Cytogenetic analysis of Triatoma pseudomaculata Corrêa and Espínola, 1964 (Hemiptera, Triatominae) from different Brazilian states.

Triatoma maculata and T. pseudomaculata are allopatric; however, it is believed that T. maculata was introduced into the Brazilian northeast by passive transportation of the nymphs between the feathers of migratory birds, followed by the speciation of T. pseudomaculata. T. pseudomaculata is the second most epidemiologically important species in the northeast of Brazil after T. brasiliensis. Therefore, given the broad range of T. pseudomaculata, the objective of the present study was to cytogenetically analyze different populations of T. pseudomaculata from different Brazilian states in order to investigate possible intraspecific chromosomal variation between them. Three adult T. pseudomaculata males from each population (Pernambuco, Ceará, Paraíba, Bahia, Rio Grande do Norte, and Piauí) were analyzed by lacto-acetic orcein and C-banding. All of the specimens analyzed exhibited the same cytogenetic characteristics, i.e., 22 chromosomes (20 autosomes and XY), a chromocenter formed by the X and Y sex chromosomes and one pair of autosomes, and heterochromatin blocks in three or four pairs of autosomes. These data confirm that all of the populations analyzed were T. pseudomaculata, and although they may be subject to different selection pressures they have maintained the genetic integrity that characterizes the species.

Description of the diploid chromosome set of Triatoma pintodiasi (Hemiptera, Triatominae).

Triatoma pintodiasi has been described and recently grouped in the Rubrovaria subcomplex. T. pintodiasi was initially compared to T. carcavalloi by staining and subsequently identified as T. circummaculata. However, after thorough examination, it was observed to be a cryptic species of T. circummaculata, and was described based on morphological features, morphometric data, and biochemical patterns of hemolymph. Thus, this paper aims to describe the karyotype of, and spermatogenesis in, T. pintodiasi, in order to elucidate the reproductive biology and taxonomy of the species. Sex chromosomes of T. pintodiasi formed a heteropyknotic chromocenter, and compaction of chromatin was observed during prophase. However, in contrast to observations in T. carcavalloi and T. circummaculata, in T. pintodiasi it was observed individualization of the sex chromosomes. The diploid chromosome set of the species 2n = 22 (20A + XY) is described through analysis of metaphases I and II. Initial cytogenetic characteristics of T. pintodiasi are described and the observed differences in the chromocenter are suggested as a possible cytotaxonomic tool. To gain a better understanding of the specific status of this cryptic species, however, we emphasize the need for further cytogenetic, molecular, biological, and biogeographical analysis, in addition to experimental hybrid crosses with other species of the Rubrovaria subcomplex.

Differentiation between Triatoma arthurneivai and Triatoma wygodzinskyi (Hemiptera: Reduviidae: Triatominae) using cytotaxonomy.

Using classic morphometric techniques to examine the head and thorax of Triatoma specimens, researchers identified a possible taxonomic problem involving T. arthurneivai (Lent & Martins) and T. wygodzinskyi (Lent). A recent geometric morphometric study indicated that the insects captured outside the Serra do Cipó region, State of Minas Gerais, Brazil, were T. wygodzinskyi. The misidentification of T. arthurneivai as T. wygodzinskyi could result in several problems associated with entoepidemiological lifting, the biological characterization of the species, and phylogenetic reconstruction. For the first time, we describe the use of cytogenetic analysis as a tool for differentiation between T. arthurneivai and T. wygodzinskyi. The results indicated that both species had the same number of chromosomes 2n = 22 (20A + XY). However, analyses of spermatocytes during early prophase indicated that it was possible to differentiate T. arthurneivai and T. wygodzinskyi, because only T. arthurneivai exhibited heteropycnotic blocks distributed in the chromatin. Therefore, we highlight the analysis of spermatocytes as a taxonomic tool for the characterization of T. arthurneivai and T. wygodzinskyi, and suggest that the technique can be used for entoepidemiological lifting in vector control programs. Thus, the results presented here, in conjunction with morphometric analyses, are of utmost taxonomic and epidemiological importance for the identification of T. arthurneivai and T. wygodzinskyi specimens.

Endocrine and regenerative cells in the midgut of Chagas' disease vector Triatoma vitticeps during different starvation periods.

