Trypanosoma cruzi diversity in naturally infected nonhuman primates in Louisiana assessed by deep sequencing of the mini-exon gene

Trypanosoma cruzi, causative agent of Chagas disease, and T. rangeli are kinetoplastid parasites endemic to Latin America. Although closely-related to T. cruzi and capable of infecting humans, T. rangeli is non-pathogenic. Both parasite species are transmitted by triatomine bugs, and the presence of T. rangeli constitutes a confounding factor in the study of Chagas prevalence and transmission dynamics. T. cruzi possesses high molecular heterogeneity: six discrete typing units (DTUs) are currently recognized. In Ecuador, TcI predominates while other DTUs are seldom reported. Here, infection by T. cruzi and/or T. rangeli in triatomine bugs from two communities of southern Ecuador was evaluated via PCR product size polymorphism of kinetoplast-minicircle sequences and the non-transcribed spacer region of the mini-exon gene. Additionally, mini-exon amplicons were deep-sequenced to analyze single nucleotide polymorphisms and mixed infections. As a control, mini-exon products from several monoclonal reference strains were included. T. cruzi genetic diversity was significantly greater in adult vectors than in nymphal stage V vectors. Among infected triatomines, deep sequencing revealed one T. rangeli infection (3%), 9 T. cruzi infections (27.3%) and 23 T. cruzi / T. rangeli mixed infections (69.7%), suggesting that T. rangeli prevalence has been largely underestimated in the region. Furthermore, deep-sequencing detected TcIV sequences in six samples (first TcIV record in southern Ecuador). Our data indicate that amplicon size analysis alone is not reliable for parasite identification/typing in mixed infections containing both T. cruzi and T. rangeli, or when multiple T. cruzi DTUs are present. Additionally, our analysis showed extensive overlap among the parasite populations present in the two studied localities (ca. 28 km apart); suggesting active parasite dispersal over the study area. Our results highlight the value of amplicon sequencing methodologies to clarify the population dynamics of kinetoplastid parasites in endemic regions and inform control campaigns in southern Ecuador.

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Deep Sequencing of the Trypanosoma cruzi GP63 Surface Proteases Reveals Diversity and Diversifying Selection among Chronic and Congenital Chagas Disease Patients

PLoS Neglected Tropical Diseases ◽

10.1371/journal.pntd.0003458 ◽

2015 ◽

Vol 9 (4) ◽

pp. e0003458 ◽

Cited By ~ 31

Author(s):

Martin S. Llewellyn ◽

Louisa A. Messenger ◽

Alejandro O. Luquetti ◽

Lineth Garcia ◽

Faustino Torrico ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Chagas Disease ◽

Deep Sequencing ◽

Diversifying Selection

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Trypanosoma cruzi Transmission Among Captive Nonhuman Primates, Wildlife, and Vectors

EcoHealth ◽

10.1007/s10393-018-1318-5 ◽

2018 ◽

Vol 15 (2) ◽

pp. 426-436 ◽

Cited By ~ 11

Author(s):

Carolyn L. Hodo ◽

Gregory K. Wilkerson ◽

Elise C. Birkner ◽

Stanton B. Gray ◽

Sarah A. Hamer

Keyword(s):

Trypanosoma Cruzi ◽

Nonhuman Primates

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An Improved Approach to Trypanosoma cruzi Molecular Genotyping by Next-Generation Sequencing of the Mini-exon Gene

Methods in Molecular Biology - T. cruzi Infection ◽

10.1007/978-1-4939-9148-8_4 ◽

2019 ◽

pp. 47-60 ◽

Cited By ~ 5

Author(s):

Alicia Majeau ◽

Claudia Herrera ◽

Eric Dumonteil

Keyword(s):

Next Generation Sequencing ◽

Trypanosoma Cruzi ◽

Next Generation ◽

Molecular Genotyping ◽

Generation Sequencing ◽

Exon Gene

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Deep sequencing reveals multiclonality and new discrete typing units of Trypanosoma cruzi in rodents from the southern United States

Journal of Microbiology Immunology and Infection ◽

10.1016/j.jmii.2018.12.004 ◽

2020 ◽

Vol 53 (4) ◽

pp. 622-633 ◽

Cited By ~ 11

Author(s):

Henry Pronovost ◽

Anna C. Peterson ◽

Bruno Ghersi Chavez ◽

Michael J. Blum ◽

Eric Dumonteil ◽

...

