Yago Visinho dos Reis

: Triatomines are insects of great importance for public health, as they are vectors of protozoan Trypanosoma cruzi, the etiological agent of Chagas disease. The genus Triatoma is the most representative in number of species and one of the most important from an epidemiological point of view. The Triatoma pseudomaculata subcomplex (composed of the species Triatoma arthurneivai, T. pseudomaculata, and T. wygodzinskyi [from the old subcomplex Triatoma maculata] and Triatoma baratai, Triatoma costalimai, Triatoma deaneorum, Triatoma jatai, and Triatoma williami [from the old Triatoma matogrossensis subcomplex]) was recently proposed based on chromosomal data. Thus, considering that the experimental crossings helped with systematic questions about triatomines, we performed crosses between the species of the T. pseudomaculata subcomplex. Thus, we crossed T. pseudomaculata and Triatoma costalimai to assess the possible presence of interspecific genomic and reproductive compatibility. In addition, based on the molecular data available on GenBank, we performed a phylogenetic systematics study to assess the evolutionary relationship of the species of the T. pseudomaculata subcomplex. Experimental crosses did not result in hybrids, demonstrating that there is a prezygotic barrier established between these species. Based on the above, we confirmed the specific status of T. pseudomaculata and T. costalimai based on the biological species concept. Furthermore, the reproductive incompatibility associated with the phylogenetic systematic analyses demonstrate that T. costalimai does not belong to the T. pseudomaculata subcomplex. Thus, we suggest that the sister species T. costalimai and T. jatai should be grouped into a new subcomplex called T. costalimai.

Species of the Rhodnius genus have a complex taxonomy because the events of phenotypic plasticity and cryptic speciation make it difficult to correctly classify these vectors. During the taxonomic history of the genus, five synonymization events occurred. Additionally, some authors suggest that R. milesi possibly represent only phenotypic polymorphisms of R. neglectus. Thus, we analyzed the specific status of R. milesi in relation to R. neglectus using phylogenetic studies with the mitochondrial gene cytochrome B and the study of reproductive barriers. The phylogenetic reconstruction grouped R. milesi together with R. neglectus from different localities, demonstrating that these taxa represent the same species based on the phylogenetic species concept. Experimental crosses demonstrate the absence of pre- and postzygotic barriers under laboratory conditions. Additionally, when the hatch rates of crosses are compared to intraspecific crosses, it can be noted that they are high and very similar. Finally, the mortality rate of the hybrids does not indicate hybrid inviability, the absence of chromosome pairing errors does not indicate hybrid sterility, and the proportion between male and female hybrids demonstrates that Haldane’s rule was not acting. Therefore, we perform the formal synonymization of R. milesi with R. neglectus.

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.

Considering that health professionals at Brazil had doubts about the entomoepidemiological issues of Chagas disease (CD), and that many of them highlighted not feeling totally safe for clinical care, the knowledge of 281 Brazilian medical students was evaluated through a cross-sectional, descriptive, prospective, and quantitative research. Most students demonstrated that they knew about the etiological agent of CD (Trypanosoma cruzi), since 279 students answered the questionnaire correctly. Furthermore, the medical students demonstrated knowledge of the main form of transmission of the parasite, as 278 students associated CD transmission with triatomines. On the other hand, approximately 25 students did not associate CD transmission with triatomine feces. Besides that, these future health professionals had difficulties in relation to the treatment of CD, as more than half of the students (176) wrongly answered that CD “is not curable” or “is curable in the chronic phase”. Based on the results obtained that point out the difficulties medical students have with CD, there is a need for undergraduate medical courses to address the neglected diseases holistically because the National Curriculum Guidelines for the medical course require the training of competent health professionals capable of integrating the biological, psychological, social, and environmental dimensions.

Several cytogenetic studies have already been performed in Triatominae, such that different karyotypes could be characterized (ranging from 2n = 21 to 25 chromosomes), being the changes in the number of chromosomes related mainly to fusion and fission events. These changes have been associated with reproductive isolation and speciation events in other insect groups. Thus, we evaluated whether different karyotypes could act in the reproductive isolation of triatomines and we analyzed how the events of karyotypic evolution occurred along the diversification of these vectors. For this, experimental crosses were carried out between triatomine species with different karyotypes. Furthermore, based on a phylogeny with 88 triatomine taxa (developed with different molecular markers), a reconstruction of ancestral karyotypes and of anagenetic and cladogenetic events related to karyotypic alterations was performed through the ChromoSSE chromosomal evolution model. All crosses performed did not result in hybrids (prezygotic isolation in both directions). Our modeling results suggest that during Triatominae diversification, at least nine cladogenetic events may be associated with karyotype change. Thus, we emphasize that these alterations in the number of chromosomes can act as a prezygotic barrier in Triatominae (karyotypic isolation), being important evolutionary events during the diversification of the species of Chagas disease vectors.

Triatoma bassolsae, T. longipennis, T. mazzottii, T. pallidipennis, T. phyllosoma and T. picturata are species that have great epidemiological importance in the transmission of Chagas disease in Mexico. However, there is no consensus regarding the specific status of these species, since they appear in various articles as species, subspecies and even subgenera. Thus, we revisited genetic, taxonomic and evolutionary data that allowed us to assess and discuss the specific status of these six species of the T. phyllosoma subcomplex. Phylogenetic studies were performed with nuclear (18S, 28S, ITS-2) and mitochondrial (16S, cytb, COI, COII, 12S) markers deposited in GenBank. In addition, data from experimental crosses were pooled and the genetic distance to the cytb gene was calculated. The phylogenetic reconstruction enabled us to rescue the six species as independent lineages. Post-zygotic reproductive isolation barriers (sterility and/or hybrid collapse) were observed for some experimental crosses. Although the other experimental crosses did not allow us to characterize reproductive barriers, these species showed high genetic distances in relation to the cytb gene (ranging from 4.6% to 14.9%). Thus, based on the revisited literature data, we confirmed the specific status of these six species of the T. phyllosoma subcomplex based on the phylogenetic and biological concepts of the species.

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.

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). Graphical