The External Morphology of the Eggs of Culex (Culex) saltanensis (Diptera: Culicidae) Under Scanning Electron Microscopy.

The aim of the present study was to describe the morphology of the eggs of Culex (Culex) saltanensis Dyar that occurs in the Neotropical region. Eggs of the Cx. (Cux.) saltanensis were collected at the Mata Atlântica FIOCRUZ campus, fixed in 1% osmium tetroxide, prepared for mounting on metal supports, observed under a scanning electron microscope, and described morphologically. The eggs had a coniform shape with a length of approximately 0.5 mm (505-510 µm) and a width in the median portion of 117 µm (113-123 µm). Upper portion is lined with tubers of irregular shape and varying sizes (0.64-1.31 µm), located on a cross-linked matrix forming bands observed under optical microscopy. The micropyle is encased in a necklace of approximately 6.6-µm plates arranged in a flower-like shape. Comparing Cx. (Cux.) saltanensis eggs with several species of different genera, important divergent characteristics can be observed. However, this study points to the need for new descriptions of eggs of species belonging to the same subgenus in order to analyze if there will be differences between them. Culex (Cux.) saltanensis eggs have particular characteristics not observed in eggs of other Culicidae genera.

Morphobiological, morphometric and ultrastructural characterization of sylvatic Trypanosoma cruzi isolates from Rio de Janeiro state, Brazil / Caracterização morfobiológica, morfométrica e ultraestrutural de isolados silvestres de Trypanosoma cruzi do estado do Rio de Janeiro, Brasil

Abstract Triatoma vitticeps is a triatomine with geographic distribution restrict to Brazil, which exhibits high prevalence of Trypanosoma cruzi natural infection. Of special epidemiologic concern, this species often invades households in the states of Rio de Janeiro, Minas Gerais and Espírito Santo. The objective of this study was to evaluate morphological and ultrastructural parameters on three T. cruzi isolates obtained from wild T. vitticeps specimens. The growth and cell differentiation of the parasite was evaluated through epimastigote and trypomastigote forms obtained in the growth curves for three distinct isolates. The maximum growth showed differences at the 20th day of the curve. Our in vitro results show a heterogeneity, regarding these features for samples cultivated under the same conditions. Morphometric analyzes based on the shape of epimastigotes and trypomastigotes corroborated such differentiation. These results highlight the need of better understanding the meaning of this diversity under an eco-epidemiological perspective.

Resumo Triatoma vitticeps é um triatomíneo com distribuição geográfica restrita ao território brasileiro, apresentando alta prevalência de infecção natural pelo Trypanosoma cruzi. Esta espécie é relevante sob o ponto de vista epidemiológico por invadir domicílios com frequência nos estados do Rio de Janeiro, Minas Gerais e Espírito Santo. O objetivo deste estudo foi avaliar parâmetros morfológicos e ultraestruturais, em três isolados de T. cruzi obtidos a partir de triatomíneos silvestres. O crescimento e a diferenciação celular do parasita foi avaliado através das formas epimastigotas e tripomastigotas obtidas nas curvas de crescimento para os três isolados. O crescimento máximo mostrou diferenças no 20º dia da curva. Nossos resultados in vitro mostram uma heterogeneidade, em relação a essas características para amostras cultivadas nas mesmas condições. As análises morfométricas baseadas na conformação de epimastigotas e trypomastigotes corroboraram essa diferenciação. Estes resultados ressaltam a necessidade de uma melhor compreensão do significado desta diversidade sob uma perspectiva eco-epidemiológica.

Morphobiological, morphometric and ultrastructural characterization of sylvatic Trypanosoma cruzi isolates from Rio de Janeiro state, Brazil.

Triatoma vitticeps is a triatomine with geographic distribution restrict to Brazil, which exhibits high prevalence of Trypanosoma cruzi natural infection. Of special epidemiologic concern, this species often invades households in the states of Rio de Janeiro, Minas Gerais and Espírito Santo. The objective of this study was to evaluate morphological and ultrastructural parameters on three T. cruzi isolates obtained from wild T. vitticeps specimens. The growth and cell differentiation of the parasite was evaluated through epimastigote and trypomastigote forms obtained in the growth curves for three distinct isolates. The maximum growth showed differences at the 20th day of the curve. Our in vitro results show a heterogeneity, regarding these features for samples cultivated under the same conditions. Morphometric analyzes based on the shape of epimastigotes and trypomastigotes corroborated such differentiation. These results highlight the need of better understanding the meaning of this diversity under an eco-epidemiological perspective.

