Transmission ecology of Trypanosoma cruzi by Rhodnius prolixus (Reduviidae: Triatominae) infesting palm-tree species in the Colombian Orinoco, indicates risks to human populations.

BACKGROUND: Chagas disease, affecting approximately eight million individuals in tropical regions, is primarily transmitted by vectors. Rhodnius prolixus, a triatomine vector, commonly inhabits in ecotopes with diverse palm tree species, creating optimal conditions for vector proliferation. This study aims to explore the transmission ecology of Trypanosoma cruzi, the causative parasite of Chagas disease, by investigating the feeding patterns and natural infection rates of R. prolixus specimens collected from various wild palm species in the Colombian Orinoco region. MATERIALS AND METHODS: To achieve this objective, we sampled 35 individuals from three palm species (Attalea butyracea, Acrocomia aculeata, and Mauritia flexuosa) in a riparian forest in the Casanare department of eastern Colombia, totaling 105 sampled palm trees. DNA was extracted and analyzed from 115 R. prolixus specimens at different developmental stages using quantitative PCR (qPCR) for T. cruzi detection and identification of discrete typing units. Feeding preferences were determined by sequencing the 12S rRNA gene amplicon through next-generation sequencing. RESULTS: A total of 676 R. prolixus specimens were collected from the sampled palms. The study revealed variation in population densities and developmental stages of R. prolixus among palm tree species, with higher densities observed in A. butyracea and lower densities in M. flexuosa. TcI was the exclusive T. cruzi discrete typing unit (DTU) found, with infection frequency positively correlated with R. prolixus abundance. Insects captured in A. butyracea exhibited higher abundance and infection rates than those from other palm species. The feeding sources comprised 13 mammal species, showing no significant differences between palm species in terms of blood sources. However, Didelphis marsupialis and Homo sapiens were present in all examined R. prolixus, and Dasypus novemcinctus was found in 89.47% of the insects. CONCLUSION: This study highlights the significance of wild palms, particularly A. butyracea, as a substantial risk factor for T. cruzi transmission to humans in these environments. High population densities and infection rates of R. prolixus were observed in each examined palm tree species.

Characterizing the blood microbiota of omnivorous and frugivorous bats (Chiroptera: Phyllostomidae) in Casanare, eastern Colombia.

Bats are known reservoirs of seemingly-innocuous pathogenic microorganisms (including viruses, bacteria, fungi, and protozoa), which are associated with triggering disease in other zoonotic groups. The taxonomic diversity of the bats' microbiome is likely associated with species-specific phenotypic, metabolic, and immunogenic capacities. To date, few studies have described the diversity of bat blood microbial communities. Then, this study used amplicon-based next generation sequencing of the V4 hypervariable region of the 16S-rRNA gene in blood samples from omnivorous (n = 16) and frugivorous (n = 9) bats from the department of Casanare in eastern Colombia. We found the blood microbiota in bats to be composed of, among others, Bartonella and Mycoplasma bacterial genera which are associated with various disease phenotypes in other mammals. Furthermore, our results suggest that the bats' dietary habits might determine the composition and the persistence of some pathogens over others in their bloodstream. This study is among the first to describe the blood microbiota in bats, to reflect on co-infection rates of multiple pathogens in the same individual, and to consider the influence of diet as a factor affecting the animal's endogenous microbial community.

Trypanosoma cruzi Parasite Burdens of Several Triatomine Species in Colombia.

Trypanosoma cruzi, the causal agent of Chagas disease, is mainly transmitted by insects of the Triatominae subfamily. In Colombia, there are 26 triatomine species, and 16 of them are naturally infected with the parasite. The parasite loads of naturally infected vectors can be significant in targeting specific species that can affect the epidemiology of the disease. Studying their ecology and behavior is vital to understand their role in T. cruzi transmission dynamics. We evaluated the parasite loads of 182 field-collected triatomines corresponding to 10 species in 13 departments across Colombia. We standardized a methodology to quantify T. cruzi DNA in these insects. We obtained a LOD (limit of detection) of 3.05 p-eq/mL. The 82% of triatomines we evaluated were positive for T. cruzi infection, with loads ranging from hundreds to millions of equivalent parasites per milliliter. Panstrongylus geniculatus, Rhodnius prolixus, and Triatoma dimidiata were the species with the highest loads of T. cruzi; however, other species whose role as vectors is still unknown were also found with high loads of parasites. Our results suggest the relevance of secondary species for T. cruzi transmission in Colombia. We hope our data can help improve entomological surveillance and vector control programs in the country and the region.

