Identification of blood-feeding sources in Panstrongylus , Psammolestes , Rhodnius and Triatoma using amplicon-based nex
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RESEARCH
Identification of blood‑feeding sources in Panstrongylus, Psammolestes, Rhodnius and Triatoma using amplicon‑based next‑generation sequencing Luisa M. Arias‑Giraldo1, Marina Muñoz1, Carolina Hernández1, Giovanny Herrera1, Natalia Velásquez‑Ortiz1, Omar Cantillo‑Barraza2, Plutarco Urbano3, Andrés Cuervo4 and Juan David Ramírez1*
Abstract 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 con‑ ducted 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 con‑ ventional 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 per‑ formed 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 feed‑ ing 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. Keywords: Chagas disease, Trypanosoma cruzi, Triatominae, Feeding sources, NGS, Colombia *Correspondence: [email protected] 1 Grupo de Investigaciones Microbiológicas‑UR (GIMUR), Departamento de Biología, Facultad de Ciencias Naturales, Universidad del Rosario, Bogotá, Colombia Full list of author information is available at the end of the article © The Author(s) 2
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