Keyword(s):

United States ◽

Trypanosoma Cruzi ◽

Deep Sequencing ◽

Southern United States

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Enzootic transmission of Trypanosoma cruzi and T. rangeli in the Federal District of Brazil

Revista do Instituto de Medicina Tropical de São Paulo ◽

10.1590/s0036-46652004000600005 ◽

2004 ◽

Vol 46 (6) ◽

pp. 323-330 ◽

Cited By ~ 30

Author(s):

Rodrigo Gurgel-Gonçalves ◽

Eduardo Dias Ramalho ◽

Marco Antônio Duarte ◽

Alexandre Ramlo Torre Palma ◽

Fernando Abad-Franch ◽

...

Keyword(s):

Trypanosoma Cruzi ◽

Disease Transmission ◽

Federal District ◽

Kinetoplast Dna ◽

Mauritia Flexuosa ◽

Gallery Forests ◽

Parasitological Survey ◽

Low Prevalence ◽

First Time ◽

Exon Gene

The Federal District of Brazil (DF) lies within the Cerrado biome, where open shrubland (savannas) is interspersed with riverside gallery forests and permanent swamps (veredas). Trypanosoma cruzi-infected native triatomines occur in the area, but the enzootic transmission of trypanosomatids remains poorly characterized. A parasitological survey involving sylvatic triatomines (166 Rhodnius neglectus collected from Mauritia flexuosa palms) and small mammals (98 marsupials and 70 rodents, totaling 18 species) was conducted in 18 sites (mainly gallery forests and veredas) of the DF. Parasites were isolated, morphologically identified, and characterized by PCR of nuclear (mini-exon gene) and kinetoplast DNA (kDNA). Six R. neglectus, seven Didelphis albiventris and one Akodon cursor were infected by trypanosomes; wild reservoir infection is documented for the first time in the DF. kDNA PCR detected T. cruzi in five R. neglectus and mini-exon gene PCR revealed T. cruzi I in isolates from D. albiventris. Parasites infecting one bug yielded T. rangeli KP1+ kDNA amplicons. In spite of the occurrence of T. cruzi-infected D. albiventris (an important wild and peridomestic reservoir) and R. neglectus (a secondary vector displaying synanthropic behavior), a low-risk of human Chagas disease transmission could be expected in the DF, considering the low prevalence infection recorded in this work. The detection of T. rangeli KP1+ associated with R. neglectus in the DF widens the known range of this parasite in Brazil and reinforces the hypothesis of adaptation of T. rangeli populations (KP1+ and KP1-) to distinct evolutionary Rhodnius lineages.

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Shelter cats host infections with multiple Trypanosoma cruzi discrete typing units in southern Louisiana

Veterinary Research ◽

10.1186/s13567-021-00923-z ◽

2021 ◽

Vol 52 (1) ◽

Author(s):

Eric Dumonteil ◽

Hans Desale ◽

Weihong Tu ◽

Brandy Duhon ◽

Wendy Wolfson ◽

...

Keyword(s):

Genetic Diversity ◽

Trypanosoma Cruzi ◽

Deep Sequencing ◽

Limited Information ◽

Convenience Sample ◽

Seropositivity Rate ◽

Southern Us ◽

Zoonotic Parasite ◽

Local Transmission ◽

Southern Louisiana

AbstractTrypanosoma cruzi is a zoonotic parasite endemic in the southern US and the Americas, which may frequently infect dogs, but limited information is available about infections in cats. We surveyed a convenience sample of 284 shelter cats from Southern Louisiana to evaluate T. cruzi infection using serological and PCR tests. Parasites from PCR positive cats were also genotyped by PCR and deep sequencing to assess their genetic diversity. We detected a seropositivity rate for T. cruzi of at least 7.3% (17/234), and 24.6% of cats (70/284) were PCR positive for the parasite. Seropositivity increased with cat age (R2 = 0.91, P = 0.011), corresponding to an incidence of 7.2% ± 1.3 per year, while PCR positivity decreased with age (R2 = 0.93, P = 0.007). Cats were predominantly infected with parasites from TcI and TcVI DTUs, and to a lesser extent from TcIV and TcV DTUs, in agreement with the circulation of these parasite DTUs in local transmission cycles. These results indicate that veterinarians should have a greater awareness of T. cruzi infection in pets and that it would be important to better evaluate the risk for spillover infections in humans.

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