Analysis of the maternal inheritance hypothesis of the exochorium in eggs from hybrids of Chagas disease vectors.

Morphological studies applied to the taxonomy of the Triatominae cover various structures (head, wing, thorax, genitalia, and eggs). Exochorial structures of hybrid eggs were characterized and compared with the parents, demonstrating that hybrids presented characteristics identical to the exochorial pattern observed in the females of the crosses, which resulted in the hypothesis that the pattern of triatomine eggs is possibly a characteristic inherited from females. Thus, we characterized the exochorium of the eggs of several triatomine hybrids and compared them with the parents, to assess the pattern of segregation and test the hypothesis of maternal inheritance. Hybrids were obtained in at least one direction from all crosses. The analysis of the exochorium of the eggs of the hybrids showed different patterns of segregation: "exclusively paternal", "predominantly maternal", "predominantly paternal", "mutual", and "differential". Curiously, none of the hybrids evaluated presented characteristics that segregated exclusively from the female parental species. Thus, we demonstrate that the hypothesis of maternal inheritance of the exochorium pattern of eggs is not valid and we emphasize the importance of alternative/combined tools (such as integrative taxonomy) for the correct identification of these insect vectors (mainly in view of possible natural hybridization events due to climate and environmental changes).

First evidence of gonadal hybrid dysgenesis in Chagas disease vectors (Hemiptera, Triatominae): gonad atrophy prevents events of interspecific gene flow and introgression.

BACKGROUND: Hybridization events between Triatoma spp. have been observed under both natural and laboratory conditions. The ability to produce hybrids can influence different aspects of the parent species, and may even result in events of introgression, speciation and extinction. Hybrid sterility is caused by unviable gametes (due to errors in chromosomal pairing [meiosis]) or by gonadal dysgenesis (GD). All of the triatomine hybrids analyzed so far have not presented GD. We describe here for the first time GD events in triatomine hybrids and highlight these taxonomic and evolutionary implications of these events. METHODS: Reciprocal experimental crosses were performed between Triatoma longipennis and Triatoma mopan. Intercrosses were also performed between the hybrids, and backcrosses were performed between the hybrids and the parent species. In addition, morphological and cytological analyzes were performed on the atrophied gonads of the hybrids. RESULTS: Hybrids were obtained only for the crosses T. mopan♀ × T. longipennis♂. Intercrosses and backcrosses did not result in offspring. Morphological analyses of the male gonads of the hybrids confirmed that the phenomenon that resulted in sterility of the hybrid was bilateral GD (the gonads of the hybrids were completely atrophied). Cytological analyses of the testes of the hybrids also confirmed GD, with no germ cells observed (only somatic cells, which make up the peritoneal sheath). CONCLUSIONS: The observations made during this study allowed us to characterize, for the first time, GD in triatomines and demonstrated that gametogenesis does not occur in atrophied gonads. The characterization of GD in male hybrids resulting from the crossing of T. mopan♀ × T. longipennis♂ highlights the importance of evaluating both the morphology and the cytology of the gonads to confirm which event resulted in the sterility of the hybrid: GD (which results in no gamete production) or meiotic errors (which results in non-viable gametes).

Karyotype Evolution in Triatominae (Hemiptera, Reduviidae): The Role of Chromosomal Rearrangements in the Diversification of Chagas Disease Vectors.

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.

Trends in evolution of the Triatomini tribe (Hemiptera, Triatominae): reproductive incompatibility between four species of geniculatus clade.

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).

Morphological, Cytological and Molecular Studies and Feeding and Defecation Pattern of Hybrids from Experimental Crosses between Triatoma sordida and T. rosai (Hemiptera, Triatominae).

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.

Do not judge a book by its cover: would Triatoma tibiamaculata (Pinto, 1926) belong to Triatoma Laporte, 1832, or to Panstrongylus Berg, 1879, with misleading homoplasies?

