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The stridulation in the subfamily Triatominae has been identified as a means of communication between species, produced by the friction of the proboscis on the prosternal stridulatory groove. Despite its biological significance, this phenomenon remains understudied, with the signal's production seemingly contingent upon the morphology of the stridulatory groove. In this study, we examined the morphology of stridulatory grooves in females and males of five species and two subspecies of Mexican triatomines using morphometric and scanning electron microscopical analysis. Our findings reveal that all analyzed species exhibit triangular-shaped stridulatory grooves with parallel ridges covering the entire groove, bordered on each side, and covered with setae. Surprisingly, we observed noticeable differences in the number of ridges and inter-ridge distance between the species Triatoma lecticularia and Triatoma rubida (p < 0.001 and p < 0.009, respectively), indicating sexual dimorphism in this aspect, a phenomenon not previously reported in the morphology of this structure. Our findings shed light on the intricate morphology of the stridulatory groove in Mexican triatomines, suggesting potential implications for their behavior and intra-specific communication.

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Chagas disease (CD) is a neglected illness affecting approximately seven million individuals, with vector transmission occurring via triatomine bugs. The Rhodniini tribe comprises 24 species, grouped into the Rhodnius and Psammolestes genera. Given the importance of accurately identifying CD vectors, the taxonomy of Psammolestes spp. was revisited using morphological and morphometric data. Specimens of P. tertius, P. coreodes, and P. arthuri were collected, and the morphological characteristics of the head, thorax, abdomen, and eggs were analyzed. Morphometric studies of eggs were also conducted. Dichotomous keys allowing for the differentiation of Psammolestes spp. were elaborated based on adult insect and egg morphological characteristics. Through these studies, it was possible to differentiate the three Psammolestes species and confirm that this genus should not be classified under the Rhodnius genus, contributing to Rhodniini taxonomy.

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Triatoma pallidipennis is an exclusive and widely distributed species in Mexico and one of the three main vectors that transmit Chagas disease in the country. The state of Hidalgo is an endemic area for Chagas disease where the presence of several species of triatomines has been reported. The objective of our work was to describe the morphology, colonization process, and reproductive behavior of T. pallidipennis in Guadalupe, Tecozautla, two years after the first collection of a specimen in this region. A total of 28 specimens was collected at both domicile and peridomicile, showing a 17.8% infection rate. The main collection site was a woodshed, and despite the collection of adults in the dwelling, we did not find eggs, exuviae, or nymphs. One female monitored from collection day until death laid 566 eggs, with a hatching rate of 95%, showing an increase of oviposition when cohabited with a male. The results showed the capacity that T. pallidipennis has to infest areas (mainly human dwellings) when it settles down, which would imply a risk for the population that lives in the locality.

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Chagas disease affects about eight million people. In view of the issues related to the influence of anthropogenic changes in the dynamics of the distribution and reproductive interaction of triatomines, we performed experimental crosses between species of the Rhodniini tribe in order to evaluate interspecific reproductive interactions and hybrid production capacity. Reciprocal crossing experiments were conducted among Rhodnius brethesi × R. pictipes, R. colombiensis × R. ecuadoriensis, R. neivai × R. prolixus, R. robustus × R. prolixus, R. montenegrensis × R. marabaensis; R. montenegrensis × R. robustus, R. prolixus × R. nasutus and R. neglectus × R. milesi. With the exception of crosses between R. pictipes ♀ × R. brethesi ♂, R. ecuadoriensis ♀ × R. colombiensis ♂ and R. prolixus ♀ × R. neivai ♂, all experimental crosses resulted in hybrids. Our results demonstrate that both allopatric and sympatric species produce hybrids, which can generate concern for public health agencies in the face of current anthropogenic events. Thus, we demonstrate that species of the Rhodniini tribe are capable of producing hybrids under laboratory conditions. These results are of great epidemiological importance and raise an important discussion about the influence of climatic and environmental interactions on Chagas disease dynamics.

