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Trypanosoma cruzi strain TcIV infects raccoons from Illinois.

ABSTRACT: The northern limits of Trypanosoma cruzi across the territory of the United States remain unknown. The known vectors Triatoma sanguisuga and T. lecticularia find their northernmost limits in Illinois; yet, earlier screenings of those insects did not reveal the presence of the pathogen, which has not been reported in vectors or reservoir hosts in this state.Five species of medium-sized mammals were screened for the presence of T. cruzi.Genomic DNA was isolated from heart, spleen and skeletal muscle of bobcats (Lynx rufus, n = 60), raccoons (Procyon lotor, n = 37), nine-banded armadillos (Dasypus novemcinctus, n = 5), Virginia opossums (Didelphis virginiana, n = 3), and a red fox (Vulpes vulpes). Infections were detected targeting DNA from the kinetoplast DNA minicircle (kDNA) and satellite DNA (satDNA). The discrete typing unit (DTU) was determined by amplifying two gene regions: the Spliced Leader Intergenic Region (SL), via a multiplex polymerase chain reaction, and the 24S? ribosomal DNA via a heminested reaction. Resulting sequences were used to calculate their genetic distance against reference DTUs.18.9% of raccoons were positive for strain TcIV; the rest of mammals tested negative.These results confirm for the first time the presence of T. cruzi in wildlife from Illinois, suggesting that a sylvatic life cycle is likely to occur in the region. The analyses of sequences of SL suggest that amplicons resulting from a commonly used multiplex reaction may yield non-homologous fragments.

SUBMITTER: Vandermark C

PROVIDER: S-EPMC5719540 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Trypanosoma cruzi strain TcIV infects raccoons from Illinois.

Vandermark Cailey C Zieman Elliott E Boyles Esmarie E Nielsen Clayton K CK Davis Cheryl C Jiménez Francisco Agustín FA

Memorias do Instituto Oswaldo Cruz 20180101 1

<h4>Background</h4>The northern limits of Trypanosoma cruzi across the territory of the United States remain unknown. The known vectors Triatoma sanguisuga and T. lecticularia find their northernmost limits in Illinois; yet, earlier screenings of those insects did not reveal the presence of the pathogen, which has not been reported in vectors or reservoir hosts in this state.<h4>Objectives</h4>Five species of medium-sized mammals were screened for the presence of T. cruzi.<h4>Methods</h4>Genomic ...[more]

PMID: 29211106

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Project description:Chagas disease has a diverse pathology caused by the parasite Trypanosoma cruzi, and is indigenous to Central and South America. A pronounced feature of the trypanosomes is the kinetoplast, which is comprised of catenated maxicircles and minicircles that provide the transcripts involved in uridine insertion/deletion RNA editing. T. cruzi exchange genetic material through a hybridization event. Extant strains are grouped into six discrete typing units by nuclear markers, and three clades, A, B, and C, based on maxicircle gene analysis. Clades A and B are the more closely related. Representative clade B and C maxicircles are known in their entirety, and portions of A, B, and C clades from multiple strains show intra-strain heterogeneity with the potential for maxicircle taxonomic markers that may correlate with clinical presentation.To perform a genome-wide analysis of the three maxicircle clades, the coding region of clade A representative strain Sylvio X10 (a.k.a. Silvio X10) was sequenced by PCR amplification of specific fragments followed by assembly and comparison with the known CL Brener and Esmeraldo maxicircle sequences. The clade A rRNA and protein coding region maintained synteny with clades B and C. Amino acid analysis of non-edited and 5'-edited genes for Sylvio X10 showed the anticipated gene sequences, with notable frameshifts in the non-edited regions of Cyb and ND4. Comparisons of genes that undergo extensive uridine insertion and deletion display a high number of insertion/deletion mutations that are likely permissible due to the post-transcriptional activity of RNA editing.Phylogenetic analysis of the entire maxicircle coding region supports the closer evolutionary relationship of clade B to A, consistent with uniparental mitochondrial inheritance from a discrete typing unit TcI parental strain and studies on smaller fragments of the mitochondrial genome. Gene variance that can be corrected by RNA editing hints at an unusual depth for maxicircle taxonomic markers, which will aid in the ability to distinguish strains, their corresponding symptoms, and further our understanding of the T. cruzi population structure. The prevalence of apparently compromised coding regions outside of normally edited regions hints at undescribed but active mechanisms of genetic exchange.

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Trypanosoma cruzi strain TcIV infects raccoons from Illinois.

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Trypanosoma cruzi strain TcIV infects raccoons from Illinois.

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