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Sample records for da leishmania amazonensis

  1. Histopatologia da forma localizada de leishmaniose cutânea por Leishmania (Leishmania amazonensis Histopathology of the localized form of cutaneous leishmaniasis due to Leishmania (Leishmania amazonensis

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    Mário A. P. Moraes

    1994-10-01

    Full Text Available São descritas as alterações microscópicas presentes na forma localizada (ulcerada da Leishmaniose cutânea produzida por Leishmania (Leishmania amazonensis. Nesse tipo de manifestação, menos conhecido do que a forma anérgica ou difusa devida ao mesmo agente, as lesões são clinicamente idênticas às de leishmaniose cutânea causada por espécies outras de Leishmania, pertencentes ao subgênero Viannia. Na infecção localizada por L. (L. amazonensis, entretanto, há um aspecto peculiar, só recentemente conhecido, ou seja, cerca de 50% dos indivíduos atingidos não reagem ao teste de Montenegro. A principal característica histológica observada foi a acumulação na derme, quase sempre focal, de numerosos macrófagos contendo no citoplasma um grande vacúolo cheio de amastigotas. O quadro é semelhante ao da forma difusa, porém sem o aspecto histiocitomatóide, próprio da última. Afora esses grupos de macrófagos, vêem-se também, na forma localizada, muitas células mononucleares da inflamação, principalmente plasmócitos e macrófagos não parasitados. Os acúmulos de macrófagos com amastigotas, quando volumosos, podem sofrer necrose na parte central; os parasitos, contidos nas células, são destruídos com elas ou liberados, e sua eliminação através da úlcera deve contribuir para a cura do processo. Esse tipo de necrose nunca foi descrito em casos da forma difusa. Não houve grande diferença, no quadro histológico, entre pacientes Montenegro-negativos e positivos. Apenas em alguns casos, do grupo Montenegro-positivo, havia granulomas formados por histiócitos epitelióides sem parasitos. Quanto à persistência das células com parasitos nas lesões, observou-se que aos seis meses ou mais de evolução, em ambos os grupos, ainda estavam elas presentes. Tal achado não é comum na leishmaniose tegumentar por L. (V. braziliensis.The microscopic changes found in the localized form of the human cutaneous leishmaniasis due to Leishmania (Leishmania amazonensis are reported. In this form, less known than the diffuse one caused by the same species, the clinical manifestations are identical to those produced by other Leishmania species of the subgenus Viannia. There is, however, in the localized infection by L (L. amazonensis, a peculiar feature, only recently discovered: about 50% of the affected individuals are Montenegro-negatives. The main histologic change observed in the skin sections was the presence of groups of macrophages with a large vacuole in the cytoplasm, containing many amastigotes. The microscopic picture is similar to that found in the diffuse form of the disease, the difference being only quantitative. When in large numbers, the macrophages suffers necrosis, which generally starts at the center of the groups. First, in this process, the membrane of the parasitized cells ruptures, and the amastigotes become free; later, both cells and parasites are destroyed. The picture can be seen either in Montenegro-negative or in Montenegro-positive patients. The macrophages with amastigotes may persist in tissues for as long as 6-7 months, while in the infections due to L (V. braziliensis the parasites usually disappear in a few weeks.

  2. Histopatologia da forma localizada de leishmaniose cutânea por Leishmania (Leishmania) amazonensis / Histopathology of the localized form of cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis

    Scientific Electronic Library Online (English)

    Mário A. P., Moraes; Fernando T., Silveira.

