The stabilizing effects of the acquired immunity on the schistosomiasis transmission modeling - the sensitivity Analysis

A mathematical model is proposed to analyze the effects of acquired immunity on the transmission of schistosomiasis in the human host. From this model the prevalence curve dependent on four parameters can be obtained. These parameters were estimated fitting the data by the maximum likelihood method. The model showed a good retrieving capacity of real data from two endemic areas of schistosomiasis: Touros, Brazil (Schistosoma mansoni) and Misungwi, Tanzania (S. haematobium). Also, the average worm burden per person and the dispersion of parasite per person in the community can be obtained from the model. In this paper, the stabilizing effects of the acquired immunity assumption in the model are assessed in terms of the epidemiological variables as follows. Regarded to the prevalence curve, we calculate the confidence interval, and related to the average worm burden and the worm dispersion in the community, the sensitivity analysis (the range of the variation) of both variables with respect to their parameters is performed.

Molecular Modeling Approaches for Determining Gene Function: application to a Putative Poly-A Binding Protein from Leishmania amazonensis (LaPABP)

The great expansion in the number of genome sequencing projects has revealed the importance of computational methods to speed up the characterization of unknown genes. These studies have been improved by the use of three dimensional information from the predicted proteins generated by molecular modeling techniques. In this work, we disclose the structure-function relationship of a gene product from Leishmania amazonensis by applying molecular modeling and bioinformatics techniques. The analyzed sequence encodes a 159 aminoacids polypeptide (estimated 18 kDa) and was denoted LaPABP for its high homology with poly-A binding proteins from trypanosomatids. The domain structure, clustering analysis and a three dimensional model of LaPABP, basically obtained by homology modeling on the structure of the human poly-A binding protein, are described. Based on the analysis of the electrostatic potential mapped on the model's surface and conservation of intramolecular contacts responsible for folding stabilization we hypothesize that this protein may have less avidity to RNA than it's L. major counterpart but still account for a significant functional activity in the parasite. The model obtained will help in the design of mutagenesis experiments aimed to elucidate the mechanism of gene expression in trypanosomatids and serve as a starting point for its exploration as a potential source of targets for a rational chemotherapy.

Human mixed infections of Leishmania spp. and Leishmania-Trypanosoma cruzi in a sub Andean Bolivian area: identification by polymerase chain reaction/hybridization and isoenzyme

Parasites belonging to Leishmania braziliensis, Leishmania donovani, Leishmania mexicana complexes and Trypanosoma cruzi (clones 20 and 39) were searched in blood, lesions and strains collected from 28 patients with active cutaneous leishmaniasis and one patient with visceral leishmaniasis. PCR-hybridization with specific probes of Leishmania complexes (L. braziliensis, L. donovani and L. mexicana) and T. cruzi clones was applied to the different DNA samples. Over 29 patients, 8 (27.6%) presented a mixed infection Leishmania complex species, 17 (58.6%) a mixed infection Leishmania-T. cruzi, and 4 (13.8%) a multi Leishmania-T. cruzi infection. Several patients were infected by the two Bolivian major clones 20 and 39 of T. cruzi (44.8%). The L. braziliensis complex was more frequently detected in lesions than in blood and a reverse result was observed for L. mexicana complex. The polymerase chain reaction-hybridization design offers new arguments supporting the idea of an underestimated rate of visceral leishmanisis in Bolivia. Parasites were isolated by culture from the blood of two patients and lesions of 10 patients. The UPGMA (unweighted pair-group method with arithmetic averages) dendrogram computed from Jaccard's distances obtained from 11 isoenzyme loci data confirmed the presence of the three Leishmania complexes and undoubtedly identified human infections by L. (V.) braziliensis, L. (L.) chagasi and L. (L.) mexicana species. Additional evidence of parasite mixtures was visualized through mixed isoenzyme profiles, L. (V.) braziliensis-L. (L.) mexicana and Leishmania spp.-T. cruzi.The epidemiological profile in the studied area appeared more complex than currently known. This is the first report of parasitological evidence of Bolivian patients with trypanosomatidae multi infections and consequences on the diseases' control and patient treatments are discussed.

