TriatoScore: an entomological-risk score for Chagas disease vector control-surveillance.

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.

Trends in taxonomy of Triatomini (Hemiptera, Reduviidae, Triatominae): reproductive compatibility reinforces the synonymization of Meccus Stål, 1859 with Triatoma Laporte, 1832.

BACKGROUND: Meccus' taxonomy has been quite complex since the first species of this genus was described by Burmeister in 1835 as Conorhinus phyllosoma. In 1859 the species was transferred to the genus Meccus and in 1930 to Triatoma. However, in the twentieth century, the Meccus genus was revalidated (alteration corroborated by molecular studies) and, in the twenty-first century, through a comprehensive study including more sophisticated phylogenetic reconstruction methods, Meccus was again synonymous with Triatoma. Events of natural hybridization with production of fertile offspring have already been reported among sympatric species of the T. phyllosoma subcomplex, and experimental crosses demonstrated reproductive viability among practically all species of the T. phyllosoma subcomplex that were considered as belonging to the genus Meccus, as well as between these species and species of Triatoma. Based on the above, we carried out experimental crosses between T. longipennis (considered M. longipennis in some literature) and T. mopan (always considered as belonging to Triatoma) to evaluate the reproductive compatibility between species of the T. phyllosoma complex. In addition, we have grouped our results with information from the literature regarding crosses between species that were grouped in the genus Meccus with Triatoma, in order to discuss the importance of experimental crosses to confirm the generic reorganization of species. RESULTS: The crosses between T. mopan female and T. longipennis male resulted in viable offspring. The hatching of hybrids, even if only in one direction and/or at low frequency, demonstrates reproductive compatibility and homeology between the genomes of the parents. CONCLUSION: Considering that intergeneric crosses usually do not result in viable offspring in Triatominae, the reproductive compatibility observed between the T. phyllosoma subcomplex species considered in the Meccus genus with species of the Triatoma genus shows that there is "intergeneric" genomic compatibility, which corroborates the generic reorganization of Meccus in Triatoma.

TriatoDex, an electronic identification key to the Triatominae (Hemiptera: Reduviidae), vectors of Chagas disease: Development, description, and performance.

Correct identification of triatomine bugs is crucial for Chagas disease surveillance, yet available taxonomic keys are outdated, incomplete, or both. Here we present TriatoDex, an Android app-based pictorial, annotated, polytomous key to the Triatominae. TriatoDex was developed using Android Studio and tested by 27 Brazilian users. Each user received a box with pinned, number-labeled, adult triatomines (33 species in total) and was asked to identify each bug to the species level. We used generalized linear mixed models (with user- and species-ID random effects) and information-theoretic model evaluation/averaging to investigate TriatoDex performance. TriatoDex encompasses 79 questions and 554 images of the 150 triatomine-bug species described worldwide up to 2017. TriatoDex-based identification was correct in 78.9% of 824 tasks. TriatoDex performed better in the hands of trained taxonomists (93.3% vs. 72.7% correct identifications; model-averaged, adjusted odds ratio 5.96, 95% confidence interval [CI] 3.09-11.48). In contrast, user age, gender, primary job (including academic research/teaching or disease surveillance), workplace (including universities, a reference laboratory for triatomine-bug taxonomy, or disease-surveillance units), and basic training (from high school to biology) all had negligible effects on TriatoDex performance. Our analyses also suggest that, as TriatoDex results accrue to cover more taxa, they may help pinpoint triatomine-bug species that are consistently harder (than average) to identify. In a pilot comparison with a standard, printed key (370 tasks by seven users), TriatoDex performed similarly (84.5% correct assignments, CI 68.9-94.0%), but identification was 32.8% (CI 24.7-40.1%) faster on average-for a mean absolute saving of ~2.3 minutes per bug-identification task. TriatoDex holds much promise as a handy, flexible, and reliable tool for triatomine-bug identification; an updated iOS/Android version is under development. We expect that, with continuous refinement derived from evolving knowledge and user feedback, TriatoDex will substantially help strengthen both entomological surveillance and research on Chagas disease vectors.

