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Mepraia is a genre composed of three species endemic to Chile. Climate projections demonstrate that these triatomines could modify their potential geographical range and does not rule out that hybridization events could be occurring. Furthermore, recent phylogenomic studies suggested possible ancestral events of hybridization and gene flow and, based on the most recent speciation model that divides Mepraia in two geographically distinct groups (the North, composed of M. garjadoi and M. parapatrica , and the South, composed of M. spinolai ), was also suggested an alternative model that may be compatible with ancient hybridization events or continued gene flow. Thus, we conducted a comprehensive study of the capacity to produce hybrids and to reproductive barriers present between Mepraia spp., with the aim of evaluating the possibility of hybridization and interspecific gene flow be occurring in natural conditions and confirming the specific status of the three Mepraia species. Even though hybrids have been obtained in all directions, the combination of different evolutionary events demonstrated here, such as low hatching rate and high mortality rate of hybrids, as well as the presence of post-zygotic reproductive barriers and possible action of Haldane’s rule indicate that there is no continuous interspecific gene flow between Mepraia spp. and confirms the specific status of species based on the biological species concept.

Triatomines are vectors that transmit the protozoan haemoflagellate Trypanosoma cruzi, the causative agent of Chagas disease. The aim of the current review is to provide a synthesis of the immune mechanisms of triatomines against bacteria, viruses, fungi and parasites to provide clues for areas of further research including biological control. Regarding bacteria, the triatomine immune response includes antimicrobial peptides (AMPs) such as defensins, lysozymes, attacins and cecropins, whose sites of synthesis are principally the fat body and haemocytes. These peptides are used against pathogenic bacteria (especially during ecdysis and feeding), and also attack symbiotic bacteria. In relation to viruses, Triatoma virus is the only one known to attack and kill triatomines. Although the immune response to this virus is unknown, we hypothesize that haemocytes, phenoloxidase (PO) and nitric oxide (NO) could be activated. Different fungal species have been described in a few triatomines and some immune components against these pathogens are PO and proPO. In relation to parasites, triatomines respond with AMPs, including PO, NO and lectin. In the case of T. cruzi this may be effective, but Trypanosoma rangeli seems to evade and suppress PO response. Although it is clear that three parasite-killing processes are used by triatomines – phagocytosis, nodule formation and encapsulation – the precise immune mechanisms of triatomines against invading agents, including trypanosomes, are as yet unknown. The signalling processes used in triatomine immune response are IMD, Toll and Jak-STAT. Based on the information compiled, we propose some lines of research that include strategic approaches of biological control.

Background Insects operate complex humoral and cellular immune strategies to fend against invading microorganisms. The majority of these have been characterized in Drosophila and other dipterans. Information on hemipterans, including Triatominae vectors of Chagas disease remains incomplete and fractionated. Results We identified putative immune-related homologs of three Triatominae vectors of Chagas disease , Triatoma pallidipennis , T. dimidiata and T. infestans (TTTs), using comparative transcriptomics based on established immune response gene references, in conjunction with the predicted proteomes of Rhodnius prolixus , Cimex lecticularis and Acyrthosiphon pisum hemimetabolous. We present a compressive description of the humoral and cellular innate immune components of these TTTs and extend the immune information of other related hemipterans. Key homologs of the constitutive and induced immunity genes were identified in all the studied hemipterans. Conclusions Our results in the TTTs extend previous observations in other hemipterans lacking several components of the Imd signaling pathway. Comparison with other hexapods, using published data, revealed that the absence of various Imd canonical components is common in several hemimetabolous species.