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The blood-sucking bug, Triatoma infestans, is one of the main vectors of Chagas disease in America. It is usually controlled with pyrethroids, but the emergence of resistance to these insecticides creates the need to look for alternative products. Eugenol, menthol and menthyl acetate are botanical monoterpenes, which produce lethal and sublethal effects on insects. The purpose of this work was to determine what type of toxicological interactions occur when binary mixtures, formed by the pyrethroid permethrin and sublehtal doses of eugenol, menthol or menthyl acetate, are applied to T. infestans. First instar nymphs were exposed to filter papers impregnated with the insecticides. The number of knocked down insects was registered at different times and Knock Down Time 50% (KT50) values were calculated. The following KT50 values with their corresponding 95% Confidence Intervals were obtained: permethrin, 47.29 (39.92 - 56.32) min; permethrin + eugenol, 34.08 (29.60 - 39.01) min; permethrin + menthol, 27.54 (23.28 - 32.55) min; permethrin + menthyl acetate, 43.62 (39.99 - 47.59) min. Eugenol and menthol increased the speed of action of permethrin (synergism), but menthyl acetate had no effect on it (additivity). These results provide the basis to further explore interactions between conventional insecticides and plant monoterpenes as potential tools for controlling T. infestans.

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Under laboratory conditions, Triatoma rosai and T. sordida are able to cross and produce hybrids. In the face of climate and environmental changes, the study of hybrids of triatomines has evolutionary and epidemiological implications. Therefore, we performed morphological, cytological and molecular studies and characterized the feeding and defecation pattern of hybrids from crosses between T. sordida and T. rosai. The morphological characterization of the female genitalia of the hybrids showed that characteristics of both parental species segregated in the hybrids. Cytogenetic analyzes of hybrids showed regular metaphases. According to molecular studies, the mitochondrial marker Cytochrome B (CytB) related the hybrids with T. sordida and the nuclear marker Internal Transcribed Spacer 1 (ITS-1) related the hybrids with T. rosai. Both parents and hybrids defecated during the blood meal. Thus, the hybrids resulting from the cross between T. sordida and T. rosai presented segregation of phenotypic characters of both parental species, 100% homeology between homeologous chromosomes, phylogenetic relationship with T sordida and with T. rosai (with CytB and ITS-1, respectively), and, finally, feeding and defecation patterns similar to the parents.

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BACKGROUND: The geniculatus clade, composed by the rufotuberculatus, lignarius, geniculatus and megistus groups, relates evolutionarily the species of the genus Panstrongylus and Nesotriatoma. Several studies have shown that triatomine hybrids can play an important role in the transmission of Chagas disease. Natural hybrids between species of the geniculatus clade have never been reported to our knowledge. Thus, carrying out experimental crosses between species of the geniculatus clade can help to elucidate the taxonomic issues as well as contribute to the epidemiological knowledge of this group. METHODS: Experimental crosses were carried out between species of the megistus and lignarius groups to evaluate the reproductive compatibility between them. A phylogenetic reconstruction was also performed with data available in GenBank for the species of the geniculatus clade to show the relationships among the crossed species. RESULTS: Phylogenetic analysis grouped the species of the geniculatus clade into four groups, as previously reported. In the interspecific crosses performed there was no hatching of eggs, demonstrating the presence of prezygotic barriers between the crossed species and confirming their specific status. CONCLUSIONS: In contrast to the other groups of the Triatomini tribe, as well as the Rhodniini, there are prezygotic barriers that prevent the formation of hybrids between species of the megistus and lignarius groups. Thus, the geniculatus clade may represent an important evolutionary model for Triatominae, highlighting the need for further studies with greater sample efforts for this clade (grouping the 17 species of Panstrongylus and the three of Nesotriatoma).

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Panstrongylusnoireaui sp. nov. from Bolivia is described based on male and female specimens. Although morphologically almost indistinguishable from Panstrongylusrufotuberculatus (Champion, 1899), the new species shows remarkable chromosome and molecular features, which are very distinctive among all others Panstrongylus species. The new species is also separated by some characteristics of the processes of the endosoma of the male genitalia. An updated key for species of Panstrongylus is provided.

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The subfamily Triatominae (Hemiptera-Reduviidae) includes more than 150 blood-sucking species, potential vectors of the protozoan Trypanosoma cruzi, causative agent of Chagas disease. A distinctive cytogenetic characteristic of this group is the presence of extremely stable chromosome numbers. Unexpectedly, the analyses of the chromosomal location of ribosomal gene clusters and other repetitive sequences place Triatominae as a significantly diverse hemipteran subfamily. Here, we advance the understanding of Triatominae chromosomal evolution through the analysis of the 45S rDNA cluster chromosomal location in 92 Triatominae species. We found the 45S rDNA clusters in one to four loci per haploid genome with different chromosomal patterns: On one or two autosomes, on one, two or three sex chromosomes, on the X chromosome plus one to three autosomes. The movement of 45S rDNA clusters is discussed in an evolutionary context. Our results illustrate that rDNA mobility has been relatively common in the past and in recent evolutionary history of the group. The high frequency of rDNA patterns involving autosomes and sex chromosomes among closely related species could affect genetic recombination and the viability of hybrid populations, which suggests that the mobility of rDNA clusters could be a driver of species diversification.