Under pressure: phenotypic divergence and convergence associated with microhabitat adaptations in Triatominae.

BACKGROUND: Triatomine bugs, the vectors of Chagas disease, associate with vertebrate hosts in highly diverse ecotopes. It has been proposed that occupation of new microhabitats may trigger selection for distinct phenotypic variants in these blood-sucking bugs. Although understanding phenotypic variation is key to the study of adaptive evolution and central to phenotype-based taxonomy, the drivers of phenotypic change and diversity in triatomines remain poorly understood. METHODS/RESULTS: We combined a detailed phenotypic appraisal (including morphology and morphometrics) with mitochondrial cytb and nuclear ITS2 DNA sequence analyses to study Rhodnius ecuadoriensis populations from across the species' range. We found three major, naked-eye phenotypic variants. Southern-Andean bugs primarily from vertebrate-nest microhabitats (Ecuador/Peru) are typical, light-colored, small bugs with short heads/wings. Northern-Andean bugs from wet-forest palms (Ecuador) are dark, large bugs with long heads/wings. Finally, northern-lowland bugs primarily from dry-forest palms (Ecuador) are light-colored and medium-sized. Wing and (size-free) head shapes are similar across Ecuadorian populations, regardless of habitat or phenotype, but distinct in Peruvian bugs. Bayesian phylogenetic and multispecies-coalescent DNA sequence analyses strongly suggest that Ecuadorian and Peruvian populations are two independently evolving lineages, with little within-lineage phylogeographic structuring or differentiation. CONCLUSIONS: We report sharp naked-eye phenotypic divergence of genetically similar Ecuadorian R. ecuadoriensis (nest-dwelling southern-Andean vs palm-dwelling northern bugs; and palm-dwelling Andean vs lowland), and sharp naked-eye phenotypic similarity of typical, yet genetically distinct, southern-Andean bugs primarily from vertebrate-nest (but not palm) microhabitats. This remarkable phenotypic diversity within a single nominal species likely stems from microhabitat adaptations possibly involving predator-driven selection (yielding substrate-matching camouflage coloration) and a shift from palm-crown to vertebrate-nest microhabitats (yielding smaller bodies and shorter and stouter heads). These findings shed new light on the origins of phenotypic diversity in triatomines, warn against excess reliance on phenotype-based triatomine-bug taxonomy, and confirm the Triatominae as an informative model system for the study of phenotypic change under ecological pressure .

Detection of Trypanosoma cruzi DNA in false negative samples of collected triatomines, xenodiagnosis material, and biopsies of experimentally infected animals.

Direct test over the gut material from triatomine vectors and xenodiagnosis over mammalian hosts are classical techniques for Trypanosoma cruzi parasitological diagnosis. Nevertheless, negative results can be a source of uncertainty. Experimental models have allowed evaluating the tissue invasion of different strains of T. cruzi, but conventional techniques for tissue biopsies involve time-consuming and elaborated procedures and have low sensitivity. Gut material of collected triatomines (microscopically negative) (n = 114), material of mammal xenodiagnoses (microscopically negative) (n = 138), and biopsy material (microscopically negative) from experimentally infected animals (n = 34) with isolates from endemic areas of Chagas' disease from Venezuela were used for DNA extraction and PCR for the amplification of kinetoplast DNA (kDNA) and satellite DNA (sDNA) of T. cruzi. Positive PCR was observed in 53.6% of collected triatomine material, 15.8% of parasitological negative xenodiagnosis material, and 70.6% in biopsies, revealing underestimation by the parasitological tests and the valour of this analysis with preserved material. Anzoátegui was the state with the highest percentage of infection, and the triatomine species Rhodnius prolixus and Panstrongylus geniculatus had the highest percentages of infection. Didelphis marsupialis and Canis familiaris were the most infected by T. cruzi revealed by PCR of xenodiagnosis material. In addition, the PCR technique allowed demonstrating the invasion of T. cruzi in all tissues analyzed, constituting a molecular marker of tissue invasion.

