Molecular epidemiology of Trypanosoma cruzi and Triatoma dimidiata in costal Ecuador

Wong, Yim Yan; Macias, Karen Jeniffer Sornosa; Martínez, Doris Guale; Solorzano, Luis; Ramírez-Sierra, María Jesús; Herrera, Claudia; Dumonteil, Eric

doi:10.1016/j.meegid.2016.04.001

Cited by 14 publications

(12 citation statements)

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“…TcI is the predominant DTU in Ecuador [13][14][15]23]. As expected, TcI-like haplotypes were widely represented in the deep sequencing analysis of the mini-exon gene performed for the Ecuadorian samples.…”

Section: Trypanosoma Rangeli and T Cruzi Genetic Diversitysupporting

confidence: 69%

“…Subsequently, a wide variety of molecular markers have been developed and employed to demonstrate the existence of seven lineages or discrete typing units (DTUs), termed TcI-TcVI and Tcbat [6][7][8][9][10][11][12]. In Ecuador, TcI predominates in the central coast [13,14] and southern highlands (Loja Province) [15]. Only two previous reports suggest the occurrence of parasites belonging to DTUs other than TcI, in triatomines and in patients of CD [16,17].…”

Section: Introductionmentioning

confidence: 99%

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Remarkable genetic diversity of Trypanosoma cruzi and Trypanosoma rangeli in two localities of southern Ecuador identified via deep sequencing of mini-exon gene amplicons

et al. 2020

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Background: Trypanosoma cruzi, the 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 disease prevalence and transmission dynamics. Trypanosoma cruzi possesses high molecular heterogeneity: seven discrete typing units (DTUs) are currently recognized. In Ecuador, T. cruzi TcI and T. rangeli KP1(-) predominate, while other genetic lineages are seldom reported. Methods: Infection by T. cruzi and/or T. rangeli in different developmental stages of triatomine bugs from two communities of southern Ecuador was evaluated via polymerase chain reaction product size polymorphism of kinetoplast minicircle sequences and the non-transcribed spacer region of the mini-exon gene (n = 48). Forty-three mini-exon amplicons were also deep sequenced to analyze single-nucleotide polymorphisms within single and mixed infections. Mini-exon products from ten monoclonal reference strains were included as controls. Results: Trypanosoma cruzi genetic richness and diversity was not significantly greater in adult vectors than in nymphal stages III and V. In contrast, instar V individuals showed significantly higher T. rangeli richness when compared with other developmental stages. Among infected triatomines, deep sequencing revealed one T. rangeli infection (3%), 8 T. cruzi infections (23.5%) and 25 T. cruzi + T. rangeli co-infections (73.5%), suggesting that T. rangeli prevalence has been largely underestimated in the region. Furthermore, deep sequencing detected TcIV sequences in nine samples; this DTU had not previously been reported in Loja Province. Conclusions: Our data indicate that deep sequencing allows for better parasite identification/typing than amplicon size analysis alone for 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

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Section: Trypanosoma Rangeli and T Cruzi Genetic Diversitysupporting

confidence: 69%

Section: Introductionmentioning

confidence: 99%

Remarkable genetic diversity of Trypanosoma cruzi and Trypanosoma rangeli in two localities of southern Ecuador identified via deep sequencing of mini-exon gene amplicons

et al. 2020

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“…We used triatomine Internal Transcribed Spacer (ITS)-2 nuclear marker for T. dimidiata genotyping and sibling species identification, as before 37 – 39 . Briefly, a 320 bp sequence was amplified (35 cycles) using the previously reported primers ITS2_200F (5′-TCGYATCTAGGCATTGTCTG-3′) and ITS2_200R (5′-CTCGCAGCTACTAAGGGAATCC-3′) and PCR conditions 40 , 41 .…”

Section: Methodsmentioning

confidence: 99%

“…For the identification of vertebrate blood meals, a 215 bp fragment of the 12 S rRNA gene was amplified (35 cycles) with the primers L1085 (5′-CCCAAACTGGGATTAGATACCC-3′ and H1259 (5′-GTTTGCTGAAGATGGCGGTA-3′) as described before 5 , 41 , 43 .…”

Section: Methodsmentioning

confidence: 99%

Detailed ecological associations of triatomines revealed by metabarcoding and next-generation sequencing: implications for triatomine behavior and Trypanosoma cruzi transmission cycles

Dumonteil¹,

Ramírez-Sierra²,

Pérez-Carrillo³

et al. 2018

Sci Rep

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Trypanosoma cruzi is the agent of Chagas disease, transmitted by hematophagous triatomine vectors. Establishing transmission cycles is key to understand the epidemiology of the disease, but integrative assessments of ecological interactions shaping parasite transmission are still limited. Current approaches also lack sensitivity to assess the full extent of this ecological diversity. Here we developed a metabarcoding approach based on next-generation sequencing to identify triatomine gut microbiome, vertebrate feeding hosts, and parasite diversity and their potential interactions. We detected a dynamic microbiome in Triatoma dimidiata, including 23 bacterial orders, which differed according to blood sources. Fourteen vertebrate species served as blood sources, corresponding to domestic, synantropic and sylvatic species, although four (human, dog, cow and mice) accounted for over 50% of blood sources. Importantly, bugs fed on multiple hosts, with up to 11 hosts identified per bug, indicating very frequent host-switching. A high clonal diversity of T. cruzi was detected, with up to 20 haplotypes per bug. This analysis provided much greater sensitivity to detect multiple blood meals and multiclonal infections with T. cruzi, which should be taken into account to develop transmission networks, and characterize the risk for human infection, eventually leading to a better control of disease transmission.

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“…In Colombia, 15 triatomine species are found to be naturally infected with T. cruzi , with Rhodnius prolixus , Triatoma dimidiata and Panstrongylus geniculatus being the most relevant [ 23 ]. The infection rate within these triatomines can surpass 40% [ 8 , 24 – 27 ], making it higher than that within other triatomines [ 28 , 29 ], and explains the relevance of these particular triatomine species, along with their capability of colonizing human dwellings [ 23 ]. The feeding preferences of these insects include humans; arboreal mammals, such as New World monkeys, sloths, opossums, and coatis; domestic mammals, such as cats, dogs, cows and rodents, as well as other animals such as reptiles and bats [ 7 – 10 , 30 ].…”

Section: Introductionmentioning

confidence: 99%

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

et al. 2020

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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 Simpsonʼs 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.

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Molecular epidemiology of Trypanosoma cruzi and Triatoma dimidiata in costal Ecuador

Cited by 14 publications

References 57 publications

Remarkable genetic diversity of Trypanosoma cruzi and Trypanosoma rangeli in two localities of southern Ecuador identified via deep sequencing of mini-exon gene amplicons

Remarkable genetic diversity of Trypanosoma cruzi and Trypanosoma rangeli in two localities of southern Ecuador identified via deep sequencing of mini-exon gene amplicons

Detailed ecological associations of triatomines revealed by metabarcoding and next-generation sequencing: implications for triatomine behavior and Trypanosoma cruzi transmission cycles

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

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