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Molecular characterization of Trypanosoma cruzi DTUs of the triatomine species in a Chagas disease endemic area.

ABSTRACT: Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected tropical infection with great public health importance. This protozoan has triatomine insects as vector but may also be transmitted through blood transfusion, organ transplants, ingestion of contaminated food, or congenitally. It has a heterogeneous population classified into Discrete Typing Units (DTUs), TcI-TcVI and TcBat. The aim of this study was to molecularly characterize the DTUs of T. cruzi in triatomines from a Chagas disease endemic area in Northeastern Brazil. Triatomines were collected and the gut content was microscopically analyzed to investigate the presence of trypanosomatid flagellates. In addition, digestive tracts of some specimens were dissected and molecularly analyzed through PCR for Trypanosoma spp. and sequencing. PCR positive samples were further submitted to a multiplex PCR for DTUs of T. cruzi. A total of 117 triatomines were collected, 93.16% being in intradomicile and 6.84% in peridomicile environments. Insects were identified as Panstrongylus lutzi (37.60%), Triatoma pseudomaculata (26.50%), Triatoma brasiliensis (23.08%) and Panstrongylus megistus (12.82%). The specimens herein analyzed presented infection rates by T. cruzi of 5.49% and 12.09% in parasitological and molecular examinations, respectively. Multiplex PCR screening revealed 70.59% of the TcI genotype, detected in all triatomine species identified in this study and 29.41% of the DTU TcIII/TcIV detected in P. megistus and P. lutzi. T. cruzi infect triatomines in intradomicile and peridomicile environments, which brings attention to the risk of human infections and to the importance of the implementation of surveillance and entomological control actions.

SUBMITTER: Silva TRM

PROVIDER: S-EPMC8901897 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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Molecular characterization of Trypanosoma cruzi DTUs of the triatomine species in a Chagas disease endemic area.

Silva Tatiene Rossana Móta TRM Rios Thaynádia Gomes TG do Nascimento Ramos Carlos Alberto CA Scofield Alessandra A Lima Thiago Antonio Rodrigues Freire TARF Alves Leucio Câmara LC Ramos Rafael Antonio Nascimento RAN de Carvalho Gílcia Aparecida GA

Journal of parasitic diseases : official organ of the Indian Society for Parasitology 20210725 1

Trypanosoma cruzi is the etiological agent of Chagas disease, a neglected tropical infection with great public health importance. This protozoan has triatomine insects as vector but may also be transmitted through blood transfusion, organ transplants, ingestion of contaminated food, or congenitally. It has a heterogeneous population classified into Discrete Typing Units (DTUs), TcI-TcVI and TcBat. The aim of this study was to molecularly characterize the DTUs of T. cruzi in triatom ...[more]

PMID: 35299926

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Project description:BackgroundTrypanosoma cruzi, a hemoflagellate protozoan parasite and the etiological agent of Chagas disease (CD), exhibits great genetic and biological diversity. Infected individuals may present clinical manifestations with different levels of severity. Several hypotheses have been proposed to attempt to correlate the diversity of clinical signs and symptoms to the genetic variability of T. cruzi. This work aimed to investigate the differential expression of proteins from two distinct genetic groups of T. cruzi (discrete typing units TcI and TcII), isolated from chronically infected individuals displaying the cardiac form of CD. For this purpose, epimastigote forms of the two isolates were cultured in vitro and the cells recovered for protein extraction. Comparative two-dimensional (2D) gel electrophoreses were performed and differentially expressed spots selected for identification by mass spectrometry, followed by database searching and protein categorization.ResultsThe 2D electrophoretic profiles revealed the complex composition of the T. cruzi extracted proteome. Protein spots were distributed along the entire pH and molecular mass ranges attesting for the integrity of the protein preparations. In total, 46 differentially expressed proteins were identified present in 40 distinct spots found in the comparative gel analyses. Of these, 16 displayed upregulation in the gel from TcI-typed parasites and 24 appeared overexpressed in the gel from TcII-typed parasites. Functional characterization of differentially expressed proteins revealed major alterations associated with stress response, lipid and amino acid metabolism in parasites of the TcII isolate, whilst those proteins upregulated in the TcI sample were primarily linked to central metabolic pathways.ConclusionsThe comparative 2D-gel electrophoresis allowed detection of major differences in protein expression between two T. cruzi isolates, belonging to the TcI and TcII genotypes. Our findings suggest that patients displaying the cardiac form of the disease harbor parasites capable of exhibiting distinct proteomic profiles. This should be of relevance to disease prognosis and treatment.

Project description:BackgroundThe DNA barcoding system using the cytochrome c oxidase subunit 1 mitochondrial gene (cox1 or COI) is highly efficient for discriminating vertebrate and invertebrate species. In the present study, we examined the suitability of cox1 as a marker for Trypanosoma cruzi identification from other closely related species. Additionally, we combined the sequences of cox1 and the nuclear gene glucose-6-phosphate isomerase (GPI) to evaluate the occurrence of mitochondrial introgression and the presence of hybrid genotypes.MethodsSixty-two isolates of Trypanosoma spp. obtained from five of the six Brazilian biomes (Amazon Forest, Atlantic Forest, Caatinga, Cerrado and Pantanal) were sequenced for cox1 and GPI gene fragments. Phylogenetic trees were reconstructed using neighbor-joining, maximum likelihood, parsimony and Bayesian inference methods. Molecular species delimitation was evaluated through pairwise intraspecific and interspecific distances, Automatic Barcode Gap Discovery, single-rate Poisson Tree Processes and multi-rate Poisson Tree Processes.ResultsBoth cox1 and GPI genes recognized and differentiated T. cruzi, Trypanosoma cruzi marinkellei, Trypanosoma dionisii and Trypanosoma rangeli. Cox1 discriminated Tcbat, TcI, TcII, TcIII and TcIV. Additionally, TcV and TcVI were identified as a single group. Cox1 also demonstrated diversity in the discrete typing units (DTUs) TcI, TcII and TcIII and in T. c. marinkellei and T. rangeli. Cox1 and GPI demonstrated TcI and TcII as the most genetically distant branches, and the position of the other T. cruzi DTUs differed according to the molecular marker. The tree reconstructed with concatenated cox1 and GPI sequences confirmed the separation of the subgenus Trypanosoma (Schizotrypanum) sp. and the T. cruzi DTUs TcI, TcII, TcIII and TcIV. The evaluation of single nucleotide polymorphisms (SNPs) was informative for DTU differentiation using both genes. In the cox1 analysis, one SNP differentiated heterozygous hybrids from TcIV sequences. In the GPI analysis one SNP discriminated Tcbat from TcI, while another SNP distinguished TcI from TcIII.ConclusionsDNA barcoding using the cox1 gene is a reliable tool to distinguish T. cruzi from T. c. marinkellei, T. dionisii and T. rangeli and identify the main T. cruzi genotypes.

Dataset Information

Molecular characterization of Trypanosoma cruzi DTUs of the triatomine species in a Chagas disease endemic area.

Publications

Molecular characterization of <i>Trypanosoma cruzi</i> DTUs of the triatomine species in a Chagas disease endemic area.

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