Species-dependent variation of the gut bacterial communities across Trypanosoma cruzi insect vectors.

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.

Ontogeny, species identity, and environment dominate microbiome dynamics in wild populations of kissing bugs (Triatominae).

BACKGROUND: Kissing bugs (Triatominae) are blood-feeding insects best known as the vectors of Trypanosoma cruzi, the causative agent of Chagas' disease. Considering the high epidemiological relevance of these vectors, their biology and bacterial symbiosis remains surprisingly understudied. While previous investigations revealed generally low individual complexity but high among-individual variability of the triatomine microbiomes, any consistent microbiome determinants have not yet been identified across multiple Triatominae species. METHODS: To obtain a more comprehensive view of triatomine microbiomes, we investigated the host-microbiome relationship of five Triatoma species sampled from white-throated woodrat (Neotoma albigula) nests in multiple locations across the USA. We applied optimised 16S rRNA gene metabarcoding with a novel 18S rRNA gene blocking primer to a set of 170 T. cruzi-negative individuals across all six instars. RESULTS: Triatomine gut microbiome composition is strongly influenced by three principal factors: ontogeny, species identity, and the environment. The microbiomes are characterised by significant loss in bacterial diversity throughout ontogenetic development. First instars possess the highest bacterial diversity while adult microbiomes are routinely dominated by a single taxon. Primarily, the bacterial genus Dietzia dominates late-stage nymphs and adults of T. rubida, T. protracta, and T. lecticularia but is not present in the phylogenetically more distant T. gerstaeckeri and T. sanguisuga. Species-specific microbiome composition, particularly pronounced in early instars, is further modulated by locality-specific effects. In addition, pathogenic bacteria of the genus Bartonella, acquired from the vertebrate hosts, are an abundant component of Triatoma microbiomes. CONCLUSION: Our study is the first to demonstrate deterministic patterns in microbiome composition among all life stages and multiple Triatoma species. We hypothesise that triatomine microbiome assemblages are produced by species- and life stage-dependent uptake of environmental bacteria and multiple indirect transmission strategies that promote bacterial transfer between individuals. Altogether, our study highlights the complexity of Triatominae symbiosis with bacteria and warrant further investigation to understand microbiome function in these important vectors. Video abstract.

Spatial distribution of synanthropic triatomines in Piaui State, Northeastern Brazil.

This study aimed to describe the spatial distribution and assess entomological indicators of synanthropic triatomines in Piaui State, Northeastern Brazil. We used surveillance data on the detection, identification and assessment of natural infection with trypanosomatids from triatomines in the State from 2014 to 2017. The State was divided into four macroregions. In relation to the dispersion rates of triatomines, they were much lower in the North, when compared to Southwest, Southeast and Central North macroregions. Infestation rates were higher in the Southwest and Southeast and intradomicile infestation rates varied during the study period, reaching high values in all regions. Insects belonging to the species Triatoma brasiliensis complex, Triatoma pseudomaculata, Triatoma sordida, and to the genus Panstrongylus spp. and Rhodnius spp. were collected during this period. T. brasiliensis was collected from all four regions of the State, but more frequently in those located in the Southeast. A similar pattern was observed for T. pseudomaculata. T. sordida was detected in the municipalities in the Southeast and Southwest regions, and less frequently in the Central North municipalities. Rhodnius spp. was detected in the Central North and North regions, and Panstrongylus spp. in the Central North and Southeast regions. The highest trypanosomatid-positivity rate of T. brasiliensis and Panstrongylus spp. was in the Southeast region. A significant proportion of the municipalities of Piaui State presents entomological parameters that indicate a risk of Chagas disease by vector transmission.

New report of Eratyrus cuspidatus Stål, 1859 (Hemiptera: Reduviidae: Triatominae) in the State of Campeche, Mexico.

