Molecular ecology of Triatoma dimidiata in southern Belize reveals risk for human infection and the local differentiation of Trypanosoma cruzi parasites.

In Belize, the main vector for Trypanosoma cruzi, the agent of Chagas disease, is Triatoma dimidiata, but transmission cycles and the risk for human infection are unclear. Therefore, the aim of this study was to identify T. dimidiata blood feeding sources and its parasite and microbial diversity, in order to reconstruct T. cruzi parasite transmission ecology in southern Belize. A metabarcoding approach based on deep sequencing of markers was used for bug taxonomy, blood meal sources, T. cruzi genotypes, and microbiota composition. Bugs were collected in 13 villages of Toledo district. Bugs fed on at least 13 species, from domestic hosts such as humans, dogs, cows, and pigs, to synanthropic species such as mice, rats, and opossums, and sylvatic species such as deer, peccary, and kinkajou, in agreement with an opportunistic feeding behavior. Nonetheless, most feeding focused on a few species, including humans. Infection with T. cruzi was detected in 24 of 39 bugs (62%), and the analysis of 242 T. cruzi mini-exon sequences (average 10 ± 5 haplotypes per bug) indicated the presence of TcI and TcIV parasite discrete typing units (DTUs). However, for both DTUs, sequences from Belize mostly clustered apart from sequences from North and South America, suggesting the local differentiation of parasites. T. dimidiata also harbored a diverse bacterial microbiota, with ontogenic changes suggesting microbiota maturation during nymphal development. Together, these results indicate a significant risk for T. cruzi infection in humans. They also highlight the need to better characterize the diversity of T. cruzi strains in the region and its impact on disease epidemiology.

Polonio R ，López-Domínguez J ，Herrera C ，Dumonteil E ... -  《-》

Disentangling Trypanosoma cruzi transmission cycle dynamics through the identification of blood meal sources of natural populations of Triatoma dimidiata in Yucatán, Mexico.

In the Yucatán Peninsula, Mexico, Triatoma dimidiata is the main vector of Trypanosoma cruzi, the causative agent of Chagas disease. Little effort has been made to identify blood meal sources of T. dimidiata in natural conditions in this region, although this provides key information to disentangle T. cruzi transmission cycles and dynamics and guide the development of more effective control strategies. We identified the blood meals of a large sample of T. dimidiata bugs collected in different ecotopes simultaneously with the assessment of bug infection with T. cruzi, to disentangle the dynamics of T. cruzi transmission in the region. A sample of 248 T. dimidiata bugs collected in three rural villages and in the sylvatic habitat surrounding these villages was used. DNA from each bug midgut was extracted and bug infection with T. cruzi was assessed by PCR. For blood meal identification, we used a molecular assay based on cloning and sequencing following PCR amplification with vertebrate universal primers, and allowing the detection of multiple blood meals in a single bug. Overall, 28.7% of the bugs were infected with T. cruzi, with no statistical difference between bugs from the villages or from sylvatic ecotopes. Sixteen vertebrate species including domestic, synanthropic and sylvatic animals, were identified as blood meal sources for T. dimidiata. Human, dog and cow were the three main species identified, in bugs collected in the villages as well as in sylvatic ecotopes. Importantly, dog was highlighted as the main blood meal source after human. Dog was also the most frequently identified animal together with human within single bugs, and tended to be associated with the infection of the bugs. Dog, human and cow were identified as the main mammals involved in the connection of sylvatic and domestic transmission cycles in the Yucatán Peninsula, Mexico. Dog appeared as the most important animal in the transmission pathway of T. cruzi to humans, but other domestic and synanthropic animals, which most were previously reported as important hosts of T. cruzi in the region, were evidenced and should be taken into account as part of integrated control strategies aimed at disrupting parasite transmission.

Moo-Millan JI ，Arnal A ，Pérez-Carrillo S ，Hernandez-Andrade A ，Ramírez-Sierra MJ ，Rosado-Vallado M ，Dumonteil E ，Waleckx E ... -  《Parasites & Vectors》

Diversity of Trypanosoma cruzi parasites infecting Triatoma dimidiata in Central Veracruz, Mexico, and their One Health ecological interactions.

