Socio-environmental factors associated with the occurrence of triatomines (Hemiptera: Reduviidae) in an endemic municipality in northern Minas Gerais, Brazil.

Triatomines are the vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. The study aimed to evaluate the association between sociodemographic and environmental factors, and changes in land use and cover, with the occurrence and abundance of triatomines by census sectors in an endemic municipality of northern Minas Gerais, Brazil. The study was conducted in Montes Claros, located in the north of Minas Gerais, Brazil. The entomological data used in the study were collected by active surveillance in the rural area from 2015 to 2019 and by passive surveillance in the urban area from 2009 to 2019. Data on sociodemographic and environmental factors and changes in land use and land cover were obtained from the urban and rural census sectors. A total of 1404 triatomines, belonging to eight species, were captured in domiciles in the rural area (2015-2019) and 277 triatomines in domiciles in the urban area (2009-2019) of the municipality of Montes Claros. The variables the number of domiciles, household economic income, pavement, NDVI, deforestation, unchanged, and anthropic proved to be positively associated with the occurrence and/or number of triatomines in census sectors, within the models. The occurrence of triatomines in the domestic environment of the municipality of Montes Claros should be considered a public health problem, as it suggests a potential risk of establishment and transmission of T. cruzi to domestic animals, farm animals, and humans.

High Parasitic Loads Quantified in Sylvatic Triatoma melanica, a Chagas Disease Vector.

Triatoma melanica is a sylvatic vector species in Brazil. In We aimed to characterize the Trypanosoma cruzi discrete typing units (DTUs), the parasitic loads, and the blood meal sources of insects collected in rocky outcrops in rural areas in the state of Minas Gerais. An optical microscope (OM) and kDNA-PCR were used to examine natural infection by T. cruzi, and positive samples were genotyped by conventional multilocus PCR. Quantification of the T. cruzi load was performed using qPCR, and the blood meal sources were identified by Sanger sequencing the 12S rRNA gene. A total of 141 T. melanica were captured. Of these, ~55% (61/111) and ~91% (63/69) were positive by OM and KDNA-PCR, respectively. We genotyped ~89% (56/63) of the T. cruzi-positive triatomines, with TcI (~55%, 31/56) being the most prevalent DTU, followed by TcIII (~20%, 11/56) and TcII (~7%, 4/56). Only TcI+TcIII mixed infections were detected in 10 (~18%) specimens. A wide range of variation in the parasitic loads of T. melanica was observed, with an overall median value of 104 parasites/intestine, with females having higher T. cruzi loads than N2, N4, and N5. TcII showed lower parasitic loads compared to TcI and TcIII. The OM positive diagnosis odds ratio between T. cruzi infection when the parasite load is 107 compared to 103 was approximately 29.1. The most frequent blood meal source was Kerodon rupestris (~58%), followed by Thrichomys apereoides (~18%), Wiedomys cerradensis (~8%), Galactis cuja (~8%) and Gallus gallus (~8%). Our findings characterize biological and epidemiological aspects of the sylvatic population of T. melanica in the study area, highlighting the need to extend surveillance and control to this vector.

Trypanosoma cruzi iron superoxide dismutases: insights from phylogenetics to chemotherapeutic target assessment.

BACKGROUND: Components of the antioxidant defense system in Trypanosoma cruzi are potential targets for new drug development. Superoxide dismutases (SODs) constitute key components of antioxidant defense systems, removing excess superoxide anions by converting them into oxygen and hydrogen peroxide. The main goal of the present study was to investigate the genes coding for iron superoxide dismutase (FeSOD) in T. cruzi strains from an evolutionary perspective. METHODS: In this study, molecular biology methods and phylogenetic studies were combined with drug assays. The FeSOD-A and FeSOD-B genes of 35 T. cruzi strains, belonging to six discrete typing units (Tcl-TcVI), from different hosts and geographical regions were amplified by PCR and sequenced using the Sanger method. Evolutionary trees were reconstructed based on Bayesian inference and maximum likelihood methods. Drugs that potentially interacted with T. cruzi FeSODs were identified and tested against the parasites. RESULTS: Our results suggest that T. cruzi FeSOD types are members of distinct families. Gene copies of FeSOD-A (n = 2), FeSOD-B (n = 4) and FeSOD-C (n = 4) were identified in the genome of the T. cruzi reference clone CL Brener. Phylogenetic inference supported the presence of two functional variants of each FeSOD type across the T. cruzi strains. Phylogenetic trees revealed a monophyletic group of FeSOD genes of T. cruzi TcIV strains in both distinct genes. Altogether, our results support the hypothesis that gene duplication followed by divergence shaped the evolution of T. cruzi FeSODs. Two drugs, mangafodipir and polaprezinc, that potentially interact with T. cruzi FeSODs were identified and tested in vitro against amastigotes and trypomastigotes: mangafodipir had a low trypanocidal effect and polaprezinc was inactive. CONCLUSIONS: Our study contributes to a better understanding of the molecular biodiversity of T. cruzi FeSODs. Herein we provide a successful approach to the study of gene/protein families as potential drug targets.

