Coleção de Vetores de Tripanosomatídeos (Fiocruz/COLVET) held at the institution Fiocruz Minas in Brazil: diversity of Triatominae (Hemiptera, Reduviidae) and relevance for research, education, and entomological surveillance.

The Coleção de Vetores de Tripanosomatídeos (Fiocruz/COLVET), Minas Gerais, Brazil, stands out as one of the most important collections of blood-sucking triatomines, the vectors of Trypanosomacruzi that causes Chagas disease. The aim is to describe the collection and the services it provides to support scientific research, educational activities, and entomological surveillance between 2013-2019.The data associated with the specimens held in Fiocruz/COLVET is available from the Sistema de Informação sobre a Biodiversidade Brasileira (SiBBr). These specimen metadata were analyzed and either tabulated or plotted on graph and maps. The records of services provided by the collection between 2013-2019 were also categorized and analyzed. There are 12,568 triatomine specimens deposited in the collection that belong to 77 species and 11 genera, from 15 American countries. Of the ~ 65 species of triatomines found in Brazil, 38 (57.6%) are present in the collection, including specimens from all biomes and all but three Brazilian states. The occurrence of Triatomacostalimai, Triatomalenti, Rhodniusnasutus, and Panstrongyluslenti apparently collected beyond their known distribution ranges are reported and discussed. The collection provided 168 services, supporting educational activities (41.7%), scientific research (35.7%), and regional/national entomological surveillance of triatomines (22.6%). Between the years 2014 and 2020, the number of biological specimens deposited in the Fiocruz/COLVET repository increased from 4,778 to 12,568 triatomine specimens. In addition to its great value to biodiversity conservation, the collection is of great importance because of its support of research and educational activities, and contributions to entomological surveillance, and, therefore, to public health.

Transcriptome-based molecular systematics: Rhodnius montenegrensis (Triatominae) and its position within the Rhodnius prolixus-Rhodnius robustus cryptic-species complex.

BACKGROUND: Rhodnius montenegrensis (Triatominae), a potential vector of Chagas disease, was described after R. robustus-like bugs from southwestern Amazonia. Mitochondrial cytb sequence near-identity with sympatric R. robustus (genotype II) raised doubts about the taxonomic status of R. montenegrensis, but comparative studies have reported fairly clear morphological and genetic differences between R. montenegrensis and laboratory stocks identified as R. robustus. Here, we use a transcriptome-based approach to investigate this apparent paradox. RESULTS: We retrieved publicly-available transcriptome sequence-reads from R. montenegrensis and from the R. robustus stocks used as the taxonomic benchmark in comparative studies. We (i) aligned transcriptome sequence-reads to mitochondrial (cytb) and nuclear (ITS2, D2-28S and AmpG) query sequences (47 overall) from members of the R. prolixus-R. robustus cryptic-species complex and related taxa; (ii) computed breadth- and depth-coverage for the 259 consensus sequences generated by these alignments; and, for each locus, (iii) appraised query sequences and full-breadth-coverage consensus sequences in terms of nucleotide-sequence polymorphism and phylogenetic relations. We found evidence confirming that R. montenegrensis and R. robustus genotype II are genetically indistinguishable and, hence, implying that they are, in all likelihood, the same species. Furthermore, we found compelling genetic evidence that the benchmark 'R. robustus' stocks used in R. montenegrensis description and in later transcriptome-based comparisons are in fact R. prolixus, although likely mixed to some degree with R. robustus (probably genotype II, a.k.a. R. montenegrensis). CONCLUSIONS: We illustrate how public-domain genetic/transcriptomic data can help address challenging issues in disease-vector systematics. In our case-study, taxonomic confusion apparently stemmed from the misinterpretation of sequence-data analyses and misidentification of taxonomic-benchmark stocks. More generally, and together with previous reports of mixed and/or misidentified Rhodnius spp. laboratory colonies, our results call into question the conclusions of many studies (on morphology, genetics, physiology, behavior, bionomics or interactions with microorganisms including trypanosomes) based on non-genotyped 'R. prolixus' or 'R. robustus' stocks. Correct species identification is a prerequisite for investigating the factors that underlie the physiological, behavioral or ecological differences between primary domestic vectors of Chagas disease, such as R. prolixus, and their sylvatic, medically less-relevant relatives such as R. robustus (s.l.) including R. montenegrensis.

Dehydration-Stress Resistance in Two Sister, Cryptic Rhodnius Species-Rhodnius prolixus and Rhodnius robustus Genotype I (Hemiptera: Reduviidae).