INTRODUCTION: Triatomine bugs are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. METHODS: Triatomine bugs were collected and identified following established protocols. In addition, infection with T. cruzi was detected by microscopic and molecular analysis. RESULTS: We captured an adult male specimen of the Eratyrus cuspidatus species that has not been reported in the state of Campeche. CONCLUSIONS: This finding provides new information on the distribution of E. cuspidatus in Mexico. However, more studies are needed to determine their epidemiological significance.

Alien Invasion: The Case of the Western Conifer-Seed Bug (Heteroptera: Coreidae) in Chile, Overreaction, and Misidentifications.

Leptoglossus occidentalis (Heidemann) is an invasive species introduced in Europe, Asia, and most recently South America. In the present study, we report the overreaction situation caused by this bug in Chile, as it has been confused with kissing bugs (Reduviidae: Triatominae), Chagas' disease vectors. During 2018 and first months of 2019, we received 74 alleged cases of kissing bugs to confirm identification. From these, a total of 63 were identified as L. occidentalis, representing a 85% of the total denounces. Additionally, the first bite case in a human is described. The situation caused by L. occidentalis in Chile is discussed, and an illustrated table is provided to correctly identify this species and tell it apart from Triatomines. It is concluded that L. occidentalis is well established in Chile, and it is necessary to educate the population on recognition of the bug and to avoid overreaction as the species can inflict painful bites, but not transmit any disease.

New report of Eratyrus cuspidatus Stål, 1859 (Hemiptera: Reduviidae: Triatominae) in the State of Campeche, Mexico

Abstract INTRODUCTION: Triatomine bugs are vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. METHODS: Triatomine bugs were collected and identified following established protocols. In addition, infection with T. cruzi was detected by microscopic and molecular analysis. RESULTS: We captured an adult male specimen of the Eratyrus cuspidatus species that has not been reported in the state of Campeche. CONCLUSIONS: This finding provides new information on the distribution of E. cuspidatus in Mexico. However, more studies are needed to determine their epidemiological significance.

Wide distribution of Trypanosoma cruzi-infected triatomines in the State of Bahia, Brazil.

BACKGROUND: The identification of Trypanosoma cruzi and blood-meal sources in synanthropic triatomines is important to assess the potential risk of Chagas disease transmission. We identified T. cruzi infection and blood-meal sources of triatomines caught in and around houses in the state of Bahia, northeastern Brazil, and mapped the occurrence of infected triatomines that fed on humans and domestic animals. METHODS: Triatominae bugs were manually captured by trained agents from the Epidemiologic Surveillance team of Bahia State Health Service between 2013 and 2014. We applied conventional PCR to detect T. cruzi and blood-meal sources (dog, cat, human and bird) in a randomized sample of triatomines. We mapped triatomine distribution and analyzed vector hotspots with kernel density spatial analysis. RESULTS: In total, 5906 triatomines comprising 15 species were collected from 127 out of 417 municipalities in Bahia. The molecular analyses of 695 triatomines revealed a ~10% T. cruzi infection rate, which was highest in the T. brasiliensis species complex. Most bugs were found to have fed on birds (74.2%), and other blood-meal sources included dogs (6%), cats (0.6%) and humans (1%). Trypanosoma cruzi-infected triatomines that fed on humans were detected inside houses. Spatial analysis showed a wide distribution of T. cruzi-infected triatomines throughout Bahia; triatomines that fed on dogs, humans, and cats were observed mainly in the northeast region. CONCLUSIONS: Synanthropic triatomines have a wide distribution and maintain the potential risk of T. cruzi transmission to humans and domestic animals in Bahia. Ten species were recorded inside houses, mainly Triatoma sordida, T. pseudomaculata, and the T. brasiliensis species complex. Molecular and spatial analysis are useful to reveal T. cruzi infection and blood-meal sources in synanthropic triatomines, identifying areas with ongoing threat for parasite transmission and improving entomological surveillance strategies.

Fragment detection of Coleopteran and Triatomine insects in experimentally contaminated acai pulp and sugarcane juice.