Comparative studies on the biology and filarial susceptibility of selected blood-feeding and autogenous Aedes togoi sub-colonies

Blood-feeding and autogenous sub-colonies were selected from a laboratory, stock colony of Aedes togoi, which was originally collected from Koh Nom Sao, Chanthaburi province, Southeast Thailand. Comparative biology and filarial susceptibility between the two sub-colonies (blood-feeding: F11, F13; autogeny: F38, F40) were investigated to evaluate their viability and vectorial capacity. The results of comparison on biology revealed intraspecific differences, i.e., the average egg deposition/gravid female (F11/F38; F13/F40), embryonation rate (F13/F40), hatchability rate (F11/F38; F13/F40), egg width (F11/F38), wing length of females (F13/F40), and wing length and width of males (F11/F38) in the blood-feeding sub-colony were significantly greater than that in the autogenous sub-colony; and egg length (F11/F38) and width (F13/F40), and mean longevity of adult females (F11/F38) and males (F13/F40) in the blood-feeding sub-colony were significantly less than that in the autogenous sub-colony. The results of comparison on filarial susceptibility demonstrated that both sub-colonies yielded similar susceptibilities to Brugia malayi [blood-feeding/autogeny = 56.7% (F11)/53.3%(F38), 60%(F13)/83.3%(F40)] and Dirofilaria immitis [blood-feeding/autogeny = 85.7%(F11)/75%(F38), 45%(F13)/29.4%(F40)], suggesting autogenous Ae. togoi sub-colony was an efficient laboratory vector in study of filariasis.

Uncertainties regarding dengue modeling in Rio de Janeiro, Brazil

Dengue fever is currently the most important arthropod-borne viral disease in Brazil. Mathematical modeling of disease dynamics is a very useful tool for the evaluation of control measures. To be used in decision-making, however, a mathematical model must be carefully parameterized and validated with epidemiological and entomological data. In this work, we developed a simple dengue model to answer three questions: (i) which parameters are worth pursuing in the field in order to develop a dengue transmission model for Brazilian cities; (ii) how vector density spatial heterogeneity influences control efforts; (iii) with a degree of uncertainty, what is the invasion potential of dengue virus type 4 (DEN-4) in Rio de Janeiro city. Our model consists of an expression for the basic reproductive number (R0) that incorporates vector density spatial heterogeneity. To deal with the uncertainty regarding parameter values, we parameterized the model using a priori probability density functions covering a range of plausible values for each parameter. Using the Latin Hypercube Sampling procedure, values for the parameters were generated. We conclude that, even in the presence of vector spatial heterogeneity, the two most important entomological parameters to be estimated in the field are the mortality rate and the extrinsic incubation period. The spatial heterogeneity of the vector population increases the risk of epidemics and makes the control strategies more complex. At last, we conclude that Rio de Janeiro is at risk of a DEN-4 invasion. Finally, we stress the point that epidemiologists, mathematicians, and entomologists need to interact more to find better approaches to the measuring and interpretation of the transmission dynamics of arthropod-borne diseases.

Suppression of Brugia malayi (sub-periodic) larval development in Aedes aegypti (Liverpool strain) fed on blood of animals immunized with microfilariae

Preliminary studies were carried out to investigate the role of filarial specific antibodies, raised in an animal model against the filarial parasite, Brugia malayi (sub-periodic), in blocking their early development in an experimental mosquito host, Aedes aegypti (Liverpool strain). In order to generate filarial specific antibodies, Mongolian gerbils, Meriones unguiculatus, were immunized either with live microfilariae (mf) of B. malayi or their homogenate. Mf were harvested from the peritoneal cavity of Mongolian gerbils with patent infection of B. malayi and fed to A. aegypti along with the blood from immunized animals. Development of the parasite in infected mosquitoes was monitored until they reached infective stage larvae (L3). Fewer number of parasites developed to first stage (L1) and subsequently to L2 and L3 in mosquitoes fed with blood of immunized animals, when compared to those fed with blood of control animals. The results thus indicated that filarial parasite specific antibodies present in the blood of the immunized animals resulted in the reduction of number of larvae of B. malayi developing in the mosquito host.

