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Trypanosoma cruzi, the causative agent of Chagas disease (American trypanosomiasis), is a highly complex zoonosis that is present throughout South America, Central America, and Mexico. The transmission of this disease is influenced by various factors, including human activities like deforestation and land use changes, which may have altered the natural transmission cycles and their connection to the environment. In this study conducted in the Argentine Chaco region, we examined the transmission dynamics of T. cruzi by collecting blood samples from wild and domestic animals, as well as triatomine bugs from human dwellings, across five sites of varying anthropic intervention. Samples were analyzed for T. cruzi infection via qPCR, and we additionally examined triatomines for bloodmeal analysis via NGS amplicon sequencing. Our analysis revealed a 15.3% infection rate among 20 wild species (n = 123) and no T. cruzi presence in 9 species of domestic animals (n = 1359) or collected triatomines via qPCR. Additionally, we found chicken (34.28%), human (21.59%), and goat (19.36%) as the predominant bloodmeal sources across all sites. These findings suggest that anthropic intervention and other variables analyzed may have directly impacted the spillover dynamics of T. cruzi’s sylvatic cycle and potentially reduced its prevalence in human habitats.

Abstract Background Chagas disease is considered to be endemic in up to 40% of the territory of Colombia, and to date 27 triatomine species have been reported the country. The purpose of this study was to update the geographical distribution of triatomine species in Colombia and assess the species richness patterns and their altitudinal distribution. Methods Occurrence data were compiled between 2007 and 2020, including from reports of entomological surveillance from the Instituto Nacional de Salud (INS), the Centro de Investigaciones en Microbiología y Parasitología Tropical (CIMPAT) at Universidad de Los Andes and a review of the literature. Geographic Information Systems (GIS) were used to describe general species richness patterns of the Triatominae subfamily. To establish the altitudinal distribution of the triatomine species, ranges were obtained from reports with unique elevation values. A generalized linear model was fitted, based on a Poisson distribution, to test the relation between triatomine species richness and Chagas disease cases (2012–2019). Results An updated geographical and altitudinal distribution for triatomine species in Colombia was established, with 507 municipalities added to the previously known distributions. The greatest triatomine richness in Colombia was found to be concentrated in the northeastern region of the country, extending towards the center to the departments of Arauca, Casanare and Meta. Regarding the altitudinal distribution, the study revealed that the species Rhodnius prolixus and Triatoma dimidiata have the greatest altitudinal ranges. The data also suggest a positive relation between species richness and number of Chagas disease cases reported per department. Conclusions Altitudinal ranges for 17 triatomine species found in Colombia are presented. Species richness and species composition patterns are also described, and areas with a higher risk of transmission based on the relation found with Chagas disease cases are highlighted. This updated distribution reveals that Panstrongylus geniculatus is the triatomine with the largest presence by municipalities in Colombia, being reported in 284 municipalities, followed by Rhodnius prolixus in 277 municipalities. Graphical Abstract

Metacyclogenesis is one of the most important processes in the life cycle of Trypanosoma cruzi. In this stage, noninfective epimastigotes become infective metacyclic trypomastigotes. However, the transcriptomic changes that occur during this transformation remain uncertain. Illumina RNA-sequencing of epimastigotes and metacyclic trypomastigotes belonging to T. cruzi DTU I was undertaken. Sequencing reads were aligned and mapped against the reference genome, differentially expressed genes between the two life cycle stages were identified, and metabolic pathways were reconstructed. Gene expression differed significantly between epimastigotes and metacyclic trypomastigotes. The cellular pathways that were mostly downregulated during metacyclogenesis involved glucose energy metabolism (glycolysis, pyruvate metabolism, the Krebs cycle, and oxidative phosphorylation), amino acid metabolism, and DNA replication. By contrast, the processes where an increase in gene expression was observed included those related to autophagy (particularly Atg7 and Atg8 transcripts), corroborating its importance during metacyclogenesis, endocytosis, by an increase in the expression of the AP-2 complex subunit alpha, protein processing in the endoplasmic reticulum and meiosis. Study findings indicate that in T. cruzi metacyclic trypomastigotes, metabolic processes are decreased, and expression of genes involved in specific cell cycle processes is increased to facilitate transformation to this infective stage.

Abstract Background Severe changes in temperature can affect the behavior and ecology of some infectious agents. Trypanosoma cruzi is a protozoan that causes Chagas disease. This parasite has high genetic variability and can be divided into six discrete typing units (DTUs). Trypanosoma cruzi also has a complex life-cycle, which includes the process of metacyclogenesis when non-infective epimastigote forms are differentiated into infective metacyclic trypomastigotes (MT). Studies in triatomines have shown that changes in temperature also affect the number and viability of MT. Methods The objective of this study was to evaluate how temperature affects the transcriptional profiles of T. cruzi I and II (TcI and TcII) MT by exposing parasites to two temperatures (27 °C and 28 °C) and comparing those to normal culture conditions at 26 °C. Subsequently, RNA-seq was conducted and differentially expressed genes were quantified and associated to metabolic pathways. Results A statistically significant difference was observed in the number of MT between the temperatures evaluated and the control, TcII DTU was not strongly affected to exposure to high temperatures compared to TcI. Similar results were found when we analyzed gene expression in this DTU, with the greatest number of differentially expressed genes being observed at 28 °C, which could indicate a dysregulation of different signaling pathways under this temperature. Chromosome analysis indicated that chromosome 1 harbored the highest number of changes for both DTUs for all thermal treatments. Finally, gene ontology (GO) analyses showed a decrease in the coding RNAs involved in the regulation of processes related to the metabolism of lipids and carbohydrates, the evasion of oxidative stress, and proteolysis and phosphorylation processes, and a decrease in RNAs coding to ribosomal proteins in TcI and TcII, along with an increase in the expression of surface metalloprotease GP63 in TcII. Conclusions Slight temperature shifts lead to increased cell death of metacyclic trypomastigotes because of the deregulation of gene expression of different processes essential for the TcI and TcII DTUs of T. cruzi.

Abstract Background A question of epidemiological relevance in Chagas disease studies is to understand Trypanosoma cruzi transmission cycles and trace the origins of (re)emerging cases in areas under vector or disease surveillance. Conventional parasitological methods lack sensitivity whereas molecular approaches can fill in this gap, provided that an adequate sample can be collected and processed and a nucleic acid amplification method can be developed and standardized. We developed a duplex qPCR assay for accurate detection and quantification of T. cruzi satellite DNA (satDNA) sequence in samples from domestic and sylvatic mammalian reservoirs. The method incorporates amplification of the gene encoding for the interphotoreceptor retinoid-binding protein (IRBP), highly conserved among mammalian species, as endogenous internal amplification control (eIAC), allowing distinction of false negative PCR findings due to inadequate sample conditions, DNA degradation and/or PCR interfering substances. Results The novel TaqMan probe and corresponding primers employed in this study improved the analytical sensitivity of the assay to 0.01 par.eq/ml, greater than that attained by previous assays for Tc I and Tc IV strains. The assay was tested in 152 specimens, 35 from 15 different wild reservoir species and 117 from 7 domestic reservoir species, captured in endemic regions of Argentina, Colombia and Mexico and thus potentially infected with different parasite discrete typing units. The eIACs amplified in all samples from domestic reservoirs from Argentina and Mexico, such as Canis familiaris, Felis catus, Sus scrofa, Ovis aries, Equus caballus, Bos taurus and Capra hircus with quantification cycles (Cq’s) between 23 and 25. Additionally, the eIACs amplified from samples obtained from wild mammals, such as small rodents Akodon toba, Galea leucoblephara, Rattus rattus, the opossums Didelphis virginiana, D. marsupialis and Marmosa murina, the bats Tadarida brasiliensis, Promops nasutus and Desmodus rotundus, as well as in Conepatus chinga, Lagostomus maximus, Leopardus geoffroyi, Lepus europaeus, Mazama gouazoubira and Lycalopex gymnocercus, rendering Cq’s between 24 and 33. Conclusions This duplex qPCR assay provides an accurate laboratory tool for screening and quantification of T. cruzi infection in a vast repertoire of domestic and wild mammalian reservoir species, contributing to improve molecular epidemiology studies of T. cruzi transmission cycles.

