Susceptibilidade de Rhodnius neglectus, Rhodnius robustus e Triatoma infestans (Hemiptera, Reduviidae, Triatominae) à infecção por duas cepas de Trypanosoma cruzi (Kinetoplastidae, Trypanosomatidae) utilizando xenodiagnóstico artificial

A susceptibilidade de ninfas de 3º estádio de Rhodnius neglectus, R. robustus e Triatoma infestans às cepas Y e AMJM de Trypanosoma cruzi foi verificada utilizando xenodiagnóstico artificial. Para a leitura do xenodiagnóstico, as fezes dos triatomíneos foram examinadas a cada dois dias, a partir do 5º até o 31º dia pós infecção, pela técnica de compressão abdominal. Os resultados mostraram diferenças na susceptibilidade dos triatomíneos para as duas cepas estudadas e o período ótimo de leitura variou do 11º ao 19º dias para a cepa Y e do 11º ao 15º dias para a cepa AMJM. Também, pôde-se concluir que para a cepa Y, as três espécies de triatomíneos demonstraram boa susceptibilidade, enquanto para a cepa AMJM, a melhor susceptibilidade foi observada com R. neglectus, seguida pelo T. infestans e R. robustus.

Description of a newly discovered Triatoma infestans (Hemiptera: Reduviidae) Foci in Ibipeba, State of Bahia, Brazil

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

A short proregion of trialysin, a pore-forming protein of Triatoma infestans salivary glands, controls activity by folding the N-terminal lytic motif

Triatoma infestans (Hemiptera: Reduviidae) is a hematophagous insect that transmits the protozoan parasite Trypanosoma cruzi, the etiological agent of Chagas` disease. Its saliva contains trialysin, a protein that forms pores in membranes. Peptides based on the N-terminus of trialysin lyse cells and fold into alpha-helical amphipathic segments resembling antimicrobial peptides. Using a specific antiserum against trialysin, we show here that trialysin is synthesized as a precursor that is less active than the protein released after saliva secretion. A synthetic peptide flanked by a fluorophore and a quencher including the acidic proregion and the lytic N-terminus of the protein is also less active against cells and liposomes, increasing activity upon proteolysis. Activation changes the peptide conformation as observed by fluorescence increase and CD spectroscopy. This mechanism of activation could provide a way to impair the toxic effects of trialysin inside the salivary glands, thus restricting damaging lytic activity to the bite site.

Inoculation of Triatoma Virus (Dicistroviridae: Cripavirus) elicits a non-infective immune response in mice

Background: Dicistroviridae is a new family of small, non-enveloped, +ssRNA viruses pathogenic to both beneficial arthropods and insect pests. Little is known about the dicistrovirus replication mechanism or gene function, and any knowledge on these subjects comes mainly from comparisons with mammalian viruses from the Picornaviridae family. Due to its peculiar genome organization and characteristics of the per os viral transmission route, dicistroviruses make good candidates for use as biopesticides. Triatoma virus (TrV) is a pathogen of Triatoma infestans (Hemiptera: Reduviidae), one of the main vectors of the human trypanosomiasis disease called Chagas disease. TrV was postulated as a potential control agent against Chagas' vectors. Although there is no evidence that TrV nor other dicistroviruses replicate in species outside the Insecta class, the innocuousness of these viruses in humans and animals needs to be ascertained. Methods: In this study, RT-PCR and ELISA were used to detect the infectivity of this virus in Mus musculus BALB/c mice. Results: In this study we have observed that there is no significant difference in the ratio IgG2a/IgG1 in sera from animals inoculated with TrV when compared with non-inoculated animals or mice inoculated only with non-infective TrV protein capsids. Conclusions: We conclude that, under our experimental conditions, TrV is unable to replicate inmice. This study constitutes the first test to evaluate the infectivity of a dicistrovirus in a vertebrate animal model.

Molecular Techniques for Dicistrovirus Detection without RNA Extraction or Purification

Dicistroviridae is a new family of small, nonenveloped, and +ssRNA viruses pathogenic to both beneficial arthropods and insect pests as well. Triatoma virus (TrV), a dicistrovirus, is a pathogen of Triatoma infestans (Hemiptera: Reduviidae), one of the main vectors of Chagas disease. In this work, we report a single-step method to identify TrV, a dicistrovirus, isolated from fecal samples of triatomines. The identification method proved to be quite sensitive, even without the extraction and purification of RNA virus.

Seroprevalence of Triatoma virus (Dicistroviridae: Cripaviridae) antibodies in Chagas disease patients

Background: Chagas disease is caused by Trypanosoma cruzi, and humans acquire the parasite by exposure to contaminated feces from hematophagous insect vectors known as triatomines. Triatoma virus (TrV) is the sole viral pathogen of triatomines, and is transmitted among insects through the fecal-oral route and, as it happens with T. cruzi, the infected insects release the virus when defecating during or after blood uptake. Methods: In this work, we analysed the occurrence of anti-TrV antibodies in human sera from Chagas disease endemic and non-endemic countries, and developed a mathematical model to estimate the transmission probability of TrV from insects to man, which ranged between 0.00053 and 0.0015. Results: Our results confirm that people with Chagas disease living in Bolivia, Argentina and Mexico have been exposed to TrV, and that TrV is unable to replicate in human hosts. Conclusions: We presented the first experimental evidence of antibodies against TrV structural proteins in human sera.