INTRODUCTION: Oral transmission of acute Chagas disease is an emerging public health concern. This study aimed to detect insect fragments in experimentally contaminated food, by comparing triatomines with other insects. METHODS: Food samples were experimentally contaminated with insects, processed to recover their fragments by light filth, and analyzed by microscopy and Polymerase Chain Reaction (PCR). RESULTS: Morphological differences between coleopteran and triatomine insects were observed in microscopic images. PCR was efficient in amplifying Triatominae DNA in the experimentally contaminated food. CONCLUSIONS: This methodology could be utilized by food analysts to identify possible insect contamination in food samples.

Spatial distribution of triatomine bugs in a Chagas disease endemic region in Brazil.

INTRODUCTION: Trypanosoma cruzi is the etiological agent of Chagas disease (CD), a zoonotic infection transmitted by triatomine bug vectors to human beings. Although the story of this parasitic infection was born in Brazil and here this has made major step forward information, the same cannot be said about the actual distribution of the triatomine vector in several areas of this country. The aim of this study was to assess the occurrence of triatomine species in an endemic region for CD in Northeastern Brazil. METHODS: A retrospective study was performed using data obtained from 2008 to 2017. All information was provided by the V Gerência Regional de Saúde of the state of Pernambuco. The spatial distribution of triatomine species was analyzed by drawing a map using the Quantum geographic information system. RESULTS: A total of 4,694 triatomine specimens (469.4 ± 221.2 per year) were collected during the period 2008-2017, with 94.5% (4,434/4,694) at the intradomicile and 5.5% (260/4,694) at peridomicile environment. Of all arthropods collected, 92.5% (4,340/4,694) and 7.5% (354/4,694) were adults and nymphs, respectively. The species most frequently detected were Panstrongylus lutzi (30.36%), Triatoma brasiliensis (26.12%), Triatoma pseudomaculata (22.43%), and Panstrongylus megistus (20.54%). CONCLUSIONS: These data contribute to a better understanding of the distribution of T. cruzi infection in the Northeastern region of Brazil. Preventive measures based on vector control should be implemented in the study area in order to reduce the burden this neglected tropical disease.

Triatomine dispersion rates and their association with socioeconomic and environmental conditions in Northeastern Brazil, from 2009 to 2013.

Triatomines are vectors of Trypanosoma cruzi , the etiological agent of Chagas disease, which affects between 6 and 12 million people in Latin America, with an incidence rate of 12 thousand cases per year. In the Ceara State, the predominance of the caatinga biome, coupled with a large rural area with precarious human habitations, provides several s shelters for these insects. In this study, we determined the spatiotemporal distribution of triatomine dispersion rates in the Cariri region, Southern Ceara and ascertained the possible association between these rates with socioeconomic and environmental factors. Dispersion rates (number of positive localities/number of searched localities × 100) were analyzed regarding 13 municipalities from the Ceara State, from 2009 to 2013. Socioeconomic and environmental variables collected from national research institutes were associated with the dispersion rates and their local empirical Bayesian estimates. All the municipalities recorded dispersion rates over 10% in all years, and 11 municipalities had average rates over 40% for the period of study. Significant differences were observed among the municipality means. The highest rates were observed in Antonina do Norte and Potengi. According to the correlation analysis, the proportion between the occupied population and the total population showed a significant negative correlation, as well as the percentage of the population who lives under adequate sanitary conditions. Both, the percentage of revenues from external sources and the percentage of urban households in reforested blocks had a significant positive correlation. Our results show that socioeconomic and environmental variables can be factors that contribute to both, the maintenance and the reduction of the elevated dispersion rates observed in the study area. Similar researches that encompass more municipalities from that region may reinforce Chagas disease surveillance and control in the Northeast of Brazil.

Synanthropic triatomines (Hemiptera: Reduviidae): infestation, colonization, and natural infection by trypanosomatids in the State of Rio Grande do Norte, Brazil.