The midgut of an insect has three main cell types, viz., digestive, endocrine and regenerative cells, and a relationship between cell morphology and blood meal has been suggested. This study evaluated the occurrence of the regenerative and endocrine cells in different regions of the midgut of female and male Triatoma vitticeps during different periods after a blood meal. Adults of both sexes were dissected at 4, 72, 120, 168, 288, 360 and 600 hours after feeding and the midgut was pulled apart. The midgut was divided into the anterior, middle and posterior regions and the fragments were analyzed under a light microscope. The results showed the presence of endocrine cells of the open type and positive for FMRFamide as well as regenerative cell nests in the midgut of T. vitticeps. An association was observed between the starvation period and frequency of FMRFamide positive and regenerative cells in the midgut of T. vitticeps. Differences in the pattern of distribution of these cells between the males and females as well as in the different regions of the midgut were also discussed.

Cytogenetic study in a mutant of Triatoma infestans (Hemiptera: Reduviidae) carrying a spontaneous autosomal fusion and an extra chromosome.

Triatomainfestans (2n = 20 A + XY, male) is a blood-sucking bug and the most important vector of Chagas disease in the Southern Cone countries. A cytogenetic analysis of 14 individuals from the Argentine Gran Chaco has revealed the presence of a naturally heterozygous for an autosomal fusion. The fusion heterozygote (2n = 19 A + 1 extra chromosome + XY, male) presented an autosomal trivalent, 8 bivalents, the X and Y sex univalents, and a minute extra chromosome at meiosis I. The autosomal trivalent divided equationally at first anaphase. At metaphase II, cells had 8 autosomes, X and Y sex chromosomes, and an autosomal pseudo-trivalent composed by 3 different-sized chromatids. The orientation of this pseudo-trivalent led to a reductional segregation. The meiotic behaviour of this new chromosome complement was highly regular. The extra chromosome did not affect the segregation of autosomes and sex chromosomes during both meiotic divisions. We propose that the extra chromosome was originated as a product of an autosomal fusion, and it might become a B chromosome. Many authors suggest that karyotype evolution in Heteroptera has proceeded mainly by fusions and fragmentations. The fact that this rearrangement has been found in a natural population of T. infestans and that it shows a regular meiotic behaviour seems to support the suggested hypothesis.

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

Cytogenetic studies in triatomines have described the occurrence of holokinetic chromosomes, heterochromatin distribution and the location of rDNA (ribosomal DNA) sites, but few aspects of nuclear organization in this group have been discussed. We have focused on ultrastructural and cytogenetic features and differences in cystic cells of seminiferous tubules between five species of Triatoma. Cystic cells showed evidence of polyploidy events and heterochromatic blocks appeared predominantly in the central region of the nuclei. Cytogenetic analyses showed that there was variation in chromocenter number between species, and that the central regions were AT-rich [DAPI+ (4',6-diamidino-2-phenylindole+)], whereas the periphery was CG-rich (CMA+). Another characteristic was the distribution of 45S rDNA, which differed according to the chromosomal location of this sequence. In all we have compared aspects of nuclear organization, polyploidy, heterochromatin, rDNA site distribution and methylation levels, as well as the relationships between five species of Triatoma from a cystic cell perspective.

Karyotype and spermatogenesis in Triatoma lenti (Hemiptera: Triatominae), a potential Chagas vector.

All species of Triatominae are susceptible to infection by Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) and consequently, potential insect vectors of Chagas disease. Currently, there are 140 known species of triatomine bugs, which can be grouped into specific species complexes. The species Triatoma lenti (Hemiptera: Triatominae) is found only in Brazil and is considered a potential vector of Chagas disease. We karyotyped male T. lenti and examined its spermatogenesis in detail. The karyotype was found to be 2n = 20A + XY, demonstrating that this organism has the modal chromosome set found in triatomines. This new information concerning males of this species contributed to biological data that will be useful for understanding this potentially important Chagas disease vector.

Effects of sodium valproate on the chromatin of Triatoma infestans (Klug, 1834) (Hemiptera, Reduviidae) under in vitro culture conditions.