Cytochemical characterization of Triatoma infestans and Panstrongylus megistus salivary gland cells (Hemiptera, Reduviidae, Triatominae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Sexual distinction between 5th instar nymphs of six 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. matogrosensis 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 female nymphs, while in five species; tergites in female nymphs are broad and in male are narrow.

Characterization of the external female genitalia of six species of Triatominae (Hemiptera: Reduviidade) by scanning electron microscopy

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

A study of the scutellum in eight Chagas disease vector species from genus Triatoma (Hemiptera, Reduviidae) using optical and scanning electron microscopy

The aim of this study was to analyze the external morphology of the scutellum through optical microscopy and scanning electron microscopy (SEM) in male specimens of Triatoma costalimai, T. delpontei, T. eratyrusiformis, T. matogrossensis, T. infestans melanosoma, T. sherlocki, T. tibiamaculata, and T. vandae. A total of 30 photographs of the scutellum were made. Magnification varied from 50X to 750X. Regarding depth and forms of the central depression, the heart-shaped form was predominant, with some exceptions, so that this shape appears to be a common characteristic for species of genus Triatoma Laporte, 1832. In T. eratyrusiformis, a kind of sensillum with important taxonomic value was observed. The different sizes and shapes of the designs found on the posterior process of the scutellum were also of important taxonomic interest. The study of the scutellum based on SEM showed valuable characteristics, allowing the use of this structure to aid the diagnosis of triatomine species. Thus, more specimens in subsequent studies and analyses of morphometric parameters should contribute to agreement on phylogenetic aspects in this genus. A Key to eight species of Triatoma based on male scutellar morphology is presented.

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.

Genetic Variability and Geographical Diversity of the Main Chagas' Disease Vector Panstrongylus megistus (Hemiptera: Triatominae) in Brazil Based on Ribosomal DNA Intergenic Sequences

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise da relação filogenética entre Triatoma carcavalloi Jurberg, Rocha; Lent, 1998; Triatoma circummaculata Stal, 1859; Triatoma klugi Carcavallo, Jurberg, Lent; Galvão, 2001 e Triatoma rubrovaria Blanchard, 1843 (Hemiptera, Reduviidae) baseada no sequenciamento de genes do DNA mitocondrial e nuclear

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

Contribution of AT-, GC-, and methylated cytidine-rich DNA to chromatin composition in Malpighian tubule cell nuclei of Panstrongylus megistus (Hemiptera, Reduviidae)

The Malpighian tubule cell nuclei of male Panstrongylus megistus, a vector of Chagas disease, contain one chromocenter, which is composed solely of the Y chromosome. Considering that different chromosomes contribute to the composition of chromocenters in different triatomini species, the aim of this study was to determine the contribution of AT-, GC-, and methylated cytidine-rich DNA in the chromocenter as well as in euchromatin of Malpighian tubule cell nuclei of P. megistus in comparison with published data for Triatoma infestans. Staining with 4',6-diamidino-2-phenylindole/actinomycin D and chromomycin A(3)/distamycin, immunodetection of 5-methylcytidine and AgNOR test were used. The results revealed AT-rich/GC-poor DNA in the male chromocenter, but equally distributed AT and GC DNA sequences in male and female euchromatin, like in T. infestans. Accumulation of argyrophilic proteins encircling the chromocenter did not always correlate with that of GC-rich DNA. Methylated DNA identified by immunodetection was found sparsely distributed in the euchromatin of both sexes and at some points around the chromocenter edge, but it could not be considered responsible for chromatin condensation in the chromocenter, like in T. infestans. However, unlike in T. infestans, no correlation between the chromocenter AT-rich DNA and nucleolus organizing region (NOR) DNA was found in P. megistus. (c) 2011 Elsevier GmbH. All rights reserved.

Seasonal activity and abundance of Orosius orientalis (Hemiptera : Cicadellidae) at agricultural sites in Southeastern Australia

Orosius orientalis is a leafhopper vector of several viruses and phytoplasmas affecting a broad range of agricultural crops. Sweep net, yellow pan trap and yellow sticky trap collection techniques were evaluated. Seasonal distribution of O. orientalis was surveyed over two successive growing seasons around the borders of commercially grown tobacco crops. Orosius orientalis seasonal activity as assessed using pan and sticky traps was characterised by a trimodal peak and relative abundance as assessed using sweep nets differed between field sites with peak activity occurring in spring and summer months. Yellow pan traps consistently trapped a higher number of O. orientalis than yellow sticky traps.