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

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.

Hydrogen peroxide induces a specific DNA base change profile in the presence of the iron chelator 2, 2’ dipyridyl in Escherichia coli

Pretreatment of Escherichia coli cultures with the iron chelator 2,2’-dipyridyl (1 mM) protects against the lethal effects of low concentrations of hydrogen peroxide (<15 mM). However, at H2O2 concentrations equal to or greater than 15 mM, dipyridyl pretreatment increases lethality and mutagenesis, which is attributed to the formation of different types of DNA lesions. We show here that pretreatment with dipyridyl (1 mM) prior to challenge with high H2O2 concentrations (≥15 mM) induced mainly G:C→A:T transitions (more than 100X with 15 mM and more than 250X with 20 mM over the spontaneous mutagenesis rate) in E. coli. In contrast, high H2O2 concentrations in the absence of dipyridyl preferentially induced A:T→T:A transversions (more than 1800X and more than 300X over spontaneous mutagenesis for 15 and 20 mM, respectively). We also show that in the fpg nth double mutant, the rpoB gene mutation (RifS-RifR) induced by 20 mM H2O2 alone (20X higher) was increased in 20 mM H2O2 and dipyridyl-treated cultures (110X higher), suggesting additional and/or different lesions in cells treated with H2O2 under iron deprivation. It is suggested that, upon iron deprivation, cytosine may be the main damaged base and the origin of the pre-mutagenic lesions induced by H2O2.

Natural history of Ctenus medius Keyserling, 1891 (Aranae, Ctenidae) II: life cycle and aspects of reproductive behavior under laboratory conditions

Ctenus medius Keyserling, 1891 is a wandering spider common in the Brazilian Atlantic Forest. It has been the subject of few studies. Thus, this work aims to elucidate aspects of its natural history, such as the life cycle and reproductive behavior of this species, through laboratory and field observations. Two females with egg sacs were observed in the laboratory and one was observed in field (Barra Mansa, 22º32'S and 44º10'W) until the emergence of the spiderlings. For observation of the immature stage development, a portion of the spiderlings from the same hatch were taken to the laboratory and watched until sexual maturity. In the field, the period between the oviposition and the emergence of spiderlings was of 36 days. The female selects a site for egg sac deposition and stays there until the spiderlings emerge. Seven days after the emergence, the female abandoned the site where the egg sac was made, concomitant to the spiderlings dispersion from observation's place and until the moment that the spiderlings started to eat. For the spiderlings kept under laboratory conditions, cannibalism was not observed in the first instars (1-4th) when sufficient food was offered. Sexual maturity happened in the 14th or 15th instars, with an average of 309.2 to 344.5 days until the last/sexual molt, respectively. Until the date of sexual maturity, there was a mortality rate of 85 percent. This species is very fragile in captivity. This hampered deductions concerning longevity. Both females and males collected in the field were induced to mate in the laboratory. Courtship movements of males were registered, but the females did not permit the mating. These data may assist in initial biological studies of Ctenus genus and offer comparative parameters for studies of other related species

Effect of Plantago australis leaves on different gastric ulcer models

The anti-ulcerogenic effect of the crude ethanolic extract (CEE) of Plantago australis leaves was tested against ethanol-, indomethacin-, and cold restrain-induced stress ulcers. The CEE (500 and 1000 mg/kg) reduced the lesion index (LI) and the ulcer index in ethanol-induced ulcers, and the dose of 1000 mg/kg increased the amount of mucous. The highest dose of the CEE reduced the LI of cold restraint-induced stress ulcers when compared to the control group. The indomethacin-induced ulcers were not affected by this extract.

Natural history of Ctenus medius Keyserling, 1891 (Araneae, Ctenidae) I: observations on habitats and the development of chromatic patterns

Ctenus medius Keyserling, 1891 is a common species in several spots of Mata Atlântica, however there is a great lack of studies in all aspects of its natural history. This work aims to elucidate aspects of ecotope preference compared to large spiders, and to provide data on the development of chromatic patterns during its life cycle. The observations on the behavior of C. medius were done in the campus of Centro Universitário de Barra Mansa (UBM) by means of observations and nocturnal collections using cap lamps. For observations on the development of chromatic patterns, spiderlings raised in laboratory, hatched from an oviposition of a female from campus of UBM, and others spiderlings collected in field were used. The field observations indicate that: C. medius seems to prefer ecotopes characterized by dense shrub vegetation or herbal undergrowth; Lycosa erythrognatha and L. nordeskioldii seems to prefer open sites; Phoneutria nigriventer seems to prefer shrub vegetation and anthropogenic ecotopes as rubbish hills; Ancylometes sp. seems to prefer ecotopes near streams. Concerning chromatic patterns, it was observed that males and females show well distinct patterns during the last two instars, allowing distinction by sex without the use of a microscope. Through chromatic patterns it was also possible to draw a distinction between C. medius and C. ornatus longer that 3 mm cephalothorax width. 69 specimens of C. medius (males and females) collected in the campus of UBM did not show a striking polymorphism in chromatic pattern, but one among 7 adult females collected in National Park of Itatiaia, showed a distinct chromatic pattern

Comparação entre os protocolos IAEA/TRS-277 e AAPM/TG-39 com o novo protocolo IAEA/TRS-381 na dosimetria em feixes de elétrons / Comparison of the IAEA/TRS-277 and the AAPM/TG-39 protocols with the new IAEA/TRS-381 for electrons dosimetry

Este trabalho apresenta uma comparação dos resultados obtidos na determinação da dose absorvida na água em feixe de elétrons usando os formalismos da AAPM/TG-39 (1994) e o da IAEA/TRS-277 (1987), com o novo protocolo IAEA/TRS-381 (1997). As medidas foram realizadas no acelerador linear modelo Therac 20, que produz feixes de elétrons de 6, 9, 13, 17 e 20 MeV, instalado no Instituto Nacional de Câncer/RJ. A curva de percentual de ionização para cada energia foi obtida, utilizando uma câmara de ionização de placas paralelas, tipo NACP, em um fantoma de água, com a finalidade de especificar a qualidade dos feixes. A calibração da câmara utilizada foi feita através da comparação com uma câmara cilíndrica tipo FARMER, com fator de calibração em termos da grandeza kerma no ar, Nk, usando o feixe de 20 MeV, de acordo com o formalisma de cada um dos protocolos. Após sua calibração, a câmara de ionização de placas paralelas foi utilizada para determinar a dose absorvida na água em cada um dos feixes. A maior diferença encontrada quando comparada às respostas obtidas foi entre os protocolos IAEA/TRS-381 e AAPM/TG-39, para o feixe de 13 MeV, em torno de 1,1 por cento.

Lethal interaction between hydrogen peroxide and o-phenanthroline in Escherichia coli

The iron chelator o-phenanthroline enhances the lethal effect of H2O2 about four hundred times in Escherichia coli when both substances are added simultaneously to the culture mediu. If o-phenanthroline is added for increasing periods of time prior to the addition of H2O2, there is a shift from this lethal interaction to protection by the chelator about seven hundred times. It is known that the Fe2+ -o-phenanthroline(I) and Fe2+ -o-phenanthroline(II) complexes are formed quickly whereas the final and more stable Fe2+ -o-phenanthroline(III) complex is formed slowly, Moreover, the mono and bis complexes react with H2O2 to produce OH., whereas the tris complex is stable towards H2O2. Therefore, the lethal effect could be explained by the kinetics of reaction of o-phenanthroline with intracellular Fe2+, i.e., the mono and bis complexes are more reactive than intracellular Fe2+