Physiological changes in Biomphalaria glabrata say, 1818 (Pulmonata: Planorbidae) caused by sub-lethal concentrations of the latex of Euphorbia splendens var. hislopii N.E.B (Euphorbiaceae)

Molluscides have been used as one of the strategies to control schistosomiasis. Many plant extracts with molluscidal effects have been tested, but the action of the latex of Euphorbia splendens var. hislopii is considered the most promising because it meets the recommendations of the World Health Organization (WHO). The objective of this study was to determine the lethal dose and identify the effects of the different doses of latex of E. splendens var. hislopii on the physiology of Biomphalaria glabrata submitted to treatment for 24 h. The concentrations of glucose, uric acid and total proteins in the hemolymph and of glycogen in the digestive gland and cephalopodal mass were determined. The LD50 value was 1 mg/l. The highest escape index was found to be at a concentration of 0.6 mg/l. The results showed that the latex of E. splendens var. hislopii caused a sharp reduction in the reserves of glycogen in the digestive gland and elevation of the protein content in the hemolymph of B. glabrata.

Localization of Brugia malayi (sub-periodic) adults in different organs of Mastomys coucha and its influence on microfilaraemia and host antibody response

Lymphatic filariasis caused by nematode parasites Wuchereria bancrofti or Brugia malayi is a spectral disease and produces wide range of immune responses and varying levels ofmicrofilaraemia in infected individuals. The relationship between the immune response of host and the developmental stage of the parasite as well as the microfilariae (mf) density and specific location of the adult worms is yet to be understood. As an experimental model, B. malayi adapted in the experimental animal Mastomys coucha has been used widely for various studies in filariasis. The present study was to assess microfilaraemia as well as the humoral immune response of M. coucha during various stages of B. malayi development and their localization in different organs. The result showed that the density of mf in the circulating blood of the experimental animal depended upon the number of female worms as well as the location and co-existence of male and female worms. The mf density in the blood increased with the increase in the number of females. The clearance of inoculated infective stage (L3) or single sex infection or segregation of male and female to different organs of infected host resulted in amicrofilaraemic condition. With respect to antibody response, those animals cleared L3 after inoculation and those with adult worm as well as mf showed low antibody levels. But those with developmental fourth stage and/or adult worms without mf showed significantly higher antibody levels.

Schistosoma mansoni major egg antigen Smp40: molecular modeling and potential immunoreactivity for anti-pathology vaccine development

The pathogenesis of Schistosoma mansoni infection is largely determined by host T-cell mediated immune responses such as the granulomatous response to tissue deposited eggs and subsequent fibrosis. The major egg antigens have a valuable role in desensitizing the CD4+ Th cells that mediate granuloma formation, which may prevent or ameliorate clinical signs of schistosomiasis.S. mansoni major egg antigen Smp40 was expressed and completely purified. It was found that the expressed Smp40 reacts specifically with anti-Smp40 monoclonal antibody in Western blotting. Three-dimensional structure was elucidated based on the similarity of Smp40 with the small heat shock protein coded in the protein database as 1SHS as a template in the molecular modeling. It was figured out that the C-terminal of the Smp40 protein (residues 130 onward) contains two alpha crystallin domains. The fold consists of eight beta strands sandwiched in two sheets forming Greek key. The purified Smp40 was used for in vitro stimulation of peripheral blood mononuclear cells from patients infected with S. mansoni using phytohemagglutinin mitogen as a positive control. The obtained results showed that there is no statistical difference in interferon-g, interleukin (IL)-4 and IL-13 levels obtained with Smp40 stimulation compared with the control group (P > 0.05 for each). On the other hand, there were significant differences after Smp40 stimulation in IL-5 (P = 0.006) and IL-10 levels (P < 0.001) compared with the control group. Gaining the knowledge by reviewing the literature, it was found that the overall pattern of cytokine profile obtained with Smp40 stimulation is reported to be associated with reduced collagen deposition, decreased fibrosis, and granuloma formation inhibition. This may reflect its future prospect as a leading anti-pathology schistosomal vaccine candidate.

