. ecuadoriensis. Random amplified polymorphic DNA (RAPD) analysis showed the same two main groups and sub-groups supporting strict T. rangeli genotypes' association with Rhodnius species. Combined with other studies, these results suggest a possible co-evolutionary association between T. rangeli genotypes and their vectors.

Trypanosoma rangeli genotypes association with Rhodnius prolixus and R. pallescens allopatric distribution in Central America.

Previous kDNA polymorphism-based reports have revealed the existence of two Trypanosoma rangeli genotypes (KP1+ and KP1-): SL and SSU rRNA gene polymorphism-based studies have revealed that five genotypes (A-E) are distributed throughout different Latin-American countries. Some evidence has shown that the genotypes' biogeographical distribution is associated with sympatric Rhodnius species. 12 T. rangeli isolates from humans and reservoirs from El Salvador, Guatemala, Honduras, Costa Rica and Panama were characterised by kDNA and mini-exon gene intergene spacer analysis and compared to 12 previously characterised isolates from humans and vectors from Colombia, Guatemala, Honduras and Venezuela. Central American isolates corresponded to genotypes called KP1(+) or lineage A and KP1(-) or lineage C. Such dimorphism was corroborated by randomly amplified polymorphic DNA (RAPD) in 22 selected isolates; a dendrogram was thus produced having two defined branches. One branch grouped KP1(-) or lineage C strains isolated from Rhodnius colombiensis (Colombia), humans (Panama), Procyon lotor and Choloepus hoffmanni (Costa Rica). The other group was formed by KP1(+) or lineage A strains isolated from Rhodnius prolixus (Colombia, Venezuela) and humans (El Salvador, Guatemala, Honduras). These results present evidence that both groups infect different mammals (humans, domestic and silvatic animals) having no association with any particular vertebrate species; however, T. rangeli KP1(+) or (A) strains have been isolated in Central America in areas where R. prolixus circulate (Honduras, El Salvador and Guatemala) and KP1(-) or (C) strains have been isolated in areas where Rhodnius pallescens is the main vector (Panama and Costa Rica) indicating a parasite-vector association. The same lineages circulate in Andean countries (Colombia, Venezuela, Ecuador and Peru), KP1+ being associated with members of the prolixus group (R. prolixus and Rhodnius robustus) and KP1- with members of the pallescens group (R. pallescens, R. colombiensis and Rhodnius ecuadoriensis).

[Trypanosoma rangeli parasite-vector-vertebrate interactions and their relationship to the systematics and epidemiology of American trypanosomiasis]. / Interacción tripanosoma-vector-vertebrado y su relación con la sistemática y la epidemiología de la tripanosomiasis Americana.

INTRODUCTION: Trypanosoma rangeli is a species of trypanosome second to T. cruzi, that is infective to humans in Latin America. Variability in the biological, biochemical and molecular characteristics between different isolates isolates of this parasite have been recorded. OBJECTIVE: Morphological and molecular characteristics were recorded from strains of T. rangeli that were isolated from different species of Rhodnius and maintained in different vertebrate species. MATERIALS AND METHODS: Nineteen strains of T. rangeli were isolated from R. prolixus, R. pallescens and R. colombiensis in Colombia, R. ecuadoriensis in Peru and R. pallescens in Panama. Polymorphism of blood trypomastigotes in ICR mice was evaluated and pleomorphism of P53 strain of T. rangeli KP1(-) inoculated in mouse, marsupial and canine was studied. RAPD analysis (randomly amplified polymorphic DNA analysis) of 12 strains isolated from four species of Rhodnius was performed. RESULT: Based on the total length of blood trypomastigotes, three discrete groups were observed. The P53 strain showed significant differences in the size of blood trypomastigotes in mouse, marsupial and canine. RAPD analysis showed that the strains segregated into two branches corresponding to strains of T. rangeli KP1(+) and T. rangeli KP1(-). All strains of T. rangeli KP1(-) clustered according to the species of Rhodnius from which they were isolated. CONCLUSION: These data reveal, for the first time, a close association amongst T. rangeli strains and Rhodnius species, confirming that each species of Rhodnius transmits to vertebrate hosts a parasite population with clear phenotypic and genotypic differences. This is further evidence that supports the concept of clonal evolution of these parasites.