Sodium valproate (VPA) is a classic anticonvulsive, a histone deacetylase inhibitor, and a chromatin remodeling inducer. When injected into specimens of Triatoma infestans, a vector of Chagas disease, VPA affects the chromatin supraorganization of chromocenter heterochromatin in only a few cells of the Malpighian tubules. To test whether this result was explained by the inaccessibility of all of the organ's cells to the drug, we investigated the nuclear phenotypes and global acetylation of lysine 9 in histone H3 (H3K9ac) in Malpighian tubules cultivated in vitro for 1-24 h in the presence of 0.05 mM-1 mM VPA. The present results revealed that the chromatin decondensation event in the chromocenter body, which was detected only under low VPA concentrations up to a 4-h treatment, was not frequent during organ culture, similar to the results for injected insects. Cultivation of T. infestans Malpighian tubules in vitro for 24 h revealed inadequate for cell preservation even in the absence of the drug. Immunofluorescence signals for H3K9ac following VPA treatment showed a slightly increased intensity in the euchromatin, but were never detected in the chromocenter bodies, except with great intensity at their periphery, where the 18S rDNA is located. In conclusion, when VPA affects the chromocenter heterochromatin in this animal cell model, it occurs through a pathway that excludes a classic global H3K9ac mark. Investigation of nonhistone proteins associated with histone methylation marks is still required to further explain the differential response of T. infestans chromatin to VPA.

Spermatological Morphology of Triatoma Species (Hemiptera: Reduviidae: Triatominae).

Spermatozoa are among the most diverse cell types, and their morphologies often provide data that can be used to reliably evaluate phylogenetic relationships. They can also help to clarify the nature of 'specific complexes', which are common among triatomines. In the present study, we evaluated the copulation behavior of Triatoma rubrovaria Blanchard 1843 (Hemiptera: Reduviidae: Triatominae) and the structural morphology of sperm from T. carcavalloi Jurberg Rocha & Lent, 1998, T. infestans Klug, 1834, T. pintodiasi Jurberg Cunha & Rocha, 2013, and T. rubrovaria. Copulatory behavior was described from the moment males and females genitalia joined until they separated. Insemination was confirmed by the presence of a spermatophore in the female's bursa copulatrix. To measure their sperm, males were dissected and their seminal vesicles were removed, squashed on glass slides, and then spread, fixed, and observed under a photomicroscope. The images obtained were analyzed to measure the sperm. Seminal vesicles were also prepared for transmission electron microscopy. We performed K-means clustering separately for each species to group their sperm based on morphology. The differences in spermatozoa length among species of Triatominae, sperm types, and the interaction between species and sperm type were assessed with a two-way analysis of variance (ANOVA). The copulation time in T. rubrovaria was 3 to 5 min, which was sufficiently long for spermatophore transfer. All taxa showed polymorphic (short and long) sperm, with significant differences in the lengths of sperm among taxa. Using electron microscopy, the sperm cells of the four taxa examined were found to have similar ultrastructural morphology, confirming the hypothesized synapomorphies of sperm within the suborder Heteroptera (Hemiptera).

Morphology of the spermatheca of Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859).

Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) is a potential vector of Chagas's disease and the comprehension of its reproductive biology is an important tool to control this insect. In the reproductive tract of female insects, the spermatheca plays a crucial role storing male spermatozoa after mating. Whithin insects the spermatheca shows a wide morphological diversity and the analysis of this characteristic can contribute to understand the reproductive biology of the species. This study describes the histology and histochemistry of the spermatheca of T. lecticularia. Females have a pair of elongated spermathecal reservoirs without associated accessory gland. The reservoir opens into the common oviduct via a narrow muscular duct. The reservoir epithelium has single layer of columnar secretory cells. The control of the release of spermatozoa from the spermatheca occurs via the muscular duct. The anatomical features of the spermatheca of T. lecticularia resemble those described of other Reduviidae. However, the histological and histochemical features of spermatheca observed in T. lecticularia were important to explain the maintenance of the viability of the spermatozoa stored.

Localization of cholecystokinin-like immunoreactivity in the central nervous system of Triatoma infestans (Insecta: Heteroptera).