An overview of the trypanosomatid (Kinetoplastida: Trypanosomatidae) parasites infecting several mammal species in Colombia.

BACKGROUND: Trypanosomatids are among the most critical parasites for public health due to their impact on human, animal, and plant health. Diseases associated with these pathogens manifest mainly in poor and vulnerable populations, where social, environmental, and biological factors modulate the case incidence and geographical distribution. METHODS: We used Sanger and amplicon-based next-generation sequencing (NGS) in samples from different mammals to identify trypanosomatid infections in several departments in Colombia. A total of 174 DNA samples (18 humans, 83 dogs, and 73 wild mammals) were analyzed by conventional PCR using a fragment of the heat shock protein 70 (Hsp70) gene and Sanger sequenced the positive samples. Twenty-seven samples were sent for amplicon-based NGS using the same gene fragment. Data obtained were used to perform diversity analyses. RESULTS: One hundred and thirteen samples were positive for PCR by Hsp70 fragment; these corresponded to 22.1% Leishmania spp., 18.6% L. amazonensis, 9.7% L. braziliensis, 14.2% L. infantum, 8% L. panamensis, and 27.4% Trypanosoma cruzi. Comparison of the identified species by the two sequencing technologies used resulted in 97% concordance. Alpha and beta diversity indices were significant, mainly for dogs; there was an interesting index of coinfection events in the analyzed samples: different Leishmania species and the simultaneous presence of T. cruzi and even T. rangeli in one of the samples analyzed. Moreover, a low presence of L. braziliensis was observed in samples from wild mammals. Interestingly, to our knowledge, this is the first report of Leishmania detection in Hydrochaeris hydrochaeris (capybara) in Colombia. CONCLUSIONS: The Hsp70 fragment used in this study is an optimal molecular marker for trypanosomatid identification in many hosts and allows the identification of different species in the same sample when amplicon-based sequencing is used. However, the use of this fragment for molecular diagnosis through conventional PCR should be carefully interpreted because of this same capacity to identify several parasites. This point is of pivotal importance in highly endemic countries across South America because of the co-circulation of different genera from the Trypanosomatidae family. The findings show an interesting starting point for One Health approaches in which coevolution and vector-host interactions can be studied.

Towards environmental detection of Chagas disease vectors and pathogen.

Chagas disease vector control relies on prompt, accurate identification of houses infested with triatomine bugs for targeted insecticide spraying. However, most current detection methods are laborious, lack standardization, have substantial operational costs and limited sensitivity, especially when triatomine bug densities are low or highly focal. We evaluated the use of FTA cards or cotton-tipped swabs to develop a low-technology, non-invasive method of detecting environmental DNA (eDNA) from both triatomine bugs and Trypanosoma cruzi for use in household surveillance in eastern Colombia, an endemic region for Chagas disease. Study findings demonstrated that Rhodnius prolixus eDNA, collected on FTA cards, can be detected at temperatures between 21 and 32 °C, when deposited by individual, recently blood-fed nymphs. Additionally, cotton-tipped swabs are a feasible tool for field sampling of both T. cruzi and R. prolixus eDNA in infested households and may be preferable due to their lower cost. eDNA detection should not yet replace current surveillance tools, but instead be evaluated in parallel as a more sensitive, higher-throughput, lower cost alternative. eDNA collection requires virtually no skills or resources in situ and therefore has the potential to be implemented in endemic communities as part of citizen science initiatives to control Chagas disease transmission.

Phylogenetic relationships and evolutionary patterns of the genus Psammolestes Bergroth, 1911 (Hemiptera: Reduviidae: Triatominae).