Chagas disease, caused by the parasite Trypanosoma cruzi, is the most important vector-borne disease in Latin America. The vectors are insects belonging to the Triatominae (Hemiptera, Reduviidae), and are widely distributed in the Americas. Here, we assess the implications of climatic projections for 2050 on the geographical footprint of two of the main Chagas disease vectors: Rhodnius prolixus (tropical species) and Triatoma infestans (temperate species). We estimated the epidemiological implications of current to future transitions in the climatic niche in terms of changes in the force of infection (FOI) on the rural population of two countries: Venezuela (tropical) and Argentina (temperate). The climatic projections for 2050 showed heterogeneous impact on the climatic niches of both vector species, with a decreasing trend of suitability of areas that are currently at high-to-moderate transmission risk. Consequently, climatic projections affected differently the FOI for Chagas disease in Venezuela and Argentina. Despite the heterogeneous results, our main conclusions point out a decreasing trend in the number of new cases of Tr. cruzi human infections per year between current and future conditions using a climatic niche approach.

Background Triatomine bugs transmit Chagas disease across Latin America, where vector control-surveillance is increasingly decentralized. Locally run systems often deal with highly diverse native-vector faunas—plus, in some areas, domestic populations of non-native species. Flexible entomological-risk indicators that cover native and non-native vectors and can support local decision-making are therefore needed. Methods We present a local-scale entomological-risk score (“TriatoScore”) that leverages and builds upon information on the ecology-behavior and distribution-biogeography of individual triatomine bug species. We illustrate our approach by calculating TriatoScores for the 417 municipalities of Bahia state, Brazil. For this, we (i) listed all triatomine bug species recorded statewide; (ii) derived a “species relevance score” reflecting whether each species is native/non-native and, if native, whether/how often it invades/colonizes dwellings; (iii) mapped each species’ presence by municipality; (iv) for native vectors, weighted presence by the proportion of municipal territory within ecoregions occupied by each species; (v) multiplied “species relevance score” × “weighted presence” to get species-specific “weighted scores”; and (vi) summed “weighted scores” across species to get municipal TriatoScores. Using standardized TriatoScores, we then grouped municipalities into high/moderate/low entomological-risk strata. Results TriatoScores were higher in municipalities dominated by dry-to-semiarid ecoregions than in those dominated by savanna-grassland or, especially, moist-forest ecoregions. Bahia’s native triatomines can maintain high to moderate risk of vector-borne Chagas disease in 318 (76.3%) municipalities. Historical elimination of Triatoma infestans from 125 municipalities reduced TriatoScores by ~ 27% (range, 20–44%); eight municipalities reported T. infestans since Bahia was certified free of Trypanosoma cruzi transmission by this non-native species. Entomological-risk strata based on TriatoScores agreed well with Bahia’s official disease-risk strata, but TriatoScores suggest that the official classification likely underestimates risk in 42 municipalities. Of 152 municipalities failing to report triatomines in 2006–2019, two and 71 had TriatoScores corresponding to, respectively, high and moderate entomological risk. Conclusions TriatoScore can help control-surveillance managers to flexibly assess and stratify the entomological risk of Chagas disease at operationally relevant scales. Integrating eco-epidemiological, demographic, socioeconomic, or operational data (on, e.g., local-scale dwelling-infestation or vector-infection frequencies, land-use change and urbanization, housing conditions, poverty, or the functioning of control-surveillance systems) is also straightforward. TriatoScore may thus become a useful addition to the triatomine bug control-surveillance toolbox. Graphical abstract

In this study, we investigate the origin and diversification of Trypanosoma cruzi vectors within the Rhodniini tribe (Triatominae subfamily) through phylogenetic analyses based on eight genes from 17 species and 497 specimens—the largest sampling of this tribe to date. Our results predominantly support the paraphyly of the genus Rhodnius , with the three Psammolestes species forming a well-supported monophyletic clade nested within it. In two reconstructions, however, Psammolestes and Rhodnius are recovered as reciprocally monophyletic, each with strong support. In Rhodnius , we find monophyletic pallescens and pictipes groups, but a paraphyletic prolixus group, with persistent phylogenetic discordances underscoring uncertainties in species placements. Divergence estimates suggest Rhodniini originated around 5.26 million years ago, notably more recent than previously thought. Evolution within the tribe appears shaped by geography, gene flow, and incomplete lineage sorting rather than traditional taxonomy. Only four species— P. arthuri , R. ecuadoriensis , R. neivai , and R. neglectus —are consistently supported across analyses, likely diversifying during Pleistocene climate changes. Other Rhodniini species may represent a panmictic population with minor structuring influenced by the Andes uplift. This study underscores the need for integrative research combining genetic, ecological, and biogeographical data to fully understand Rhodniini speciation and diversification.