The distribution of cholecystokinin-like immunoreactivity was studied in the central nervous system of the heteropteran insect Triatoma infestans using high-sensitivity immunocytochemistry. In the protocerebrum, CCK-IR somata were observed in the anteromedial, anterolateral and posterior cell-body layers. The neuropils displayed different densities of immunoreactive neurites. Few immunoreactive somata were found in the optic lobe in both the medial and lateral soma rinds, as well as in the proximal optic lobe. Immunoreactive fibers were present in the medulla and lobula neuropils. The sensory deutocerebrum contained a higher number of immunopositive perikarya than the antennal mechanosensory and motor center. The antennal lobe glomeruli displayed a moderate density of immunoreactive fibers. With regard to the subesophageal ganglion, numerous CCK-IR somata were found close to the root of the mandibular nerve; others were present in the soma rind of the remaining neuromeres. CCK-IR perikarya were present in both thoracic ganglia, with the abdominal neuromeres containing the highest number of positive somata. The neuropils of both ganglia showed moderate densities of immunopositive processes. The distribution of CCK-LI in somata and neuropils of central nervous system of T. infestans is widespread suggesting that a CCK-like peptide may act mainly as a neuromodulator in the integration of information from distinct sensory receptors.

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.

The modern morphometric approach to identify eggs of Triatominae.

BACKGROUND: Egg morphometrics in the Triatominae has proved to be informative for distinguishing tribes or genera, and has been based generally on traditional morphometrics. However, more resolution is required, allowing species or even population recognition, because the presence of eggs in the domicile could be related to the species ability to colonize human dwellings, suggesting its importance as a vector. RESULTS: We explored the resolution of modern morphometric methods to distinguish not only tribes and genera, but also species or geographic populations in some important Triatominae. Four species were considered, representing two tribes and three genera: Panstrongylus chinai and P. howardi, Triatoma carrioni and Rhodnius ecuadoriensis. Within R. ecuadoriensis, two geographical populations of Ecuador were compared. For these comparisons, we selected the most suitable day of egg development, as well as the possible best position of the egg for data capture. The shape of the eggs in the Triatominae does not offer true anatomical landmarks as the ones used in landmark-based morphometrics, except for the egg cap, especially in eggs with an evident "neck", such as those of the Rhodniini. To capture the operculum shape variation, we used the landmark- and semilandmark-based method. The results obtained from the metric properties of the operculum were compared with the ones provided by the simple contour of the whole egg, as analyzed by the Elliptic Fourier Analysis. Clear differences could be disclosed between the genera, between the species - among which two very close species (P. chinai and P. howardi), as well as between two allopatric, conspecific populations. The whole egg contour (including the operculum) produced reclassification scores much more satisfactory than the ones obtained using the operculum only. CONCLUSIONS: We propose the outline-based approach as the most convenient characterization tool to identify unknown eggs at the species or population levels.

Histone epigenetic marks in heterochromatin and euchromatin of the Chagas' disease vector, Triatoma infestans.

Triatoma infestans, a vector of Chagas' disease, shows several particular cell biology characteristics, including the presence of conspicuous heterochromatic bodies (chromocenters) where DNA methylation has not been previously detected. Whether histone modifications contribute to the condensed state of these bodies has not yet been studied. Here, we investigated epigenetic modifications of histones H3 and H4 and presence of the non-histone heterochromatin protein (HP1-α) in the chromocenters and euchromatin of T. infestans cell nuclei, using immunocytochemistry. The effect of different concentrations of the histone deacetylase inhibitors valproic acid (VPA) and sodium butyrate (NaBt) on chromocenter condensation was visually examined; in VPA-treated specimens, this effect was also analyzed by image analysis. Trimethylated H3K9 signals, which were revealed in chromocenter and non-chromocenter areas, were strongest in chromocenters, whereas selected acetylated histone marks and mono- and dimethylated H3K9 and H4K20 signals were detected only in euchromatin. Weak trimethylated H4K20 signals and variable distribution of HP1-α were detected in chromocenters of part of the cellular population analyzed. Although specific VPA and NaBt treatment conditions affected the heterochromatin condensation pattern, they did not induce a decrease in survival and molting rates of the T. infestans nymphs. The VPA-induced chromatin remodeling was not accompanied by induction of H3K9 acetylation in chromocenters. Present findings regarding histone modifications and effects following VPA or NaBt treatments did not yet solve the question of which factors are responsible for maintenance of the condensed state of chromocenters in T. infestans. A possibility requiring further investigation remains on histone methylation marks and/or non-histone proteins.

The fine structure of the ocelli of Triatoma infestans (Hemiptera: Reduviidae).