BACKGROUND: Triatoma tibiamaculata is a species distributed in ten Brazilian states which has epidemiological importance as it has already been found infecting household areas. The taxonomy of this triatomine has been quite unstable: it was initially described as Eutriatoma tibiamaculata. Later, the species was transferred from the genus Eutriatoma to Triatoma. Although included in the genus Triatoma, the phylogenetic position of T. tibiamaculata in relation to other species of this genus has always been uncertain once this triatomine was grouped in all phylogenies with the genus Panstrongylus, rescuing T. tibiamaculata and P. megistus as sister species. Thus, we evaluated the generic status of T. tibiamaculata using phylogenetic and chromosomal analysis. METHODS: Chromosomal (karyotype) and phylogenetic (with mitochondrial and nuclear markers) analyses were performed to assess the relationship between T. tibiamaculata and Panstrongylus spp. RESULTS: The chromosomal and phylogenetic relationship of T. tibiamaculata and Panstrongylus spp. confirms the transfer of the species to Panstrongylus with the new combination: Panstrongylus tibiamaculatus. CONCLUSIONS: Based on chromosomal and phylogenetic characteristics, we state that P. tibiamaculatus comb. nov. belongs to the genus Panstrongylus and that the morphological features shared with Triatoma spp. represent homoplasies.

Triatoma sordida (Hemiptera, Triatominae) from La Paz, Bolivia: an incipient species or an intraspecific chromosomal polymorphism?

BACKGROUND: Triatoma sordida is one of the main Chagas disease vectors in Brazil. In addition to Brazil, this species has already been reported in Bolivia, Argentina, Paraguay, and Uruguay. It is hypothesized that the insects currently identified as T. sordida are a species subcomplex formed by three cytotypes (T. sordida sensu stricto [s.s.], T. sordida La Paz, and T. sordida Argentina). With the recent description of T. rosai from the Argentinean specimens, it became necessary to assess the taxonomic status of T. sordida from La Paz, Bolivia, since it was suggested that it may represent a new species, which has taxonomic, evolutionary, and epidemiological implications. Based on the above, we carried out molecular and experimental crossover studies to assess the specific status of T. sordida La Paz. METHODS: To evaluate the pre- and postzygotic barriers between T. sordida La Paz and T. sordida s.s., experimental crosses and intercrosses between F1 hybrids and between F2 hybrids were conducted. In addition, cytogenetic analyses of the F1 and F2 hybrids were applied with an emphasis on the degree of pairing between the homeologous chromosomes, and morphological analyses of the male gonads were performed to evaluate the presence of gonadal dysgenesis. Lastly, the genetic distance between T. sordida La Paz and T. sordida s.s. was calculated for the CYTB, ND1, and ITS1 genes. RESULTS: Regardless of the gene used, T. sordida La Paz showed low genetic distance compared to T. sordida s.s. (below 2%). Experimental crosses resulted in offspring for both directions, demonstrating that there are no prezygotic barriers installed between these allopatric populations. Furthermore, postzygotic barriers were not observed either (since the F1 × F1 and F2 × F2 intercrosses resulted in viable offspring). Morphological and cytogenetic analyses of the male gonads of the F1 and F2 offspring demonstrated that the testes were not atrophied and did not show chromosome pairing errors. CONCLUSION: Based on the low genetic distance (which configures intraspecific variation), associated with the absence of prezygotic and postzygotic reproductive barriers, we confirm that T. sordida La Paz represents only a chromosomal polymorphism of T. sordida s.s.

Trends in evolution of the Rhodniini tribe (Hemiptera, Triatominae): experimental crosses between Psammolestes tertius Lent & Jurberg, 1965 and P. coreodes Bergroth, 1911 and analysis of the reproductive isolating mechanisms.

BACKGROUND: The tribe Rhodniini is a monophyletic group composed of 24 species grouped into two genera: Rhodnius and Psammolestes. The genus Psammolestes includes only three species, namely P. coreodes, P. tertius and P. arthuri. Natural hybridization events have been reported for the Rhodniini tribe (for genus Rhodnius specifically). Information obtained from hybridization studies can improve our understanding of the taxonomy and systematics of species. Here we report the results from experimental crosses performed between P. tertius and P. coreodes and from subsequent analyses of the reproductive and morphological aspects of the hybrids. METHODS: Crossing experiments were conducted between P. tertius and P. coreodes to evaluate the pre- and post-zygotic barriers between species of the Rhodniini tribe. We also performed cytogenetic analyses of the F1 hybrids, with a focus on the degree of pairing between the homeologous chromosomes, and morphology studies of the male gonads to evaluate the presence of gonadal dysgenesis. Lastly, we analyzed the segregation of phenotypic characteristics. RESULTS: Interspecific experimental crosses demonstrated intrageneric genomic compatibility since hybrids were produced in both directions. However, these hybrids showed a high mortality rate, suggesting a post-zygotic barrier resulting in hybrid unviability. The F1 hybrids that reached adulthood presented the dominant phenotypic segregation pattern for P. tertius in both directions. These insects were then intercrossed; the hybrids were used in the cross between P. tertius ♀ × P. coreodes ♂ died before oviposition, and the F1 hybrids of P. coreodes ♀ x P. tertius ♂ oviposited and their F2 hybrids hatched (however, all specimens died after hatching, still in first-generation nymph stage, pointing to a hybrid collapse event). Morphological analyses of male gonads from F1 hybrids showed that they did not have gonadal dysgenesis. Cytogenetic analyses of these triatomines showed that there were metaphases with 100% pairing between homeologous chromosomes and metaphases with pairing errors. CONCLUSION: The results of this study demonstrate that Psammolestes spp. have intrageneric genomic compatibility and that post-zygotic barriers, namely unviability of hybrid and hybrid collapse, resulted in the breakdown of the hybrids of P. tertius and P. coreodes, confirming the specific status of species based on the biological concept of species.

