Study of the trypanomicidal action of crude extract of Mandevilla velutína in mice infected with Trypanosotna cruzi

Chagas disease is one of the most important parasitic infections in several regions of Latin America, including Brazil, due to its high incidence, prevalence and socio-economic repercussions that this endemic causes to countries. Despite efforts, a very small number of trypanomycin drugs have been indicated for the specific treatment of the disease, all with side effects that limit use. In the Uberlândia region, there are popular reports that the rhizome extract of Mandevilla velutina is indicated for patients with this disease. This work was developed in order to investigate the action of this extract on the parasitemia and mortality of mice infected with 5x104 blood forms of the T. cruzi Y strain. The extract was obtained by macerating fragments of the rhizome in 15% cereal alcohol, which was lyophilized (EBL) and resuspended in distilled water. The EBL administered to the mice, orally, presented a low toxicity in the organism of these animals. When EBL was administered in increasing doses (up to 200mg / kg) in mice infected with T. cruzi, a reduction in parasitemia and an increase in survival were observed, which were dose dependent. The previous treatment of the animals, with a dose of 200mg / kg, proved to be effective in reducing parasitemia, being more efficient when the treatment was carried out 24 hours before infection. When infected animals were treated with two daily doses of EBL (71.5 and 143 mg / kg) for 30 days and compared with animals treated with Benzonidazole (two daily doses of 50 mg / kg), it was found that EBL is more effective than Benzonidazole in the doses administered. To investigate the possible involvement of the NO System in the mechanism of action of the EBL, infected animals were treated with EBL and NOS inhibitor (NOARG or L-NAME) where an attenuation of the effects of the EBL was observed. When measuring NO2 and NO3 in the serum of normal animals and treated with EBL, there was a significant increase in NO2 and NO3 concentrations. When comparing parasitemia and serum NO3 levels of infected animals treated and not treated with EBL, it was found that the untreated animals showed higher values ​​of parasitemia and NO3, between the 8th and 14th day post-infection. The trypanomicidal action promoted by the EBL, was also tested in vitro in the blood containing T. cruzi, incubated with EBL (24 and 48 hours), resulting in the reduction of the trypomastigote count. In the evaluation of the genotoxic potential of the EBL, the results suggest that the EBL has protective effects on the formation of 'onucleated erythrocytes. In the phytochemical screening, the presence of several auímic classes was verified, including the tannins and the terpenoids, which may present anti-parasitic activity. This work suggests that the crude M. velutina extract has a component (s) capable of acting on the parasitemia of mice infected with T. cruzi, significantly reducing the number of parasites during infection, and this reduction may be correlated with the Nitric Oxide System.Dissertação (Mestrado)A doença de Chagas é uma das mais importantes infecções parasitárias em várias regiões da América Latina, incluindo o Brasil, em virtude da sua alta incidência, prevalência e repercussão sócio-econômica que esta endemia causa aos países. Apesar dos esforços, um número muito reduzido de drogas tripanomicidas tem sido indicadas para o tratamento específico da doença, todas com efeitos colaterais que limitam a utilização. Na região de Uberlândia, existem relatos populares de que o extrato do rizoma de Mandevilla velutina é indicado aos pacientes portadores desta doença. Este trabalho foi desenvolvido com o intuito de investigar a ação deste extrato sobre a parasitemia e mortalidade de camundongos infectados com 5x104 formas sanguíneas da cepa Y de T. cruzi. O extrato foi obtido pela maceração de fragmentos do rizoma em álcool de cereais a 15%, o qual foi liofilizado (EBL) e ressuspendido em água destilada. O EBL administrado aos camundongos, por via oral, apresentou uma baixa toxicidade no organismo destes animais. Quando o EBL foi administrado em doses crescentes (até 200mg/kg) em camundongos infectados com T. cruzi, observou-se redução da parasitemia e aumento da sobrevivência, os quais foram dose dependentes. O tratamento prévio dos animais, com uma dose de 200mg/kg, mostrou-se eficaz na diminuição da parasitemia, sendo mais eficiente quando o tratamento foi realizado 24 horas antes da infecção. Quando os animais infectados foram tratados com duas doses diárias de EBL (71,5 e 143 mg/kg) durante 30 dias e comparados com os animais tratados com Benzonidazol (duas doses diárias de 50mg/kg), verificou-se que o EBL é mais eficaz do que o Benzonidazol nas doses administradas. Para investigar o possível envolvimento do Sistema NO no mecanismo de ação do EBL, animais infectados foram tratados com EBL e inibidor da NOS (NOARG ou L-NAME) onde se observou uma atenuação dos efeitos do EBL. Ao realizar a dosagem de NO2 e de NO3 no soro de animais normais e tratados com EBL verificou-se nestes um aumento significante nas concentrações de NO2 e NO3. Ouando se comparou as parasitemias e os níveis séricos de NO3 dos animais infectados tratados e não tratados com EBL, verificou-se que os animais não tratados apresentaram maiores valores de parasitemia e NO3, entre 8o e 14° dia pós-infecção. A acão tripanomicida promovida pelo EBL, foi testada também in vitro no sangue contendo T. cruzi, incubado com EBL (24 e 48 horas), resultando na redução da contagem de tripomastigotas. Na avaliação do potencial genotóxico do EBL, os resultados sugerem que o EBL possui efeitos protetores na formação de eritrócitos ’ onucleados Na triagem fitoquímica verificou-se a presença de várias classes auímicas incluindo os taninos e os terpenóides que podem apresentar atividade antinarasitária Os resultados obtidos neste trabalho sugerem que 0 extrato bruto de M. velutina possui componente(s) capaz(es) de atuar sobre a parasitemia de camundongos infectados com T. cruzi, reduzindo significantemente 0 número de parasitos durante a infecção, sendo que esta redução pode estar correlacionada com o Sistema Óxido Nítrico