The morphology and fine structure of the ocelli of Triatoma infestans have been analyzed by means of light and electron microscopy. The two dorsal ocelli of this species are located behind the compound eyes, looking dorsally and frontally. Externally, the ocelli are marked by the corneal lenses virtually spherical in form and limited internally by a cuticular apodeme. The lens focuses the incoming rays beyond the retina. A single layer of corneagen cells lies below the cuticular lens. The corneagen cells and photoreceptors are arranged in a cup-like fashion beneath the cuticular lens. A distal retinal zone comprises the rhabdoms, which are laterally connected in an hexagonal meshwork. A middle retinal zone comprises the receptor cell segment free of rhabdom, and a proximal zone their axons. In the middle zone, the oviform nuclei and spheroids are located. Screening pigment granules are present within the retinal cell. Spherical mitochondria are homogeneously distributed in the cytoplasm of the cell body. In the axonal zone, mitochondria are found in the peripheral region. Axons from receptor cells extend into the ocellar neuropile at the base of the ocelli, to synapse with second order neurons. The large axons of second order neurons are bundled by glial cells. The ocellar plexus exhibits a high diversity of synaptic unions (i.e. axo-dendritic, axo-axonic, dendro-axonic, and dendro-dendritic).

Differentiation and morphogenesis of Triatoma infestans (Klug 1834) female gonads. I--post embryonic development.

The post-embryonic development of the female gonads in Triatoma infestans (Hemiptera, Heteroptera), insects of importance in health affairs as harbors and vectors of different tripanosomatidea flagellates, is presented in a complete follow-up since insect hatches from the egg up to the last molt in the fifth instar stage. The detailed description of the morphological changes which occur in each instar as well as careful measurements evaluating its size increase have been analyzed by stereomicroscopy, phase contrast, dark field, and oblique illumination, in order to optimize the observations as well the photographic register of gonad morphology and structure. The analysis was performed on gonad specimens obtained from broods no less than twenty (20) nymph bugs, reared at constant temperature and fed-up regularly. According to the results of our study we can assert that gonad differentiation takes place in early phases of the insect development. Such is the case that first instars nymph's present absolutely and easily recognizably male and female gonads. From the third instar on beside the filament region, the three zones in each ovariole body is distinguished, a differentiation that is more noticeable during the fourth instar where a definite organization is present at the vitelarium. Such a clear cut zone development continues intensively during the fifth instar. Finally at the end of such fifth nymph stage and when the last molt toward adults is prepared, clear signs of ovariole maturation take place since oocytes in early vitelogenesis are found.

Hydrophobic attachment of Trypanosoma cruzi to the rectal cuticle of Triatoma infestans and its influence on metacyclogenesis - a review.

Trypanosoma cruzi colonizes mainly the rectum of the vector, especially the rectal glands. We investigated the basic architecture of the rectal cuticle of Triatoma infestans and the mode of attachment of T. cruzi in the small intestine and the rectum. In addition, we determined the capacity of culture-derived epimastigotes to attach to artificial substrates and the influence of attachment on metacyclogenesis. After incubation of the rectum with wheat germ lectin (WGA) coupled to gold particles, the procuticle contained chitin, but the two layers of the epicuticle and the superficial layer bordering the rectal lumen did not. The specific fluorochrom Nile Red stained the entire rectal cuticle green, indicating the waxy composition of the superficial layer. In electron microscopic analysis the parasites were attached to the hydrophobic superficial wax layer but not to the epithelium of the midgut. In vitro culture-derived epimastigotes attached with a high affinity to all hydrophobic substrates tested, whereas hydrophilic substrates did not permit attachment. Emulsified hexadecane localized the attachment molecules to the terminal part of the flagellum. Inhibition of attachment by coating the culture tubes with hydrophilic agarose and constant agitation decreased the rate of epimastigote to trypomastigote transformation, whereas wax coating enhanced metacyclogenesis.

Morphology of the spermatheca of Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) / Morfologia da espermateca de Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859)

Abstract Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) is a potential vector of Chagas's disease and the comprehension of its reproductive biology is an important tool to control this insect. In the reproductive tract of female insects, the spermatheca plays a crucial role storing male spermatozoa after mating. Whithin insects the spermatheca shows a wide morphological diversity and the analysis of this characteristic can contribute to understand the reproductive biology of the species. This study describes the histology and histochemistry of the spermatheca of T. lecticularia. Females have a pair of elongated spermathecal reservoirs without associated accessory gland. The reservoir opens into the common oviduct via a narrow muscular duct. The reservoir epithelium has single layer of columnar secretory cells. The control of the release of spermatozoa from the spermatheca occurs via the muscular duct. The anatomical features of the spermatheca of T. lecticularia resemble those described of other Reduviidae. However, the histological and histochemical features of spermatheca observed in T. lecticularia were important to explain the maintenance of the viability of the spermatozoa stored.

