Epidemiologia da leishmaniose visceral no município de Fortaleza, Ceará / Epidemiology of visceral leishmaniasis in Fortaleza, Ceará, Brazil

O objetivo do trabalho foi analisar os dados relacionados à transmissão da leishmaniose visceral (LV), no município de Fortaleza, e discutir a respeito da distribuição do vetor, reservatório doméstico e casos humanos ocorridos no período de 2009 a 2013. O presente estudo é do tipo descritivo realizado por meio de levantamento de dados secundários. A correlação entre casos humanos, caninos e número de flebotomíneos foi feita pelo teste de correlação de Spearman, com nível de significância de 5%. No período de 2009 a 2013 foram confirmados 941 casos e 55 óbitos. A letalidade média no período foi de 5,84%. Na distribuição por sexo, houve uma maior proporção de casos no sexo masculino em todos os anos analisados. A faixa etária de 1 a 4 anos apresentou a maior porcentagem de casos, destacando-se o ano de 2010 com 31,5% dos casos. A distribuição média de casos por regional demonstrou um predomínio nas regionais I, V e VI. Em relação aos flebotomíneos, a espécie encontrada em maior abundância foi Lutzomyia (Lutzomyia) longipalpis, seguido de Lutzomyia (Lutzomyia) migonei e Lutzomyia (Lutzomyia) lenti. Segundo os registros da Secretaria Municipal de Saúde de Fortaleza, 39.626 cães foram soro reagentes para LV nos anos de 2009 a 2013, destes 14.313 foram eutanasiados. O ano de 2013 destacou-se com 17.808 cães soro reagentes, sendo 1.273 da SER (Secretaria Executiva Regional) III, 2.572 da SER V e 1.909 da SER VI. Não foi observada correlação significativa entre o número de flebotomíneos capturados e o número de casos caninos de LV(p>0,05). Houve correlação negativa entre casos humanos e caninos (r=-0,0388) e correlação positiva entre casos humanos e número de flebotomíneos (r=0,7469). Os achados criam perspectivas para a identificação de outros fatores que podem influenciar a incidência de casos humanos e caninos, como a participação de outros possíveis reservatórios e vetores na cadeia de transmissão da leishmaniose visceral no município de Fortaleza.(AU)

The objective was to analyze the data relating to the transmission of visceral leishmaniasis (LV) in Fortaleza municipality, and discuss about the vector distribution, domestic reservoir and human cases from 2009 to 2013.This study is descriptive conducted through a survey of secondary data. The correlation between human and canine cases and sandflies number was taken by Spearman correlation test, with 5% significance level. In the period 2009 to 2013, 941 cases and 55 deaths were confirmed. The mortality rate for the period was 5.84%. There was a higher proportion of cases among men in all the years analyzed. The age group 1-4 years old had the highest percentage of cases, highlighting the year 2010 with 31.5% of the cases. The average distribution of cases by SER demonstrated a predominance in SER I, V and VI. With regard to the vector species found in greater abundance was Lutzomyia (Lutzomyia) longipalpis, followed by Lutzomyia (Lutzomyia) migonei and Lutzomyia (Lutzomyia) lenti. According to the records, 39,626 dogs were serum reagents for LV in the years 2009-2013, these 14,313 were killed. The year 2013 stood out with 17,808 dogs serum reagents, and 1,273 of SER III, 2572 of SER V and 1909 of SER VI. There was no significant correlation between the number of sand flies and the number of canine VL cases (p?γτ;0.05). There was a negative correlation between human and canine cases (r = -0.0388) and correlation was observed between human cases and number of sand flies (r= 0.7469).Our findings create prospects for the identification of other factors that may influence the incidence of human and canine cases, such as the participation of other possible reservoirs and vectors in the transmission of the visceral leishmaniasis in Fortaleza.(AU)

Leishmanicidal and cholinesterase inhibiting activities of phenolic compounds of Dimorphandra gardneriana and Platymiscium floribundum, native plants from Caatinga biome / Atividade inibitória Leishmanicida e Colinesterásica de Compostos Fenólicos de Dimorphandra gardneriana e Platymiscium fliribunbum, plantas nativas do bioma Caatinga

In recent years, the Brazilian Health Ministry and the World Health Organization have supported research into new technologies that may contribute to the surveillance, new treatments, and control of visceral leishmaniasis within the country. In light of this, the aim of this study was to isolate compounds from plants of the Caatinga biome, and to investigate their toxicity against promastigote and amastigote forms of Leishmania infantum chagasi, the main responsible parasite for South American visceral leishmaniasis, and evaluate their ability to inhibit acetylcholinesterase enzyme (AChE). A screen assay using luciferase-expressing promastigote form and an in situ ELISA assay were used to measure the viability of promastigote and amastigote forms, respectively, after exposure to these substances. The MTT colorimetric assay was performed to determine the toxicity of these compounds in murine monocytic RAW 264.7 cell line. All compounds were tested in vitro for their anti-cholinesterase properties. A coumarin, scoparone, was isolated from Platymiscium floribundum stems, and the flavonoids rutin and quercetin were isolated from Dimorphandra gardneriana beans. These compounds were purified using silica gel column chromatography, eluted with organic solvents in mixtures of increasing polarity, and identified by spectral analysis. In the leishmanicidal assays, the compounds showed dose-dependent efficacy against the extracellular promastigote forms, with an EC50 for scoporone of 21.4µg/mL, quercetin and rutin 26 and 30.3µg/mL, respectively. The flavonoids presented comparable results to the positive control drug, amphotericin B, against the amastigote forms with EC50 for quercetin and rutin of 10.6 and 43.3µg/mL, respectively. All compounds inhibited AChE with inhibition zones varying from 0.8 to 0.6, indicating a possible mechanism of action for leishmacicidal activity.