Serologic evidence of the recent circulation of Saint Louis encephalitis virus and high prevalence of equine encephalitis viruses in horses in the Nhecolândia sub-region in South Pantanal, Central-West Brazil

As in humans, sub-clinical infection by arboviruses in domestic animals is common; however, its detection only occurs during epizootics and the silent circulation of some arboviruses may remain undetected. The objective of the present paper was to assess the current circulation of arboviruses in the Nhecolândia sub-region of South Pantanal, Brazil. Sera from a total of 135 horses, of which 75 were immunized with bivalent vaccine composed of inactive Eastern equine encephalitis virus (EEEV) and Western equine encephalitis virus(WEEV) and 60 were unvaccinated, were submitted to thorough viral isolation, reverse transcriptase polymerase chain reaction (RT-PCR) and neutralization tests for Saint Louis encephalitis virus (SLEV), EEEV, WEEV and Mayaro virus (MAYV). No virus was isolated and viral nucleic-acid detection by RT-PCR was also negative. Nevertheless, the prevalence of neutralizing antibodies in horses older than seven months was 43.7% for SLEV in equines regardless of vaccine status, and 36.4% for WEEV and 47.7% for EEEV in unvaccinated horses. There was no evidence of MAYV infections. The serologic evidence of circulation of arboviruses responsible for equine and human encephalitis, without recent official reports of clinical infections in the area, suggests that the Nhecolândia sub-region in South Pantanal is an important area for detection of silent activity of arboviruses in Brazil.

The effect of sub-lethal doses of Azadirachta indica (Meliaceae) oil on the midgut of Spodoptera frugiperda (Lepidoptera, Noctuidae)

The fall armyworm, Spodoptera frugiperda, is one of the major field pests for maize production. It is mainly controlled by means of synthetic, and more recently by resistant cultivar of maize expressing Bt toxins. The neem tree, Azadirachta indica, is a plant that can potentially control insects with the advantage of being food and environmental safe. The aim of this study was to assess the effect of neem oil on the development and survival of S. frugiperda caterpillars by assessing histological alterations caused on their midgut. Newly hatched caterpillars were submitted to three neem oil concentrations: 0.006; 0.05; 0.4%, which were added to their artificial diet. Ten 3rd instar caterpillars, taken from each treatment, were submitted to histological analysis. The alimentary canals from the specimens were fixed in Baker for 12 hours, desiccated and diaphanized in alcohol/xylol (1:1) and xylol. After placing the samples in paraffin, they were sliced in 8 µm sections and stained with hematoxylin-eosin stain. The neem oil added to the diet of S. frugiperda caused total mortality at dose of 0.4% whilst still in the first instars, prolonged the larval and pupal stages, and reduced the pupal weight. Histo-physiological alterations such as degeneration of the epithelial lining of the midgut and in the peritrophic matrix were found at all concentrations of neem oil.

Aptidão para reflorestamento das sub-bacias dos canais do Mangue e do Cunha, município do Rio de Janeiro

Avaliou-se a aptidão para reflorestamento das terras das partes não edificadas da vertente norte do maciço da Tijuca, sub-bacias dos canais do Mangue e do Cunha, com o intuito de subsidiar ações do Programa Mutirão Reflorestamento da Secretaria Municipal de Meio Ambiente do Rio de Janeiro. A avaliação da aptidão das terras estimou graus de limitação dos parâmetros: deficiência de nutrientes, deficiência de água, susceptibilidade à erosão e impedimentos ao manejo. Estes graus de limitação foram estimados para os componentes das unidades de mapeamento de solos, considerando as informações de solos e paisagens do mapeamento existente. Para a digitalização e organização das informações geradas, foram utilizados sistemas de informações geográficas. As seguintes classes de aptidão para reflorestamento foram determinadas: 11,2 % de Regular, 81,5 % de Restrita e 7,3 % de Inapta. A declividade representa o fator limitante de maior importância para as terras da classe de aptidão restrita, seguida da presença de rochosidade.