Triatomine and Trypanosoma cruzi discrete typing units distribution in a semi-arid area of northeastern Brazil.

The occurrence of triatomine species, their bloodmeal sources and the discrete typing units (DTUs) of Trypanosoma cruzi isolated from them were determined in different municipalities of the state of Rio Grande do Norte, Brazil. Triatomine captures were carried out in the rural areas of 23 municipalities. The genotyping of T. cruzi isolates was performed using the mitochondrial cytochrome c oxidase subunit 2 (coii) gene, the D7 region of the 24Sα rDNA, and the spliced leader intergenic region (SL-IR). Five triatomine species were captured, and the most frequent was Triatoma brasiliensis (84.3%; 916/1086), which was found in 16 of the 23 municipalities surveyed, and infested all types of environment investigated. The TcI DTU was found in all mesoregions surveyed in 51.5% (17/33) of the culture-positive samples. In contrast, TcII (9.1%; 3/33) was detected in the Central mesoregion, while TcIII (27.3%; 9/33) was found in all mesoregions. The geographic distribution and spatial overlap of different DTUs was inferred using the superposition of the radius of occurrence of isolates and using ecological niche distribution modelling. Triatoma brasiliensis was found infected in all mesoregions and with all three T. cruzi DTUs, including mixed infections. With regard to bloodmeal sources, the DNA of rodents was found in triatomines infected with either TcI or TcIII, while that of domestic animals and humans was associated with both single and mixed infections. Our findings demonstrate that different DTUs of T. cruzi are widely dispersed among triatomines in our study area. The association of T. brasiliensis with several different mammalian hosts, as well as overlapping areas with different DTUs, suggests that this triatomine species may have an important role as a vector in both anthropic and sylvatic environments.

TriatoDex, an electronic identification key to the Triatominae (Hemiptera: Reduviidae), vectors of Chagas disease: Development, description, and performance.

Correct identification of triatomine bugs is crucial for Chagas disease surveillance, yet available taxonomic keys are outdated, incomplete, or both. Here we present TriatoDex, an Android app-based pictorial, annotated, polytomous key to the Triatominae. TriatoDex was developed using Android Studio and tested by 27 Brazilian users. Each user received a box with pinned, number-labeled, adult triatomines (33 species in total) and was asked to identify each bug to the species level. We used generalized linear mixed models (with user- and species-ID random effects) and information-theoretic model evaluation/averaging to investigate TriatoDex performance. TriatoDex encompasses 79 questions and 554 images of the 150 triatomine-bug species described worldwide up to 2017. TriatoDex-based identification was correct in 78.9% of 824 tasks. TriatoDex performed better in the hands of trained taxonomists (93.3% vs. 72.7% correct identifications; model-averaged, adjusted odds ratio 5.96, 95% confidence interval [CI] 3.09-11.48). In contrast, user age, gender, primary job (including academic research/teaching or disease surveillance), workplace (including universities, a reference laboratory for triatomine-bug taxonomy, or disease-surveillance units), and basic training (from high school to biology) all had negligible effects on TriatoDex performance. Our analyses also suggest that, as TriatoDex results accrue to cover more taxa, they may help pinpoint triatomine-bug species that are consistently harder (than average) to identify. In a pilot comparison with a standard, printed key (370 tasks by seven users), TriatoDex performed similarly (84.5% correct assignments, CI 68.9-94.0%), but identification was 32.8% (CI 24.7-40.1%) faster on average-for a mean absolute saving of ~2.3 minutes per bug-identification task. TriatoDex holds much promise as a handy, flexible, and reliable tool for triatomine-bug identification; an updated iOS/Android version is under development. We expect that, with continuous refinement derived from evolving knowledge and user feedback, TriatoDex will substantially help strengthen both entomological surveillance and research on Chagas disease vectors.