Triatoma dimidiata is the main vector of Trypanosoma cruzi parasites in Veracruz, Mexico, and its association with human housing appears variable. Also, in spite of a high seroprevalence of T. cruzi infection in humans, parasite transmission remains poorly understood. Therefore, we aimed to identify T. dimidiata blood feeding sources and its parasite and microbial diversity to reconstruct T. cruzi parasite transmission ecology in central Veracruz, Mexico, within a One Health/Ecohealth framework. We used a metabarcoding and deep sequencing approach of specific markers for the simultaneous identification of T. dimidiata haplogroup (ITS-2), vertebrate blood meals (12 s gene), T. cruzi parasites (mini-exon gene), and gut microbiota (bacterial 16 s). Twelve species of domestic/synanthropic animals and humans were identified as blood sources, with multiple feeding on 4.2 ± 0.4 hosts per bug. The feeding/parasite transmission network was strongly centered on humans, emphasizing a significant risk of infection. We also unambiguously confirmed the presence of TcI, TcII, TcV and TcVI DTUs in T. dimidiata, and sequences from Veracruz tended to cluster apart from parasites from other regions, suggesting some level of local differentiation. Analysis of T. dimidiata microbiota suggested that several bacterial families may be associated with the presence/absence of T. cruzi, and some of these associations may also be parasite DTU-specific. Such integrative approaches within the EcoHealth/One Health framework provide key insights on T. cruzi transmission and potential novel strategies for disease control.

Murillo-Solano C ，Ramos-Ligonio A ，López-Monteon A ，Guzmán-Gómez D ，Torres-Montero J ，Herrera C ，Dumonteil E ... -  《-》

Uncovering vector, parasite, blood meal and microbiome patterns from mixed-DNA specimens of the Chagas disease vector Triatoma dimidiata.

Chagas disease, considered a neglected disease by the World Health Organization, is caused by the protozoan parasite Trypanosoma cruzi, and transmitted by >140 triatomine species across the Americas. In Central America, the main vector is Triatoma dimidiata, an opportunistic blood meal feeder inhabiting both domestic and sylvatic ecotopes. Given the diversity of interacting biological agents involved in the epidemiology of Chagas disease, having simultaneous information on the dynamics of the parasite, vector, the gut microbiome of the vector, and the blood meal source would facilitate identifying key biotic factors associated with the risk of T. cruzi transmission. In this study, we developed a RADseq-based analysis pipeline to study mixed-species DNA extracted from T. dimidiata abdomens. To evaluate the efficacy of the method across spatial scales, we used a nested spatial sampling design that spanned from individual villages within Guatemala to major biogeographic regions of Central America. Information from each biotic source was distinguished with bioinformatics tools and used to evaluate the prevalence of T. cruzi infection and predominant Discrete Typing Units (DTUs) in the region, the population genetic structure of T. dimidiata, gut microbial diversity, and the blood meal history. An average of 3.25 million reads per specimen were obtained, with approximately 1% assigned to the parasite, 20% to the vector, 11% to bacteria, and 4% to putative blood meals. Using a total of 6,405 T. cruzi SNPs, we detected nine infected vectors harboring two distinct DTUs: TcI and a second unidentified strain, possibly TcIV. Vector specimens were sufficiently variable for population genomic analyses, with a total of 25,710 T. dimidiata SNPs across all samples that were sufficient to detect geographic genetic structure at both local and regional scales. We observed a diverse microbiotic community, with significantly higher bacterial species richness in infected T. dimidiata abdomens than those that were not infected. Unifrac analysis suggests a common assemblage of bacteria associated with infection, which co-occurs with the typical gut microbial community derived from the local environment. We identified vertebrate blood meals from five T. dimidiata abdomens, including chicken, dog, duck and human; however, additional detection methods would be necessary to confidently identify blood meal sources from most specimens. Overall, our study shows this method is effective for simultaneously generating genetic data on vectors and their associated parasites, along with ecological information on feeding patterns and microbial interactions that may be followed up with complementary approaches such as PCR-based parasite detection, 18S eukaryotic and 16S bacterial barcoding.

Orantes LC ，Monroy C ，Dorn PL ，Stevens L ，Rizzo DM ，Morrissey L ，Hanley JP ，Rodas AG ，Richards B ，Wallin KF ，Helms Cahan S ... -  《PLoS Neglected Tropical Diseases》

The diversity of the Chagas parasite, Trypanosoma cruzi, infecting the main Central American vector, Triatoma dimidiata, from Mexico to Colombia.

Dorn PL ，McClure AG ，Gallaspy MD ，Waleckx E ，Woods AS ，Monroy MC ，Stevens L ... -  《PLoS Neglected Tropical Diseases》