Resumo Triatoma lecticularia (Hemiptera: Reduviidae) (Stal, 1859) é um potencial vetor da doença de Chagas e a compreensão de sua biologia reprodutiva é um importante fator para seu controle populacional. No aparelho reprodutor feminino dos insetos, a espermateca desempenha a importante funcão de armazenar os espermatozoides após cópula. Nos insetos, a espermateca apresenta uma ampla diversidade morfológica e a análise destas características pode contribuir com o entendimento da biologia reprodutiva das espécies. Este estudo descreve histológica e histoquimicamente a espermateca de T. lecticularia. As fêmeas tem um par de espermatecas alongadas sem glândulas acessórias associadas. O reservatório conecta-se ao oviduto comum através de um ducto muscular curto que controla a liberação dos espermatozoides. O epitélio do reservatório possui uma camada de células secretoras colunares. As características anatômicas da espermateca de T. lecticularia são semelhantes às encontradas em outros Reduviidae. Entretanto, as características histológicas e histoquímicas observadas na espermateca são importantes para explicar a manutenção da viabilidade dos espermatozoides armazenados.

Spatial distribution of AT- and GC-rich DNA within interphase cell nuclei of Triatoma infestans Klug.

Heterochromatin bodies in single- and multichromocentered interphase cell nuclei of Triatoma infestans, a vector of Chagas disease, have been suggested to contain AT-rich DNA, based on their positive response to Q-banding and Hoechst 33248 treatment. No information exists on whether GC-rich DNA is also present in these nuclei and whether it plays a role on chromatin condensation. Considering that methodologies more precise than those previously used to determine DNA base composition in situ are currently available, and that the spatial distribution of chromatin areas differing in composition in interphase cell nuclei of different species is a matter of interest, the localization of AT- and GC-rich DNA in T. infestans nuclei is revisited here. The methodologies used included DAPI/AMD and CMA(3)/Distamycin differential staining, Feulgen-DNA image analysis following Msp I and Hpa II enzymatic digestion, 5-methylcytidine immunodetection, AgNOR response, confocal microscopy, and the 5-aza-2'-deoxycytidine (5-AZA) demethylation assay. The results identified the presence of AT-rich/GC-poor DNA in chromocenters and evenly distributed AT and GC sequences in euchromatin. A GC-rich DNA zone encircling the chromocenters was also found but it could not be associated with NOR regions. To corroborate the DNA AT-richness in T. infestans nuclei, bioinformatic analyses were also performed. Methylated cytosine was evident at some points of the chromocenters' edge in single- and multichromocentered nuclei and at the euchromatin of multichromocentered nuclei and could be transiently affected by the 5-AZA treatment. The present results suggest that in the particular case of chromocenters of the hemipteran T. infestans, cytosine methylation is not a relevant factor involved in chromatin condensation.

Digestive cells in the midgut of Triatoma vitticeps (Stal, 1859) in different starvation periods.

Triatoma vitticeps (Stal, 1859) is a hematophagous Hemiptera that, although being considered wild, can be found in households, being a potential Chagas' disease vector. This work describes the histology and ultrastructure of the midgut of T. vitticeps under different starvation periods. Fifteen adults of both sexes starved for 3, 7, 20 and 25 days were studied. In general, digestive cells had apical microvilli, basal plasma membrane infoldings and central nucleus. The perimicrovillar membrane was found in all insects examined. Digestive cells of anterior midgut had lipid droplets, glycogen granules, developed basal labyrinth associated with mitochondria suggesting their role in nutrient storage and in fluid and ion transport. The cells of median and posterior regions of the midgut were rich in rough endoplasmic reticulum, lysosomes, vesicles and granules with different electron-densities. Moreover, cells of the posterior portion of the midgut had hemozoyn granules and mitochondria in the apical cytoplasm close to microvilli, suggesting their role in blood digestion and active nutrient absorption. The midgut of T. vitticeps showed differences in digestive cells associated with the time after feeding, and the increase of vesicles amount in long starvation periods, which suggests enzyme storage, which is readily used after a blood meal.