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.

Segregation of phenotypic characteristics in hybrids of Triatoma brasiliensis species complex (Hemiptera, Reduviidae, Triatominae).

There are currently 156 species, grouped into 18 genera and five tribes included in the subfamily Triatominae. All of them are potential vectors of Chagas disease. Triatoma is paraphyletic and the species of this genus have been grouped into complexes and subcomplexes. Triatoma brasiliensis complex is a monophyletic group composed of eight taxa: T. b. brasiliensis, T. b. macromelanosoma, T. juazeirensis, T. sherlocki, T. petrocchiae, T. lenti, T. bahiensis, T. melanica. Experimental crosses have helped in systematic, taxonomic and evolutionary issues of these vectors. Based on this, we carried out experimental crosses between T. lenti and four other species of the T. brasiliensis complex and analyzed the segregation pattern of phenotypic characteristics of T. lenti, T. b. brasiliensis, T. b. macromelasoma, T. juazeirensis and T. melanica in the hybrids. The hybrids resulting from the crosses between T. b. brasiliensis ♀ x T. lenti ♂, T. juazeirensis ♀ x T. lenti ♂, and T. melanica ♀ x T. lenti ♂ showed segregation of characteristics of both parental species. On the other hand, the hybrids between T. lenti ♀ x T. juazeirensis ♂, T. b. macromelasoma ♀ x T. lenti ♂, and T. lenti ♀ x T. melanica ♂ showed a specific pattern of T. lenti, T. lenti and T. melanica, respectively. Thus, the pattern of segregation of morphological characteristics between species of the T. brasiliensis complex was characterized. These results highlight the importance of integrative taxonomy for the correct identification of Chagas disease vectors grouped in the T. brasiliensis complex if natural hybridization events occur.

Triatoma rosai sp. nov. (Hemiptera, Triatominae): A New Species of Argentinian Chagas Disease Vector Described Based on Integrative Taxonomy.

Chagas disease is the most prevalent neglected tropical disease in the Americas and makes an important contribution to morbidity and mortality rates in countries where it is endemic since 30 to 40% of patients develop cardiac diseases, gastrointestinal disorders, or both. In this paper, a new species of the genus Triatoma is described based on specimens collected in the Department San Miguel, Province of Corrientes, Argentina. Triatoma rosai sp. nov. is closely related to T. sordida (Stål, 1859), and was characterized based on integrative taxonomy using morphological, morphometric, molecular data, and experimental crosses. These analyses, combined with data from the literature (cytogenetics, electrophoresis pattern, molecular analyses, cuticular hydrocarbons pattern, geometric morphometry, cycle, and average time of life as well as geographic distribution) confirm the specific status of T. rosai sp. nov. Natural Trypanosoma cruzi infection, coupled with its presence mostly in peridomestic habitats, indicates that this species can be considered as an important Chagas disease vector from Argentina.

Parasite × vector relationship in Chagas disease: does Trypanosoma cruzi (Chagas, 1909) infection affect the spermatogenesis of Triatoma infestans (Klug, 1834)?

The parasite-vector interaction of Chagas disease is still poorly understood and the understanding of this relationship can help in the development of new strategies to control Trypanosoma cruzi transmission, which is the etiological agent of this disease. Considering the need to know if T. cruzi can cause some pathology in the reproductive system of the Chagas disease vectors, we investigated the spermatogenesis of Triatoma infestans infected by T. cruzi through histological and cytogenetic analysis. Trypanosoma cruzi Bolivia strain infection was not pathogenic for the reproductive system of T. infestans, because all the analyzed males had normal spermatogenesis, with all phases (spermatocytogenesis, meiosis and spermiogenesis) happening without any change. Thus, we demonstrated that the presence of T. cruzi Bolivia strain does not have influence in the spermatogenesis of T. infestans and we suggest that the influences on reproductive system observed for other species were a result of the action of the parasite on gametogenesis of females.