BACKGROUND: The evolutionary history of biodiversity in South America has been poorly studied in the seasonal dry tropical forest (SDTF). Species diversification in this ecosystem may have a twofold explanation. First, intermittent connections in the middle and late Pleistocene promoted species dispersal and/or genetic connectivity between lineages isolated in disjunct patches of forest. Second, allopatric speciation proceeded immediately after the formation and colonization of the SDTF in the Neogene. Here we studied the diversification of Psammolestes, a genus endemic of the SDTF and naturally infected with Trypanosoma cruzi (agent of Chagas disease), using a combination of phylogenetic, population genetics and niche model methods, and evaluated the reliability of the three morphospecies currently recognized. RESULTS: Our multilocus analyses recovered P. coreodes and P. tertius in a monophyletic clade sister to P. arthuri. Species delimitation tests recovered these lineages as different species despite the shared genetic variation observed between P. coreodes and P. tertius in five genes. Also, genetic variation of the genus clustered in three groups that were consistent with the three morphospecies. Our demographic model predicted a scenario of divergence in absence of gene flow, suggesting that mixed haplotypes may be the result of shared ancestral variation since the divergence of the subtropical-temperate species P. coreodes and P. tertius. In contrast, the tropical species P. arthuri was highly differentiated from the other two in all tests of genetic structure, and consistently, the Monmonier's algorithm identified a clear geographical barrier that separates this species from P. coreodes and P. tertius. CONCLUSIONS: We found three genetically structured lineages within Psammolestes that diverged in absence of gene flow in the late Miocene. This result supports a scenario of species formation driven by geographical isolation rather than by divergence in the face of gene flow associated with climatic oscillations in the Pleistocene. Also, we identified the Amazon basin as a climatic barrier that separates tropical from subtropical-temperate species, thus promoting allopatric speciation after long range dispersion. Finally, each species of Psammolestes occupies different climatic niches suggesting that niche conservatism is not crucial for species differentiation. These findings influence the current vector surveillance programs of Chagas disease in the region.

Development of an Amplicon-Based Next-Generation Sequencing Protocol to Identify Leishmania Species and Other Trypanosomatids in Leishmaniasis Endemic Areas.

Trypanosomatid infections are an important public health threat affecting many low-income countries across the tropics, particularly in the Americas. Trypanosomatids can infect many vertebrate, invertebrate, and plant species and play an important role as human pathogens. Among these clinically relevant pathogens are species from the genera Leishmania and Trypanosoma. Mixed trypanosomatid infections remain a largely unexplored phenomenon. Herein, we describe the application of an amplicon-based next-generation sequencing (NGS) assay to detect and identify trypanosomatid species in mammalian reservoirs, human patients, and sand fly vectors throughout regions of Leishmania endemicity. Sixty-five samples from different departments of Colombia, including two samples from Venezuela, were analyzed: 49 samples from cutaneous leishmaniasis (CL) patients, 8 from sand flies, 2 from domestic reservoirs (Canis familiaris), and 6 from wild reservoirs (Phyllostomus hastatus). DNA from each sample served to identify the presence of trypanosomatids through conventional PCR using heat shock protein 70 (HSP70) gene as the target. PCR products underwent sequencing by Sanger sequencing and NGS, and trypanosomatid species were identified by using BLASTn against a reference database built from trypanosomatid-derived HSP70 sequences. The alpha and beta diversity indexes of amplicon sequence variants were calculated for each group. The results revealed the presence of mixed infections with more than two Leishmania species in 34% of CL samples analyzed. Trypanosoma cruzi was identified in samples from wild reservoirs, as well as in sand fly vectors. Coinfection events with three different Leishmania species were identified in domestic reservoirs. These findings depose the traditional paradigm of leishmaniasis as being a single-species-driven infection and redraw the choreography of host-pathogen interaction in the context of multiparasitism. Further research is needed to decipher how coinfections may influence disease progression. This knowledge is key to developing an integrated approach for diagnosis and treatment. IMPORTANCE Traditionally, there has been a frequent, yet incorrect assumption that phlebotomine vectors, animal reservoirs, and human hosts are susceptible to Leishmania infection by a single parasite species. However, current evidence supports that these new vector-parasite-reservoir associations lend vectors and reservoirs greater permissiveness to certain Leishmania species, thus promoting the appearance of coinfection events, particularly in disease-endemic regions. The present study describes the application of an amplicon-based next-generation sequencing (NGS) assay to detect and identify trypanosomatid species in mammalian reservoirs, human patients, and sand fly vectors from regions of endemicity for leishmaniasis. This changes our understanding of the clinical course of leishmaniasis in areas of endemicity.

