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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 localities ( c. 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.

parasites and vectors

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