Belo Horizonte, Brazil
Belo Horizonte, Brazil

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Valderrama A.,Federal University of Viçosa | Valderrama A.,Instituto Conmemorativo Gorgas Of Estudios Of La Salud | Tavares M.G.,Federal University of Viçosa | Filho J.D.A.,Instituto Rene Rachou Fiocruz
Memorias do Instituto Oswaldo Cruz | Year: 2011

In Panama, species of the genus Lutzomyia are vectors of American cutaneous leishmaniasis (ACL). There is no recent ecological information that may be used to develop tools for the control of this disease. Thus, the goal of this study was to determine the composition, distribution and diversity of Lutzomyia species that serve as vectors of ACL. Sandfly sampling was conducted in forests, fragmented forests and rural environments, in locations with records of ACL. Lutzomyia gomezi, Lutzomyia panamensis and Lutzomyia trapidoi were the most widely distributed and prevalent species. Analysis of each sampling point showed that the species abundance and diversity were greatest at points located in the fragmented forest landscape. However, when the samples were grouped according to the landscape characteristics of the locations, there was a greater diversity of species in the rural environment locations. The Kruskal Wallis analysis of species abundance found that Lu. gomezi and Lu. trapidoi were associated with fragmented environments, while Lu. panamensis, Lutzomyia olmeca bicolor and Lutzomyia ylephiletor were associated with forested environments. Therefore, we suggest that human activity influences the distribution, composition and diversity of the vector species responsible for leishmaniasis in Panama.

Martins G.F.,Federal University of Viçosa | Serrao J.E.,Federal University of Viçosa | Ramalho-Ortigao J.M.,Kansas State University | Pimenta P.F.P.,Instituto Rene Rachou Fiocruz
Memorias do Instituto Oswaldo Cruz | Year: 2011

The insect fat body plays major roles in the intermediary metabolism, in the storage and transport of haemolymph compounds and in the innate immunity. Here, the overall structure of the fat body of five species of mosquitoes (Aedes albopictus, Aedes fluviatilis, Culex quinquefasciatus, Anopheles aquasalis and Anopheles darlingi) was compared through light, scanning and transmission electron microscopy. Generally for mosquitoes, the fat body consists of lobes projecting into the haemocoel and is formed by great cell masses consisting of trophocytes and oenocytes. Trophocytes are rich in lipid droplets and protein granules. Interestingly, brown pigment granules, likely ommochromes, were found exclusively in the trophocytes located within the thorax and near the dorsal integument of Anopheles, which is suggestive of the role these cells play in detoxification via ommochrome storage. This study provides a detailed comparative analysis of the fat body in five different mosquito species and represents a significant contribution towards the understanding of the structural-functional relationships associated with this organ.

Martins G.F.,Federal University of Viçosa | Ramalho-Ortigao J.M.,Kansas State University | Lobo N.F.,University of Notre Dame | Severson D.W.,University of Notre Dame | And 2 more authors.
Memorias do Instituto Oswaldo Cruz | Year: 2011

Oenocytes are ectodermic cells present in the fat body of several insect species and these cells are considered to be analogous to the mammalian liver, based on their role in lipid storage, metabolism and secretion. Although oenocytes were identified over a century ago, little is known about their messenger RNA expression profiles. In this study, we investigated the transcriptome of Aedes aegypti oenocytes. We constructed a cDNA library from Ae. aegypti MOYO-R strain oenocytes collected from pupae and randomly sequenced 687 clones. After sequences editing and assembly, 326 high-quality contigs were generated. The most abundant transcripts identified corresponded to the cytochrome P450 superfamily, whose members have roles primarily related to detoxification and lipid metabolism. In addition, we identified 18 other transcripts with putative functions associated with lipid metabolism. One such transcript, a fatty acid synthase, is highly represented in the cDNA library of oenocytes. Moreover, oenocytes expressed several immunity-related genes and the majority of these genes were lysozymes. The transcriptional profile suggests that oenocytes play diverse roles, such as detoxification and lipid metabolism, and increase our understanding of the importance of oenocytes in Ae. aegypti homeostasis and immune competence.

