Interaction between Trypanosoma rangeli and the Rhodnius prolixus salivary gland depends on the phosphotyrosine ecto-phosphatase activity of the parasite

Trypanosoma rangeli is the trypanosomatid that colonizes the salivary gland of its insect vector, with a profound impact on the feeding capacity of the insect. In this study we investigated the role of the phosphotyrosine (P-Tyr) ecto-phosphatase activity of T. rangeli in its interaction with Rhodnius prolixus salivary glands. Long but not short epimastigotes adhered to the gland cells and the strength of interaction correlated with the enzyme activity levels in different strains. Differential interference contrast microscopy demonstrated that clusters of parasites are formed in most cases, suggesting cooperative interaction in the adhesion process. The tightness of the correlation was evidenced by modulating the P-Tyr ecto-phosphatase activity with various concentrations of inhibitors. Sodium orthovanadate, ammonium molybdate and zinc chloride decreased the interaction between T. rangeli and R. prolixus salivary glands in parallel. Levamisole, an inhibitor of alkaline phosphatases, affected neither process. EDTA strongly inhibited adhesion and P-Tyr ecto-phosphatase activity to the same extent, an effect that was no longer seen if the parasites were pre-incubated with the chelator and then washed. When the P-Tyr ecto-phosphatase of living T. ranged epimastigotes was irreversibly inactivated with sodium orthovanadate and the parasite cells were then injected into the insect thorax, colonization of the salivary glands was greatly depressed for several days after blood feeding. Addition of P-Tyr ecto-phosphatase substrates such as p-nitrophenyl phosphate (pNPP) and P-Tyr inhibited the adhesion of T. rangeli to salivary glands, but P-Ser, P-Thr and beta-glycerophosphate were completely ineffective. Immunoassays using anti-P-Tyr-residues revealed a large number of P-Tyr-proteins in extracts of R. prolixus salivary glands, which could be potentially targeted by T. rangeli during adhesion. These results indicate that dephosphorylation of structural P-Tyr residues on the gland cell surfaces, mediated by a P-Tyr ecto-phosphatase of the parasite, is a key event in the interaction between T. rangeli and R. prolixus salivary glands. (C) 2012 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.

Description of Rhodnius montenegrensis n. sp (Hemiptera: Reduviidae: Triatominae) from the state of Rondonia, Brazil

We present here a multisource approach that takes advantage of several disciplines to address a taxonomic issue. A triatomine related to Rhodnius robustus Larrousse, 1927 was recently found in the state of Rondonia, Brazil. The name Rhodnius montenegrensis n. sp. is suggested because it was found in the municipality of Monte Negro. The main differences between these two species can be detected in the female and male genitalia, but there are also noticeable differences in their eggs. Molecular analysis using PCR-RFLP technique and Bayesian inferences based on a fragment of the Cytochrome b (Cyt b) gene corroborated the morphological findings. We used this integrative approach to address the taxonomic decision for a new Rhodnius species and its relationship with others of this genus. Results obtained herein stress that morphology must be used as the major approach for obtaining phenotypic information, and molecular data should be taken as a complementary tool.

Identifying health centers in Honduras infested with Rhodnius prolixus using the seroprevalence of Chagas disease in children younger than 13 years

The objective of this study is to determine if a Chagas disease protocol starting with a serological survey is as reliable at identifying insect-infested areas as one using the gold standard entomological survey. The study found that health center areas infested with Rhodnius prolixus were identified using a threshold seroprevalence of 0.1%. The serological survey took half the time and was 30% less expensive than the entomological survey. Developing countries with limited resources may find this strategy useful in combating Chagas disease. This strategy also identifies seropositive children, which facilitates their treatment.

Estrutura de populações de Rhodnius neglectus Lent e Psammolestes tertius Lent & Jurberg (Hemiptera, Reduviidae) em ninhos de pássaros (Furnariidae) presentes na palmeira Mauritia flexuosa no Distrito Federal, Brasil

Rhodnius neglectus Lent, 1954 {Rn}e Psammolestes tertius Lent & Jurberg, 1965 {Pt} são triatomíneos que ocorrem em ninhos de aves, principalmente da família Furnariidae. O ciclo biológico dessas espécies é conhecido em condições de laboratório, sendo poucos estudos em ecótopos silvestres. Para analisar a infestação e estrutura de populações de Rn e Pt em ninhos de aves presentes na palmeira Mauritia flexuosa Linnaeus, em duas estações climáticas do Brasil Central, foram amostradas 41 palmeiras com evidências de nidificação dePhacellodomus ruber Vieillot, 1817 (22 na estação chuvosa e 19 na estação seca) em quatro áreas do Distrito Federal. Os insetos foram capturados usando-se coleta manual na copa da palmeira, identificados morfologicamente, separados por sexo e estádio ninfal. Fezes e glândulas salivares de Rn foram examinadas para verificar infecção por Trypanosoma cruzi Chagas, 1909 e/ou T. rangeli Tejera, 1920. Trinta e cinco palmeiras com ninhos de P. ruber estavam infestadas por Rn (85%) e 22 por Pt (53%). 442 indivíduos foram coletados na estação seca (200 Rn e 242 Pt) e 267 na estação chuvosa (136 Rn e 131 Pt). O único fator relacionado significativamente com a densidade de triatomíneos nas palmeiras foi a área. A estrutura etária das populações mostrou: a) maior abundância de adultos nas populações de Pt, b) maior abundância de machos em ambas as espécies e c) presença de fêmeas ovipondo em ambas as estações. Nenhum dos 177 triatomíneos examinados estava infectado por T. cruzi ou T. rangeli. A estrutura etária das populações de Rn e Pt não diferiu significativamente entre as estações amostradas, indicando ausência de marcada sazonalidade para essas espécies. _______________________________________________________________________________________ ABSTRACT