    1994-10-01

    Full Text Available São descritas as alterações microscópicas presentes na forma localizada (ulcerada) da Leishmaniose cutânea produzida por Leishmania (Leishmania) amazonensis. Nesse tipo de manifestação, menos conhecido do que a forma anérgica ou difusa devida ao mesmo agente, as lesões são clinicamente idênticas às [...] de leishmaniose cutânea causada por espécies outras de Leishmania, pertencentes ao subgênero Viannia. Na infecção localizada por L. (L.) amazonensis, entretanto, há um aspecto peculiar, só recentemente conhecido, ou seja, cerca de 50% dos indivíduos atingidos não reagem ao teste de Montenegro. A principal característica histológica observada foi a acumulação na derme, quase sempre focal, de numerosos macrófagos contendo no citoplasma um grande vacúolo cheio de amastigotas. O quadro é semelhante ao da forma difusa, porém sem o aspecto histiocitomatóide, próprio da última. Afora esses grupos de macrófagos, vêem-se também, na forma localizada, muitas células mononucleares da inflamação, principalmente plasmócitos e macrófagos não parasitados. Os acúmulos de macrófagos com amastigotas, quando volumosos, podem sofrer necrose na parte central; os parasitos, contidos nas células, são destruídos com elas ou liberados, e sua eliminação através da úlcera deve contribuir para a cura do processo. Esse tipo de necrose nunca foi descrito em casos da forma difusa. Não houve grande diferença, no quadro histológico, entre pacientes Montenegro-negativos e positivos. Apenas em alguns casos, do grupo Montenegro-positivo, havia granulomas formados por histiócitos epitelióides sem parasitos. Quanto à persistência das células com parasitos nas lesões, observou-se que aos seis meses ou mais de evolução, em ambos os grupos, ainda estavam elas presentes. Tal achado não é comum na leishmaniose tegumentar por L. (V.) braziliensis. Abstract in english The microscopic changes found in the localized form of the human cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis are reported. In this form, less known than the diffuse one caused by the same species, the clinical manifestations are identical to those produced by other Leishmania [...] species of the subgenus Viannia. There is, however, in the localized infection by L (L.) amazonensis, a peculiar feature, only recently discovered: about 50% of the affected individuals are Montenegro-negatives. The main histologic change observed in the skin sections was the presence of groups of macrophages with a large vacuole in the cytoplasm, containing many amastigotes. The microscopic picture is similar to that found in the diffuse form of the disease, the difference being only quantitative. When in large numbers, the macrophages suffers necrosis, which generally starts at the center of the groups. First, in this process, the membrane of the parasitized cells ruptures, and the amastigotes become free; later, both cells and parasites are destroyed. The picture can be seen either in Montenegro-negative or in Montenegro-positive patients. The macrophages with amastigotes may persist in tissues for as long as 6-7 months, while in the infections due to L (V.) braziliensis the parasites usually disappear in a few weeks.

  3. Clonaje de la proteína de choque térmico de 20 kDa de Leishmania amazonensis / Cloning of 20 kDa heat shock protein of Leishmania amazonensis

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    Ana Margarita, Montalvo Álvarez; Cristina, Folgueira Fernández; José María, Requena Rolanía.

    2009-08-01

    Full Text Available INTRODUCCIÓN: la inducción de las proteínas de choque térmico constituyen un mecanismo homeostático que protege a las células del efecto destructivo del calor u otras condiciones de estrés ambiental, paralelamente, ellas cumplen importantes funciones celulares. La proteína de choque térmico de 20 kD [...] a se reportó recientemente en Leishmania amazonensis. OBJETIVO: describir la metodología utilizada para realizar el clonaje de las proteínas de choque térmico, lo que permitió acometer estudios de algunas propiedades biológicas. MÉTODOS: la región codificante del gen hsp20 se amplificó mediante la reacción en cadena de la polimerasa con cebadores adecuados. El producto amplificado se clonó inicialmente en el vector pCR2.1 (Invitrogen) y después en el vector de expresión en procariotas pET-28b (Novagen), para obtener proteína recombinante. De manera paralela, el mismo fragmento se clonó en el vector de expresión en eucariotas pcDNA3 (Invitrogen) para obtener un posible preparado vacunal de ADN. Se realizó la secuenciación nucleotídica de los clones obtenidos, con la finalidad de verificar su fidelidad. RESULTADOS: se obtuvieron plásmidos recombinantes que codifican la HSP20 de Leishmania, y permiten la obtención de proteína recombinante y de ADN en forma masiva. CONCLUSIONES: ambos plásmidos fueron útiles para estudiar algunas de las propiedades biológicas de esta proteína. Este acercamiento puede ser de interés en otros trabajos de esta índole y constituir una guía metodológica. Abstract in english INTRODUCTION: the induction of heat shock proteins is a homeostatic mechanism that protects cells from the deleterious effects of thermal and other environmental stresses. In addition, they have important cell functions. The 20kDa heat shock protein in Leishmania amazonensis was recently reported. O [...] BJECTIVE: to describe the methodology used for cloning of heat shock proteins, which allowed the study of some biological properties. METHODS: the hsp20 gene coding region was amplified by polymerase chain reaction using adequate primers. The amplified product was initially cloned in pCR2.1 vector (Invitrogen) and then in pET-28b vector (Novagen), to obtain recombinant protein. The same fragment was cloned also in the eukariote expression vector pcDNA3 (Invitrogen). The nucleotidic sequencing of the different clones was made, in order to verify their fidelity. RESULTS: the recombinant plasmids that encode HSP20 protein in Leishmania and allow obtaining massively recombinant protein and DNA were produced. CONCLUSIONS: both plasmids were useful to study some of the biological properties of this protein. This approach could be useful for similar research and represent a suitable methodological guideline.