Succinate dehydrogenase gene as a marker for studying Blastocystis genetic diversity.

Blastocystis has been reported as the most common eukaryotic microorganism residing in the intestines of both humans and animals, with a prevalence of up to 100% in some populations. Since this is a cryptic species, sequence polymorphism are the single strategy to analyses its genetic diversity, being traditionally used the analysis of ssu rRNA gene sequence to determine alleles and subtypes (STs) for this species. This multicopy gene has shown high diversity among different STs, making necessary to explore other genes to assess intraspecific diversity. This study evaluated the use of a novel genetic marker, succinate dehydrogenase (SDHA), for the typing and evaluation of the genetic diversity and genetic population structure of Blastocystis. In total, 375 human fecal samples were collected and subjected to PCR, subtyped using the ssu rRNA marker, and then the SDHA gene was amplified via PCR for 117 samples. We found some incongruences between tree topologies for both molecular markers. However, the clustering by ST previously established for Blastocystis was congruent in the concatenated sequence. SDHA showed lower reticulation (The origination of a lineage through the partial merging of two ancestor lineages) signals and better intra ST clustering ability. Clusters with geographical associations were observed intra ST. The genetic diversity was lower in the marker evaluated compared to that of the ssu rRNA gene (nucleotide diversity = 0.03344 and 0.16986, respectively) and the sequences analyzed showed population expansion with genetic differentiation principally among STs. The ssu rRNA gene was useful to explore interspecific diversity but together with the SDHA gene the resolution power to evaluate intra ST diversity was higher. These results showed the potential of the SDHA marker for studying the intra ST genetic diversity of Blastocystis related with geographical location and the inter ST diversity using the concatenated sequences.

Species-dependent variation of the gut bacterial communities across Trypanosoma cruzi insect vectors.

Triatomines (Hemiptera: Reduviidae) are the insect vectors of Trypanosoma cruzi, the causative agent of Chagas disease. The gut bacterial communities affect the development of T. cruzi inside the vector, making the characterization of its composition important in the understanding of infection development. We collected 54 triatomine bugs corresponding to four genera in different departments of Colombia. DNA extraction and PCR were performed to evaluate T. cruzi presence and to determine the discrete typing unit (DTU) of the parasite. PCR products of the bacterial 16S rRNA gene were pooled and sequenced. Resulting reads were denoised and QIIME 2 was used for the identification of amplicon sequence variants (ASVs). Diversity (alpha and beta diversity) and richness analyses, Circos plots, and principal component analysis (PCA) were also performed. The overall T. cruzi infection frequency was 75.9%, with TcI being the predominant DTU. Approximately 500,000 sequences were analyzed and 27 bacterial phyla were identified. The most abundant phyla were Proteobacteria (33.9%), Actinobacteria (32.4%), Firmicutes (19.6%), and Bacteroidetes (7.6%), which together accounted for over 90% of the gut communities identified in this study. Genera were identified for these main bacterial phyla, revealing the presence of important bacteria such as Rhodococcus, Serratia, and Wolbachia. The composition of bacterial phyla in the gut of the insects was significantly different between triatomine species, whereas no significant difference was seen between the state of T. cruzi infection. We suggest further investigation with the evaluation of additional variables and a larger sample size. To our knowledge, this study is the first characterization of the gut bacterial structure of the main triatomine genera in Colombia.

Identification of blood-feeding sources in Panstrongylus, Psammolestes, Rhodnius and Triatoma using amplicon-based next-generation sequencing.