Abstract Background Oil palm plantation establishment in Colombia has the potential to impact Chagas disease transmission by increasing the distribution range of Rhodnius prolixus. In fact, previous studies have reported Trypanosoma cruzi natural infection in R. prolixus captured in oil palms (Elaeis guineensis) in the Orinoco region, Colombia. The aim of this study is to understand T. cruzi infection in vectors in oil palm plantations relative to community composition and host dietary specialization by analyzing vector blood meals and comparing these results to vectors captured in a native palm tree species, Attalea butyracea. Methods Rhodnius prolixus nymphs (n = 316) were collected from A. butyracea and E. guineensis palms in Tauramena, Casanare, Colombia. Vector blood meals from these nymphs were determined by amplifying and sequencing a vertebrate-specific 12S rRNA gene fragment. Results Eighteen vertebrate species were identified and pigs (Sus scrofa) made up the highest proportion of blood meals in both habitats, followed by house mouse (Mus musculus) and opossum (Didelphis marsupialis). Individual bugs feeding only from generalist mammal species had the highest predicted vector infection rate, suggesting that generalist mammalian species are more competent hosts for T. cruzi infection . Conclusions Oil palm plantations and A. butyracea palms found in altered areas provide a similar quality habitat for R. prolixus populations in terms of blood meal availability. Both habitats showed similarities in vector infection rate and potential host species, representing a single T. cruzi transmission scenario at the introduced oil palm plantation and native Attalea palm interface.

Trypanosoma cruzi, a zoonotic kinetoplastid protozoan parasite, is the causative agent of American trypanosomiasis (Chagas disease). Having a very plastic, repetitive and complex genome, the parasite displays a highly diverse repertoire of surface molecules, with pivotal roles in cell invasion, immune evasion and pathogenesis. Before 2016, the complexity of the genomic regions containing these genes impaired the assembly of a genome at chromosomal level, making it impossible to study the structure and function of the several thousand repetitive genes encoding the surface molecules of the parasite. We here describe the genome assembly of the Sylvio X10/1 genome sequence, which since 2016 has been used as a reference genome sequence for T. cruzi clade I (TcI), produced using high coverage PacBio single-molecule sequencing. It was used to analyze deep Illumina sequence data from 34 T. cruzi TcI isolates and clones from different geographic locations, sample sources and clinical outcomes. Resolution of the surface molecule gene distribution showed the unusual duality in the organization of the parasite genome, a synteny of the core genomic region with related protozoa flanked by unique and highly plastic multigene family clusters encoding surface antigens. The presence of abundant interspersed retrotransposons in these multigene family clusters suggests that these elements are involved in a recombination mechanism for the generation of antigenic variation and evasion of the host immune response on these TcI strains. The comparative genomic analysis of the cohort of TcI strains revealed multiple cases of such recombination events involving surface molecule genes and has provided new insights into T. cruzi population structure.

Panstrongylus geniculatus (Latreille, 1811) is the triatomine with the largest geographic distribution in Latin America. It has been reported in 18 countries from southern Mexico to northern Argentina, including the Caribbean islands. Although most reports indicate that P. geniculatus has wild habitats, this species has intrusive habits regarding human dwellings mainly located in intermediate deforested areas. It is attracted by artificial light from urban and rural buildings, raising the risk of transmission of Trypanosoma cruzi. Despite the wide body of published information on P. geniculatus, many knowledge gaps exist about its biology and epidemiological potential. For this reason, we analysed the literature for P. geniculatus in Scopus, PubMed, Scielo, Google Scholar and the BibTriv3.0 databases to update existing knowledge and provide better information on its geographic distribution, life cycle, genetic diversity, evidence of intrusion and domiciliation, vector-related circulating discrete taxonomic units, possible role in oral T. cruzi transmission, and the effect of climate change on its biology and epidemiology.

Trypanosoma cruzi is a flagellated protozoan that causes Chagas disease; it presents a complex life cycle comprising four morphological stages: epimastigote (EP), metacyclic trypomastigote (MT), cell-derived trypomastigote (CDT) and amastigote (AM). Previous transcriptomic studies on three stages (EPs, CDTs and AMs) have demonstrated differences in gene expressions among them; however, to the best of our knowledge, no studies have reported on gene expressions in MTs. Therefore, the present study compared differentially expressed genes (DEGs), and signaling pathway reconstruction in EPs, MTs, AMs and CDTs. The results revealed differences in gene expressions in the stages evaluated; these differences were greater between MTs and AMs-PTs. The signaling pathway that presented the highest number of DEGs in all the stages was associated with ribosomes protein profiles, whereas the other related pathways activated were processes related to energy metabolism from glucose, amino acid metabolism, or RNA regulation. However, the role of autophagy in the entire life cycle of T. cruzi and the presence of processes such as meiosis and homologous recombination in MTs (where the expressions of SPO11 and Rad51 plays a role) are crucial. These findings represent an important step towards the full understanding of the molecular basis during the life cycle of T. cruzi.

Abstract Background Ethnoichthyology studies the knowledge of human communities about the uses, perception, and behavioral characteristics of fish, based on their cosmology, cosmogony, and culture. This study focused on the Cubeo (Pâmiwâ) ethnic group that inhabits the Cuduyarí River (Vaupés, Colombia) provides information on fishing activities, use of fishes, its ecological distributions (occurrence in rivers, pools, and/or streams) and seasonality (occurrence in low and/or high waters). Additionally, names in the Central Tukano language (Cubeo) are provided. Methods The compilation of information was based on a visual catalog prepared from rapid ecological evaluations, previously carried out by the Amazon Institute of Scientific Research SINCHI. Workshops with the indigenous people were held in communities in the lower, middle, and upper sections of the basin to collect information. Network analyses were done to determine the fishing gear selectivity of the most common species and gears used regarding ecosystems. The coherence in the perception of fish resources among the three communities was assessed via multivariate analyses. Results The results showed that a large percentage (89%) of the species are consumed. There is a generalized perception of resource decline, and there is no coherence regarding the ecological and seasonal distributions of fish reported by the three communities. These differences may be due to an effect of the structural characteristics of the basin in the different sectors where the communities are located. Conclusions Species consumed by the Cubeo in the Cuduyarí do not represent a high commercial value at the Amazon River basin scale. Nevertheless, locally, they are the main source of animal protein and most of the time, the sole source of income. The importance of these species must be recognized and must be a priority for conservation. Knowledge related to the fish resource by indigenous groups, including basic information and its usage, is essential to guide and implement management and conservation strategies specific to each region under study.

We report the characterization of Trypanosoma cruzi of southern South American origin among humans, domestic vectors, and peridomestic hosts in Colombia using high-resolution nuclear and mitochondrial genotyping. Expanding our understanding of the geographic range of lineage TcVI, which is associated with severe Chagas disease, will help clarify risk of human infection for improved disease control.

The Triatominae subfamily includes 151 extant and three fossil species. Several species can transmit the protozoan parasite Trypanosoma cruzi, the causative agent of Chagas disease, significantly impacting public health in Latin American countries. The Triatominae can be classified into five tribes, of which the Rhodniini is very important because of its large vector capacity and wide geographical distribution. The Rhodniini tribe comprises 23 (without R. taquarussuensis) species and although several studies have addressed their taxonomy using morphological, morphometric, cytogenetic, and molecular techniques, their evolutionary relationships remain unclear, resulting in inconsistencies at the classification level. Conflicting hypotheses have been proposed regarding the origin, diversification, and identification of these species in Latin America, muddying our understanding of their dispersion and current geographic distribution. Clarifying these factors can help for the design of vector control strategies. The aim of this review is to depict the different approaches used for taxonomy of the Rhodniini and to shed light on their evolution and biogeography.

The increase in the global land temperature, expected under predictions of climate change, can directly affect the transmission of some infectious diseases, including Chagas disease, an anthropozoonosis caused by Trypanosoma cruzi and transmitted by arthropod vectors of the subfamily Triatominae. This work seeks to study the effects of temperature on the development of the life cycle, fertility and fecundity of the insect vector Rhodnius prolixus and on the metacyclogenesis of T. cruzi. All of the variables were subjected to 3 temperatures: 26°C, 28°C and 30°C. Hatching time was evaluated, along with time to fifth instar, time to adult, fecundity studied using the e-value, and egg viability during the first 3 reproductive cycles. In addition, the amounts of metacyclic trypomastigotes of the TcI and TcII DTUs in R. prolixus were evaluated from days 2 to 20 at two-day intervals and from weeks 6 to 8 post-infection. Decreases were observed in time to hatching (15-10 days on average) and in time to fifth instar (70-60 days on average) and transition to adult (100-85 days on average). No significant differences in egg viability were observed in any of the reproductive cycles evaluated, but an increase in fecundity was observed at 30°C during the third reproductive cycle. At 30°C, there was also an increase in the number of infective forms and a decrease in the time at which metacyclic trypomastigotes were detected in the rectal ampulla of the insects for both TcI and TcII. According to these results, the expected temperature increase under climate change would cause an increase in the number of insects and a greater probability of infection of the parasite, which affects the transmission of Chagas disease.