Prezygotic isolation confirms the exclusion of Triatoma melanocephala, T. vitticeps and T. tibiamaculata of the T. brasiliensis subcomplex (Hemiptera, Triatominae).

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.

Taxonomical over splitting in the Rhodnius prolixus (Insecta: Hemiptera: Reduviidae) clade: Are R. taquarussuensis (da Rosa et al., 2017) and R. neglectus (Lent, 1954) the same species?

The use of subtle features as species diagnostic traits in taxa with high morphological similarity sometimes fails in discriminating intraspecific variation from interspecific differences, leading to an incorrect species delimitation. A clear assessment of species boundaries is particularly relevant in disease vector organisms in order to understand epidemiological and evolutionary processes that affect transmission capacity. Here, we assess the validity of the recently described Rhodnius taquarussuensis (da Rosa et al., 2017) using interspecific crosses and molecular markers. We did not detect differences in hatching rates in interspecific crosses between R. taquarussuensis and R. neglectus (Lent, 1954). Furthermore, genetic divergence and species delimitation analyses show that R. taquarussuensis is not an independent lineage in the R. prolixus group. These results suggest that R. taquarussuensis is a phenotypic form of R. neglectus instead of a distinct species. We would like to stress that different sources of evidence are needed to correctly delimit species. We consider this is an important step in understanding vectorial Chagas disease spread and transmission.

Triatoma vitticeps (Stal, 1859) (Hemiptera, Triatominae): A Chagas Disease Vector or a Complex of Vectors?

Triatoma vitticeps is a Chagas disease vector that was found infected with Trypanosoma cruzi in homes. As this species is endemic from Brazil (Bahia, Espírito Santo, Minas Gerais, and Rio de Janeiro) and no study comparing the specimens from different Brazilian states was conducted, we analyzed the genetic distance (16S rDNA, Cyt b, and COI mitochondrial genes) and the chromosomal characteristics for T. vitticeps from Minas Gerais, Rio de Janeiro, and Espírito Santo. All specimens showed the same cytogenetic characteristics. On the other hand, the different mitochondrial genes demonstrated high intraspecific variation between the genetic distances of T. vitticeps from different states ranging from 2.3% to 7.2%. Based on this, our results suggest that possibly what is characterized as T. vitticeps is a complex of cryptic species (or subspecies).

Karyotype Evolution of Chagas Disease Vectors (Hemiptera, Triatominae).

The Triatominae subfamily is composed of 153 hematophagous species that are potential vectors of Trypanosoma cruzi, the etiological agent of Chagas disease. Karyotypic studies in triatomines were initiated in 1909. There are 92 karyotypes described, all grouped into the tribes Rhodniini and Triatomini. Recently, a phylogenetic study of the triatomines that combines molecular data with geological changes was performed. We now discuss how the karyotype evolved with the diversification of the triatomines.

Triatoma vitticeps subcomplex (Hemiptera, Reduviidae, Triatominae): a new grouping of Chagas disease vectors from South America.

BACKGROUND: Triatomines have been grouped into complexes and subcomplexes based largely on morphological and geographical distribution. Although these groupings are not formally recognised as taxonomic ranks, they are likely monophyletic. However, recent studies have demonstrated that some subcomplexes from South America did not form monophyletic groups, and reorganisations have been suggested. One suggested reorganisation is to exclude Triatoma vitticeps, T. melanocephala, and T. tibiamaculata from the T. brasiliensis subcomplex. However, T. vitticeps and T. melanocephala exhibit several similar characteristics, including morphologic, cytogenetic, and phylogenetic features, a factor which supports the creation of a new subcomplex. Thus, this study aimed to describe the T. vitticeps subcomplex. RESULTS: T. vitticeps and T. melanocephala are sister species and share a phylogenetic relationship, several similar morphological characteristics, the same composition of constitutive heterochromatin (Xs CG-rich and Y AT-rich), the same karyotype (2n = 20A + X1X2X3Y), and the same meiotic behaviour during spermatogenesis. Based on karyosystematics, for example, the T. vitticeps subcomplex may differ from all of the other subcomplexes from South America, as well as from the Rhodniini tribe and the genus Panstrongylus. We argue that the case of agmatoploidy involving the X chromosome was responsible for the karyotype divergence of this subcomplex in relation to the other South America subcomplexes. CONCLUSIONS: Based on the phenotypic characteristics (morphology) and genotypes (cytogenetics and molecular features), we propose the creation of the monophyletic T. vitticeps subcomplex, which we believe is distinct from all other subcomplexes from South America.