INTRODUCTION: The ecoepidemiological situation in the State of Rio Grande do Norte, Brazil is characterized by frequent invasion and colonization of domiciliary units (DUs) by several triatomine species, with high rates of natural infection by Trypanosoma cruzi. METHODS: We evaluated the possibility of vector transmission of T. cruzi based on records of the occurrence of domiciled triatomines collected by the Secretariat of State for Public Health from 2005 to 2015. During this period, 67.7% (113/167) of municipalities conducted at least one active search and 110 recorded the presence of insects in DUs. These activities were more frequent in municipalities considered to have a high and medium-level risk of T. cruzi transmission. RESULTS: Of 51,569 captured triatomines, the most common species were Triatoma brasiliensis (47.2%) and T. pseudomaculata (40.2%). Colonies of T. brasiliensis, T. pseudomaculata, T. petrocchiae, Panstrongylus lutzi, and Rhodnius nasutus were also recorded in the intradomicile and peridomicile. Natural infection by trypanosomatids was detected in 1,153 specimens; the highest rate was found in R. nasutus (3.5%), followed by T. brasiliensis (2.5%) and T. pseudomaculata (2.4%). There have been high levels of colonization over the years; however, not all infested DUs have been sprayed. CONCLUSIONS: This is the first report of intradomicile and peridomicile colonization by P. lutzi. These results demonstrate the risk of new cases of infection by T. cruzi and reinforce the need for continuous entomological surveillance in the State of Rio Grande do Norte.

Distinguishing two species of Cavernicola (Hemiptera, Reduviidae, Triatominae) with matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

The blood-sucking insects of the subfamily Triatominae are vectors of Chagas disease, the most impairing protozoan parasitic infection in Latin America. Among the five tribes known in the subfamily, one of the least studied is Cavernicolini. It has only two species within Cavernicola genus (Barber, 1937), little is known about the biochemistry of the species of this genus, therefore, using MALDI-TOF MS we provide a better understanding of the two species and differentiates them. The distinction was made by the different spectral profile of the species, where C. lenti presents unique signals in many regions, while the C. pilosa shows high-intensity signals and m/z in high bands. The application of digital mass spectrometry combined with chemometric methods was able to accurately distinguish two species of the genus Cavernicola.

Occurrence of triatomines (Hemiptera: Reduviidae) in domestic and natural environments in Novo Remanso, Itacoatiara, Amazonas, Brazil.

INTRODUCTION: The present study reports the presence of triatomines in natural, peridomestic, and intradomicile environments in Itacoatiara municipality, state of Amazonas, a non-endemic region for Chagas disease. METHODS: Active search was performed inside tree trunks, and palm trees, residences, and peridomiciles localized near the forest area. RESULTS: Twenty adults and ten triatomines nymphs were collected, fifteen of which were from natural forests, thirteen from intradomiciles, and two from peridomicile areas. CONCLUSIONS: The new records of adults and nymphs of triatomines in the intra- and peridomiciles suggest the adoption of prophylactic measures for vector surveillance in the study area.

Deep Learning Algorithms Improve Automated Identification of Chagas Disease Vectors.

Vector-borne Chagas disease is endemic to the Americas and imposes significant economic and social burdens on public health. In a previous contribution, we presented an automated identification system that was able to discriminate among 12 Mexican and 39 Brazilian triatomine (Hemiptera: Reduviidae) species from digital images. To explore the same data more deeply using machine-learning approaches, hoping for improvements in classification, we employed TensorFlow, an open-source software platform for a deep learning algorithm. We trained the algorithm based on 405 images for Mexican triatomine species and 1,584 images for Brazilian triatomine species. Our system achieved 83.0 and 86.7% correct identification rates across all Mexican and Brazilian species, respectively, an improvement over comparable rates from statistical classifiers (80.3 and 83.9%, respectively). Incorporating distributional information to reduce numbers of species in analyses improved identification rates to 95.8% for Mexican species and 98.9% for Brazilian species. Given the 'taxonomic impediment' and difficulties in providing entomological expertise necessary to control such diseases, automating the identification process offers a potential partial solution to crucial challenges.