Abstract Background Control initiatives and continuous surveillance of vector-borne transmission have proved to be effective measures for diminishing the incidence of Chagas disease in endemic countries. However, the active dispersal of infected sylvatic adult triatomines by flight represents one of the main obstacles to eliminating domestic transmission. Methods In order to determine the risk that active dispersal of sylvatic adult triatomines represents in Colombian northeastern plains, we quantified the distribution and abundance of triatomines in palm trees (primarily Attalea butyracea) using live bait traps. Directional light traps were used to estimate the frequency of sylvatic triatomine dispersal and their possible origin. Finally, the effect of environmental parameters and artificial light sources on the take-off of sylvatic Rhodnius prolixus was evaluated in field experiments. Results R. prolixus was found in 90 % of the palm trees that densely aggregated toward the northern portion of the study area. R. prolixus, and three other sylvatic triatomine species were found to actively disperse and were attracted to the directional light traps (Triatoma maculata, Panstrongylus geniculatus and Psammolestes arthuri). Temperature, relative humidity, wind speed and night luminosity did not affect the active dispersal of the triatomines which is higher the first two hours after sunset. Artificial lights from houses at 60 and 110 m played a key role in the directionality of the R. prolixus take-offs. Trypanosoma cruzi was isolated from R. prolixus, T. maculata and P. geniculatus and was genotyped as T. cruzi I, III and IV. Conclusions Our results highlight the potential risk in Colombian northeastern plains of actively dispersing sylvatic triatomines and their role in the domestic introduction of Discrete Typing Units of T. cruzi associated to sylvatic foci of Chagas disease transmission.

Rhodnius prolixus, a blood-sucking triatomine with domiciliary anthropophilic habits, is the main vector of Chagas disease. The current paradigm of Trypanosoma cruzi transmission in Columbia includes a sylvatic and domiciliary cycle co-existing with domestic and sylvatic populations of reservoirs. The aim of this study is to evaluate the population densities and relative abundance of triatomines and mammals that may be involved in the sylvatic cycle of Chagas disease to clarify the epidemiological scenario in an endemic area in the province of Casanare. Insect vectors on Attalea butyracea palms were captured using both manual searches and bait traps. The capture of mammals was performed using Sherman and Tomahawk traps. We report an infestation index of 88.5% in 148 palms and an index of T. cruzi natural infection of 60.2% in 269 dissected insects and 11.9% in 160 captured mammals. High population densities of triatomines were observed in the sylvatic environment and there was a high relative abundance of reservoirs in the area, suggesting a stable enzootic cycle. We found no evidence of insect domiciliation. Taken together, these observations suggest that eco-epidemiological factors shape the transmission dynamics of T. cruzi, creating diverse scenarios of disease transmission.

This opinion piece presents an approach to standardisation of an important aspect of Chagas disease drug discovery and development: selecting Trypanosoma cruzi strains for in vitro screening. We discuss the rationale for strain selection representing T. cruzi diversity and provide recommendations on the preferred parasite stage for drug discovery, T. cruzi discrete typing units to include in the panel of strains and the number of strains/clones for primary screens and lead compounds. We also consider experimental approaches for in vitro drug assays. The Figure illustrates the current Chagas disease drug-discovery and development landscape.

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.

La enfermedad de Chagas causada por el parásito Trypanosoma cruzi es una zoonosis compleja, ampliamente distribuida en el continente americano. La infección puede ser adquirida a través de las heces de insectos triatominos, transfusión de sangre, trasplante de órganos, vía oral, por transmisión congénita y por accidentes de laboratorio. El completo entendimiento de la etiología y epidemiología de la enfermedad de Chagas a través de su distribución geográfica es complejo y permanece bajo intensa investigación hasta la actualidad. Los recientes estudios sobre la variabilidad genética del parásito han dado nuevas luces de los diferentes escenarios de los ciclos de transmisión de la enfermedad y su patogénesis en humanos. El propósito principal para la caracterización molecular de T.cruzi y sus múltiples genotipos está dirigido hacia su asociación con la clínica y la patogenesis de la enfermedad, así como al esclarecimiento de los diferentes escenarios de transmisión y los aspectos coevolutivos relacionados con reservorios e insectos vectores. La caracterización molecular de los diferentes aislamientos a partir de humanos, insectos y reservorios, ha permitido identificar la amplia variabilidad genética del parásito, abriendo nuevos caminos hacia la búsqueda de nuevos blancos terapéuticos y pruebas diagnósticas más específicas que contribuyan a mitigar la enfermedad de Chagas.

Introducción. La enfermedad de Chagas, cuyo agente causal es Trypanosoma cruzi, constituye una antropozoonosis ampliamente distribuida en América Latina. Los estudios moleculares y los perfiles genéticos han demostrado que el parásito presenta una gran variabilidad y han permitido la clasificación de T. cruzi en seis unidades discretas de tipificación (I-VI), de las cuales, TcII a TcVI han sido ampliamente caracterizadas por distintos marcadores moleculares. Objetivo. Evaluar la variabilidad genética de TcI, mediante el uso de marcadores de microsatélites. Materiales y métodos. Se evaluaron siete microsatélites en quince aislamientos colombianos obtenidos mediante PCR a partir de reservorios, vectores y humanos, y se analizaron en un gel para desnaturalizar de poliacrilamida de un secuenciador láser fluorescente automático (ALF). Los datos se analizaron en un software para análisis de genética de poblaciones (Arlequin® 3.1 y Microsat®). Resultados. Los resultados demostraron variabilidad dentro de TcI al obtener 24 alelos, de los cuales, 12 se reportan por primera vez. Aunque se encontraron genotipos asociados a la infección humana y al ciclo selvático de transmisión, ningún locus permitió comprobar la presencia de los genotipos previamente reportados. Los parámetros de heterocigocidad observada y esperada, permitieron determinar la presencia de dos poblaciones (aislamientos domésticos y selváticos); asimismo, el desequilibrio de ligamento facilitó la creación de dos mapas físicos para los loci analizados. Conclusiones. Se corrobora la gran variabilidad genética presente en TcI, lo que sugiere un patrón de variación intraespecífica en Colombia.Introduction: Chagas disease which is caused by the protozoan Trypanosoma cruzi, is a major public health problem in Latin American countries with a different distribution of the parasite across the continent in which nearly 15 million people are infected and 28 million are at risk. Genetic profiling of T. cruzi has shown great diversity and variability of the parasite allowing its classification into six discrete typing units (DTUs I-VI) in which TcII to TcVI are well characterized by different molecular markers. However, the presence of four subgroups according to the transmission cycle has only been reported in DTU T. cruzi I. Objective: To evaluate the genetic variability evidenced within TCI by the use of microsatellite markers. Materials and methods: Seven microsatellite loci were tested in fifteen Colombian isolates from vectors, reservoirs, and humans by means of PCR and automatic laser fluorescent sequencer (ALF). Data were analyzed using a population genetic data analysis software (Arlequin® 3.1 and Microsat®). Results: Variability among the isolates was demonstrated with 24 alleles, from which twelve had never been reported before. However, none of the microsatellite loci were able to support the idea of genotypes within TcI strains. The parameters of expected and observed heterozygocity allowed us to determine presence of two populations (domestic and sylvatic isolates); likewise, the linkage disequilibrium helped with the construction of two physical maps for the loci analyzed. Conclusions: We corroborated the high genetic variability displayed by TcI populations suggesting a pattern of intraspecific variation in Colombia.

The breeding sites of Culicoides pachymerus are described for the first time in western Boyacá Province, Colombia, where this species is a public health problem. In addition to being a nuisance due to its enormous density and its high biting rates, C. pachymerus cause dermatological problems in the human population. Analysis of microhabitats by the sugar flotation technique and the use of emergence traps allowed us to recover 155 larvae of Culicoides spp and 65 adults of C. pachymerus from peridomiciliary muddy substrates formed by springs of water and constant rainwater accumulation. These important findings could aid in the design of integrated control meas-ures against this pest.