Domestic Rhodnius ecuadoriensis (Hemiptera, Reduviidae) infestation in Northern Peru: a comparative trial of detection methods during a six-month follow-up

Two passive methods in the assessment of intradomiciliary infestation by Rhodnius ecuadoriensis were tested: (i) the Gomes Nuñez sensor box (GN), (ii) sheets of white typing paper and (iii) one active timed manual method. The study was carried out in the Alto Chicama River Valley, Province of Gran Chimú, Department of La Libertad. The study design consisted of an initial searching of triatomines inside of the domestic environment by the manual capture active procedure (man/hour) covering all the studied houses. Then, matched pairs of GN boxes and paper sheets were simultaneously installed in the bedrooms of 207 households distributed in 19 localities. A comparative prospective trial of these passive detection devices were monitored at 2, 4 and, finally 6 months follow-up. Parasitological Trypanosoma rangeli and/or T. cruzi infections were investigated in two houses with high level of infestation by R. ecuadoriensis.16.9% of the 207 households investigated by an initial active manual method were infested with R. ecuadoriensis. The proportion of infested houses fluctuated from 6.2 to 55.5% amongst the 19 localities investigated. T. rangeli natural infection was detected in R.ecuadoriensis specimens collected in two households. Parasite rates in the bugs ranged from 16.6 to 21.7% respectively. The most striking fact was an average rate of salivary gland infection ranging from 7.4 to 8.3%. At the end of the sixth month period, a cumulative incidence of 31.4% of positive GN boxes against 15.9% for paper sheets was recorded. All three methods combined detected domestic infestation in 129 (62.3%) of the 207 houses studied in the 19 localities. The range of houses infested varies from 6.7% to 92.9%. In areas with low bug density infestation rates, the methodology experienced in our studies, seems to be the best choice for investigations on domestic R. ecuadoriensis populations.

Biochar a partir de biomassa de Miscanthus e Quebracho, produzido em forno tubular em sistema aberto e sob fluxo de nitrogênio.

2015

Administración en la cadena de suministro y su relación con el desempeño de las PYME del sector agroindustrial

En la actualidad las organizaciones están en búsqueda de una mejora continua, esto lleva a que las empresas hagan una revisión de estrategias que permitan alcanzar una posición de líderes en los diferentes sectores en los que se desempeñan. Una de las estrategias para lograr un alto posicionamiento en las nuevas empresas es la adecuada gestión que hacen de la cadena de suministro. El sector agroindustrial tiene un amplia cadena de suministro desde la obtención de materias primas hasta la llegada al cliente final, por su gran magnitud requiere de una adecuada administración de procesos que permitan ser eficaces y eficientes para alcanzar logros propuestos, aprovechar los recursos limitados con los que cuentan para su cumplimiento y, por último, la capacidad propia para la transformación de sus recursos. Para lograr que las empresas obtengan ventajas competitivas, es necesario que los distintos eslabones de la cadena de suministros cuenten con una capacidad de interacción que les permita, tanto agilizar como asegurar el éxito durante la incorporación de los nuevos productos en el mercado. Por medio de esta investigación se busca esclarecer y analizar la importancia de la administración en la cadena de suministro y la relación con el desempeño, basándose en las Pymes del sector agroindustrial. La intención es evaluar la información de diversos autores quienes han hablado de la relación que existe entre la cadena de suministro y su desempeño, teniendo en cuenta variables como el sector agroindustrial y las Pymes.

Evaluación de la validez de la escala llanto para el dolor en neonatos y menores de cinco años

Introducción: La escala LLANTO para dolor es una escala que hasta la fecha ha sido solo validada en población infantil española, actualmente no se conocen datos en población colombiana. Se pretende validar la escala de dolor LLANTO en pacientes neonatos y menores de 5 años, a través de su aplicación en pacientes atendidos en una de tres instituciones, además comparándola con las escalas FLACC y PIPP dependiendo de edad del paciente. Metodología: Se incluyeron niños con cualquier tipo de dolor, clasificándolos en dos grupos por edad: 1) neonatos y 2) niños entre 1 mes y 5 años de edad, que asistieron a la Fundación Cardioinfantil, Clínica Infantil Colsubsidio o al Hospital Universitario Mayor. Las escalas fueron aplicadas por dos residentes de pediatría y una enfermera especializada en el cuidado de población infantil. Para la prueba piloto se diseñó un cuestionario determinar dificultades en la aplicación de la escala LLANTO. Una vez corregidos los problemas identificados se procederá a la validación de la escala. Resultados: Se presentan los datos de la prueba piloto. Se incluyeron 8 neonatos y 8 niños entre 1 mes y 5 años, esta muestra fue obtenida en un periodo de un mes, con la encuesta se evaluó la aceptación y entendimiento de la escala LLANTO por parte de los evaluadores. La prueba piloto mostró resultados favorables en el 100% de los encuestados. Discusión: Se considera que la escala LLANTO no requiere cambios para continuar con su validación.