BACKGROUND: The Triatomini and Rhodniini (Hemiptera: Reduviidae) tribes include the most diverse Chagas disease vectors; however, the phylogenetic relationships within the tribes remain obscure. This study provides the most comprehensive phylogeny of Triatomini reported to date. METHODS: The relationships between all of the Triatomini genera and representatives of the three Rhodniini species groups were examined in a novel molecular phylogenetic analysis based on the following six molecular markers: the mitochondrial 16S; Cytochrome Oxidase I and II (COI and COII) and Cytochrome B (Cyt B); and the nuclear 18S and 28S. RESULTS: Our results show that the Rhodnius prolixus and R. pictipes groups are more closely related to each other than to the R. pallescens group. For Triatomini, we demonstrate that the large complexes within the paraphyletic Triatoma genus are closely associated with their geographical distribution. Additionally, we observe that the divergence within the spinolai and flavida complex clades are higher than in the other Triatoma complexes. CONCLUSIONS: We propose that the spinolai and flavida complexes should be ranked under the genera Mepraia and Nesotriatoma. Finally, we conclude that a thorough morphological investigation of the paraphyletic genera Triatoma and Panstrongylus is required to accurately assign queries to natural genera.

Triatomines (Hemiptera: Reduviidae) are the insect vectors of Trypanosoma cruzi , the causative agent of Chagas disease. The gut bacterial communities affect the development of T . cruzi inside the vector, making the characterization of its composition important in the understanding of infection development. We collected 54 triatomine bugs corresponding to four genera in different departments of Colombia. DNA extraction and PCR were performed to evaluate T . cruzi presence and to determine the discrete typing unit (DTU) of the parasite. PCR products of the bacterial 16S rRNA gene were pooled and sequenced. Resulting reads were denoised and QIIME 2 was used for the identification of amplicon sequence variants (ASVs). Diversity (alpha and beta diversity) and richness analyses, Circos plots, and principal component analysis (PCA) were also performed. The overall T . cruzi infection frequency was 75.9%, with TcI being the predominant DTU. Approximately 500,000 sequences were analyzed and 27 bacterial phyla were identified. The most abundant phyla were Proteobacteria (33.9%), Actinobacteria (32.4%), Firmicutes (19.6%), and Bacteroidetes (7.6%), which together accounted for over 90% of the gut communities identified in this study. Genera were identified for these main bacterial phyla, revealing the presence of important bacteria such as Rhodococcus , Serratia , and Wolbachia . The composition of bacterial phyla in the gut of the insects was significantly different between triatomine species, whereas no significant difference was seen between the state of T . cruzi infection. We suggest further investigation with the evaluation of additional variables and a larger sample size. To our knowledge, this study is the first characterization of the gut bacterial structure of the main triatomine genera in Colombia.

Triatomines are haematophagous insects involved in the transmission of Trypanosoma cruzi, the aetiological agent of Chagas disease. The vector competence of these arthropods can be highly variable, depending on the species. A precise identification is therefore crucial for the epidemiological surveillance of T. cruzi and the determination of at-risk human populations. To circumvent the difficulties of morphological identification and the lack of comprehensiveness of the GenBank database, we hereby propose an alternative method for triatomine identification. The femurs of the median legs of triatomines from eight different species from French Guiana were subjected to matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) analysis. Method evaluation was performed on fresh specimens and was applied to dry specimens collected between 1991 and 2003. Femur-derived protein extracts provided reproducible spectra within the same species along with significant interspecies heterogeneity. Validation of the study by blind test analysis provided 100% correct identification of the specimens in terms of the species, sex and developmental stage. MALDI-TOF mass spectrometry appears to be a powerful tool for triatomine identification, which is a major step forward in the fight against Chagas disease.

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.