Over the last 10 years, Uruguay, Chile and Brazil have been certified as being free from disease transmission by Triatoma infestans, the main domiciliated vector for Chagas disease in the Southern Cone countries. This demonstrates that programmes addressing the vector for the disease's transmission are effective. These programmes have resulted in a dramatic decrease in the incidence of Chagas disease in Latin America. Guatemala was certified a few months ago as being free from disease transmission by Rhodnius prolixus, the main domiciliated vector for Chagas disease in Central American countries. However, the main concern for different countries' current control programmes is the continuity and sustainability of future vector control actions. The prevalence and incidence figures for individuals infected by Trypanosoma cruzi in Mexico and Andean and Central American countries highlights the need for broadened strategies in the struggle against the disease and its vectors. A number of triatomine insects are parasite vectors, each with a different life history. Therefore, it is important that new vector control strategies be proposed, keeping in mind that some species are found in peridomiciliary areas and wild ecotopes. The only viable control strategy is to reduce human interactions with vector insects so that the re-infestation and re-colonisation of human habitats will not take place.

Constituye para mí en calidad de miembro de las dos Academias, de Medicina y de Ciencias Exactas, Físicas y Naturales, un verdadero honor dirigirme a ustedes en una ocasión memorable, cuando se celebran los 200 años del natalicio de Carlos Darwin y 150 años de la publicación de su libro acerca del origen de las especies.He seleccionado el tema de Darwin en Suramérica teniendo en cuenta que durante el breve intervalo del llamado siglo de los exploradores de 1735 a 1859, el continente suramericano jugó un papel fundamental para el desarrollo de la ciencia adelantado por los científicos y naturalistas de la época.En realidad fueron los naturalistas quienes verdaderamente descubrieron el continente a través de sus exploradores y revelaron al mundo novedosos hallazgos que sirvieron de base para el desarrollo posterior de todas las ciencias exactas, físicas y naturales. Voy a enfocar esta presentación principalmente a personajes que influyeron en la vida y obra de Darwin y que le permitieron finalmente concretar su colosal obra.En 1734 la Academia de Ciencias de París tomó la resolución que habría de tener resultados trascendentales para el mundo entero: enviar a Suramérica la primera expedición científica encabezada por el joven Charles-Marie de La Condamine. Su propósito era lograr la medición exacta de un grado de latitud, para resolver la amarga y larga controversia, entre newtonianos, que, desde hacía mucho tiempo agitaba el seno de la Academia.

The bionomics of sand flies (Diptera: Phlebotominae) was studied monthly for two consecutive years in alluvial gallery forests in the department of Casanare, Northeastern Colombia. A total of 2,365 specimens and 10 species were captured using CDC light traps and Shannon traps, and from diurnal resting places, and human landing collections. Lutzomyia fairtigi Martins (55%), Lutzomyia micropyga (Mangabeira) (20.9%), and Lutzomyia antunesi (Coutinho) (13.5%) were the predominant species in the region. Lutzomyia flaviscutellata and Lutzomyia panamensis, potential vectors of Leishmania in Colombia and neighboring countries, were also collected, but in low numbers. Lu. fairtigi is an endemic species to Colombia, and minimal data are available on its biology and distribution. The present study provides additional information about Lu. fairtigi, such as the diurnal activity displayed by females on cloudy days, the greater density during the rainy season (April to October), marked anthropophilia, and the presence of flagellates in the midgut of one female.

The Andean Countries' Initiative (ACI) for controlling Chagas disease was officially created in 1997 within the framework of the Hipolito Unanue Agreement (UNANUE) between the Ministries of Health of Colombia, Ecuador, Peru, and Venezuela. Its objective was to interrupt transmission via vector and transfusion in the region, taking into account that there are 12.5 million people at risk in the four Andean countries forming the initiative in the area and around 3 million people are infected by Trypanosoma cruzi. The progress of control activities for the vector species present in the Andean sub-region, for different reasons, has been slow and control interventions have still not been installed in all geographical areas occupied by the target species. This has been partly due to lack of knowledge about these vector populations' biological characteristics, and consequent uncertainty about which are the appropriate control measures and strategies to be implemented in the region. The main vector species present important similarities in Venezuela and Colombia and in Ecuador and Northern Peru and they can be approached in a similar way throughout the whole regions, basing approaches on and adapting them to the current strategies being developed in Venezuela during the 1960s which have been progressively adopted in the Southern Cone and Central-American region. Additional measures are needed for keeping endemic areas free from Rhodnius prolixus silvatic populations, widely spread in the Orinoco region in Colombia and Venezuela. Regarding aetiological treatment, it is worth mentioning that (with the exception of Colombia) none of the other countries forming the ACI have registered medicaments available for treating infected young people. There are no suitable follow-up programmes in the sub-region or for treating cases of congenital Chagas disease. An integral and integrated programme encompassing all the aspects including transmission by transfusion which seems to have achieved extremely encouraging results in all countries, are urgently needed.

Introducción. La electroforesis de hemeproteínas salivares en geles de almidón es una técnica de reciente aplicación para diferenciar especies del género Rhodnius que poseen gran similaridad fenotípica o para diferenciar subpoblaciones de la misma especie, separadas geográficamente. De las 15 especies de Rhodnius descritas en latinoamérica, por lo menos ocho han sido reportadas en el territorio colombiano. Objetivo. Utilizar la electroforesis de hemeproteínas salivares para diferenciar R. prolixus de R. colombiensis, dos especies con similaridad fenotípica cuyos ciclos doméstico y selvático se superponen en la cuenca alta del río Magdalena en la región Central de Colombia. Materiales y Métodos. El contenido de las glándulas salivares de cada insecto se sometió a electroforesis en gel de almidón utilizando buffer de glicina y revelado de las bandas con 3,3',5,5'-tetrametilbenzidina. Los patrones de bandeo fueron registrados fotográficamente. Resultados. Usando esta técnica se observaron los perfiles electroforéticos de las hemeproteínas salivares de R. prolixus y R. colombiensis los cuales permitieron su inequívoca diferenciación. Conclusión. Estos resultados demuestran la utilidad de la técnica para la diferenciación entre R. prolixus y R. colombiensis en adición a métodos morfométricos y moleculares previamente utilizados para diferenciar estas dos especies.

La presente publicación recopila la información de registros de triatominos y datos sobre infección natural con tripanosomátidos a nivel departamental y municipal, publicada hasta la fecha así como la reportada por los servicios departamentales de salud e institutos de investigación. Se presentan figuras elaboradas de acuerdo a la información suministrada por los registros y una clasificación de la fauna triatomínica de acuerdo a las condiciones ecoepidemiológicas del país, teniendo en cuenta la altitud como factor determinante en la distribución de estos insectos. Teniendo en cuenta la frecuencia con que se reportan en el domicilio y peridomicilio, se consideran las siguientes especies como las de mayor riesgo de transmisión en Colombia: Rhodnius prolixus, Triatoma dimidiata, Triatoma maculata y Triatoma venosa. Se resalta la importancia de la vigilancia entomológica como herramienta indispensable para reforzar las estrategias de control de la transmisión de la enfermedad de Chagas, permitiendo también la evaluación del riesgo que representan las especies de triatominos silvestres en Colombia.

Introducción. Trypanosoma rangeli es la segunda especie de tripanosoma que infecta al hombre en América Latina. Se ha observado variabilidad en las características biológicas, bioquímicas y moleculares en diferentes aislamientos de este parásito. Objetivo. Estudiar las características morfológicas y moleculares de cepas de T. rangeli aisladas de diferentes especies de Rhodnius e inoculadas en diferentes especies de vertebrados. Materiales y métodos. Se utilizaron 19 cepas de T. rangeli aisladas de R. prolixus, R. pallescens y R. colombiensis en Colombia, R. ecuadoriensis en Perú y R. pallescens en Panamá. Se evaluó el polimorfismo de los tripomastigotes sanguíneos en ratones ICR y se estudió el pleomorfismo de la cepa P53 de T. rangeli KP1(-) inoculada en ratón, marsupial y canino. Se efectuó análisis de ADN polimorfo amplificado aleatorio en 12 cepas aisladas de cuatro especies de Rhodnius. Resultados. Se observaron tres grupos discretos en la longitud total de los tripomastigotes sanguíneos y la cepa P53 presentó diferencias significativas en el tamaño de los tripomastigotes sanguíneos en ratón, marsupial y canino. El análisis de ADN polimorfo amplificado aleatorio mostró segregación de las cepas en dos ramas correspondientes a las cepas de T. rangeli KP1(+) y T. rangeli KP1(-). De otra parte todos las cepas de T. rangeli KP1(-) se agruparon de acuerdo con las especies de Rhodnius de las cuales fueron aisladas. Conclusión. Este es el primer estudio que revela una estrecha asociación entre cepas de T. rangeli y las especies de Rhodnius, confirmando que cada especie de Rhodnius transmite al hospedero vertebrado poblaciones del parásito con claras diferencias fenotípicas y genotípicas, lo cual soporta la evolución clonal de estas poblaciones.