Genes of cathepsin L-like proteases in Trypanosoma rangeli isolates: Markers for diagnosis, genotyping and phylogenetic relationships

We have sequenced genes encoding cathepsin L-like (CatL-like) cysteine proteases from isolates of Trypanosoma rangeli from humans, wild mammals and Rhodnius species of Central and South America. Phylogenetic trees of sequences encoding mature CatL-like enzymes of T rangeli and homologous genes from other trypanosomes, Leishmania spp. and bodonids positioned sequences of T rangeli (rangelipain) closest to T cruzi (cruzipain). Phylogenetic tree of kinetoplastids based on sequences of CatL-like was totally congruent with those derived from SSU rRNA and gGAPDH genes. Analysis of sequences from the CatL-like catalytic domains of 17 isolates representative of the overall phylogenetic diversity and geographical range of T rangeli supported all the lineages (A-D) previously defined using ribosomal and spliced leader genes. Comparison of the proteolytic activities of T rangeli isolates revealed heterogeneous banding profiles of cysteine proteases in gelatin gels, with differences even among isolates of the same lineage. CatL-like sequences proved to be excellent targets for diagnosis and genotyping of T rangeli by PCR. Data from CatL-like encoding genes agreed with results from previous studies of kDNA markers, and ribosomal and spliced leader genes, thereby corroborating clonal evolution, independent transmission cycles and the divergence of T rangeli lineages associated with sympatric species of Rhodnius. (c) 2009 Elsevier B.V. All rights reserved.

Trypanosoma rangeli isolates of bats from Central Brazil: Genotyping and phylogenetic analysis enable description of a new lineage using spliced-leader gene sequences

Trypanosoma rangeli infects several mammalian orders but has never confidently been described in Chiroptera, which are commonly parasitized by many trypanosome species. Here, we described trypanosomes from bats captured in Central Brazil identified as T rangeli,.T. dionisii, T cruzimarinkellei and T cruzi. Two isolates, Tra643 from Platyrrhinus lineatus and Tra1719 from Artibeus plamirostris were identified as T rangeli by morphological, biological and molecular methods, and confirmed by phylogenetic analyses. Analysis using SSU rDNA sequences clustered these bat trypanosomes together with T rangeli from other hosts, and separated them from other trypanosomes from bats. Genotyping based on length and sequence polymorphism of PCR-amplified intergenic spliced-leader gene sequences assigned Tra1719 to the lineage A whereas Tra643 was shown to be a new genotype and was assigned to the new lineage E. To our knowledge, these two isolates are the earliest T rangeli from bats and the first isolates from Central Brazil molecularly characterized. Rhodnius stali captured for this study was found infected by T rangeli and T cruzi. (c) 2008 Elsevier B.V. All rights reserved.

Purification and partial characterization of a midgut membrane-bound alpha-glucosidase from Quesada gigas (Hemiptera: Cicadidae)

Adults of Quesada gigas (Hemiptera: Cicadidae) have a major alpha-glucosidase bound to the perimicrovillar membranes, which are lipoprotein membranes that surround the midgut cell microvilli in Hemiptera and Thysanoptera. Determination of the spatial distribution of alpha-glucosidases in Q. gigas midgut showed that this activity is not equally distributed between soluble and membrane-bound isoforms. The major membrane-bound enzyme was solubilized in the detergent Triton X-100 and purified to homogeneity by means of gel filtration on Sephacryl S-100, and ion-exchange on High Q and Mono Q columns. The purified alpha-glucosidase is a protein with a pH optimum of 6.0 against the synthetic substrate p-nitrophenyl alpha-D-glucoside and M(r) of 61,000 (SDS-PAGE). Taking into account V(Max)/K(M) ratios, the enzyme is more active on maltose than sucrose and prefers oligomaltodextrins up to maltopentaose, with lower efficiency for longer chain maltodextrins. The Q gigas alpha-glucosidase was immunolocalized in perimicrovillar membranes by using a monospecific polyclonal antibody raised against the purified enzyme from Dysdercus peruvianus. The role of this enzyme in xylem fluid digestion and its possible involvement in osmoregulation is discussed. (C) 2009 Elsevier Inc. All rights reserved.

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.

Acid phosphatase activity distribution in salivary glands of triatomines (Heteroptera, Reduviidae, Triatominae)

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

Ultrastructural analysis of the nucleolar aspects at spermiogenesis in triatomines (Heteroptera, Triatominae)

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

Study of the nucleolar cycle and ribosomal RNA distribution during meiosis in triatomines (Triatominae, Heteroptera)

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