Diagnóstico parasitológico de horticultures no monitoramento da contaminação parasitária em ambientes rurais

The objective of this study was to detect possible carriers of parasites and/or commensals between the horticulturists of Fair of the Producer from Upper Paranaiba, Minas Gerais. A total of 30 horticulturists were instructed to collect three stool samples on alternate days during the months of August and September 2007. Horticulturists were positive 40% for one or more parasite and / or dinner, and found: Giardia lamblia (3.5%), Entamoeba histolytica / E. dispar (7%), Entamoeba coli (13%), Endolimax nana (13%) and Ascaris lumbricoides (3.5%). Stool of gardeners is of importance in monitoring parasite sanitary conditions of vegetables sold in urban environments.O objetivo deste trabalho foi diagnosticar possíveis portadores de parasitos e ou comensais intestinais entre os horticultores da Feira do Produtor da região do Alto Paranaíba, Minas Gerais. Um total de 30 horticultores foi instruído a coletar três amostras de fezes em dias alternados durante os meses de agosto e setembro de 2007. Horticultores apresentaram positividade de 40% para um ou mais parasito e/ou comensal, tendo sido encontrados: Giardia lamblia (3,5%), Entamoeba histolytica/E. dispar (7%), Entamoeba coli (13%), Endolimax nana (13%) e Ascaris lumbricoides (3,5%). O diagnóstico parasitológico de horticultores é de grande importância no monitoramento parasitário das condições higiênicas de hortaliças comercializadas em ambientes urbanos

Evaluation of allelopathic potential of leaf extract of kielmeyera coriacea on lactuca sativa l

Kielmeyera coriacea Mart. (Clusiaceae), popularly known as “pau-santo”, is a typical Brazilian cerrado tree known due to its varied secondary metabolites. This study aimed to determine the allelopathic potential of the hydroalcoholic extract of leaves of K. coriacea through bioassays of seed germination, seedling growth and mitotic index of Lactuca sativa L. (lettuce). In addition it was done the tetrazolium assay and a phytochemical screening. The extract concentrations caused alterations in germination parameters, in root growth and in the mitotic index. The phytochemical screening showed the presence of triterpenes, coumarins, steroids, flavonoids and condensed tannins, compounds known to confer allelopathic characteristics upon other species. These data indicate that K. coriacea presents an allelopathic potential because its leaf extracts interfere with germination and growth without any interference of pH and osmotic potential in the results.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Antigenotoxic effects of Mandevilla velutina (Gentianales, Apocynaceae) crude extract on cyclophosphamide-induced micronuclei in Swiss mice and urethane-induced somatic mutation and recombination in Drosophila melanogaster