Introducción. Rhodnius colombiensis ocasionalmente ingresa a las viviendas humanas por lo cual se ha sugerido que podría desempeñar un importante papel en la transmisión de la tripanosomiasis americana. Objetivo. Definir el desempeño de R. colombiensis como vector, comparando los patrones de alimentación y defecación con los de R. prolixus, el principal vector domiciliado de Trypanosoma cruzi en Colombia. Materiales y métodos. Para cada estadio de R. colombiensis y R. prolixus se estudió el tiempo promedio para iniciar la picada, el tiempo para alcanzar la repleción, el número de interrupciones y defecaciones durante la comida, el tiempo transcurrido entre el fin de la alimentación y la primera defecación, el número de defecaciones durante 10, 60 y 95 min después de la comida y la cantidad de sangre ingerida. Resultados. El tiempo promedio para iniciar la picada de las ninfas N5, machos y hembras presentaron diferencias significativas entre las dos especies. El promedio de defecaciones por insecto durante los 10 min después de la alimentación fue mayor para cada estadio de R. prolixus y presentó diferencias significativas con R. colombiensis. Por otro lado, el peso promedio de sangre ingerida por R. colombiensis y R. prolixus en cada estadio presentó diferencias significativas en N1, N2, N5 y hembras. Conclusión. R. colombiensis presenta menor número de defecaciones que R. prolixus durante la comida. Un mayor porcentaje de R. prolixus defecan durante los 10, 60 y 95 min después de la alimentación. Sin embargo, R. colombiensis permanece mayor tiempo asociado al hospedero vertebrado, lo cual aumentaría la probabilidad de transmisión teniendo en cuenta el ingreso ocasional de los adultos a las viviendas humanas y sus elevadas prevalencias con T. cruzi y T. rangeli.

Introducción. Rhodnius colombiensis es un triatomino silvestre asociado a palmas de vino (Attalea butyracea) en la cuenca alta del río Magdalena (Colombia). La frecuente invasión de estos vectores al domicilio y las elevadas prevalencias de Trypanosoma cruzi en infecciones naturales constituyen un riesgo en la transmisión de la enfermedad de Chagas. Objetivo. Comparar la duración del ciclo de vida de R. colombiensis y R. prolixus en condiciones de laboratorio. Materiales y métodos. Se utilizaron 92 insectos de cada especie. Se estudió la duración de cada estadio, el número de comidas por estadio, el porcentaje de mortalidad, la causa de muerte, el promedio de huevos puestos por cada hembra, el número de huevos fértiles y la longevidad de los adultos. Resultados. Los promedios de duración de todos los estadios de desarrollo de R. colombiensis fueron mayores que en R. prolixus, presentando diferencias significativas en el tiempo transcurrido desde huevo hasta adulto. Los promedios de huevos puestos y huevos fértiles, presentaron diferencias significativas, siendo mayores en R. prolixus que en R. colombiensis. El porcentaje total de mortalidad de R. colombiensis fue de 31,5% y de 6,5% para R. prolixus. La longevidad en las hembras fue mayor en R. prolixus. Conclusiones. Los estadios de R. prolixus tienen menor duración, las ninfas en general realizan menor número de comidas que R. colombiensis, los adultos de R. prolixus toman mayor número de comidas y ovipositan mayor número de huevos fértiles, presentando hembras con mayor longevidad. Todos estos parámetros indican que R. prolixus presenta mejor éxito reproductivo que R. colombiensis, en las condiciones experimentales utilizadas. Los datos inéditos sobre el ciclo de vida de R. colombiensis serán de utilidad para el mantenimiento de las colonias en el laboratorio.

Introducción. Rhodnius prolixus es el vector más importante de la enfermedad de Chagas en Colombia. La caracterización genética de esta especie resulta útil para comprender su potencial de dispersión. La distribución del vector y la estructura genética de sus poblaciones, son factores importantes para implementar de manera adecuada las estrategias de control y vigilancia epidemiológica de la enfermedad de Chagas. Objetivo. Establecer relaciones genéticas entre poblaciones de R. prolixus capturadas en diferentes hábitat y áreas geográficas de Colombia, para dilucidar la estructura genética y dispersión del vector en el territorio colombiano. Materiales y métodos. Se analizaron tres poblaciones domiciliadas de R. prolixus provenientes de Tolima, Cundinamarca y la Sierra Nevada de Santa Marta; y una población silvestre procedente de Casanare. Se emplearon dos técnicas moleculares para evaluar la estructura genética de las poblaciones: análisis del ITS-2 del ADN ribosomal por PCR/RFLP y RAPDs. Resultados. R. prolixus presenta variabilidad genética moderada (Fst 0,057-0,15), entre las poblaciones domiciliadas se encontraron tasas de migración (Nm>1) que revelan flujo genético. Se encontró diferenciación genética de moderada-alta entre la población silvestre de Casanare y las poblaciones domésticas del centro del país (Tolima y Cundinamarca). Conclusión. Las poblaciones domiciliadas de R. prolixus son homogéneas debido a que existe flujo genético entre éstas; lo cual es favorable para el control químico, mientras que la población silvestre agrupa aparte de las domiciliadas. Se evidencia la necesidad de estudiar la estructura genética de los focos silvestres, sus posibles rutas de dispersión y el riesgo epidemiológico que representan.

BACKGROUND:Chagas disease is an anthropozoonosis caused by Trypanosoma cruzi. Two drugs are currently used for the etiological treatment of the disease: Nifurtimox (Lampit) and Benznidazole. This study presents a quasi-experimental trial (non-control group) of sixty-two patients who were treated for Chagas disease with Nifurtimox (Lampit), and were then followed for 30 months post-treatment. The safety of Nifurtimox (Lampit) for Chagas disease in this group of children primarily between 4 and 19 years old was also evaluated. MATERIALS AND METHODS:The 62 patients included in the study were selected when resulted seropositive for two out of three fundamentally different serological tests. All children were treated during two months according to protocols established by WHO. Monitoring was performed every twenty days to evaluate treatment safety. In 43 patients, two different serological tests: ELISA and IFAT; and two parasitological tests: blood culture, and real time PCR, (qPCR) were performed to assess therapeutic response, defined as post-treatment serological negativization. PRINCIPAL FINDINGS:All patients completed the treatment successfully, and six patients abandoned the post-treatment follow-up. Adverse effects occurred in 74% of patients, but only 4.8% of cases required temporary suspension to achieve 100% adherence to the 60-day treatment, and all symptoms reverted after treatment completion. Both parasite load (measured through qPCR) and antibodies (ELISA absorbance) evidenced a significant median reduction 6 months after treatment from 6.2 to 0.2 parasite equivalents/mL, and from 0.6 to 0.2 absorbance units respectively (p

Triatoma venosa presents a restricted geographical distribution in America and is considered as a secondary vector of Chagas disease in Colombia and Ecuador. A total of 120 adult insects were collected in domestic and peridomestic habitats in an endemic area of the department of Boyacá, Colombia, in order to determine their genetic structure through morphometric and molecular techniques. The head and wings of each specimen were used for the analyses of size, shape, and sexual dimorphism. A significant sexual dimorphism was found, although no differences in size among the studied groups were detected. Differences were found in the analyzed structures except for male heads. DNA was extracted from the legs in order to carry out the internal transcriber space-2 (ITS-2) amplification and the randon amplified polymorphic DNA (RAPD) analyses. Length polymorphisms were not detected in the ITS-2. Fst and Nm values were estimated (0.047 and 3.4, respectively). The high genetic flow found among the insects captured in the domicile and peridomiciliary environment does not permit a genetic differentiation, thus establishing the peridomicile as an important place for epidemiological surveillance.

Trypanosoma rangeli, a parasite generally considered non-pathogenic for man, is the second species of human trypanosome to be reported from the New World. The geographical distribution of T. rangeli often overlaps with that of T. cruzi, the same vertebrate and invertebrate hosts being infected. Their differentiation thus becomes of real, practical importance, particularly as they share approximately half the antigenic determinants recognized by the humoral response. Little is known about the life cycle of T. rangeli in the vertebrate host, although thousands of human and wild animal infections have been reported. Recent studies have revealed 2 major phylogenetic lineages in T. rangeli having different characteristics, thus leading to better understanding of the epidemiology and interactions with this parasite's vertebrate hosts and triatomine vectors. Based on further genetic characterization analysis, the authors have proposed 2 alternative hypotheses and consider that T. rangeli could have had clonal evolution or have been subjected to speciation processes.