Nanopore Sequencing Significantly Improves Genome Assembly of the Protozoan Parasite Trypanosoma cruzi.

Chagas disease was described by Carlos Chagas, who first identified the parasite Trypanosoma cruzi from a 2-year-old girl called Berenice. Many T. cruzi sequencing projects based on short reads have demonstrated that genome assembly and downstream comparative analyses are extremely challenging in this species, given that half of its genome is composed of repetitive sequences. Here, we report de novo assemblies, annotation, and comparative analyses of the Berenice strain using a combination of Illumina short reads and MinION long reads. Our work demonstrates that Nanopore sequencing improves T. cruzi assembly contiguity and increases the assembly size in ∼16 Mb. Specifically, we found that assembly improvement also refines the completeness of coding regions for both single-copy genes and repetitive transposable elements. Beyond its historical and epidemiological importance, Berenice constitutes a fundamental resource because it now constitutes a high-quality assembly available for TcII (clade C), a prevalent lineage causing human infections in South America. The availability of Berenice genome expands the known genetic diversity of these parasites and reinforces the idea that T. cruzi is intraspecifically divided in three main clades. Finally, this work represents the introduction of Nanopore technology to resolve complex protozoan genomes, supporting its subsequent application for improving trypanosomatid and other highly repetitive genomes.

Does Triatoma brasiliensis occupy the same environmental niche space as Triatoma melanica?

BACKGROUND: Triatomines (Hemiptera, Reduviidae) are vectors of Trypanosoma cruzi, the causative agent of Chagas disease, one of the most important vector-borne diseases in Latin America. This study compares the environmental niche spaces of Triatoma brasiliensis and Triatoma melanica using ecological niche modelling and reports findings on DNA barcoding and wing geometric morphometrics as tools for the identification of these species. METHODS: We compared the geographic distribution of the species using generalized linear models fitted to elevation and current data on land surface temperature, vegetation cover and rainfall recorded by earth observation satellites for northeastern Brazil. Additionally, we evaluated nucleotide sequence data from the barcode region of the mitochondrial cytochrome c oxidase subunit I (CO1) and wing geometric morphometrics as taxonomic identification tools for T. brasiliensis and T. melanica. RESULTS: The ecological niche models show that the environmental spaces currently occupied by T. brasiliensis and T. melanica are similar although not equivalent, and associated with the caatinga ecosystem. The CO1 sequence analyses based on pair wise genetic distance matrix calculated using Kimura 2-Parameter (K2P) evolutionary model, clearly separate the two species, supporting the barcoding gap. Wing size and shape analyses based on seven landmarks of 72 field specimens confirmed consistent differences between T. brasiliensis and T. melanica. CONCLUSION: Our results suggest that the separation of the two species should be attributed to a factor that does not include the current environmental conditions. However, as the caatinga is a biome that has existed in the area for at least the last 18,000 years, past conditions might have had an influence in the speciation process. The DNA Barcoding approach may be extended to these members of the subfamily Triatominae.

Analysis of the geographical distribution of Triatoma vitticeps (Stål, 1859) based on data of species occurrence in Minas Gerais, Brazil.

The purpose of this study was to determine the potential for infestation of man-made environments by Triatoma vitticeps, correlating the epidemiological importance of this species with that of others present in the Brazilian state of Minas Gerais. In addition, climatic variables that could help explain the distribution of this species and clarify its population dynamics were sought. This was done by carrying out a historical survey of the presence of T. vitticeps in artificial ecotopes, using data from the Fundação Nacional de Saúde (FUNASA). Monthly records of bioclimatic variables averaged for the period 1950-2000 and pixel size of 1kmx1km provided a reference for spatial distribution analysis. Annual rainfall and rainfall of the most humid trimester are the best indicators of the species distribution. To confirm the importance of these variables, T. vitticeps eggs were exposed to different levels of relative humidity. Hatching was found to vary significantly, and low humidity showed a significant negative effect on egg hatching. Our results demonstrate a strong association between T. vitticeps and high environmental humidity, which apparently acts as a limiting factor on the distribution of this triatomine.