A Mandevilla velutina crude extract was investigated using the mouse micronucleus test (MNT) and the Drosophila melanogaster somatic mutation and recombination test (SMART) using standard (ST) and high bioactivation (HB) crosses. The MNT used 10 mg, 20 mg or 40 mg per 100 g of body weight (bw) of extract with and without 0.2 mg per 100 g bw peritoneal cyclophosphamide. There was no genotoxicity in the negative control or extract only groups and, compared to the cyclophosphamide control, there was a significant reduction in micronucleated polychromatic erythrocytes in all the groups given extract plus cyclophosphamide. For SMART larvae were fed 5 or 10 mg mL-1 of extract for seven days with and without 0.89 mg mL-1 of urethane given on day seven. The ST and HB flies showed no significant differences in spots between the negative control and the extract only groups. The number of urethane-induced spots was reduced by the highest concentration of extract for the ST flies and by both concentrations of extract for the HB flies. The results suggest that M. velutina extract is not genotoxic but is antigenotoxic.ISSN:1415-4757ISSN:1678-468

Atividade antioxidante e determinação de fenóis totais, carotenoides, betacarotenos, licopeno e zinco em variedades branca, amarela e rosada de Manihot Esculenta Crantz

There are cassava varieties that present compounds as carotenoids, beta-carotene, lycopene and minerals important for human and animal health. The present study evaluated the antioxidant activity of the white, yellow and pinkish varieties of Manihot esculenta, by mean of the DPPH test and by the ferrous ion-chelating activity. Furthermore, the total phenols, carotenoids, beta-carotene, lycopene and zinc contents were also determined. Utilizing the DPPH test it was possible to find that extracts of boiled samples presented higher antioxidant activity (89.53% - pinkish) in comparison to the fresh samples (1.97% - white). For the ferrous ion-chelating test, the highest activity was found for the boiled pinkish variety extract (63.43%) and the lowest was for fresh yellow extract (17.34%) the white sample did not present activity. The highest concentration of total phenols and zinc content was obtained for the boiled pinkish variety extract 136.12 mg EAG/g of extract and 0,811ppm, respectively, in the concentration of 1000 µg/mL. The pinkish variety presented also higher quantity of pigments, including carotenoid (29.40 µg/g), beta-carotene (9.14 µg/100g) and lycopene (68.92%). According to the results obtained in this study it was possible to conclude that the yellow and pinkish varieties of M. esculenta present quantity of phenolic compounds and minerals sufficient to attribute the antioxidant activity and may thus contribute to reduce oxidative damage and be used as nutraceuticals or directly ingested in the diet to maintain good health.Existem variedades de mandioca que apresentam compostos como os carotenoides, beta-caroteno, licopeno e minerais importantes para a saúde humana e animal. O presente estudo avaliou a atividade antioxidante das variedades branca, amarela e rosada de Manihot esculenta, por meio de teste de DPPH e pela atividade quelante de íons ferro. Além disso, o conteúdo de fenóis totais, carotenoides, beta-caroteno, licopeno e zinco também foram determinados. Utilizando o teste de DPPH foi possível verificar que os extratos de amostras cozidas apresentaram maior atividade antioxidante (89,53% - rosada) em comparação com as amostras frescas (1,97% - branca). Para o teste de atividade quelante de íons ferro, a maior atividade foi encontrada para o extrato da variedade rosada cozida (63,43%) e a menor foi do extrato da amarela in natura (17,34%) a amostra branca não apresentou atividade. A maior concentração de fenóis e de zinco foi obtida para o extrato da variedade rosada cozida 136,12 mg EAG/g de extrato e 0,811 ppm, respectivamente, na concentração de 1000 µg/mL. A variedade rosada apresentou também maior quantidade de pigmentos, incluindo carotenoides (29,40 µg/g); beta-caroteno (9,14 µg/100g) e licopeno (68,92%). De acordo com os resultados obtidos no presente estudo, foi possível concluir que as variedades amarela e rosada de M. esculenta, apresentam quantidades de compostos fenólicos e minerais suficientes para atribuir a atividade antioxidante, podendo assim, contribuir para diminuir os danos oxidativos e serem utilizadas como nutracêuticos ou diretamente ingeridas na dieta para manter a boa saúde

Determination of heavy metals and genotoxicity of water from an artesian well in the city of Vazante- MG, Brazil

The city of Vazante-MG is of great socioeconomic and environmental interest because it is the most important zinc producer district of Brazil. The mineral processing and geochemical processes may determine high concentrations of heavy metals in water intended for human consumption. Thus, the present study aimed to quantify and evaluate the heavy metal genotoxicity of artesian water in the city by Atomic absorption spectrophotometer analysis and testing with the Allium cepa test, respectively. This study reveals a chemical contamination in well water in the city, caused by the presence of heavy metals. Therefore, it can be considered that the high levels of heavy metals found in water samples are correlated with the genotoxic events observed in root cells of A. cepa