BACKGROUND: Chagas disease, caused by infection with the protozoan Trypanosoma cruzi, remains a serious public health issue in Latin America. Genetically diverse, the species is sub-divided into six lineages, known as TcI-TcVI, which have disparate geographical and ecological distributions. TcII, TcV, and TcVI are associated with severe human disease in the Southern Cone countries, whereas TcI is associated with cardiomyopathy north of the Amazon. T. cruzi persists as a chronic infection, with cardiac and/or gastrointestinal symptoms developing years or decades after initial infection. Identifying an individual's history of T. cruzi lineage infection directly by genotyping of the parasite is complicated by the low parasitaemia and sequestration in the host tissues. METHODOLOGY/PRINCIPAL FINDINGS: We have applied here serology against lineage-specific epitopes of the T. cruzi surface antigen TSSA, as an indirect approach to allow identification of infecting lineage. Chagasic sera from chronic patients from a range of endemic countries were tested by ELISA against synthetic peptides representing lineage-specific TSSA epitopes bound to avidin-coated ELISA plates via a biotin labelled polyethylene glycol-glycine spacer to increase rotation and ensure each amino acid side chain could freely interact with their antibodies. 79/113 (70%) of samples from Brazil, Bolivia, and Argentina recognised the TSSA epitope common to lineages TcII/TcV/TcVI. Comparison with clinical information showed that a higher proportion of Brazilian TSSApep-II/V/VI responders had ECG abnormalities than non-responders (38% vs 17%; p

Chagas disease caused by the protozoan Trypanosoma cruzi is an important public health problem in Latin America. The immunological mechanisms involved in Chagas disease pathogenesis remain incompletely elucidated. The aim of this study was to explore cytokine profiles and their possible association to the infecting DTU and the pathogenesis of Chagas disease.109 sero-positive T. cruzi patients and 21 negative controls from Bolivia and Colombia, were included. Flow cytometry assays for 13 cytokines were conducted on human sera. Patients were divided into two groups: in one we compared the quantification of cytokines between patients with and without chronic cardiomyopathy; in second group we compared the levels of cytokines and the genetic variability of T. cruzi.Significant difference in anti-inflammatory and pro-inflammatory cytokines profiles was observed between the two groups cardiac and non-cardiac. Moreover, serum levels of IFN-γ, IL-12, IL-22 and IL-10 presented an association with the genetic variability of T.cruzi, with significant differences in TcI and mixed infections TcI/TcII.Expression of anti-inflammatory and pro-inflammatory cytokines may play a relevant role in determining the clinical presentation of chronic patients with Chagas disease and suggests the occurrence of specific immune responses, probably associated to different T. cruzi DTUs.

En 1998, se presentaron en Colombia 256.697 casos de malaria: 132.712 por Plasmodium falciparum, 121.161 por Plasmodium vivax y 2.824 infecciones mixtas. Dada esta situación, el Programa de Malaria del Laboratorio Nacional de Referencia se propuso evaluar la PCR anidada frente a la gota gruesa, como método alternativo para el diagnóstico de malaria. Se utilizó la reacción en cadena de la polimerasa (PCR) anidada para amplificar secuencias variables de genes de la subunidad ribosomal pequeña 18s (ssrARN) de P falciparum y P vivax. Para evaluar la PCR, se recolectaron 102 muestras de pacientes febriles en Quibdó (Chocó) y se determinó la presencia de parásitos tanto por PCR como por observación microscópica (gota gruesa). Los resultados de las dos pruebas se compararon entre sí. Los criterios de evaluación fueron: precisión, exactitud, limite de detección, sensibilidad, especificidad, valores predictivos positivo (VPP) y negativo (VPN) e índice kappa. La PCR anidada presentó una sensibilidad general del 100%: 96,7% para Pvivax y 100% para P falciparum. La especificidad general para P vivax y para P falciparum fue del 100%. El VPP general de la prueba fue del 90% y el VPN fue de 90.3%; para P vivax, el VPP fue de 77,0% y el VPN de 90.6%: para P falciparum, el VPP fue de 66,1% y el VPN de 94,3%. El índice kappa generaI fue de 1 y el de especie fue de 0,96. El límite de detección del ADN parasitario fue de 10 fg/ml. La PCR anidada detectó una muestra con infección mixta de P. vivax y P. falciparun que, por gota gruesa, dio resultado positivo para P. vivax. Se recomienda el uso de la PCR anidada como método de tamizaje en estudios epidemiológicos para la evaluación del Programa de Control Nacional de Malaria, en estudios orientados a detectar de manera temprana parasitemias en pacientes tratados cuya posible causa pueda ser la resistencia a los antimaláricos y como prueba de referencia para evaluar nuevos métodos diagnósticos.

Domiciliated Rhodnius prolixus and sylvatic R. colombiensis were analyzed in order to confirm their genetic divergence and verify the risk that the latter represents in the domiciliation process, and to provide tools for identifying the sources of possible reinfestation by triatomines in human dwellings allowing control programs to be undertaken. Comparison of random amplified polymorphic DNA amplification patterns and cluster analysis suggests reproductive discontinuity between the two species. The calculated statistical F value of 0.24 and effective migration rate of 0.6 individuals per generation are insufficient to maintain genetic homogeneity between them and confirm the absence of present genetic flow. R. colombiensis presents higher intrapopulation variability. Polymerase chain reaction of ribosomal DNA supports these findings. The low genetic flow between the two species implies that R. colombiensis do not represent an epidemiological risk for the domiciliary transmission of Trypanosoma cruzi in the Tolima Department. The lower variability of the domiciliated R. prolixus could result in greater susceptibility to the use of pesticides in control programs.

Se presenta la distribución geográfica y la importancia epidemiológica en la transmisión de la enfermedad de Chagas de las 23 especies de triatominos presentes en Colombia, tomando como base los registros bibliográficos existentes en el pais y las colecciones de referencia del Laboratorio de Entomologia del Instituto Nacional de Salud y del Centro de Investigaciones en Microbiologia y Parasitologia Tropical de la Universidad de los Andes; además, se realiza un análisis de las condiciones ecológicas en las cuales han sido registrados triatominos en los paises con fronteras con Colombia para poder establecer la presencia de posibles especies que aún no se han registrado en nuestro pais. De esta manera. existe la posibilidad de tener en Colombia especies con hábitos silvestres como: Alberprosenia malheiroi. Belminus laportei y Cavernicola lenti, preentes todas ellas en los departamentos amazónicos; Triatoma nigromaculata en la Cordillera Oriental y con hábitos intradomiciliarios; Panstrongylus chinai con posibilidad de encontrarse en hábitats selváticos o en las viviendas de los colonos de la Amazonia debido a su atracción por la luz y Alberprosenia goyovargasien hábitats silvestres de la frontera con Venezuela. Se resalta la importancia de la vigilancia entomológica a nivel nacional como estrategia para reforzar los estudios de triatominos silvestres en Colombia.

Como parte del estudio para el manejo y la conservación del Parque Natural Nacional Chiribiquete, desarrollado por la Fundación Puerto Rastrojo, se realizó un inventario de la fauna entomológica hematófaga en la estación biológica de Puerto Abeja (rio Mesay, municipio de Solano, departamento de Caquetá). Se calcularon los índices de similitud de Jaccard y el indice de diversidad de Shannon-Weaver y se determinaron los patrones de picadura de los dipteros diurnos para los seis tipos de bosques evaluados. Se registraron 31 especies de Culicidae, 1 de Triatominae, 1 de Ceratopogonidae, 2 de Simuliidae, 10 de Phlebotominae y 6 de Tabanidae. Se obtuvieron tres nuevos registros de especies de Culicidae para Colombia: Sabethes glaucodaemon, Trichoprosopon espini y Uranotaenia pallidoventer; y se amplió el registro de varios insectos hematófagos en el departamento de Caquetá. Se presentan los patrones de actividad de las especies diurnas y su relación con los tipos de bosque y se sugieren posibles medidas de control para evitar enfermedades transmitidas por diferentes vectores, resaltando la importancia epidemiológica de las especies capturadas que presentan capacidad vectorial.