Nucleolar Persistence: Peculiar Characteristic of Spermatogenesis of the Vectors of Chagas Disease (Hemiptera, Triatominae).

All species of triatomines are considered potential vectors of Chagas disease and the reproductive biology of these bugs has been studied by different approaches. In 1999, nucleolar persistence during meiosis was observed in the subfamily for the first time. Recently, it has been observed that all species within the genus Rhodnius exhibit the same phenomenon, suggesting that it may be a synapomorphy of the triatomines. Thus, this article aims to analyze the nucleolar behavior during spermatogenesis of 59 triatomine species. All analyzed species exhibited nucleolar persistence during meiosis. Recently, it has been suggested that nucleolar persistence may be fundamental for the spermatogenesis of these vectors, since it is related to the formation of the chromatoid body. Therefore, we emphasize that this phenomenon is a peculiarity of the Triatominae subfamily and that further studies are required to analyze whether the nucleolar material that persists is active.

Reproductive Biology of Triatoma brasiliensis (Hemiptera, Triatominae) During the Imaginal Molt.

The triatomines are vectors of the protozoan Trypanosoma cruzi, etiologic agent of Chagas disease. These insects are sexually active after the imaginal molt. Some aspects have been studied in Triatoma brasiliensis during the imaginal molt, such as autogeny in virgin females and the relationship between blood ingestion by fifth instar nymph and the realization of the imaginal molt. Thus, to aid in the understanding of reproductive biology and developmental physiology of these vectors, this article analyzes the spermatogenesis of T. brasiliensis during the imaginal molt. The analysis of the seminiferous tubules from males in the fifth instar during imaginal molt has demonstrated that T. brasiliensis has only a few spermatids and a plentiful quantity of sperm. Thus, we suggest that during imaginal molt the cell division is disrupted aiming to reduce energy costs and the differentiation into sperm is stimulated to ensure the paternity of the adult male.

Chromosomal characteristics and distribution of constitutive heterochromatin in the Matogrossensis and Rubrovaria subcomplexes.

Since 1966 the triatomines were grouped in complexes and specific subcomplexes. Although the complex and subcomplexes not have taxonomic importance, should be monophyletic groups and cytogenetic tools have proved to be of great importance to characterize these species groupings. Based on this, this paper aims to describe the chromosomal characteristics and heterochromatic pattern of Matogrossensis and Rubrovaria subcomplexes, in order to contribute to the taxonomic and evolutionary relationships of these vectors. In this study, at least three males from each species (Triatoma baratai, Triatoma costalimai, Triatoma guazu, Triatoma jurbergi, Triatoma matogrossensis, Triatoma vandae, Triatoma williami, Triatoma carcavalloi, Triatoma circummaculata, Triatoma klugi, Triatoma pintodiasi and Triatoma rubrovaria) were analyzed by means analyzed by means of cytogenetic techniques of C-banding. All species showed the same cytogenetic characteristics: 22 chromosomes, low variation in the size of autosomes, sex chromosome Y larger than X, initial prophase composed of only one heterochromatic chromocenter formed by the sex chromosomes X and Y (except for T. pintodiasi that presented the sex chromosomes individualized during all stages of prophase) and presence of constitutive heterochromatin restricted to sex chromosome Y. These characteristics, although common to Matogrossensis and Rubrovaria subcomplexes allow to distinguish these species of species grouped in most of South America subcomplexes, as Brasiliensis, Maculata, Sordida and Insfestans. Thus, the cytogenetic analysis was of extreme importance to differentiate both subcomplexes of the other subcomplexes of South America. However, probably due to evolutionary proximity existing between these subcomplexes was not possible to observar species differences that make up the Matogrossensis subcomplex of the Rubrovaria subcomplex. Therefore, we emphasize that new comparative analyzes, as experimental hybrid crosses and molecular cytogenetic analysis are necessary to clarify the evolutionary relationship between these important subcomplexes of vectors.