Cytogenetic characterization reveals differences in nuclear organization of cystic cells among Brazilian species of Triatoma (Heteroptera, Reduviidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Morfologia de ovos de Triatoma circummaculata e Triatoma rubrovaria (Hemiptera, Reduviidae)

OBJETIVO: Estudar morfologicamente ovos de T. circummaculata e T. rubrovaria. MÉTODOS: Quarenta ovos das duas espécies foram mensurados por meio de projetor de perfil Nikon modelo 6C, e utilizado o teste Student, para execução da análise estatística. As observações morfológicas foram feitas em microscópio eletrônico de varredura JEOL, modelo JSM-T330A. RESULTADOS/CONCLUSÕES: A análise estatística revelou que os ovos e as cascas de ovos de T. rubrovaria são significativamente maiores que os de T. circummaculata. Observações microscópicas mostraram que a borda corial, assim como a goteira espermática, são bem mais evidentes nos ovos de T. rubrovaria que nos de T. circummaculata. O exocório das duas espécies é formado por áreas poligonais, que são hexagonais em sua maioria. Foram observados também o momento da eclosão do embrião e os detalhes das exúvias embrionárias.

Ribosomal DNA ITS-1 intergenic spacer polymorphism in triatomines (Triatominae, Heteroptera)

The length polymorphism of ribosomal DNA ITS-1 intergenic spacer was analyzed in eight species of triatomines belonging to Triatoma, Rhodnius, and Panstrongylus genera. The analyzed species were Rhodnius domesticus, R. neivai, R. robustus, Triatoma brasiliensis, T. infestans, T. vitticeps, Panstrongylus megistus, and P. herreri. These insects are vectors of Chagas' disease, one of the most prominent public health problems among South American countries. This work allowed the differentiation between species of the Triatomini and Rhodniini tribes through the analysis of ITS-1 length polymorphism by PCR and RFLP techniques. The species of the Triatoma and Panstrongylus genera presented an amplified ITS-1 fragment between 600 and 1000 bp, whereas Rhodnius presented a less variable ITS-1 length fragment, around 300 bp, which could reflect the monophyletic origin of the Rhodniini tribe. Species belonging to this genus were further differentiated by RFLP with HaeIII and AluI endonucleases. Our results corroborate the hypothesis of polyphyletic origin in this group of insects and contribute to knowledge about evolutionary relationships in triatomines.

SEXUAL DISTINCTION BETWEEN 5TH INSTAR NYMPHS OF 6 SPECIES OF TRIATOMINAE (HEMIPTERA, REDUVIIDAE)

The aim of this paper is to present the analysis of sexual morphological differences observed in 5th instar nymphs of the following species: Panstrongylus megistus; Rhodnius neglectus; Triatoma brasiliensis; T. infestans; T. matogrossensis and T. tibiamaculata. Male and female nymphs were examined and photographed with a Scanning Electron Microscope. The 9th segment dimensions of dorsal and ventral faces were determined through a Profile Projector. Results and statistical analysis showed significant differences: the 9th sternite is significantly broader in male than in female nymphs, while in five species, tergites in female nymphs are broad and in male are narrow.

Cytogenetic study on three species of the genus Triatoma (Heteroptera: Reduviidae) with emphasis on nucleolar organizer regions

The use of banding techniques allows the recognition of chromosomal pairs and karyotypical arrangements. However, its application in Heteroptera holocentric chromosomes is limited. Thus, little is known about their structure, specially their Nucleolar Organizer Regions (NORs). A comparative analysis of the nucleolar characteristics present during spermatogenesis in Triatoma platensis, Triatoma protacta and Triatoma tibiamaculata seems to indicate that in this group of insects nucleolar fragmentation occurs after prophase I. The study of chromosomal structure of these triatomines indicates that NORs are located at some telomeric and interstitial autosome regions and at sexual chromosomes (X/X1X2).

Estudo morfológico e histológico dos ovos de seis espécies do gênero Triatoma (Hemiptera: Reduviidae)

The objective of this study was to analyze the morphological and histological characteristics of eggs from six triatomine species in order to obtain a generic and specific characterization of this group of Trypanosoma cruzi vectors and better understand their phylogenetic and taxonomic aspects. The eggs of the respective species came from the collection of the Laboratório de Triatomíneos e Culicídeos da Faculdade de Saúde Pública/USP, from the Insetário do Serviço Especial de Saúde de Araraquara - SESA and from the Triatomine insectary of the Laboratório Nacional e Internacional de Referência em Taxonomia de Triatomíneos (FIOCRUZ), in Rio de Janeiro. Morphological studies were done with optical microscopy and scanning electron microscopy. Histological analyses used only optical microscopy. All of the species analyzed showed a predominance of hexagonal cells. The exchorion of Triatoma breyeri Del Ponte, T. costalimai Verano & Galvão, and T. tibiamaculata (Pinto) consists of unornamented cells, overlapping or slightly separated, with a smooth, padded appearance. Meanwhile, in T. matogrossensis Leite & Barbosa, T. sherlocki Papa, Juberg, Carcavallo, Cerqueira & Barata, and T. williami Galvão, Souza & Lima, the exchorion cells are ornamented on their entire surface with perforations and fissures. Egg histology showed undulations, pores, and orifices. The study helps to expand the generic and specific knowledge of the Triatominae subfamily. The characteristics identified in the exchorion of the eggshells can help separate close species. These new parameters will back the elaboration of future dichotomous keys, help to determine each vector species' role and expand knowledge of the various species.