Comparing the egg ultrastructure of three Psorophora ferox (Diptera: Culicidae) populations / Comparação da ultraestrutura de ovos de três populações de Psorophora ferox (Diptera: Culicidae)

Abstract Characterising the external morphology of mosquito eggs is important, since it facilitates the identification of material from breeding sites and contributes to the understanding of species biology and group systematics. Therefore, was to compare eggs from different Psorophora ferox populations using scanning electron microscopy (SEM). Eggs were obtained from adult female of Ps. ferox collected in the Poços das Antas Biological Reserve (Reserva Biológica de Poços das Antas, Rio de Janeiro, Brazil). From each female, one portion of eggs (n = 5) was reared for identification purposes, and the second portion (n = 10) was examined using SEM for morphometric analysis. The egg morphology was then compared to that of eggs from Ps. ferox populations in Florida (USA) and Arena (Trinidad). The exochorion ornamentation of the populations differs considerably in the morphology of the tubercles of the chorionic cells, external chorionic reticulum, micropylar collar, and micropyle.

Resumo Caracterização da morfologia externa dos ovos do mosquito é importante, uma vez que facilita a identificação de materiais a partir de locais de reprodução e contribui para a compreensão da biologia das espécies e sistemática do grupo. O objetivo do presente estudo foi comparar os ovos de diferentes populações Psorophora ferox usando MEV. Os ovos foram obtidos a partir de fêmeas de Ps. ferox que foram coletadas na Reserva Biológica de Poço das Antas (Reserva Biológica de Poço das Antas, Rio de Janeiro, Brasil). A partir de cada fêmea, uma parte dos ovos (n = 5) foi criado para fins de identificação, e uma segunda parte (n = 10) foi examinado usando MEV para análise morfométrica. Foi então comparada a morfologia dos ovos de populações de Ps.ferox da Flórida (EUA) e Arena (Trinidad). A ornamentação do exocório das populações difere consideravelmente na morfologia dos tubérculos das células coriônicas, retículo coriônico externo, colar micropilar e micrópila.

Comparing the egg ultrastructure of three Psorophora ferox (Diptera: Culicidae) populations.

Characterising the external morphology of mosquito eggs is important, since it facilitates the identification of material from breeding sites and contributes to the understanding of species biology and group systematics. Therefore, was to compare eggs from different Psorophora ferox populations using scanning electron microscopy (SEM). Eggs were obtained from adult female of Ps. ferox collected in the Poços das Antas Biological Reserve (Reserva Biológica de Poços das Antas, Rio de Janeiro, Brazil). From each female, one portion of eggs (n = 5) was reared for identification purposes, and the second portion (n = 10) was examined using SEM for morphometric analysis. The egg morphology was then compared to that of eggs from Ps. ferox populations in Florida (USA) and Arena (Trinidad). The exochorion ornamentation of the populations differs considerably in the morphology of the tubercles of the chorionic cells, external chorionic reticulum, micropylar collar, and micropyle.

Sperm dimorphism in the Triatoma brasiliensis species complex and its applications.

Morphological and structural features of the sperm of the Triatoma brasiliensis Neiva, 1911 (Hemiptera: Reduviidae) species complex were examined in this first study investigating the sperm of Heteroptera and the genus Triatoma. Males were dissected and their seminal vesicles removed. For measurement, seminal vesicles were squashed on glass slides, spread, fixed and observed under a photomicroscope. The images were analysed and measures of sperm were made. Data were analysed using one-way analysis of variance and Tukey's test to detect differentiation among taxa. Furthermore, seminal vesicles were prepared for studies of transmission electron microscopy. All taxa studied showed polymorphic (short and long) sperm. The sperm of Triatoma brasiliensis macromelasoma was significantly longer (in total length) than that of the other four members of the complex, which supports the hypothesis of hybrid speciation of this member of the complex as an increase in the size of typical hybrids under heterosis was previously shown. The sperm cells of the five taxa have similar ultrastructural morphology. The ultrastructural features observed confirm the hypothesis, raised by previous studies, that they are synapomorphic to the suborder Heteroptera.