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Triatoma petrocchiae is a species morphologically similar to T. b. brasiliensis (which resulted in a synonymization event); despite this similarity, genetic, morphological, and experimental crossbreeding studies confirmed the specific status of T. petrocchiae. Considering that both species have been reported living in sympatry and that, for a long time, most species of the T. brasiliensis complex were considered only chromatic variants of T. b. brasiliensis, we carried out experimental crosses between T. b. brasiliensis and T. petrocchiae (to confirm whether these species are reproductively isolated) and between T. lenti and T. petrocchiae (to assess whether T. petrocchiae also presents prezygotic isolation with the other species of the T. brasiliensis complex). Reciprocal experimental crosses were conducted, and weekly, the eggs were collected, counted, and separated in new containers to assess the hatch rate. Neither cross resulted in hybrids, demonstrating that there are pre-zygotic reproductive barriers installed between T. petrocchiae and the other species of the T. brasiliensis complex. On the basis of the results above, we demonstrated that T. petrocchiae is reproductively isolated from T. b. brasiliensis and T. lenti. Furthermore, we suggest that T. petrocchiae is the species most derived from the T. brasiliensis complex.

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Triatoma longipennis (Usinger) is an important vector of Trypanosoma cruzi in western, central and northern Mexico, due to its wide distribution, high infection rates, and epidemiological indices. However, its population genetics has not been completely characterized. In this study, the intra-specific relationships between different T. longipennis populations were analyzed from seven states in Mexico using mitochondrial cyt B as a marker. Our results show that the population of Jalisco was the most diverse, with the highest genetic and haplotypic variation (Hd = 0.978, π = 0.099 and θ = 0.079), even 25 times higher than some other populations analyzed. Heterogeneous migration and gene flow were observed without relation to their geographical distribution, that is, nearby populations may present high values of gene flow with low migration. In contrast, remote populations have low gene flow values with high migration. Genetic isolation was apparently present in the Guanajuato population, however, Mantel's analyzes to determine when an isolation by distance is present did not show correlation between genetic (FST) and geographic (Km) distances (P = 0.064). The STRUCTURE analyzes showed that populations such as Chihuahua, Jalisco, Nayarit and Michoacán appear to show a similar population structure, suggesting a common ancestor. Our results suggest two routes of diversification of T. longipennis highly influenced by anthropogenic effects. Elucidation of the population genetic structure of T. longipennis will help to better understand the role of gene flow and migration in the dispersal of this important Chagas disease vector.

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BACKGROUND Panstrongylus rufotuberculatus (Hemiptera-Reduviidae) is a triatomine species with a wide geographic distribution and a broad phenotypic variability. In some countries, this species is found infesting and colonising domiciliary ecotopes representing an epidemiological risk factor as a vector of Trypanosoma cruzi, etiological agent of Chagas disease. In spite of this, little is known about P. rufotuberculatus genetic diversity. METHODS Cytogenetic studies and DNA sequence analyses of one nuclear (ITS-2) and two mitochondrial DNA sequences (cyt b and coI) were carried out in P. rufotuberculatus individuals collected in Bolivia, Colombia, Ecuador and Mexico. Moreover, a geometric morphometrics study was applied to Bolivian, Colombian, Ecuadorian and French Guiana samples. OBJECTIVES To explore the genetic and phenetic diversity of P. rufotuberculatus from different countries, combining chromosomal studies, DNA sequence analyses and geometric morphometric comparisons. FINDINGS We found two chromosomal groups differentiated by the number of X chromosomes and the chromosomal position of the ribosomal DNA clusters. In concordance, two main morphometric profiles were detected, clearly separating the Bolivian sample from the other ones. Phylogenetic DNA analyses showed that both chromosomal groups were closely related to each other and clearly separated from the remaining Panstrongylus species. High nucleotide divergence of cyt b and coI fragments were observed among P. rufotuberculatus samples from Bolivia, Colombia, Ecuador and Mexico (Kimura 2-parameter distances higher than 9%). MAIN CONCLUSIONS Chromosomal and molecular analyses supported that the two chromosomal groups could represent different closely related species. We propose that Bolivian individuals constitute a new Panstrongylus species, being necessary a detailed morphological study for its formal description. The clear morphometric discrimination based on the wing venation pattern suggests such morphological description might be conclusive.