La enfermedad de Chagas o tripanosomiasis americana está considerada como la cuarta causa de mortalidad en América Latina, provocando 43.000 muertes por año, principalmente debidas a la cardiopatía ocasionada por el parásito Trypanosoma cruzi cuando se anida en las fibras cardíacas. La existencia de la enfermedad de Chagas en seres humanos es un hecho puramente accidental. En la medida en que el hombre fue entrando en contacto con los focos naturales y provocó desequilibrios ecológicos, forzó a los triatomíneos infectados a ocupar viviendas humanas, llevándose a cabo el proceso de domiciliación, ya que no solamente encuentran refugio sino también suficiente alimento en la sangre humana y de animales domésticos. De esta manera, entró el hombre a formar parte activa de la cadena epidemiológica de la enfermedad de Chagas.Así pues, en esta patología se presentan ciclos silvestres sin participación del hombre y ciclos domiciliarios con su participación, los cuales son integrados e interdependientes. En el proceso de la dispersión geográfica y biológica el parásito ha evolucionado hasta el punto que existen una gran diversidad de subpoblaciones o cepas, las cuales una vez aisladas de los diversos huéspedes y estudiadas en el laboratorio exhiben características bien diferenciadas.La enfermedad asociada a la pobreza y a las malas condiciones de vivienda se transmite además de los insectos triatomíneos, a través de transfusiones de sangre contaminadas por el parásito y también congénitamente de madre infectada a hijo.Se han adelantado acciones de prevención y control en diversos países latinoamericanos promovidas por la Organización Mundial de la Salud (OMS) y la Organización Panamericana de la Salud (OPS) conjuntamente con los Ministerios de Salud de los países respectivos. En 1991 nace, por ejemplo, la así llamada Iniciativa de los Países del Cono Sur, la cual tuvo como objetivo central interrumpir la transmisión vectorial y transfusional de la enfermedad de Chagas en Argentina, Brasil, Bolivia, Chile, Paraguay y Uruguay. Las herramientas que se tienen a la mano son eficaces y económicamente abordables por los Ministerios de Salud de los países mencionados: insecticidas de efecto residual, pinturas insecticidas de lenta liberación, potes fumígenos, cajas sensoras para la detección de reinfestaciones domiciliares, nuevos materiales de construcción y pruebas de laboratorio para el tamizaje de las donaciones de sangre para transfusiones.Los estudios epidemiológicos adelantados con la estrecha colaboración de los Ministerios de Salud de los países de América Latina permiten calcular que un total de 100 millones de personas, es decir un cuarto de los habitantes de América Latina, viven en áreas endémicas y están en riesgo de adquirir la infección y que hay 18 millones de personas infectadas en todo el continente.La Iniciativa de los Países del Cono Sur fue establecida por los Ministerios de Salud de estos países en 1991 en Brasilia y ha venido trabajando a través de una Comisión Técnica Intergubernamental constituida por los responsables de los programas de control...

Situación Actual de Colombia.La enfermedad de Chagas o tripanosomiasis americana es una enfermedad parasitaria crónica causada por un protozoario flagelado el Trypanosoma cruzi, descrito por primera vez por Carlos Chagas, médico brasileño, a comienzos de este siglo y en su honor se denominó la enfermedad que lleva su nombre.Este parásito normalmente se transmite al ser humano a través de insectos triatomíneos estrictamente hematófagos de la familia Reduviidae, en el momento en que perforan la piel para succionar la sangre que los alimenta.Sin embargo, no se inocula directamente por intermedio de las estructuras bucales del insecto en el momento de la picadura como en el caso de las tripanosomiasis africanas, si no que se deposita pasivamente en la piel a través de las heces del insecto, penetrando en el cuerpo por la herida que causa la picadura u otras abrasiones de la piel o de la mucosa. El T. cruzi, también puede transmitirse por infección congénita, por transfusión de sangre contaminada o por el transplante de órganos contaminados. El ciclo vital del parásito es largo y complejo y su desarrollo tiene varias etapas, tanto en el vector triatomineo como en el huésped vertebrado .La Enfermedad de Chagas constituye una amenaza permanente para casi la cuarta parte de toda la población de América Latina. Si bien la enfermedad se encuentra presente en toda América Central y del Sur, sus manifestaciones y características epidemiológicas son altamente variables entre una y otra zona endémica. Existe una gran diversidad en las tasas de prevalencia, formas de transmisión, características parasitarias, patología clínica, vectores y reservorios.Más que cualquier otra enfermedad parasitaria, la enfermedad de Chagas se relaciona con el desarrollo económico y social de la población: los insectos triatomineos y las enfermedades que ellos transmiten existirán mientras en Latinoamérica persista la vivienda inadecuada, la migración frecuente de personas y la rápida urbanización. Hasta que no cambien estas condiciones, para lo cual habrán de transcurrir muchos años, deberá continuarse la lucha contra la enfermedad.La enfermedad de Chagas representa un problema de salud grave en 17 países Latinoamericanos, con un total estimado por la Organización Mundial de la Salud de 100 millones de personas expuestas a la enfermedad, y al rededor de 20 millones de personas infectadas (1). Fuera de los focos naturales de transmisión, se ha notificado la transmisión por transfusión sanguínea con creciente frecuencia.Se estima que cada año se infectan alrededor de 500.000 personas y de éstas 300.000 son niños. La mortalidad asciende al 10-15% durante la fase aguda de la enfermedad.En Colombia, el T. cruzi se detecta frecuentemente a todo lo largo del Valle del Río Magdalena, en la región del Catatumbo, la Sierra Nevada de Santa Marta, el piedemonte de los Llanos Orientales y la Serranía de la Macarena...

Se presentan los antecedentes de la presencia de triatominos domiciliados en Colombia desde la primera publicación hecha por el doctor César Uribe en 1929 (Editorial Minerva, Bogotá) donde se consigna el hallazgo de Trypanosoma cruzi y Trypanosoma rangeli en 15 ejemplares de Rhodnius prolixus provenientes de Prado, Tolima. A continuación se analizan las demás publicaciones en orden cronológico. Informe del Profesor Emile Brumpt (Revista de Higiene, año XX – Números 11 y 12, 1939, Distribución de los Triatominos en Colombia, Hernando Ucrós, Hernando Rocha, Marcos Duque (Antioquia Médica Vol. 21 – 1971 No. 8; Distribution of Triatomine – transmitted trypanosomamiasis in Colombia and vew records of the bugs and infections. A. D’Alessandro, Pablo Barreto and C. A. Duarte R. (1972, J. Med. Ent. Vol. 8, No. 2: 159-172; Colombian Triatominae and their infestation with Trypanosomatid Flagellates. Marinkelle Cornelis J. (1972 Mitt Inst. – Aleman Invest. Cient. 6:13-29); Nuevos registros de triatominos domiciliarios y extradomiciliarios en Colombia. Antonio D’Alessandro, Pablo Barreto y Mauricio Thomas. Colombia Médica Vol. 12 No. 2 – 1981.Basándose en la anterior información se llega a la conclusión de que los Triatominae domiciliados en Colombia se encuentran en poblaciones situadas sobre los 2.000msnm, principalmente en las zonas central y oriental del país.El Instituto Nacional de Salud en el año de 1990, como una primera etapa para tratar de determinar la magnitud del problema de la tripanosomiasis americana en Colombia, realizó una investigación para establecer la verdadera distribución de los triatominos domiciliarios en el país.El estudio se hizo usando el sistema de muestreo, en una muestra estadísticamente representativa. Se inspeccionaron las viviendas de las zonas rurales; se capturaron triatominos vectores que, posteriormente, fueron examinados sistemáticamente en búsqueda de tripanosomas y, se les practicó xenodiagnóstico a los animales domésticos con el mismo fin...

BACKGROUND: Trypanosoma cruzi, the causative agent of Chagas disease, displays significant genetic variability revealed by six Discrete Typing Units (TcI-TcVI). In this pathology, oral transmission represents an emerging epidemiological scenario where different outbreaks associated to food/beverages consumption have been reported in Argentina, Bolivia, Brazil, Ecuador and Venezuela. In Colombia, six human oral outbreaks have been reported corroborating the importance of this transmission route. Molecular epidemiology of oral outbreaks is barely known observing the incrimination of TcI, TcII, TcIV and TcV genotypes. METHODOLOGY AND PRINCIPAL FINDINGS: High-throughput molecular characterization was conducted performing MLMT (Multilocus Microsatellite Typing) and mtMLST (mitochondrial Multilocus Sequence Typing) strategies on 50 clones from ten isolates. Results allowed observing the occurrence of TcI, TcIV and mixed infection of distinct TcI genotypes. Thus, a majority of specific mitochondrial haplotypes and allelic multilocus genotypes associated to the sylvatic cycle of transmission were detected in the dataset with the foreseen presence of mitochondrial haplotypes and allelic multilocus genotypes associated to the domestic cycle of transmission. CONCLUSIONS: These findings suggest the incrimination of sylvatic genotypes in the oral outbreaks occurred in Colombia. We observed patterns of super-infection and/or co-infection with a tailored association with the severe forms of myocarditis in the acute phase of the disease. The transmission dynamics of this infection route based on molecular epidemiology evidence was unraveled and the clinical and biological implications are discussed.

BACKGROUND: A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation. METHODOLOGY/FINDINGS: An international collaborative study was launched by expert PCR laboratories from 16 countries. Currently used strategies were challenged against serial dilutions of purified DNA from stocks representing T. cruzi discrete typing units (DTU) I, IV and VI (set A), human blood spiked with parasite cells (set B) and Guanidine Hidrochloride-EDTA blood samples from 32 seropositive and 10 seronegative patients from Southern Cone countries (set C). Forty eight PCR tests were reported for set A and 44 for sets B and C; 28 targeted minicircle DNA (kDNA), 13 satellite DNA (Sat-DNA) and the remainder low copy number sequences. In set A, commercial master mixes and Sat-DNA Real Time PCR showed better specificity, but kDNA-PCR was more sensitive to detect DTU I DNA. In set B, commercial DNA extraction kits presented better specificity than solvent extraction protocols. Sat-DNA PCR tests had higher specificity, with sensitivities of 0.05-0.5 parasites/mL whereas specific kDNA tests detected 5.10(-3) par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/µl of DNA from all stocks, 5 par/mL spiked blood). The median values of sensitivities, specificities and accuracies obtained in testing the Set C samples with the 16 tests determined to be good performing by analyzing Sets A and B samples varied considerably. Out of them, four methods depicted the best performing parameters in all three sets of samples, detecting at least 10 fg/µl for each DNA stock, 0.5 par/mL and a sensitivity between 83.3-94.4%, specificity of 85-95%, accuracy of 86.8-89.5% and kappa index of 0.7-0.8 compared to consensus PCR reports of the 16 good performing tests and 63-69%, 100%, 71.4-76.2% and 0.4-0.5, respectively compared to serodiagnosis. Method LbD2 used solvent extraction followed by Sybr-Green based Real time PCR targeted to Sat-DNA; method LbD3 used solvent DNA extraction followed by conventional PCR targeted to Sat-DNA. The third method (LbF1) used glass fiber column based DNA extraction followed by TaqMan Real Time PCR targeted to Sat-DNA (cruzi 1/cruzi 2 and cruzi 3 TaqMan probe) and the fourth method (LbQ) used solvent DNA extraction followed by conventional hot-start PCR targeted to kDNA (primer pairs 121/122). These four methods were further evaluated at the coordinating laboratory in a subset of human blood samples, confirming the performance obtained by the participating laboratories. CONCLUSION/SIGNIFICANCE: This study represents a first crucial step towards international validation of PCR procedures for detection of T. cruzi in human blood samples.

Chagas disease caused by Trypanosoma cruzi is a complex disease that is endemic and an important problem in public health in Latin America. The T. cruzi parasite is classified into six discrete taxonomic units (DTUs) based on the recently proposed nomenclature (TcI, TcII, TcIII, TcIV, TcV and TcVI). The discovery of genetic variability within TcI showed the presence of five genotypes (Ia, Ib, Ic, Id and Ie) related to the transmission cycle of Chagas disease. In Colombia, TcI is more prevalent but TcII has also been reported, as has mixed infection by both TcI and TcII in the same Chagasic patient. The objectives of this study were to determine the T. cruzi DTUs that are circulating in Colombian chronic Chagasic patients and to obtain more information about the molecular epidemiology of Chagas disease in Colombia. We also assessed the presence of electrocardiographic, radiologic and echocardiographic abnormalities with the purpose of correlating T. cruzi genetic variability and cardiac disease. Molecular characterization was performed in Colombian adult chronic Chagasic patients based on the intergenic region of the mini-exon gene, the 24Sα and 18S regions of rDNA and the variable region of satellite DNA, whereby the presence of T.cruzi I, II, III and IV was detected. In our population, mixed infections also occurred, with TcI-TcII, TcI-TcIII and TcI-TcIV, as well as the existence of the TcI genotypes showing the presence of genotypes Ia and Id. Patients infected with TcI demonstrated a higher prevalence of cardiac alterations than those infected with TcII. These results corroborate the predominance of TcI in Colombia and show the first report of TcIII and TcIV in Colombian Chagasic patients. Findings also indicate that Chagas cardiomyopathy manifestations are more correlated with TcI than with TcII in Colombia.

Human Chagas disease is a purely accidental occurrence. As humans came into contact with the natural foci of infection might then have become infected as a single addition to the already extensive host range of Trypanosoma cruzi that includes other primates. Thus began a process of adaptation and domiciliation to human habitations through which the vectors had direct access to abundant food as well as protection from climatic changes and predators. Our work deals with the extraction and specific amplification by polymerase chain reaction of T. cruzi DNA obtained from mummified human tissues and the positive diagnosis of Chagas disease in a series of 4,000-year-old Pre-Hispanic human mummies from the northern coast of Chile. The area has been inhabited at least for 7,000 years, first by hunters, fishers and gatherers, and then gradually by more permanent settlements. The studied specimens belonged to the Chinchorro culture, a people inhabiting the area now occupied by the modern city of Arica. These were essentially fishers with a complex religious ideology, which accounts for the preservation of their dead in the way of mummified bodies, further enhanced by the extremely dry conditions of the desert. Chinchorro mummies are, perhaps, the oldest preserved bodies known to date.

Phylogenetic studies of Trypanosoma cruzi have identified the existence of two groups: T. cruzi I and T. cruzi II. There are aspects that still remain unknown about the genetic variability within the T. cruzi I group. Given its epidemiological importance, it is necessary to have a better understanding of T. cruzi transmission cycles. Our purpose was to corroborate the existence of haplotypes within the T. cruzi I group and to describe the genetic variability and phylogenetic relationships, based on single nucleotide polymorphisms (SNPs) found in the miniexon gene intergenic region, for the isolates from different hosts and epidemiological transmission cycles in Colombian regions. 31 T. cruzi isolates were molecularly characterized. Phylogenetic relationships within T. cruzi I isolates showed four haplotype groups (Ia–Id), associated with their transmission cycle. In previous studies, we reported that haplotype Ia is mainly associated with the domestic cycle and domiciliated Rhodnius prolixus. Haplotype Ib is associated with the domestic cycle and peridomestic cycle, haplotype Ic is closely related with the peridomestic cycle, and haplotype Id is strongly associated with the sylvatic cycle. The phylogenetic methodologies applied in this study are tools that bolster the associations among isolates and thus shed light on Chagas disease epidemiology.

Among Chagas disease triatomine vectors, the largest genus, Triatoma, includes species of high public health interest. Triatoma dimidiata, the main vector throughout Central America and up to Ecuador, presents extensive phenotypic, genotypic, and behavioral diversity in sylvatic, peridomestic and domestic habitats, and non-domiciliated populations acting as reinfestation sources. DNA sequence analyses, phylogenetic reconstruction methods, and genetic variation approaches are combined to investigate the haplotype profiling, genetic polymorphism, phylogeography, and evolutionary trends of T. dimidiata and its closest relatives within Triatoma. This is the largest interpopulational analysis performed on a triatomine species so far.Triatomines from Mexico, Guatemala, Honduras, Nicaragua, Panama, Cuba, Colombia, Ecuador, and Brazil were used. Triatoma dimidiata populations follow different evolutionary divergences in which geographical isolation appears to have had an important influence. A southern Mexican-northern Guatemalan ancestral form gave rise to two main clades. One clade remained confined to the Yucatan peninsula and northern parts of Chiapas State, Guatemala, and Honduras, with extant descendants deserving specific status. Within the second clade, extant subspecies diversity was shaped by adaptive radiation derived from Guatemalan ancestral populations. Central American populations correspond to subspecies T. d. dimidiata. A southern spread into Panama and Colombia gave the T. d. capitata forms, and a northwestern spread rising from Guatemala into Mexico gave the T. d. maculipennis forms. Triatoma hegneri appears as a subspecific insular form.The comparison with very numerous Triatoma species allows us to reach highly supported conclusions not only about T. dimidiata, but also on different, important Triatoma species groupings and their evolution. The very large intraspecific genetic variability found in T. dimidiata sensu lato has never been detected in a triatomine species before. The distinction between the five different taxa furnishes a new frame for future analyses of the different vector transmission capacities and epidemiological characteristics of Chagas disease. Results indicate that T. dimidiata will offer problems for control, although dwelling insecticide spraying might be successful against introduced populations in Ecuador.