Comparing the egg ultrastructure of three Psorophora ferox (Diptera: Culicidae) populations

Abstract Characterising the external morphology of mosquito eggs is important, since it facilitates the identification of material from breeding sites and contributes to the understanding of species biology and group systematics. Therefore, was to compare eggs from different Psorophora ferox populations using scanning electron microscopy (SEM). Eggs were obtained from adult female of Ps. ferox collected in the Poços das Antas Biological Reserve (Reserva Biológica de Poços das Antas, Rio de Janeiro, Brazil). From each female, one portion of eggs (n = 5) was reared for identification purposes, and the second portion (n = 10) was examined using SEM for morphometric analysis. The egg morphology was then compared to that of eggs from Ps. ferox populations in Florida (USA) and Arena (Trinidad). The exochorion ornamentation of the populations differs considerably in the morphology of the tubercles of the chorionic cells, external chorionic reticulum, micropylar collar, and micropyle.

Resumo Caracterização da morfologia externa dos ovos do mosquito é importante, uma vez que facilita a identificação de materiais a partir de locais de reprodução e contribui para a compreensão da biologia das espécies e sistemática do grupo. O objetivo do presente estudo foi comparar os ovos de diferentes populações Psorophora ferox usando MEV. Os ovos foram obtidos a partir de fêmeas de Ps. ferox que foram coletadas na Reserva Biológica de Poço das Antas (Reserva Biológica de Poço das Antas, Rio de Janeiro, Brasil). A partir de cada fêmea, uma parte dos ovos (n = 5) foi criado para fins de identificação, e uma segunda parte (n = 10) foi examinado usando MEV para análise morfométrica. Foi então comparada a morfologia dos ovos de populações de Ps.ferox da Flórida (EUA) e Arena (Trinidad). A ornamentação do exocório das populações difere consideravelmente na morfologia dos tubérculos das células coriônicas, retículo coriônico externo, colar micropilar e micrópila.

Morphobiological, morphometric and ultrastructural characterization of sylvatic Trypanosoma cruzi isolates from Rio de Janeiro state, Brazil

Abstract Triatoma vitticeps is a triatomine with geographic distribution restrict to Brazil, which exhibits high prevalence of Trypanosoma cruzi natural infection. Of special epidemiologic concern, this species often invades households in the states of Rio de Janeiro, Minas Gerais and Espírito Santo. The objective of this study was to evaluate morphological and ultrastructural parameters on three T. cruzi isolates obtained from wild T. vitticeps specimens. The growth and cell differentiation of the parasite was evaluated through epimastigote and trypomastigote forms obtained in the growth curves for three distinct isolates. The maximum growth showed differences at the 20th day of the curve. Our in vitro results show a heterogeneity, regarding these features for samples cultivated under the same conditions. Morphometric analyzes based on the shape of epimastigotes and trypomastigotes corroborated such differentiation. These results highlight the need of better understanding the meaning of this diversity under an eco-epidemiological perspective.

Resumo Triatoma vitticeps é um triatomíneo com distribuição geográfica restrita ao território brasileiro, apresentando alta prevalência de infecção natural pelo Trypanosoma cruzi. Esta espécie é relevante sob o ponto de vista epidemiológico por invadir domicílios com frequência nos estados do Rio de Janeiro, Minas Gerais e Espírito Santo. O objetivo deste estudo foi avaliar parâmetros morfológicos e ultraestruturais, em três isolados de T. cruzi obtidos a partir de triatomíneos silvestres. O crescimento e a diferenciação celular do parasita foi avaliado através das formas epimastigotas e tripomastigotas obtidas nas curvas de crescimento para os três isolados. O crescimento máximo mostrou diferenças no 20º dia da curva. Nossos resultados in vitro mostram uma heterogeneidade, em relação a essas características para amostras cultivadas nas mesmas condições. As análises morfométricas baseadas na conformação de epimastigotas e trypomastigotes corroboraram essa diferenciação. Estes resultados ressaltam a necessidade de uma melhor compreensão do significado desta diversidade sob uma perspectiva eco-epidemiológica.

Penial morphology in three species of Brazilian tuco-tucos, Ctenomys torquatus, C. minutus, and C. flamarioni (Rodentia: Ctenomyidae).

The present study analyses the glans penis and baculum morphology of three Brazilian tuco-tucos, Ctenomys torquatus Lichtenstein, 1830, Ctenomys minutus Nehring, 1887 and Ctenomys flamarioni Travi, 1981, in order to identify possible variations and understand some more about this taxonomically complex group. We used fixed penis from 15 previously listed adult specimens. For a more detailed baculum analysis, the penis underwent dissection and diaphanisation, whereas to analyse the glans penis surface we used Scanning Electron Microscopy (SEM). Results showed striking differences in baculum morphology among the three species. While C. minutus have a particular V-shaped proximal baculum tip, C. flamarioni baculum is thin throughout the shaft with rounded proximal and distal tips. Ctenomys torquatus have a shorter and larger baculum, similar to what has previously been described for the species. Glans penis surface microstructure analyses also revealed inter-specific differences, with penial spines varying in shape, size and, especially density. Although C. torquatus has a relatively small penis, it has the largest penial spine density, which suggests a more complex penial ornamentation in this species.