BACKGROUND: Triatomines are hematophagous insects that play an important role as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. These insects have adapted to multiple blood-feeding sources that can affect relevant aspects of their life-cycle and interactions, thereby influencing parasitic transmission dynamics. We conducted a characterization of the feeding sources of individuals from the primary circulating triatomine genera in Colombia using amplicon-based next-generation sequencing (NGS). METHODS: We used 42 triatomines collected in different departments of Colombia. DNA was extracted from the gut. The presence of T. cruzi was identified using real-time PCR, and discrete typing units (DTUs) were determined by conventional PCR. For blood-feeding source identification, PCR products of the vertebrate 12S rRNA gene were obtained and sequenced by next-generation sequencing (NGS). Blood-meal sources were inferred using blastn against a curated reference dataset containing the 12S rRNA sequences belonging to vertebrates with a distribution in South America that represent a potential feeding source for triatomine bugs. Mean and median comparison tests were performed to evaluate differences in triatomine blood-feeding sources, infection state, and geographical regions. Lastly, the inverse Simpson's diversity index was calculated. RESULTS: The overall frequency of T. cruzi infection was 83.3%. TcI was found as the most predominant DTU (65.7%). A total of 67 feeding sources were detected from the analyses of approximately 7 million reads. The predominant feeding source found was Homo sapiens (76.8%), followed by birds (10.5%), artiodactyls (4.4%), and non-human primates (3.9%). There were differences among numerous feeding sources of triatomines of different species. The diversity of feeding sources also differed depending on the presence of T. cruzi. CONCLUSIONS: To the best of our knowledge, this is the first study to employ amplicon-based NGS of the 12S rRNA gene to depict blood-feeding sources of multiple triatomine species collected in different regions of Colombia. Our findings report a striking read diversity that has not been reported previously. This is a powerful approach to unravel transmission dynamics at microgeographical levels.

Development of a Multilocus Sequence Typing Scheme for Giardia intestinalis.

Giardia intestinalis is an intestinal protozoan most commonly found in humans. It has been grouped into 8 assemblages (A-H). Markers such as the glutamate dehydrogenase gene, triose phosphate isomerase and beta-giardin (ß-giardin) have been widely used for genotyping. In addition, different genetic targets have been proposed as a valuable alternative to assess diversity and genetics of this microorganism. Thus, our objective was to evaluate new markers for the study of the diversity and intra-taxa genetic structure of G. intestinalis in silico and in DNA obtained from stool samples. We analysed nine constitutive genes in 80 complete genome sequences and in a group of 24 stool samples from Colombia. Allelic diversity was evaluated by locus and for the concatenated sequence of nine loci that could discriminate up to 53 alleles. Phylogenetic reconstructions allowed us to identify AI, AII and B assemblages. We found evidence of intra- and inter-assemblage recombination events. Population structure analysis showed genetic differentiation among the assemblages analysed.

Molecular detection and genotyping of intestinal protozoa from different biogeographical regions of Colombia.

BACKGROUND: Intestinal parasitic protozoa represent a serious problem of public health particularly in developing countries. Protozoa such as Blastocystis, Giardia intestinalis, Entamoeba histolytica and Cryptosporidium spp. are associated with diarrheal symptoms. In Colombia, there is little region-specific data on the frequency and circulating genotypes/species of these microorganisms. Therefore, the main objective of our study was to employ molecular detection and genotyping of G. intestinalis and Blastocystis, Cryptosporidium and Entamoeba spp. in samples from different biogeographical regions of Colombia. METHODS: We collected 649 human fecal samples from five biogeographical regions of Colombia: the Amazon, Andean, Caribbean, Orinoco and Pacific regions. Blastocystis, G. intestinalis, Cryptosporidium spp. and Entamoeba complex were detected by microscopy and conventional PCR. Molecular genotyping was conducted to identify Blastocystis subtypes (STs) (18s), G. intestinalis assemblages (triose phosphate isomerase and glutamate dehydrogenase) and Cryptosporidium species (18s). Genetic diversity indices were determined using dnasp.5. RESULTS: We detected G. intestinalis in 45.4% (n = 280) of samples, Blastocystis in 54.5% (n = 336) of samples, Cryptosporidium spp. in 7.3% (n = 45) of samples, Entamoeba dispar in 1.5% (n = 9) of samples, and Entamoeba moshkovskii in 0.32% (n = 2) of samples. Blastocystis STs 1-4, 8 and 9 and G. intestinalis assemblages AII, BIII, BIV, D and G were identified. The following Cryptosporidium species were identified: C. hominis, C. parvum, C. bovis, C. andersoni, C. muris, C. ubiquitum and C. felis. The Caribbean region had the highest frequency for each of the microorganisms evaluated (91.9% for G. duodenalis, 97.3% for Blastocystis, 10.8% for Cryptosporidium spp., 13.5% for E. dispar and 2.7% for E. moshkovskii). The Orinoco region had a high frequency of Blastocystis (97.2%) and the Andean region had a high frequency of G. intestinalis (69.4%). High and active transmission was apparent in several regions of the country, implying that mechanisms for prevention and control of intestinal parasitosis in different parts of the country must be improved.