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The Triatoma brasiliensis complex is composed of six species (Triatoma bahiensis, T. juazerensis, T. lenti, T. melanica, T. petrocchiae, and T. sherlocki) and two subspecies (T. brasiliensis brasiliensis and T. b. macromelasoma). Phylogenetic studies have shown that it is a monophyletic group, but the morphological relationships among the members can be better understood. We applied a tool of high resolution based on morphology to identify details that characterize the female genitals for each member of this group. The external genital structures of five to fifteen females of each taxon were analyzed. Insects had their abdomen transversely cut to separate the last abdominal segments, and representative illustrations obtained via scanning electron microscopy (SEM) are shown. Observations were performed in the dorsal and the ventral views. The morphological characterization of female external genitalia allowed a differentiation of each species/subspecies. The subspecies T. b. brasiliensis and T. b. macromelasoma were the most similar. On the other hand, dorsal view of structures evidenced some unique characteristics for T. petrocchiae and T. sherlocki. Regarding these structures for T. petrocchiae, the differentiation was in accordance with molecular studies, since this species exhibits the highest genetic distances in relation to other species of T. brasiliensis complex.

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Triatoma petrocchiae is the newly member of the Triatoma brasiliensis species complex. This species overlaps with T. brasiliensis in geographic and ecotypic occupation in the sylvatic habitat because both inhabit rocky outcrops in the semi-arid portion of Brazilian northeast. In this region T. brasiliensis is the most important Chagas disease vector because it constantly colonizes domiciles. In contrast, T. petrocchiae is rarely found in peri or intradomiciliary habitats - reason why little is known about this species. Therefore, Here, we present information for the first time on. the T. petrocchiae ecotopes, genetic diversity, Trypanosoma cruzi prevalence/genotyping in comparison to T. brasiliensis. We found T. brasilensis (N = 223) and T. petrocchiae (N = 69) in co-habitation in rocky outcrops in three Districts of Paraíba and Rio Grande do Norte states. Forty-tree T. petrocchiae insects of eleven sampling spots (composing three geographic populations) were genotyped for the mitochondrial Cyt B gene and little geographic structure was observed. Tajima's D test suggested that species is evolving toward a mutation-drift equilibrium in our collection range. Sylvatic T. petrocchiae had 4% (3/68) of infected insects by T. cruzi, whereas T. brasiliensis had 26% (59/223). Fluorescent Fragment Length Barcoding demonstrated that all three T. petrocchiae harbored TcI whereas T. brasiliensis had TcI, but also TcIII, TcII/TcVI and T. rangeli genotype A, sometimes under mixed infections. None of infected T. petrocchiae were carrying mixed infections. However, this result should be confirmed using a larger pool of infected bugs. We here presented the first documentation of T. rangeli infecting T. brasiliensis. The finding of infected T. petrocchiae calls for constant vector monitoring because the epidemiologic scenario is dynamic and sylvatic vectors are progressively found in adaptation to anthropic environments.

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The Triatoma genus is paraphyletic, and its species are grouped into complexes and subcomplexes. Given the fact that species that make up a given subcomplex generally share chromosomal traits, we analyzed the distribution of AT- and CG-rich DNA of the T. brasiliensis species subcomplex, in order to establish affinities among members of the T. brasiliensis subcomplex based on chromatin and chromosome traits and develop an identification key for the four monophyletic Triatoma subcomplexes from South America. All species exhibited a CG-rich X sex chromosome and autosomes, as well as an AT-rich Y sex chromosome. This feature can be used as a diagnostic characteristic to determine whether a given species is a member of the T. brasiliensis subcomplex, because it enables the differentiation of these species from all Triatoma of South America. Thus, we confirmed the chromosomal relationship of the T. brasiliensis species subcomplex and developed a dichotomous key based on the chromocenter to differentiate the species from this subcomplex from the other monophyletic Triatoma subcomplexes from South America.

