Journal Article

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

Hernández, Carolina, Muñoz, Marina, Cuervo, Andrés, Herrera, Giovanny, Urbano, Plutarco, Ramírez, Juan David, Arias-Giraldo, Luisa M., Velásquez-Ortiz, Natalia, Cantillo-Barraza, Omar

2020

Overview

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.

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Analysis / Animals / Biomedical and Life Sciences / Biomedicine / Birds / Birds - blood / Birds - genetics / Blood / blood meal / Chagas disease / Chagas Disease - parasitology / Chagas Disease - transmission / Colombia / data collection / Datasets / Deoxyribonucleic acid / digestive system / Disease transmission / DNA / DNA, Protozoan / Dynamics / Entomology / etiological agents / Feeding / Feeding Behavior / Feeding sources / genes / Genetic diversity / Health aspects / hematophagy / High-Throughput Nucleotide Sequencing - methods / Humans / Identification / Infections / Infectious Diseases / Insect Vectors - genetics / Insect Vectors - parasitology / Insects / Intestines / Marsupials / Next-generation sequencing / NGS / Nucleotide sequence / Panstrongylus / Panstrongylus - genetics / Panstrongylus - parasitology / Parasites / Parasitology / PCR / Polymerase chain reaction / Primates / Primates - blood / Primates - genetics / Protozoa / quantitative polymerase chain reaction / Real-Time Polymerase Chain Reaction / Regions / Rhodnius / Rhodnius - genetics / Rhodnius - parasitology / RNA / rRNA 12S / Sequencing / Species diversity / Triatoma / Triatoma - genetics / Triatoma - parasitology / Triatominae / Triatominae - genetics / Triatominae - parasitology / Tropical diseases / Tropical Medicine / Trypanosoma cruzi / Trypanosoma cruzi - genetics / Trypanosoma cruzi - isolation & purification / Vector-borne diseases / Vectors / Vertebrates / Veterinary Medicine/Veterinary Science / Virology