Male reproductive system structure and accessory glands ultrastructure of two species of Triatoma (Hemiptera, Reduviidae, Triatominae).

Morphological studies in male reproductive system of insect, and accessory glands functions, can contribute to the knowledge of reproductive aspects in Triatominae. The present study aims to describe the structure of male reproductive system under scanning electron microscopy, and the ultrastructure of accessory glands during the process of maturation, in T. brasiliensis and T. melanica of different ages. These system consist of two testis, two vas deferens, two seminal vesicles, four pair of accessory glands and one ejaculatory duct. The four mesodermal male accessory glands of T. melanica and T. brasiliensis, present secretion storage in the first days of adult life in starved males. During the maturation process of male accessory glands of T. brasiliensis and T. melanica, granules with different sizes, shapes and electron density were seen. In T. melanica small vacuoles are released into the gland lumen in one-day-old males, however, after three and five days few granules are found in the cell cytoplasm. The secretory granules in T. brasiliensis increase in size and amount, in the five days old adult. The secretory pathway in the male accessory glands of the two species seems to be merocrine and apocrine suggesting that different substances are being produced at different times. The cell microvilli in T. brasiliensis are longer and narrower, these can be absent in some regions of the cell surface of T. melanica. Maturation of male accessory glands in the triatomine species studied occurred without adult blood meal. Its function can led to new insights for the comprehension of reproductive aspects in Triatominae.

Major cysteine protease (cruzipain) in Z3 sylvatic isolates of Trypanosoma cruzi from Rio de Janeiro, Brazil.

Trypanosoma cruzi, the etiologic agent of Chagas' disease, is represented by a set of parasites which circulate between man, vectors, domestic and wild animals. Recently, our group isolated from Triatoma vitticeps strains of T. cruzi that were characterized as belonging to the Z3 phylogenetic lineage. Since very little is known about the biological and/or biochemical markers of sylvatic Z3 isolates, we have studied the protein and protease profiles of distinct Z3 isolates designated as SMM10, SMM53, SMM88, and SMM98. By means of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, both quantitative and qualitative differences were observed in the protein profiles of these strains. All strains produced an acidic cysteine protease of 45 kDa, resembling cruzipain activity. The strain SMM10 synthesized an additional 55 kDa metalloprotease. Using Western blotting and anti-cruzipain antibody to detect cruzipain-like molecules, a 40-kDa reactive molecule was identified in all strains; in the strain SMM10, an 80-kDa protein was also reacted. Studies about cruzipain isoforms from sylvatic parasites could be valuable tools in the comprehension of the genetic variability in the pathogenesis of Chagas' disease.

Ultrastructural features of the midgut epithelium of females Lutzomyia intermedia (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae).

A morphological study of the midgut of Lutzomyia intermedia, the primary vector of cutaneous leishmaniasis, in southeast Brazil, was conducted by light, scanning and transmission electron microscopy. The midgut is formed by a layer of epithelium of columnar cells on a non-cellular basal lamina, under which there is a musculature, which consists of circular and longitudinal muscular fibers. A tracheolar network is observed surrounding and penetrating in the musculature. Females were examined 12, 24, 48, 72 h and 5 days following a blood meal and were analyzed comparatively by transmission electron microscopy with starved females. In starved females, the epithelium of both the anterior and posterior sections of the midgut present whorl shaped rough endoplasmic reticulum. The posterior section does not present well-developed cellular structures such as mitochondria. Observations performed at 12, 24, 48 and 72 h after the blood meal showed morphological changes in the cellular structures in this section, and the presence of the peritrophic matrix up to 48 h after the blood meal. Digestion is almost complete and a few residues are detected in the lumen 72 h after blood feeding. Finally, on the 5th day after the blood meal all cellular structures present the original feature resembling that seen in starved sand flies. Morphometric data confirmed the morphological observations. Mitochondria, nuclei and microvilli of midgut epithelial cells are different in starved and blood fed females. The mitochondria present a similar profile in the epithelium of both the anterior and posterior section of the midgut, with higher dimension in starved females. The cell microvilli in the posterior section of the midgut of starved females are twice the size of those that had taken a blood meal. We concluded that there are changes in the midgut cellular structures of L. intermedia during the digestion of blood, which are in agreement with those described for other hematophagous diptera.