Parasitos e comensais intestinais em pacientes neoplásicos submetidos à quimioterapia

The objective of this study was to verify the prevalence of intestinal parasites and/or commensals in the neoplastic patients undergoing chemotherapy. Stool samples were analyzed by the method of Lutz (1919) and Rugai (1954), in triplicate. This work was composed of three groups, the first one (GI) formed by neoplastic patients that are not undergoing chemotherapy, the second (GII) comprised patients who were undergoing chemotherapy, and the third group (GIII) consisting of patients who completed chemotherapy. A total of 30 patients (GI-5, GII-18 and GIII-7) were screened at the Assis Regional Hospital of the Unified Health System of Assis, São Paulo.Additional information on antiparasitic treatment and tumor type were obtained by questionnaire. The positivity was 66.7% (20 cases) for intestinal parasites and/or commensals. The helminths were Ascaris lumbricoides (36.7%), Hookworms (20%) and Hymenolepis diminuta (3.3%). Among the highlights are protozoan Giardia lamblia (46.7%), Entamoeba coli (6.7%), E. histolytica /dispar (3.3%), Endolimax nana (3.3%) and Iodameba butschlii (3.3%). The high frequency of intestinal parasites and/or commensals in the neoplastic patients can be attributed to poor personal hygiene and lack of immunity to reinfection and poor knowledge of the prophylaxis of infection by protozoa and helminths. The results indicate the necessity of adopting a new criterion for neoplastic patients undergoing chemotherapy, primarily performing parasitological diagnosis, treatment and monitoring of cure of intestinal parasitic infections in this risk group.O objetivo deste trabalho foi verificar a freqüência de parasitos e/ou comensais intestinais em pacientes neoplásicos submetidos à quimioterapia. As amostras de fezes foram analisadas pelo método de Lutz (1919) e Rugai (1954), em triplicata. O trabalho foi composto por três grupos, o primeiro (GI) formado por pacientes neoplásicos não submetidos à quimioterapia, o segundo (GII) formado por pacientes que estavam em tratamento quimioterápico, e o terceiro grupo (GIII) constituído por pacientes que terminaram a quimioterapia. Um total de 30 pacientes (GI –5, GII-18 e GIII-7) foram triados no Hospital Regional de Assis do Sistema Único de Saúde da cidade de Assis, São Paulo. Dados complementares sobre tratamento antiparasitário e tipo de tumor foram obtidos por questionário. A positividade foi de 66,7% (20 casos) para parasitos e/ou comensais intestinais. Os helmintos encontrados foram: Ascaris lumbricoides (36,7%), Ancilostomídeos (20%) e Hymenolepis diminuta (3,3%). Entre os protozoários destacaram-se: Giardia lamblia (46,7%), Entamoeba coli (6,7%), E. histolytica/E. dispar (3,3%), Endolimax nana (3,3%) e Iodameba butschlii (3,3%). A elevada freqüência de parasitos e/ou comensais intestinais em pacientes neoplásicos pode ser atribuída a higiene pessoal inadequada e ausência de imunidade a re-infecções e ao pouco conhecimento da profilaxia para infecção por protozoários e helmintos. Os resultados obtidos indicam a necessidade da adoção de um novo critério para os pacientes neoplásicos submetidos à quimioterapia realizando primeiramente diagnóstico parasitológico, tratamento e acompanhamento de cura das parasitoses intestinais neste grupo de risco

Genotoxic activity and toxicity of Baccharis trimera Less. regarding the bioaccumulation of heavy metals

Baccharis trimera Less. (“carqueja”) is a native plant from Brazil, used in folk medicine preparations such as infusions and/or decoctions. The objective of this study was to evaluate the toxicity and genotoxicity of aqueous and ethanolic extracts of B. trimera and quantify heavy metals bioaccumulated in this specie. The extracts were prepared with ultra pure water. The Allium cepa test was conducted to examine the genotoxic activity of the extracts. The toxicological activity was used as bioassay of Artemia salina. Heavy metals were analyzed by atomic absorption spectrophotometery. The ethanolic extract showed the greatest effect on the mitotic index and chromosomal aberrations when treated with aqueous extract. The extracts showed activity in the toxicological concentrations. Heavy metals analyzed (Cd = 0.013, Cu = 0.089, Pb = 0.097 and Zn = 1.918 μg/ml) were measured at levels above those recommended. Therefore, B. trimera is a heavy metal bioacumulator specie, being able to promote genotoxicity activity in meristematic cells of A. cepa and toxicological activity on larvae of A. salina.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP