Comparative morphology of eggs of Rhodnius domesticus Neiva & Pinto, Rhodnius neglectus Lent and Rhodnius prolixus Stål (Hemiptera: Reduviidae: Triatominae).

Rhodnius species are potential vectors of the etiological agent of Chagas disease (CD), the protozoan Trypanosoma cruzi. CD impacts around seven million people in Latin America, resulting in approximately fourteen thousand deaths per year. Several species of Rhodnius are notable not only for their epidemiological relevance, but also for the challenging distinction between their species. Rhodnius has twenty species, each with its specific epidemiological importance. Rhodnius neglectus and Rhodnius prolixus are found with colonies in domiciliary environments. The observation of eggs in human dwellings signals the colonization process of these insects, increasing the risk of contamination of the population, since correct identification of eggs is necessary to help more effective vector control programs. Here we highlight diagnostic characters of eggs for these three species.

Triatomine vectors of Trypanosoma cruzi in an endemic area for Chagas disease in Northeast Brazil

ABSTRACT Background: We assessed the distribution of triatomines in an endemic area for Chagas disease. Methods: This retrospective study used secondary data extracted from the Official System of the National Chagas Disease Control Program (Sistema Oficial do Programa Nacional de Controle da Doença de Chagas - SisPCDCh). Results: A total of 7,257 (725.7 ± 221.7 per year) specimens were collected from 2013 to 2022. Most of them (6,792; 93.6%) were collected in the intradomicile and 465 (6.4%) in the peridomicile. A total of 513 (7.1%) triatomines tested positive for the presence of trypomastigote forms, similar to Trypanosoma cruzi. Conclusions: The spatial analysis revealed a heterogeneous distribution of triatomines across different municipalities.

Evidence that hematophagous triatomine bugs may eat plants in the wild.

Blood feeding is a secondary adaptation in hematophagous bugs. Many proteins are secreted in the saliva that are devoted to coping with the host's defense and to process the blood meal. Digestive enzymes that are no longer required for a blood meal would be expected to be eventually lost. Yet, in many strictly hematophagous arthropods, α-amylase genes, which encode the enzymes that digest starch from plants, are still present and transcribed, including in the kissing bug Rhodnius prolixus (Hemiptera, Reduviidae) and its related species, which transmit the Chagas disease. We hypothesized that retaining α-amylase could be advantageous if the bugs occasionally consume plant tissues. We first checked that the α-amylase protein of Rhodnius robustus retains normal amylolytic activity. Then we surveyed hundreds of gut DNA extracts from the sylvatic R. robustus to detect traces of plants. We found plant DNA in 8% of the samples, mainly identified as Attalea palm trees, where R. robustus are usually found. We suggest that although of secondary importance in the blood-sucking bugs, α-amylase may be needed during occasional plant feeding and thus has been retained.

Infection with Trypanosoma Cruzi Chagas and Characterization of Human Habitats of Triatoma Picturata (Usinger) in Western Mexico.

Chagas disease is one of the most important vector-borne diseases in Latin America. Triatoma picturata (Usinger), distributed in western Mexico, is one of the most important vectors of Trypanosoma cruzi Chagas. Knowing the type and materials used for building homes could lead to improved methods for controlling triatomines by focusing efforts on controlling every species of triatomine in its specific microhabitat. Houses in three studied localities of western Mexico that harbored T. picturata were characterized, as well as two entomological indices. Infestation Index values varied from 10.8% to 51.9% in the three localities, whereas Natural Infection Index values varied from 18.1% to 35.2%. Characteristics of houses invaded by T. picturata also varied. In Juan Gil Preciado, with the highest Entomological Index values, most houses were built of adobe walls with a roof of clay tiles and a dirt floor, and included the presence of domestic animals. Our data indicate that the potential risk of infection with T. cruzi differs markedly for inhabitants of the three studied localities, which demonstrates the importance of studying local populations of triatomines. Our study should contribute to decision-making regarding triatomine control efforts by providing profiles of houses that are more susceptible to triatomine invasion.

Diet of the sylvatic triatomine Mepraia spinolai: Association with Trypanosoma cruzi infection near human settlements.

The proximity between infectious disease vector populations and human settlements, and the infection prevalence of vector populations can determine the rate of encounters between vectors and humans and hence infection risk. The diet of sylvatic triatomine vectors (kissing bugs) provides evidence about the host species involved in the maintenance of the protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. Here, we characterized the diet of the Chilean endemic triatomine Mepraia spinolai using Next Generation Sequencing (NGS), and evaluated the relation between T. cruzi infection status and proximity to human settlements, with the proportion of human and human-associated (domestic and synanthropic) vertebrates in the diet. We sampled 28 M. spinolai populations, covering a latitudinal range of ∼800 km in Chile. For each population, genomic DNA was obtained from M. spinolai intestinal content. We assessed T. cruzi infection individually, and sequenced vertebrate cytochrome b to characterize the diet from infected and uninfected pooled samples. Human and human-associated animals were present in the diet of both T. cruzi-infected (13.50 %) and uninfected (10.43 %) kissing bugs. The proportion of human and human-associated vertebrates in the diet of infected M. spinolai was negatively associated with the distance from surrounding human settlements, but no relationship was detected for uninfected kissing bugs. This pattern could be related to alterations of kissing bug feeding behavior when infected by the protozoan. Our results highlight the relevance of developing a deeper knowledge of the wild transmission cycle of T. cruzi, thus advancing in the surveillance of vectors present in the natural environment near human settlements.

Differential Spreading of Microsatellites in Holocentric Chromosomes of Chagas Disease Vectors: Genomic and Evolutionary Implications.

This study focused on analyzing the distribution of microsatellites in holocentric chromosomes of the Triatominae subfamily, insect vectors of Chagas disease. We employed a non-denaturing FISH technique to determine the chromosomal distribution of sixteen microsatellites across twenty-five triatomine species, involving five genera from the two principal tribes: Triatomini and Rhodniini. Three main hybridization patterns were identified: strong signals in specific chromosomal regions, dispersed signals dependent on microsatellite abundance and the absence of signals in certain chromosomal regions or entire chromosomes. Significant variations in hybridization patterns were observed between Rhodniini and Triatomini species. Rhodniini species displayed weak and scattered hybridization signals, indicating a low abundance of microsatellites in their genomes. In contrast, Triatomini species exhibited diverse and abundant hybridization patterns, suggesting that microsatellites are a significant repetitive component in their genomes. One particularly interesting finding was the high abundance of GATA repeats, and to a lesser extent AG repeats, in the Y chromosome of all analyzed Triatomini species. In contrast, the Y chromosome of Rhodniini species did not show enrichment in GATA and AG repeats. This suggests that the richness of GATA repeats on the Y chromosome likely represents an ancestral trait specific to the Triatomini tribe. Furthermore, this information can be used to elucidate the evolutionary relationships between Triatomini and other groups of reduviids, contributing to the understanding of the subfamily's origin. Overall, this study provides a comprehensive understanding of the composition and distribution of microsatellites within Triatominae genomes, shedding light on their significance in the evolutionary processes of these species.

Resistance to deltamethrin in Triatoma infestans (Hemiptera: Reduviidae): Does it influence the phenotype of antennae, wings, and heads?

In vector control terms, insecticide resistance is the development of the capacity, of an insect population, to tolerate doses of an insecticide that are lethal to most individuals in a typical population of the same species. The genetic changes that determine resistance may have adaptive costs in the resistant phenotype or, conversely, may result in an adaptive advantage when compared to susceptible insects in the environment without insecticides. Triatoma infestans is one of the main vectors of Trypanosoma cruzi in the southern cone of South America. High insecticide resistance in T. infestans was detected in Argentina in Salta and Chaco provinces. The objective of this study was to determine the possible morphometric changes in wings, heads, and the antennal phenotype of deltamethrin-resistant T. infestans (RR) males and females compared to susceptible insects (SS), evaluating its implication in adaptive processes such as olfactory capacity, dispersion, and probability of colonizing new habitats, among others. Nine type I landmarks were marked on wings, 5 type II landmarks on heads, and 10 antennal sensilla were counted on 106 adults of both sexes (resistant and susceptible from first and second laboratory generations). Morphological divergence was observed between the two groups (RR and SS). The RR insects showed smaller sizes of wings and heads and shape compatible with lower dispersal potential and different active dispersal behaviors. Antennae also revealed sensory simplification in RR and divergence between RR and SS, although more marked in females. This study characterizes for the first time T. infestans RR and SS through wings, heads, and antennae. The results suggest a lower dispersive potential in resistant insects and the differences described lay the foundations for the identification of a resistance biomarker in triatomines.

First report of Rhodnius amazonicus Almeida, Santos & Sposina, 1973 (Hemiptera, Reduviidae, Triatominae) invading a dwelling in the state of Amapá, Brazil.

Rhodnius amazonicus has been previously reported in the Brazilian states of Amazonas and Pará, as well as in French Guiana. However, this is the first recorded presence of this species in Amapá, which is situated in the northern region of Brazil. The specimen was collected from a house in the rural area of the municipality of Porto Grande. Other triatomines, such as Panstrongylus geniculatus, Rhodnius pictipes, and Eratyrus mucronatus, were also found in the same locality in different houses. These species are vectors of Trypanosoma cruzi, which causes Chagas disease. Therefore, this report may contribute to understanding transmission in the state of Amapá, where new infections and outbreaks of Chagas disease have been recorded.

Review of Kissing Bugs (Hemiptera: Reduviidae: Triatominae) from China with Descriptions of Two New Species.

Triatominae, the only blood-sucking subfamily in Reduviidae, are the vectors of Chagas disease. The majority of them are distributed in the Americas, while the diversity in China has been underestimated, as only two species have been recorded. Here, we describe two new species from China, Triatoma picta Zhao & Cai sp. nov. and T. atrata Zhao & Cai sp. nov., and provide a redescription of T. sinica Hsiao, 1965, along with remarks on T. rubrofasciata (De Geer, 1773). To facilitate the identification, we include photos, especially of genitalia, as well as a distribution map and a key to Chinese triatomines. We calculated the pairwise genetic distances between 23 Triatoma species, which further supported the validity of these new species. We anticipate that our taxonomic review will be useful for identifying Chinese Triatominae.

Triatoma yelapensis sp. nov. (Hemiptera: Reduviidae) from Mexico, with a Key of Triatoma Species Recorded in Mexico.

Thirty-four species of Triatominae (Hemiptera, Reduviidae) are recorded in Mexico, Triatoma Laporte, 1832 the most speciose genus in this country. Here, we describe Triatoma yelapensis sp. nov. from the Pacific coast of Jalisco (Mexico). The most similar species to T. yelapensis sp. nov. is T. recurva (Stål, 1868), but they differ in head longitude, the proportion of labial segments, coloration pattern of corium and connexivum, spiracles location, and male genitalia. To provide statistical support for the morphological distinctiveness of the new species, we performed a geometric morphometric analysis of T. yelapensis sp. nov., T. dimidiata s.s. (Latreille, 1811), T. gerstaeckeri (Stål, 1859), and T. recurva (Stål, 1868), considering head morphology. We also provide an updated key of the genus Triatoma for species recorded in Mexico.

Dichotomous Keys Based on Cytogenetic Data for Triatomines Reported in Brazilian Regions with Outbreaks of Orally Transmitted Chagas Disease (Pernambuco and Rio Grande Do Norte).

Chagas disease (CD) affects about eight million people worldwide. Brazil has the highest number of estimated cases and the largest number of deaths due to CD. Considering the recent outbreaks of oral CD involving at least 27 cases of acute CD in Pernambuco (PE) as well as 18 cases and 2 deaths in the Rio Grande do Norte (RN), we developed dichotomous keys for the identification of triatomine species in these Brazilian states based on cytogenetic data. All triatomine species could be distinguished by cytogenetic characteristics, emphasizing the importance of the newly developed taxonomic keys for the correct identification of triatomes from PE and RN, particularly for species that exhibit morphological similarities, such as Triatoma brasilensis and T. petrocchiae (present in both states) and T. maculata and T. pseudomaculata (as T. pseudomaculata has been misidentified as T. maculata in PE and RN). These alternative keys are expected to provide a useful tool for the scientific community and, above all, health agents, aimed at preventing mistakes from occurring in the identification of the vectors present in PE and RN related to CD outbreaks caused by oral infection.

Making the Genome Huge: The Case of Triatoma delpontei, a Triatominae Species with More than 50% of Its Genome Full of Satellite DNA.

The genome of Triatoma delpontei Romaña & Abalos 1947 is the largest within Heteroptera, approximately two to three times greater than other evaluated Heteroptera genomes. Here, the repetitive fraction of the genome was determined and compared with its sister species Triatoma infestans Klug 1834, in order to shed light on the karyotypic and genomic evolution of these species. The T. delpontei repeatome analysis showed that the most abundant component in its genome is satellite DNA, which makes up more than half of the genome. The T. delpontei satellitome includes 160 satellite DNA families, most of them also present in T. infestans. In both species, only a few satellite DNA families are overrepresented on the genome. These families are the building blocks of the C-heterochromatic regions. Two of these satellite DNA families that form the heterochromatin are the same in both species. However, there are satellite DNA families highly amplified in the heterochromatin of one species that in the other species are in low abundance and located in the euchromatin. Therefore, the present results depicted the great impact of the satellite DNA sequences in the evolution of Triatominae genomes. Within this scenario, satellitome determination and analysis led to a hypothesis that explains how satDNA sequences have grown on T. delpontei to reach its huge genome size within true bugs.

Influence of blood meal source on the biological parameters of Triatoma pallidipennis (Heteroptera: Reduviidae) from Mexico.

Chagas disease is one of the most important vector-borne diseases in Latin America, including Mexico. Triatoma pallidipennis (Stål) (Hemiptera: Reduviidae) is a Mexican triatomine vector commonly associated with different hosts. The influence of six blood meals (rabbits, rats, mice, dogs, cats and chickens) on six biological parameters of the biology of T. pallidipennis was evaluated. A significant difference was found in the period of egg-to-adult development between the five mammalian feeds (mean 195 days) and the chicken feed (221 days). The probability of survival was significantly lower in the chicken cohort (0.285). The total number of blood meals to moult from the first instar to the adult stage was the highest in the chicken cohort (10-15). This cohort had the significantly highest rate of females at the end cycle. The mean number of eggs laid per female and the egg eclosion rate were similar among the six food sources. Most results seemed to be influenced by the higher nutritional quality of the mammalian blood compared to the bird's blood and the increased energy expenditure required for the digestion of bird blood. These results clearly show that T. pallidipennis, unlike other triatomine species, has a high reproductive capacity when feeding on different hosts.

Opportunistic or selective? Stage-dependent feeding behavior in a wild vector of Chagas disease.

The composition and contribution of different host species in the dynamics of vector-borne zoonotic parasites are particularly relevant for public health. Hence, the study of host selection by vectors is fundamental. Developmental stage and infection status are factors that may modulate vector feeding behavior. In the semi-arid Mediterranean ecosystem of South America, the transmission of Trypanosoma cruzi, the protozoan causing Chagas disease, includes the triatomine vector Mepraia spinolai and several vertebrate species. In this field study, we examined whether M. spinolai exhibits an opportunistic feeding behavior dependent upon developmental stage and/or infection status. We found that M. spinolai does not feed according to the relative availability of vertebrate species. In addition, early stage nymphs (first/second instars) fed on twice as many different species as middle (third/fourth instars) and late (fifth instars and adults) M. spinolai, with the former feeding on native rodents and lizards and the latter mostly on rabbits. Infected and uninfected M. spinolai showed similar feeding profiles. Wild triatomine species might be described as stage-dependent selective blood feeders, as a consequence of the temporal and spatial scale at which host-vector interactions occur, highlighting that all developmental stages might be infected and capable of transmitting T. cruzi.

Triatoma infestans, to be or not to be autogenic?

Autogeny, the ability to develop eggs without a meal in the adult stage, has been described in several groups of arthropods, especially hematophagous Diptera Nematocera. In obligate hematophagous hemimetabolous insects that feed on blood in all their instars, such as Triatominae, this concept gives rise to species with apparently facultative autogeny, such as Triatoma infestans. Generalized linear models were applied to explain egg production by the predictor variables molting weight as a proxy of nymphal accumulated reserves and digested blood weight as an indicator of adult reserve in fasted, incompletely fed and engorged at repletion females. The relationship between these indicators of nutritional status and egg development turned out to be a continuous function in which, with molting weights greater than 254 mg, the insects are autogenic, but for the first batch of eggs with molting weights between 132 and 253 mg, they require one adult meal of at least 202 mg, and with molting weights less than 131 mg at least two meals are required. Both molting weight and blood intake could determine oocyte production in an additive manner, thus the concept of autogeny as a switch on-off phenomenon is not directly applicable to Triatominae. Nevertheless, autogenic ability would allow Triatominae with relatively long cycles to accelerate population growth under favorable or low competition conditions during colonization or recovery after a control intervention.

Molecular and cytogenetic evidence for sibling species in the Chagas disease vector Triatoma maculata.

Triatoma maculata (Hemiptera, Reduviidae, Triatominae) occurs across dry-to-semiarid ecoregions of northern South America, where it transmits Trypanosoma cruzi, causative agent of Chagas disease. Using 207 field-caught specimens from throughout the species' range, mitochondrial(mt) DNA sequence data, and cytogenetics, we investigated inter-population genetic diversity and the phylogenetic affinities of T. maculata. Mitochondrial DNA sequence analyses (cytb and nd4) disclosed a monophyletic T. maculata clade encompassing three distinct geographic groups: Roraima formation (Guiana shield), Orinoco basin, and Magdalena basin (trans-Andean). Between-group cytb distances (11.0-12.8%) were larger than the ~7.5% expected for sister Triatoma species; the most recent common ancestor of these T. maculata groups may date back to the late Miocene. C-heterochromatin distribution and the sex-chromosome location of 45S ribosomal DNA clusters both distinguished Roraima bugs from Orinoco and Magdalena specimens. Cytb genealogies reinforced that T. maculata is not sister to Triatoma pseudomaculata and probably represents an early (middle-late Miocene) offshoot of the 'South American Triatomini lineage'. In sum, we report extensive genetic diversity and deep phylogeographic structuring in T. maculata, suggesting that it may consist of a complex of at least three sibling taxa. These findings have implications for the systematics, population biology, and perhaps medical relevance of T. maculata sensu lato.

Massive invasion on a Triatominae insectarium (Hemiptera: Heteroptera: Reduviidae) by Attagenus fasciatus (Thunberg, 1795) (Coleoptera: Dermestidae: Attageninae)

ABSTRACT Background: Triatominae are vectors of the protozoan Trypanosoma cruzi, the causative agent of Chagas disease. This study aims to report an infestation on Triatominae colonies by a beetle, previously identified as a pest. Methods: The management of these colonies should be improved to maximize their usefulness, and factors that may cause harm to them should be avoided as much as possible. Results: This is the first report on a coleopteran infestation on living Triatominae colonies worldwide. Conclusions: The present record provides an important warning to researchers who maintain insectaries in general, especially those who rear triatomines, to carry protective measures against such invasions.

Occurrence of triatomines in public spaces: An atypical case in the Southwestern Brazilian Amazon

ABSTRACT Background: Triatomines infest atypical public spaces in the Western Amazon. Methods: Frequent visitors to these spaces captured the insects in the state of Acre, Brazil (Rio Branco and Cruzeiro do Sul). Results: Six insects were found in a penitentiary, a church, a school, a university, a hospital, and a health center. Five of the insects were adults (three positive for Trypanosoma cruzi) and one was a nymph. Conclusions: This is the first report of triatomine occurrence in schools or churches. These data are important for implementing surveillance strategies and alerting individuals about possible changes in Chagas disease transmission dynamics.

Conhecimentos sobre a doença de Chagas entre escolares de dois municípios de Minas Gerais, Brasil / Knowledge about Chagas disease among schoolchildren from two municipalities of Minas Gerais, Brazil

Resumo Introdução: No Brasil, a vigilância entomológica da doença de Chagas (DCh) é respaldada na participação comunitária, que notifica a presença dos vetores nas habitações. Assim, a sustentabilidade desta medida de controle depende do conhecimento da população sobre os vetores. Objetivo: Avaliar o conhecimento de escolares sobre a doença de Chagas nos municípios mineiros de Itatiaiuçu (vigilância ativa) e Sabará (vigilância inativa). Método: Questionário semiestruturado foi aplicado para alunos matriculados no ensino médio em duas escolas de Sabará (rural e urbana) e uma em Itatiaiuçu (urbana). Resultados: Em geral, os alunos residentes em Sabará demonstraram conhecimento limitado acerca dos vetores, patógeno, transmissão e doença (sem diferenças relevantes quanto à localização entre área urbana e rural), quando comparados aos residentes em Itatiaiuçu, que apesar de ter uma vigilância mais atuante, também não apresentaram conhecimento totalmente satisfatório. Conclusões: Os resultados enfatizam a carência de informações sobre a DCh tanto em relação ao conhecimento acadêmico quanto às campanhas de educação firmadas pelo programa. Ações educativas para sensibilizar a comunidade e promover o conhecimento sobre esta doença mostram-se necessárias e fundamentais para a manutenção da vigilância ao longo do tempo e espaço.

Abstract Background: In Brazil, entomological surveillance of Chagas disease (ChD) is based on community participation, which notifies the presence of vectors inside their homes. Thus, the sustainability of this control measure depends on the knowledge of the population about the vectors. Objective: To evaluate the knowledge of schoolchildren about ChD in the municipalities of Itatiaiuçu (active surveillance) and Sabará (inactive surveillance). Method: A semi-structured questionnaire was given to students enrolled in two high schools in Sabará (rural and urban) and one in Itatiaiuçu (urban). Results: In general, students living in Sabará demonstrated only limited knowledge about vectors, pathogens, transmission, and disease (with no significant differences between the schools in urban and rural areas). Similarly, the schoolchildren in Itatiaiuçu, despite living in an area of active surveillance, also did not have a fully satisfactory knowledge of ChD. Conclusions: The results emphasize the lack of information about ChD, both in relation to academic knowledge and the education campaigns established by the program. Improvements in educational interventions to raise community awareness and promote knowledge about ChD are necessary and vital for maintaining vigilance against ChD over space and time.

Population size regulation is density-dependent in Rhodnius prolixus (Hemiptera: Reduviidae) through an irritability mechanism

BACKGROUND Physical factors can determine the level of triatomine abundance, but do not regulate their population densities, and neither do natural enemies. OBJECTIVES To identify the processes associated with density-dependent triatomine population regulation. METHODS We set-up a laboratory experiment with four interconnected boxes; the central box harbored Rhodnius prolixus bugs and one hamster. Stage 5 and adult densities of 10, 20, 30, 40, and 60 bugs per hamster, were replicated four times (except the density of 60 bugs). Hamster's irritability and several triatomine responses were measured: feeding, development time and longevity, mortality, fecundity, dispersal, and the net reproductive value (R o ). FINDINGS Density had a statistically significant effect on irritability, but not on the percent of bugs feeding. Density was significant on blood meal size ingested in bugs that did not move between boxes, but not significant when the bugs moved. Density and irritability affected the proportion of stage 5 nymphs molting, and the proportion of adult bugs dying per day and over a three-week period. There was a highly significant effect of density and irritability on R o . MAIN CONCLUSIONS We showed that a density-dependent mechanism, acting through the irritability of the host, seems the most plausible process regulating populations in triatomines.