Nos últimos anos, o Ministério da Saúde do Brasil e a Organização Mundial da Saúde tem apoiado a investigação de novas tecnologias que possam contribuir para a vigilância, novos tratamentos e controle da leishmaniose visceral no país. Assim, o objetivo deste trabalho foi isolar compostos de plantas do bioma Caatinga, e investigar a toxicidade destes compostos contra as formas promastigotas e amastigotas de Leishmania infantum chagasi, principal parasita responsável pela leishmaniose visceral na América do Sul, e avaliar a sua capacidade para inibir a enzima acetil-colinesterase (AChE). Após a exposição aos compostos em estudo, foram realizados testes utilizando a forma promastigota que expressa luciferase e ELISA in situ para medir a viabilidade das formas promastigotas e amastigota, respectivamente. O ensaio colorimétrico MTT foi realizado para determinar a toxicidade destas substâncias utilizando células monocíticas murina RAW 264.7. Todos os compostos foram testados in vitro para as sua propriedade anti-colinesterásica. Um cumarina, escoparona, foi isolada a partir de hastes de Platymiscium floribundum, e os flavonóides, rutina e quercetina, foram isolados a partir de grãos de Dimorphandra gardneriana. Estes compostos foram purificados, utilizando cromatografia em coluna gel eluída com solventes orgânicos em misturas de polaridade crescente, e identificados por análise espectral. Nos ensaios leishmanicidas, os compostos fenólicos mostraram eficácia contra as formas extracelulares promastigotas, com EC50 para escoporona de 21.4µg/mL e para quercetina e rutina 26 e 30.3µg/mL, respectivamente. Os flavonóides apresentaram resultados comparáveis à droga controle, a anfotericina B, contra as formas amastigotas com EC50 para quercetina e rutina de 10.6 e 43.3µg/mL, respectivamente. Os compostos inibiram a enzima AChE com halos de inibição variando de 0,8 a 0,6cm, indicando um possível mecanismo de ação para a atividade leishmanicida.

Leishmanicidal activity in vitro of Musa paradisiaca L. and Spondias mombin L. fractions.

Visceral leishmaniasis (VL) is a zoonotic disease characterized by infection of mononuclear phagocytes by Leishmania chagasi. The primary vector is Lutzomyia longipalpis and the dog is the main domestic reservoir. The control and current treatment of dogs using synthetic drugs have not shown effectiveness in reducing the incidence of disease in man. In attempt to find new compounds with leishmanicidal action, plant secondary metabolites have been studied in search of treatments of VL. This study aimed to evaluate the leishmanicidal activity of Musa paradisiaca (banana tree) and Spondias mombin (cajazeira) chemical constituents on promastigotes and amastigotes of L. chagasi. Phytochemical analysis by column chromatography was performed on ethanol extracts of two plants and fractions were isolated. Thin layer chromatography was used to compare the fractions and for isolation the substances to be used in vitro tests. The in vitro tests on promastigotes of L. chagasi used the MTT colorimetric method and the method of ELISA in situ was used against amastigotes besides the cytotoxicity in RAW 264.7 cells. Of the eight fractions tested, Sm1 and Sm2 from S. mombin had no action against promastigotes, but had good activity against amastigotes. The fractions Mp1 e Mp4 of M. paradisiaca were very cytotoxic to RAW 264.7 cells. The best result was obtained with the fraction Sm3 from S. mombin with IC(50) of 11.26 µg/ml against promastigotes and amastigotes of 0.27 µg/ml. The fraction Sm3 characterized as tannic acid showed the best results against both forms of Leishmania being a good candidate for evaluation in in vivo tests.

In vitro effect of Aloe vera, Coriandrum sativum and Ricinus communis fractions on Leishmania infantum and on murine monocytic cells.

In South America, visceral leishmaniasis is a zoonosis caused by the protozoan species Leishmania infantum (syn. L. chagasi) and is primarily transmitted through the bite of the female Lutzomyia longipalpis. Its main reservoir in urban areas is the dog. The application of control measures recommended by health agencies have not achieved significant results in reducing the incidence of human cases, and the lack of effective drugs to treat dogs resulted in the prohibition of this course of action in Brazil. Therefore, it is necessary to search new alternatives for the treatment of canine and human visceral leishmaniasis. The objectives of this study were to evaluate the in vitro effect of fractions from Aloe vera (aloe), Coriandrum sativum (coriander), and Ricinus communis (castor) on promastigotes and amastigotes of L. infantum and to analyze the toxicity against the murine monocytic cells RAW 264.7. To determine the viability of these substances on 50% parasites (IC50), we used a tetrazolium dye (MTT) colorimetric assay (bromide 3-4.5-dimethylthiazol-2-yl-2,5-dephenyltetrazolium), and on amastigotes we performed an in situ ELISA. All fractions were effective against L. infantum promastigotes and did not differ from the positive control pentamidine (p>0.05). However, the R. communis ethyl acetate and chloroform fractions, as well as the C. sativum methanol fraction, were the most effective against amastigotes and did not differ from the positive control amphotericin B (p>0.05). The R. communis ethyl acetate fraction was the least toxic, presenting 83.5% viability of RAW 264.7 cells, which was similar to the results obtained with amphotericin B (p>0.05). Based on these results, we intend to undertake in vivo studies with R. communis ethyl acetate fractions due the high effectiveness against amastigotes and promastigotes of L. infantum and the low cytotoxicity towards murine monocytic cells.