Tavares M.G.,Federal University of Viçosa | Filho J.D.A.,Instituto Rene Rachou Fiocruz
Parasites and Vectors | Year: 2014

Background: Lutzomyia gomezi (Nitzulescu, 1931) is one of the main Leishmania (Vianna) panamensis vectors in Panama, and despite its medical significance, there are no population genetic studies regarding this species. In this study, we used the sequences of the mitochondrial gene cytochrome b/start of NADH1 and the nuclear elongation gene -1 in order to analyze genetic variation and phylogeographic structure of the Lu. gomezi populations. Methods. A total of 86 Lu. gomezi individuals were captured in 38 locations where cutaneous leishmaniasis occurred. DNA was extracted with phenol/chloroform methods and amplification of genes was performed using PCR primers for mitochondrial and nuclear markers. Results: We found a total of 37 and 26 haplotypes of mitochondrial and nuclear genes, high haplotype diversity (h) for all three populations were detected with both molecular markers. Nucleotide diversity (π) was estimated to be high for all three populations with the mitochondrial marker, which was opposite to the estimate with the nuclear marker. In the AMOVA Φst recorded moderate (mitochondrial) and small (nuclear) population structure with statistical significance among populations. The analysis of the fixation index (Fst) used to measure the differentiation of populations showed that with the exception of the population located in the region of Bocas del Toro, the other populations presented with minor genetic differentiation. The median-Joining network of the mitochondrial marker reveled three clusters and recorded four haplotypes exclusively of localities sampled from Western Panama, demonstrating strong divergence. We found demographic population expansion with Fús Fs neutrality test. In the analysis mismatch distribution was observed as a bimodal curve. Conclusion: Lu. gomezi is a species with higher genetic pool or variability and mild population structure, due to possible capacity migration and local adaptation to environmental changes or colonization potential. Thus, knowledge of the genetic population and evolutionary history is useful to understand the implications of different population genetic structures for cutaneous leishmaniasis epidemiology. © 2014 Valderrama et al.; licensee BioMed Central Ltd.

Carvalho G.M.L.,Instituto Rene Rachou Fiocruz | Brazil R.P.,Instituto Oswaldo Cruz Fiocruz | Sanguinette C.C.,Instituto Rene Rachou Fiocruz | Filho J.D.A.,Instituto Rene Rachou Fiocruz
Memorias do Instituto Oswaldo Cruz | Year: 2010

As inhabitants of forested areas, caves and anthropic environments, the phlebotomines deserve special attention because some species are able to transmit trypanosomatids, bacteria and viruses to vertebrates. Phlebotomines are also a nuisance because they cause painful bites, which may ultimately produce allergic manifestations. The lack of information about the presence and behaviours of sand flies in caves has aroused the curiosity of researchers for a long time. In the present paper, we describe a new species of sand fly that was captured in a cave located in the municipal district of Arraias in the southeastern region of the state of Tocantins. The morphological features of this new species permit it to be included in the alphabetica group of the Martinsmyia genus.

Quaresma P.F.,Instituto Rene Rachou Fiocruz | Carvalho G.M.L.,Instituto Rene Rachou Fiocruz | Ramos M.C.N.F.,Instituto Rene Rachou Fiocruz | Filho J.D.A.,Instituto Rene Rachou Fiocruz
Memorias do Instituto Oswaldo Cruz | Year: 2012

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

da Graca G.C.,Instituto Oswaldo Cruz | Volpini A.C.,Instituto Oswaldo Cruz | Volpini A.C.,Instituto Rene Rachou Fiocruz | Romero G.A.S.,University of Brasilia | And 5 more authors.
Memorias do Instituto Oswaldo Cruz | Year: 2012

In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorfism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.

Krautz-Peterson G.,Tufts University | Simoes M.,Instituto Rene Rachou FIOCRUZ | Faghiri Z.,Tufts University | Ndegwa D.,Tufts University | And 4 more authors.
PLoS Pathogens | Year: 2010