Simulação da variabilidade espacial da erosão hídrica em uma sub-bacia hidrográfica de Latossolos no sul de Minas Gerais

A simulação da distribuição espacial da erosão do solo consiste em uma ferramenta poderosa para o planejamento conservacionista em bacias hidrográficas, sendo uma importante aplicação da Equação Universal de Perdas de Solo (EUPS) associada a princípios de interpolação espacial, principalmente a geoestatística. Este trabalho objetivou simular a distribuição espacial da erosão hídrica numa sub-bacia hidrográfica da região do Alto Rio Grande (MG), aplicando ferramentas geoestatísticas para distribuição espacial e mapeamento. Diferentes cenários de uso do solo foram analisados. A erosão foi estimada com base na EUPS aplicada a células, considerando as unidades pedológicas e diferentes usos em cada célula. O fator topográfico (LS) das células foi obtido com base no Modelo Digital de Elevação da sub-bacia, identificando comprimento e direção principal do escoamento. A erosividade média anual da região é de 8.030 MJ mm ha-1 h-1 ano-1, e a erodibilidade dos solos foi extraída da literatura. A EUPS foi aplicada a cada célula, levando-se em conta a situação atual do solo, áreas degradadas plantadas com eucalipto, pastagem plantada e plantio convencional de milho, considerando a ocupação de toda a sub-bacia hidrográfica. Na situação atual, a sub-bacia apresenta taxas de erosão inferiores aos limites de tolerância para os respectivos solos, com exceção das áreas degradadas ocupadas por eucalipto e pastagem em Cambissolo. No entanto, em todas as situações analisadas, seu lado leste apresentou as maiores perdas de solo, especialmente para os cenários de eucalipto nas condições atuais e plantio convencional de milho em Cambissolo e Latossolo Vermelho-Amarelo, sendo necessária a aplicação de técnicas conservacionistas.

Calibração de modelo para a simulação de vazão e de fósforo total nas sub-bacias dos Rios Conrado e Pinheiro - Pato Branco (PR)

O desenvolvimento de modelos hidrológicos capazes de predizer o impacto de fontes difusas de poluição e do uso e ocupação do solo na qualidade das águas superficiais e subterrâneas tem auxiliado o estudo de agroecossistemas. Com esse objetivo, foi utilizado o modelo SWAT 2005 (Soil and Water Assessment Tool) para avaliar sua sensibilidade na predição da vazão e do fluxo de massa do P total. O estudo foi realizado em duas microbacias hidrográficas contíguas, dos rios Conrado e Pinheiro, afluentes do rio Pato Branco, localizadas nos municípios de Pato Branco e Mariópolis, no Estado do Paraná. Foram utilizados dados climatológicos do período 1979/2006 e dados observados de vazão e concentração de P total dos anos 2004/2005 de duas estações de monitoramento, localizadas na parte inferior do curso principal dos rios Conrado e Pinheiro. Utilizou-se a interface AvSWAT-X, com o SIG ArcView 3.3® e a extensão Spatial Analyst 2.0®, para entrada e manipulação dos dados. As médias anuais e mensais observadas de vazão e P total foram comparadas aos dados simulados. O Coeficiente de Eficiência de Nash-Sutcliffe (COE) foi utilizado para avaliar a eficiência do modelo. A modelagem foi melhorada com a associação de análise de sensibilidade, autocalibração e calibração manual, verificando-se que, com frequência de amostragem regular, o modelo SWAT 2005 realizou de forma aceitável as simulações de vazão e de exportação de P total. Já com frequência de amostragem irregular e pequeno número de dados, os procedimentos de análise de sensibilidade e de autocalibração não foram eficientes na calibração do modelo SWAT 2005 para a simulação de vazão e exportação de P total. Foram encontrados diferentes níveis de sensibilidade entre as duas estações, refletindo as desigualdades entre as Unidades de Resposta Hidrológica. A distribuição mensal simulada das exportações de P mostrou a heterogeneidade da aplicação do nutriente ao longo do ano.