The Chagas disease domestic transmission cycle in Guatemala: Parasite-vector switches and lack of mitochondrial co-diversification between Triatoma dimidiata and Trypanosoma cruzi subpopulations suggest non-vectorial parasite dispersal across the Motagua

Parasites transmitted by insects must adapt to their vectors and reservoirs. Chagas disease, an American zoonosis caused by Trypanosoma cruzi, is transmitted by several species of triatomines. In Central America, Triatoma dimidiata is a widely dispersed vector found in sylvatic and domestic habitats, with distinct populations across the endemic region of Guatemala. Our aim was to test the strength of association between vector and parasite genetic divergence in domestic environments. Microsatellite (MS) loci were used to characterize parasites isolated from T. dimidiata (n=112) collected in domestic environments. Moderate genetic differentiation was observed between parasites north and south of the Motagua Valley, an ancient biogeographic barrier (FST 0.138, p=0.009). Slightly reduced genotypic diversity and increased heterozygosity in the north (Allelic richness (Ar)=1.00-6.05, FIS -0.03) compared to the south (Ar=1.47-6.30, FIS 0.022) suggest either a selective or demographic process during parasite dispersal. Based on parasite genotypes and geographic distribution, 15 vector specimens and their parasite isolates were selected for mitochondrial co-diversification analysis. Genetic variability and phylogenetic congruence were determined with mitochondrial DNA sequences (10 parasite maxicircle gene fragments and triatomine ND4+CYT b). A Mantel test as well as phylogenetic, network and principal coordinates analyses supported at least three T. dimidiata haplogroups separated by geographic distance across the Motagua Valley. Maxicircle sequences showed low T. cruzi genetic variability (π nucleotide diversity 0.00098) with no evidence of co-diversification with the vector, having multiple host switches across the valley. Sylvatic Didelphis marsupialis captured across the Motagua Valley were found to be infected with T. cruzi strains sharing MS genotypes with parasites isolated from domiciliated triatomines. The current parasite distribution in domestic environments can be explained by multiple parasite-host switches between vector populations and selection or bottleneck processes across the Motagua Valley, with a possible role for didelphids in domestic transmission.

Pennington PM ，Messenger LA ，Reina J ，Juárez JG ，Lawrence GG ，Dotson EM ，Llewellyn MS ，Cordón-Rosales C ... -  《-》

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》

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》

Migration and Gene Flow Among Domestic Populations of the Chagas Insect Vector Triatoma dimidiata (Hemiptera: Reduviidae) Detected by Microsatellite Loci.

Triatoma dimidiata (Latreille, 1811) is the most abundant and significant insect vector of the parasite Trypanosoma cruzi in Central America, and particularly in Guatemala. Tr. cruzi is the causative agent of Chagas disease, and successful disease control requires understanding the geographic distribution and degree of migration of vectors such as T. dimidiata that frequently re-infest houses within months following insecticide application. The population genetic structure of T. dimidiata collected from six villages in southern Guatemala was studied to gain insight into the migration patterns of the insects in this region where populations are largely domestic. This study provided insight into the likelihood of eliminating T. dimidiata by pesticide application as has been observed in some areas for other domestic triatomines such as Triatoma infestans. Genotypes of microsatellite loci for 178 insects from six villages were found to represent five genetic clusters using a Bayesian Markov Chain Monte Carlo method. Individual clusters were found in multiple villages, with multiple clusters in the same house. Although migration occurred, there was statistically significant genetic differentiation among villages (FR T = 0.05) and high genetic differentiation among houses within villages (FSR = 0.11). Relatedness of insects within houses varied from 0 to 0.25, i.e., from unrelated to half-sibs. The results suggest that T. dimidiata in southern Guatemala moves between houses and villages often enough that recolonization is likely, implying the use of insecticides alone is not sufficient for effective control of Chagas disease in this region and more sustainable solutions are required.

Stevens L ，Monroy MC ，Rodas AG ，Hicks RM ，Lucero DE ，Lyons LA ，Dorn PL ... -  《-》

Landscape ecology of Trypanosoma cruzi in the southern Yucatan Peninsula.

Landscape interactions of Trypanosoma cruzi (Tc) with Triatoma dimidiata (Td) depend on the presence and relative abundance of mammal hosts. This study analyzed a landscape adjacent to the Calakmul Biosphere Reserve, composed of conserved areas, crop and farming areas, and the human community of Zoh Laguna with reported Chagas disease cases. Sylvatic mammals of the Chiroptera, Rodentia, and Marsupialia orders were captured, and livestock and pets were sampled along with T. dimidiata in all habitats. Infection by T. cruzi was analyzed using mtDNA markers, while lineage and DTU was analyzed using the mini-exon. 303 sylvatic specimens were collected, corresponding to 19 species during the rainy season and 114 specimens of 18 species during dry season. Five bats Artibeus jamaicensis, Artibeus lituratus, Sturnira lilium, Sturnira ludovici, Dermanura phaeotis (Dp) and one rodent Heteromys gaumeri were collected in the three habitats. All but Dp, and including Carollia brevicauda and Myotis keaysi, were infected with predominately TcI in the sylvatic habitat and TcII in the ecotone. Sigmodon hispidus was the rodent with the highest prevalence of infection by T. cruzi I and II in ecotone and domestic habitats. Didelphis viginiana was infected only with TcI in both domestic and sylvatic habitats; the only two genotyped human cases were TcII. Two main clades of T. cruzi, lineages I (DTU Ia) and II (DTU VI), were found to be sympatric (all habitats and seasons) in the Zoh-Laguna landscape, suggesting that no species-specific interactions occur between the parasite and any mammal host, in any habitat. We have also found mixed infections of the two principal T. cruzi clades in individuals across modified habitats, particularly in livestock and pets, and in both haplogroups of T. dimidiata. Results are contradictory to the dilution hypothesis, although we did find that most resilient species had an important role as T. cruzi hosts. Our study detected some complex trends in parasite transmission related to lineage sorting within the matrix. Intriguingly, TcIa is dominant in terrestrial small wildlife in the sylvatic habitat and is the only parasite DTU found in D. virginiana in the domestic habitat, although its frequency remained constant in sylvatic and ecotone vectors. Bats have a key role in TcVI dispersal from the sylvatic habitat, while dogs, sheep, and humans are drivers of TcVI between domestic and ecotone habitats. Overall, our results allow us to conclude that T. cruzi transmission is dependent on host availability within a highly permeable landscape in Zoh Laguna.

López-Cancino SA ，Tun-Ku E ，De la Cruz-Felix HK ，Ibarra-Cerdeña CN ，Izeta-Alberdi A ，Pech-May A ，Mazariegos-Hidalgo CJ ，Valdez-Tah A ，Ramsey JM ... -  《-》