Temporal Variation of the Presence of Rhodnius prolixus (Hemiptera: Reduviidae) Into Rural Dwellings in the Department of Casanare, Eastern Colombia.

Rhodnius prolixus (Stål, 1859) is the major vector of Trypanosoma cruzi in Colombia and Venezuela. The species is strongly associated with high-altitude ecotopes, such as sylvatic palms (Attalea butyracea), where spatially and temporally stable infestations are established. We investigated temporal variation in regards to the presence of R. prolixus in rural dwellings in the department of Casanare (eastern Colombia) over a period of 12 mo. Thirty houses were sampled from January to December 2017 by installing Maria sensors, collecting triatomines through community entomological surveillance, and conducting a monthly search in each house. The collection of specimens from the houses varied significantly by month with the higher number of collections occurring in the low-rainfall season and the lower number of collections occurring in the months of increased precipitation. The proportions of males, females, and nymphs also varied significantly throughout the time period: nymphs (fifth instar only) were reported only during May, July, and September and significantly greater numbers of females than males were reported in the inspected dwellings in all months. Density, crowding, and colonization indices varied according to the season. A bloodmeal analysis revealed 17 different hosts. A total of 42 randomly selected R. prolixus specimens were subjected to molecular analyses for detection of T. cruzi DNA with 22 found positive (infection prevalence of 52%). In conclusion, we observed a high presence of R. prolixus (infected with T. cruzi) in dwellings close to native palm plantations. These findings indicate a high risk of vector transmission of T. cruzi for people in the study areas and challenges for the current vector control schemes in the region.

Trypanosoma cruzi infection, discrete typing units and feeding sources among Psammolestes arthuri (Reduviidae: Triatominae) collected in eastern Colombia.

BACKGROUND: Chagas disease (CD) is caused by the protozoan parasite Trypanosoma cruzi, and is transmitted by hematophagous insects of the family Reduviidae. Psammolestes arthuri is a sylvatic triatomine distributed in Colombia and Venezuela which feeds on birds and there are a few studies that have reported Ps. arthuri naturally infected with T. cruzi. In Colombia, Ps. arthuri has been found in dwellings, making it important to evaluate its possible role in the T. cruzi transmission cycle. We aimed to evaluate the presence of T. cruzi and feeding sources of Ps. arthuri to elucidate new possible scenarios of T. cruzi transmission in the country. METHODS: A total of 60 Ps. arthuri were collected in Arauca and Casanare, Colombia. We detected and genotyped T. cruzi and identified feeding sources. The frequency of the presence of T. cruzi was obtained and compared with different eco-epidemiological variables. Multiple correspondence analysis was conducted to explore associations between eco-epidemiological variables and the presence of T. cruzi; with these results, a logistic regression was used to determine statistical associations. RESULTS: The infection rate of T. cruzi was 70.7% and was mostly associated with insect stage, sex, bird nest and feeding source. Regarding discrete typing units (DTUs), TcI was found in 54.7% samples, of which 21.7% (5/23) were TcIDom, 52.1% (12/23) had mixed infection (TcIDom-TcISylv), and single infection with TcISylv was not detected. Mixed infections (TcI/TcII-TcVI) were found in 9.52% (4/42) of the samples; of these, 14.2% (6/42) were TcII-TcVI. A total of 15 feeding sources were identified and the most frequent were: Cranioleuca baroni (35.85%), Homo sapiens (26.42%), Thraupis episcopus (11.32%) and Serinus albogularis (3.77%). CONCLUSIONS: Although Ps. arthuri is mainly ornithophilic, this species may be feeding on other animals that can be infected with T. cruzi, possibly playing a role maintaining the zoonotic cycle of the parasite. Further studies with molecular techniques and wider sampling are needed to improve information regarding infection rates, ecotopes and habits with the aim of evaluating whether Ps. arthuri could be a potential T. cruzi vector.