Rhodnius prolixus Stål, a major Chagas disease vector, often colonizes in houses, whereas its sister species, Rhodnius robustus Larrousse genotype I, does not colonize in houses and has little medical relevance. Factors potentially underlying this crucial difference remain largely uncharted. The 'microclimate-adaptation hypothesis' notes that R. prolixus is adapted to the dry microclimate of small-crowned Copernicia palms, whereas R. robustus I exploits the high-moisture microclimate of large-crowned Attalea and Acrocomia. Hence, R. prolixus, but not R. robustus I, would be (pre)adapted to the relatively dry microclimate typical of man-made habitats. This hypothesis predicts that, while severe dehydration should harm both species similarly, R. prolixus should withstand moderate-to-mild dehydration stress better than R. robustus I. To test this prediction, we compared fitness metrics of genotyped R. prolixus and R. robustus I kept at 28°C and under severe (20% relative humidity, RH), moderate (40% RH), or mild dehydration stress (75% RH). Egg-hatching success increased with decreasing dehydration stress in R. robustus I (0% → 19% → 100%), but was high across treatments in R. prolixus (78% → 100% → 100%). Both species underwent high, early mortality under severe dehydration; under moderate and mild stress, R. prolixus experienced less mortality and survived longer than R. robustus I. Our results suggest that adaptation to distinct palm-crown microclimates may partly underlie the so far unexplained differences in house-colonization ability among Rhodnius Stål species. Experimental replication across additional species/populations will be required to further probe this adaptive hypothesis-which, if supported, may also provide insight into the likely responses of Chagas disease vectors to climate change.

Drivers of house invasion by sylvatic Chagas disease vectors in the Amazon-Cerrado transition: A multi-year, state-wide assessment of municipality-aggregated surveillance data.

BACKGROUND: Insecticide spraying efficiently controls house infestation by triatomine bugs, the vectors of Trypanosoma cruzi. The strategy, however, is ineffective against sylvatic triatomines, which can transmit Chagas disease by invading (without colonizing) man-made structures. Despite growing awareness of the relevance of these transmission dynamics, the drivers of house invasion by sylvatic triatomines remain poorly understood. METHODS/FINDINGS: About 12,000 sylvatic triatomines were caught during routine surveillance in houses of Tocantins state, Brazil, in 2005-2013. Using negative binomial regression, information-theoretic model evaluation/averaging, and external model validation, we investigated the effects of regional (Amazon/Cerrado), landscape (preservation/disturbance), and climate covariates (temperature, rainfall) on the municipality-aggregated numbers of house-invading Rhodnius pictipes, R. robustus, R. neglectus, and Panstrongylus geniculatus. House invasion by R. pictipes and R. robustus was overall more frequent in the Amazon biome, tended to increase in municipalities with more well-preserved land, and decreased in rainier municipalities. Across species, invasion decreased with higher landscape-disturbance levels and in hotter-day municipalities. Invasion by R. neglectus and P. geniculatus increased somewhat with more land at intermediate disturbance and peaked in average-rainfall municipalities. Temperature effects were more pronounced on P. geniculatus than on Rhodnius spp. CONCLUSIONS: We report widespread, frequent house invasion by sylvatic triatomines in the Amazon-Cerrado transition. Our analyses indicate that readily available environmental metrics may help predict the risk of contact between sylvatic triatomines and humans at coarse geographic scales, and hint at specific hypotheses about climate and deforestation effects on those vectors-with some taxon-specific responses and some seemingly general trends. Thus, our focal species appear to be quite sensitive to higher temperatures, and might be less common in more heavily-disturbed than in better-preserved environments. This study illustrates, in sum, how entomological routine-surveillance data can be efficiently used for Chagas disease risk prediction and stratification when house-colonizing vectors are absent.

Biogeography of Brazilian populations of Panstrongylus megistus (Hemiptera, Reduviidae, Triatominae) based on molecular marker and paleo-vegetational data.

In epidemiological terms, Panstrongylus megistus is one of the most important species of triatomine bug in Brazil. Samples from 11 localities were studied using the random amplification of polymorphic DNA (RAPD) technique, which was able to differentiate the study populations clearly. Biogeographical data indicate that these populations could already have arisen 18,000 years ago (C(14)), it being possible to differentiate insects from the Brazilian states of Santa Catarina (SC) in the south, Ceará (CE) in the northeast and another large intermediate block containing the remaining eight populations from five other states. These results agree with those obtained by phenograms constructed from RAPD data, in which the SC population lies opposite those of CE, consistent with the greatest geographical distance between these localities. The other eight populations (Alagoas (AL), Bahia (BA), Goiás (GO), Minas Gerais (MG) and São Paulo (SP)) are closer genetically and originated in areas whose vegetational characteristics have remained similar to each other during the last 18,000 years, thus allowing greater contact between them. The greatest divergence of this group of insects and those of Ceará appears to have occurred 8000 years ago. This more humid period gave rise to other landscape changes, allowing greater differentiation of the vegetation and consequent expansion of P. megistus populations. Formation of the Serras do Mar and Mantiqueira probably created geographical barriers that favored a certain degree of isolation and greater differentiation of the SC population. Atlantic forest remnants within the caatinga domain (created between 25 and 17,000 years ago), where the CE populations originated probably constitute ecological refugia produced by successive amplification and retraction of the most suitable habitats for this species.