Adult schistosomes live in the host's bloodstream where they import nutrients such as glucose across their body surface (the tegument). The parasite tegument is an unusual structure since it is enclosed not by the typical one but by two closely apposed lipid bilayers. Within the tegument two glucose importing proteins have been identified; these are schistosome glucose transporter (SGTP) 1 and 4. SGTP4 is present in the host interactive, apical tegumental membranes, while SGTP1 is found in the tegumental basal membrane (as well as in internal tissues). The SGTPs act by facilitated diffusion. To examine the importance of these proteins for the parasites, RNAi was employed to knockdown expression of both SGTP genes in the schistosomula and adult worm life stages. Both qRT-PCR and western blotting analysis confirmed successful gene suppression. It was found that SGTP1 or SGTP4-suppressed parasites exhibit an impaired ability to import glucose compared to control worms. In addition, parasites with both SGTP1 and SGTP4 simultaneously suppressed showed a further reduction in capacity to import glucose compared to parasites with a single suppressed SGTP gene. Despite this debility, all suppressed parasites exhibited no phenotypic distinction compared to controls when cultured in rich medium. Following prolonged incubation in glucose-depleted medium however, significantly fewer SGTP-suppressed parasites survived. Finally, SGTP-suppressed parasites showed decreased viability in vivo following infection of experimental animals. These findings provide direct evidence for the importance of SGTP1 and SGTP4 for schistosomes in importing exogenous glucose and show that these proteins are important for normal parasite development in the mammalian host. © 2010 Krautz-Peterson et al.

Periago M.V.,Instituto Rene Rachou FIOCRUZ | Bethony J.M.,George Washington University
Microbes and Infection | Year: 2012

Hookworm disease from Necator americanus and Ancylostoma duodenale affects approximately 700 million people, with N. americanus being the predominant species. Unlike other pathogens (e.g., bacterial infections), where " virulence" is described in regards to acute pathogenesis and case-fatality, hookworms are well-evolved, multicellular parasites that establish long-term infections in their human hosts with a subtle and chronic, but insidious, pathogenesis, usually in the form of iron deficiency anemia from parasite blood feeding that, over time, has devastating effects on the human host especially when it involves children or women of child bearing years. As such, many of the typical terms for " virulence factors" used in other reviews in this special edition cannot be applied to hookworm (e.g., " colonization", " invasion", " or " toxicity"); rather the virulence of hookworm infection comes in terms of their ability to maintain a chronic blood-feeding infection in the lumen of relatively healthy human hosts, an infection that is usually measured in years but can sometimes be measured in decades. In the current manuscript, we describe the routes of invasion hookworms take into their human hosts and the means by which they modulate the human immune system to maintain this long-term parasitism. Little data on hookworm infection comes from actual human infections; instead, much of the data is derived from observations of laboratory animal models, in which hookworms fail to establish this distinctive " chronic infection," either due to physiological or immunological responses of these animal models. Hence, the mode and effects of chronic immunity must be extrapolated from this very different sort of infection to humans. Herein, we aim to synthesize immunological information from both types of models in the context of immune regulation and protection in order to identify future research focuses for the development of new treatment alternatives (i.e. drugs and vaccines). © 2012 Institut Pasteur.

Dias J.C.P.,Instituto Rene Rachou FIOCRUZ
Revista da Sociedade Brasileira de Medicina Tropical | Year: 2011

Very soon Carlos Chagas took into account the need of trypanosomiasis control, considering its great social impact and geographical dispersion The vector was considered the more vulnerable target and housing improvement the basic strategy to face the disease. In parallel, it was required a more clinical visibility for the disease, as an argument for its control. The first concrete tentative ocorred in 1918 when Souza Araújo dedicating his efforts in Paraná, trying housing improvement. He was followed by Ezequiel Dias et al, in 1921, employing chemical compounds against the vector, The chemical fight will be retaken by Emmanuel Dias in 1944, assaying several old compounds, fire thrower and cyanidric gas. In 1946, DDT showed to be ineffective, but one year later Dias & Pellegrino described the insecticide gammexane, higly effective against domestic triatomines. Working with Mario Pinotti, expanded trials occurred in Minas Gerais (Triangle Region), justifying the expansion of the campaing to other endemic regions, with the rationale of continuous work in contiguous areas. In 1957 Pedreira de Freitas proposed the selective spraying, which was the model for the future strategy of program evaluation, by SUVEN and SUCAN organizations. In 1975 the national program is reorganized, launching two national surveys (entomology and serology). In 1979 the new pyrethroid compounds are tried and im 1983 the national program is expanded. Transfusion transmitted Chagas Disease was studied since the 1950 by the Nussenzweig group in S. Paulo, showing to be vulnerable to chemoprophylaxis and blood donor pre transfusional serologic screening. Nevertheless, these preventive measures only were implemented in the 1980 decade, following the emergence of HIV/AIDS pandemic. Practically, since the pioneer essays, the control of Chagas Disease transmission showed to be efficient against vector and blood bank mechanisms, depending on continuity, educative support and political will.

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