Cytogenetic analysis in the spermatogenesis of Triatoma melanosoma (Reduviidae; Heteroptera)

Triatomines are of great concern in public health because they are vectors of Chagas' disease. This study presents an analysis of the species Triatoma melanosoma. The cytogenetic characteristics of triatomines include holocentric chromosomes, post-reductional meiosis in the sex chromosomes and nucleolar fragmentation in the meiotic cycle. The methodology utilized consisted of the techniques of lacto-acetic orcein staining and silver ion impregnation. The organs analyzed were adult testicles. The results enabled to classify the chromosomes by number and size, being three large, eight medium and one small heterochromosome. The three largest chromosomes and the heterochromosomes showed heteropyknotic chromatin in meiosis. The heterochromosomes in 8.05% of the cells in metaphase I behaved as pseudobivalents, contrasting with 91.95% of the cells with individualized sex chromosomes, confirming the achiasmatic nature of these chromosomes. However, the pseudobivalents occurred prominently in metaphase II (78.38%), this fact probably is related to the post-reductional nature of the sex chromosomes. The nucleolus in T. melanosoma persisted until the diplotene phase after which it began to fragment. Nucleolar corpuscles were observed in metaphases I and II and during anaphases I and II, these characteristics being related to the phenomenon of nucleolar persistence. In the initial spermatids, peripheral silver ion impregnation occurred, which could be analogous to the pre-nucleolar corpuscles observed after fragmentation. Thus, this study extends our knowledge of the characteristics of triatomines, in particular, heteropyknotic degree, kinetic activity, formation of sex chromosome achiasmatic pseudobivalency, confirmation of the fragmentation phenomenon, and post-meiotic nucleolar reactivation. ©FUNPEC-RP.

Description of eggs and nymphal instars of Triatoma baratai Carcavallo and Jurberg, 2000 based on optical and scanning electron microscopy (Hemiptera: Reduviidae: Triatominae)

Triatoma baratai Carcavallo & Jurberg, is a wild (i.e., nonperidomestic) species found in the State of Mato Grosso do Sul (Bodoquena region, county of Bonito), Brazil. Its eggs and nymphs are described here based on optical and scanning electron microscopy. The operculum and exochorion have pentagonal, hexagonal, and heptagonal cells, with small cracks and small random pits. Differences in the eggs and five nymphal instars of T. baratai allow them to be distinguished from the sympatric species Triatoma williami Galvão, Souza & Lima, and from six of the nine members of the Triatoma oliveirai complex. The most useful differentiating characters are in the color, shape of the abdomen, head, and total body length. Keys are provided to separate the eggs and nymphal instars of six of the nine members of the Triatoma oliveirai species complex. Copyright ©2009 Magnolia Press.

Distribution of constitutive heterochromatin in two species of triatomines: Triatoma lenti Sherlock and Serafim (1967) and Triatoma sherlocki Papa, Jurberg, Carcavallo, Cerqueira & Barata (2002)

Triatoma lenti and Triatoma sherlocki are hemipterans that belong to the brasiliensis subcomplex. In triatomines, the constitutive heterochromatin pattern is species-specific and allows, in many cases, for the grouping of species. Thus, we cytogenetically analyzed T. sherlocki and T. lenti using C-banding, and we compared the results with previous ones obtained in other species of the brasiliensis subcomplex. Both species were found to have a male diploid chromosome number of 22 chromosomes (2n = 20A. +. XY) with heterochromatic blocks at one or both chromosomal ends of all autosomal pairs. During early meiotic prophase, they showed a large heteropycnotic chromocenter constituted by the association of both sex chromosomes plus two autosomal pairs and many heterochromatic blocks dispersed inside the nucleus. All of these cytogenetic characteristics are similar to those observed in other species of brasiliensis subcomplex, results which confirm the grouping of T. sherlocki and T. lenti within this subcomplex. However, we emphasize the importance of other approaches, such as molecular analysis, to confirm the placement of T. lenti within the brasiliensis subcomplex. © 2012 Elsevier B.V.

Cross-mating experiments detect reproductive compatibility between Triatoma sherlocki and other members of the Triatoma brasiliensis species complex

Phylogenetic approaches based on mitochondrial DNA variation (fragments of Cyt B and 16S ribosomal RNA) have revealed Triatoma sherlocki as the most recent species addition to the Triatoma brasiliensis species complex; a monophyletic group which includes T. brasiliensis, Triatoma melanica, and Triatoma juazeirensis. T. sherlocki is the most differentiated among all species of this complex: it is unable to fly, possesses longer legs than the other members, and has reddish tonality in some parts of its exochorion. We question whether these species are reproductively compatible because of this pronounced morphological differentiation, and therefore, we present a series of cross breeding experiments that test compatibility between T. sherlocki and other members of the T. brasiliensis complex. We extended our analyses to include crosses between T. sherlocki and Triatoma lenti, because the latter has been suggested as a possible member of this complex. T. sherlocki male. ×. T. lenti female pairs failed to produce hybrids. All other crosses of T. sherlocki and members of T. brasiliensis species complex, as well as backcrosses, produced viable offspring through the third generation. This study stresses the importance of searching for the features that may isolate members of the T. brasiliensis species complex. © 2013 Elsevier B.V.

Estudo comparativo das características citogenéticas e moleculares de Triatoma maculata e Triatoma pseudomaculata (Triatominae, Heteroptera)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Estudo citogenético comparativo entre Triatoma maculata e triatoma pseudomaculata (Triatominae, Heteroptera)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Variabilidade genética de três colônias de Triatoma rubrovaria (Blanchard, 1843), (Hemiptera, Reduviidae), oriundas do estado do Rio Grande do Sul, avaliadas por meio do seqüenciamento de genes do DNA mitocondrial e ribossomal

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)