Ecology of Trypanosoma cruzi I genotypes across Rhodnius prolixus captured in Attalea butyracea palms.

Trypanosoma cruzi, the agent of Chagas disease exhibits significant genetic diversity. This parasite is divided into six discrete typing units (DTUs) where T. cruzi I (TcI) is the most widespread in the Americas. TcI genotypes have been associated to domestic and sylvatic cycles of transmission (TcIDom and sylvatic TcI). Due to the importance of the enzootic transmission, we determined the frequency of TcI genotypes present in Rhodnius prolixus captured in different regions of the palm A. butyracea to understand the ecology of the disease and the importance of A. butyracea palms as ecotopes of R. prolixus. Forty A. butyracea palms were sampled (base crown, mid-point and crown) capturing 105 individuals identified as R. prolixus by morphological and molecular barcoding. We conducted molecular detection and typing of T. cruzi across 59 individuals. The results showed that all the insects were infected with TcI; 28.57% were sylvatic TcI, 12.38% TcIDom and 15,24% mixed infections (TcIDom/sylvatic TcI). Statistical analysis showed a similar behavior between TcIDom and mixed infections in the mid-point and at the crown of the palm, being more frequent in the crown, while sylvatic TcI does not seem to have a specific association with any of the sampled areas. These findings are consistent with other studies showing high mobility of the insect vector between different ecotopes, increasing the need to develop improvements in the programs of disease control.

Effect of the physiognomy of Attalea butyracea (Arecoideae) on population density and age distribution of Rhodnius prolixus (Triatominae).

BACKGROUND: Rhodnius prolixus Stål, 1859 is one of the main vectors of Trypanosoma (Schyzotrypanum) cruzi Chagas, 1909. In its natural forest environment, this triatomine is mainly found in palm tree crowns, where it easily establishes and develops dense populations. The aim of this study was to evaluate the effect of the physiognomy and reproductive status of Attalea butyracea on the population relative density and age structure of R. prolixus and to determine the vector's population stratification according to the vertical and horizontal profile of an A. butyracea forest. METHODS: Using live bait traps, 150 individuals of A. butyracea with different physiognomy and 40 individuals with similar physiognomy (crown size, number of leaves, palm tree height, diameter at breast height, reproductive status) were sampled for triatomines in Yopal, Casanare-Colombia. Temperature and relative humidity were measured in the crown of the palm tree. Entomological indices and natural infection rates were also determined. RESULTS: The relative population density of R. prolixus on natural A. butyracea groves is associated with the palm's height, number of leaves and crown volume. The young immature stages were present mostly at the crown's base and the advanced immature stages and adults were present mostly at the crown of the palm tree. This distribution correlates with the temperature stability and relative humidity in the base and the fluctuation of both environmental variables in the palm's crown. A higher density of R. prolixus was found as the palm tree height increased and as the distance of the palm with respect to the forest border decreased, especially towards anthropically intervened areas. A density index of 12.6 individuals per palm tree with an infestation index of 88.9% and a colonization index of 98.7% was observed. 85.2% was the infection index with T. cruzi. CONCLUSION: The physiognomy of palm trees affects the relative population density and the distribution of developmental stages of R. prolixus. Therefore, they constitute a risk factor for the potential migration of infected insects from wild environments towards residential environments and the subsequent epidemiological risk of transmission of T. cruzi to people.

Comparación de métodos para la captura de triatominos (Hemiptera: Reduviidae) en palmas Attalea butyraceaen los Llanos Orientales de Colombia / Comparison of methods for the capture of triatomines (Hemiptera: Reduviidae) in Attalea butyracea palms in the Eastern Plains of Colombia

Introducción. El estudio de los triatominos en sus ecótopos naturales tiene gran significado en la epidemiología de la enfermedad de Chagas. La recolección de insectos vivos y su establecimiento en insectarios permite realizar incontables estudios. Por ello se necesitan métodos efectivos para la captura de triatominos silvestres, especialmente especies intrusas como Rhodnius prolixus. Objetivo. Comparar cuatro métodos para la captura de triatominos en palmas para medir su efectividad. Materiales y métodos. El estudio se llevó a cabo en la localidad de Maní, departamento de Casanare, Colombia, con predominio de Attalea butyracea. Se compararon cuatro metodologías en 67 palmas: trampas con cebo vivo, búsqueda directa en palmas, tala y disección. Resultados. Los métodos fueron efectivos para recolectar ejemplares juveniles y adultos. La tala y la búsqueda manual mostraron los mayores índices de infestación (100 % y 83,33 %). La trampa Angulo mostró mayores índices de hacinamiento (4,93), densidad (3,76), rango de abundancia (1-13) e índice adulto/ninfa (0,22). La tala y la disección fue el método que significativamente recolectó más triatominos en comparación con los otros (p<0,002), seguido de la trampa Angulo. No se observaron diferencias en la captura de los estadios de ninfas entre los métodos evaluados (p>0,05). Conclusiones. Aunque la tala fue el mejor método para la recolección del mayor número de triatominos, los métodos con el cebo vivo con adhesivos o refugios para atrapar los insectos, fueron una buena alternativa. Entre ellos, la trampa Angulo mostró mejores resultados y evitó la exposición del investigador al riesgo y la intervención de ecotopos naturales.

Introduction: Studying triatominae in their natural ecotopes has great significance in the epidemiology of Chagas disease. Gathering live insects and the establishment of insectaries allows countless studies. Therefore, effective methods are needed for capturing wild triatomines, especially intrusive species such as Rhodnius prolixus. Objective: To compare four methods of capturing triatomines in palms to measure their effectiveness. Materials and methods: Our study was conducted in Maní, department of Casanare, Colombia, with a predominance of A. butyracea. We compared four methodologies in 67 palms: live-bait traps, direct search in the palms, felling and dissection. Results: The methods were effective to collect juveniles and adults. Felling and manual search showed the highest infestation rates (100% and 83.33%). Higher indexes of overcrowding (4.93), density (3.76), abundance range (1-13) and adult/nymph index (0.22) were obtained with the Angulo trap. Felling and dissection was the method which significantly collected a greater number of triatomines compared with the other methods (p <0.002); it was followed by the Angulo trap. We did not find any differences in the collection of the nymphal stages among the evaluated methods (p> 0.05).Conclusions: Even though felling was the best method for collecting the highest number of triatomines, live-bait with adhesives or shelters to trap insects were a good alternative to collect the insects in palms. Among them, the Angulo trap showed better results and it prevents the risk of exposure of the researcher and the intervention of natural ecotopes.

[Comparison of methods for the capture of triatomines ( Hemiptera: Reduviidae ) in Attalea butyracea palms in the Eastern Plains of Colombia]. / Comparación de métodos para la captura de triatominos (Hemiptera: Reduviidae) en palmas Attalea butyracea en los Llanos Orientales de Colombia.

INTRODUCTION: Studying triatominae in their natural ecotopes has great significance in the epidemiology of Chagas disease. Gathering live insects and the establishment of insectaries allows countless studies. Therefore, effective methods are needed for capturing wild triatomines, especially intrusive species such as Rhodnius prolixus . OBJECTIVE: To compare four methods of capturing triatomines in palms to measure their effectiveness. MATERIALS AND METHODS: Our study was conducted in Maní, department of Casanare, Colombia, with a predominance of A. butyracea . We compared four methodologies in 67 palms: live-bait traps, direct search in the palms, felling and dissection. RESULTS: The methods were effective to collect juveniles and adults. Felling and manual search showed the highest infestation rates (100% and 83.33%). Higher indexes of overcrowding (4.93), density (3.76), abundance range (1-13) and adult/nymph index (0.22) were obtained with the Angulo trap. Felling and dissection was the method which significantly collected a greater number of triatomines compared with the other methods (p <0.002); it was followed by the Angulo trap. We did not find any differences in the collection of the nymphal stages among the evaluated methods (p> 0.05). CONCLUSIONS: Even though felling was the best method for collecting the highest number of triatomines, live-bait with adhesives or shelters to trap insects were a good alternative to collect the insects in palms. Among them, the Angulo trap showed better results and it prevents the risk of exposure of the researcher and the intervention of natural ecotopes.