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Chagas disease is caused by the Trypanosoma cruzi and transmitted mainly by triatomines. Triatoma is a paraphyletic group and the species of this genus are grouped into complexes and subcomplexes. Morphological data and geographical distribution grouped initially T. melanocephala, T. vitticeps and T. tibiamaculata in the T. brasiliensis subcomplex. However, karyotypic and phylogenetic analysis suggested the exclusion of T. melanocephala, T. vitticeps and T. tibiamaculata from this subcomplex. Considering that studies of experimental crosses can help to understand the systematics of species, we performed experimental crosses between T. melanocephala, T. vitticeps and T. tibiamaculata with T. b. brasiliensis. No crosses resulted in hybrids. Taking into account that the species of the T. brasiliensis subcomplex do not present interspecific prezygotic barriers, the characterization of reproductive barriers shows that T. melanocephala, T. vitticeps and T. tibiamaculata do not present an evolutionary proximity relationship with the species of this subcomplex. Thus, we confirmed the exclusion of these species from the T. brasiliensis subcomplex and we emphasize the importance of experimental crosses for evolutionary studies.

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Chagas disease represents a significant public health problem in Mexico. In the state of Hidalgo, studies on the presence of triatomines are scarce and restricted to a few locations. To determine the risk of transmission in the state of Hidalgo and stratify the vector potential, the distribution of Triatominae was surveyed from 2015 to 2016 in collaboration with primary health care services and local communities. A total of 570 specimens was collected in 278 houses in 25 municipalities. The species of Triatominae detected were T. dimidiata with 391 samples, T. mexicana with 159 samples, T. gerstaeckeri with 17 samples, and T. barberi with three samples. The samples were collected in domestic and peridomestic areas. The general index of natural infection was 9.8%. Indices of colonization and crowding were determined for species and municipality and results were variable.

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Chagas disease or American trypanosomiasis affects six to seven million people worldwide, mostly in Latin America. This disease is transmitted by hematophagous insects known as "kissing bugs" (Hemiptera, Triatominae), with Triatoma infestans and Rhodnius prolixus being the two most important vector species. Despite the fact that both species present the same diploid chromosome number (2n = 22), they have remarkable differences in their total DNA content, chromosome structure and genome organization. Variations in the DNA genome size are expected to be due to differences in the amount of repetitive DNA sequences. The T. infestans genome-wide analysis revealed the existence of 42 satellite DNA families. BLAST searches of these sequences against the R. prolixus genome assembly revealed that only four of these satellite DNA families are shared between both species, suggesting a great differentiation between the Triatoma and Rhodnius genomes. Fluorescence in situ hybridization (FISH) location of these repetitive DNAs in both species showed that they are dispersed on the euchromatic regions of all autosomes and the X chromosome. Regarding the Y chromosome, these common satellite DNAs are absent in T. infestans but they are present in the R. prolixus Y chromosome. These results support a different origin and/or evolution in the Y chromosome of both species.

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The chemical control of Triatoma infestans, the major Chagas disease vector in southern South America, has been threatened in the last years by the emergence of pyrethroid-resistant bug populations. As an alternative approach, the efficacy of the entomopathogenic fungus Beauveria bassiana to control T. infestans populations (regardless their pyrethroid susceptibility) has been demonstrated. Growing research efforts on the interaction between T. infestans and B. bassiana by molecular, ecological, biochemical and behavioral traits has allowed framing such interaction as an evolutionary arms race. This review will focus on the relationships established in this particular host-pathogen system, compiling available data on the relevance of fungal pathogenesis, insect behavior, population dynamics and human intervention to favor fungal dissemination in bug populations. The current snapshot shows the fungus ahead in the evolutionary arms race and predicts a promissory landscape for the biological control of Chagas disease vectors.

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Morphological characters can be used to distinguish the vast majority of triatomine species, but the existence of high levels of phenotypic plasticity and recently diverged species can lead to erroneous determinations. To approach this problem, we analyzed the male and female morphologies of the scutella of Triatoma barberi, T. dimidiata, T. lecticularia, T. mexicana, T. recurva, T. rubida, and two sub-species, T. protracta protracta and T. protracta nahuatlae. Scutellum samples were observed by scanning electron microscopy and subjected to morphological analysis and morphometric investigation using a canonical discriminant analysis. The results revealed differences primarily in central depression shape, posterior process, and vestiture. We observed clear dimension-based differences in scutellum morphometry in all the taxa under study, providing sound evidence for species and subspecies differentiation. On the other hand, there is no difference between sexes in T. lecticularia, T. protracta protracta, and T. protracta nahuatlae. Our methodology can be implemented to differentiate species of the genus Triatoma.

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