  4. Subcellular localization of an intracellular serine protease of 68 kDa in Leishmania (Leishmania amazonensis promastigotes

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    José Andrés Morgado-Díaz

    2005-07-01

    Full Text Available Here we report the subcellular localization of an intracellular serine protease of 68 kDa in axenic promastigotes of Leishmania (Leishmania amazonensis, using subcellular fractionation, enzymatic assays, immunoblotting, and immunocytochemistry. All fractions were evaluated by transmission electron microscopy and the serine protease activity was measured during the cell fractionation procedure using a-N-r-tosyl-L-arginine methyl ester (L-TAME as substrate, phenylmethylsulphone fluoride (PMSF and L-1-tosylamino-2-phenylethylchloromethylketone (TPCK as specific inhibitors. The enzymatic activity was detected mainly in a membranous vesicular fraction (6.5-fold enrichment relative to the whole homogenate, but also in a crude plasma membrane fraction (2.0-fold. Analysis by SDS-PAGE gelatin under reducing conditions demonstrated that the major proteolytic activity was found in a 68 kDa protein in all fractions studied. A protein with identical molecular weight was also recognized in immunoblots by a polyclonal antibody against serine protease (anti-SP, with higher immunoreactivity in the vesicular fraction. Electron microscopic immunolocalization using the same polyclonal antibody showed the enzyme present at the cell surface, as well as in cytoplasmic membranous compartments of the parasite. Our findings indicate that the internal location of this serine protease in L. amazonensis is mainly restricted to the membranes of intracellular compartments resembling endocytic/exocytic elements.

  5. Leishmania mexicana amazonensis: heterogeneity in 5-nucleotidase and peroxidase activities of mononuclear phagocytes during in vivo and in vitro infection Leishmania mexicana amazonensis: heterogeneidade da 5’-Nucleotidase e da peroxidase em fagócitos mononucleares durante infecção in vivo e in vitro

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    Suzana Côrte-Real

    1988-03-01

    Full Text Available The degree of maturation of cells of the Mononuclear Phagocyte System (MPS, during in vivo and in vitro infection by Leishmania mexicana amazonenesis, was evaluated in this study. The macrophages' differentiation was assayed by cytochemical characterization at the ultrastrctural level, using two well-established markers: 5'-nucleotidase enzyme activity, for revealing the mature cells, and the peroxidase activity present in the cell granules to demonstrate immature mononuclear phagocytes. only a few mcrophages, demonstrating 5'-nucleotidase positive reaction in both the plasma membrane and within their cytoplasmic vesicles, were found scattered in the chronic inflammation at the L. m. amazonensis lesions in albino mice. However, by the peroxidase activity analysis, we were also able to demonstrate the presence of immature MPS cells, which predominate, together with parasitized vacuolated macrophages, in chronic lesions induced in this systemby L. m. amazonensis. The implications of these results on the pathogenesis of murine cutaneous leishmaniasis are discussed.Um estudo sobre o grau de maturação das células do Sistema Fagocítico Mononuclear foi realizado durante a infecção in vivo e in vitro com a Leishmania mexicana amazonensis. A caracterização da diferenciação das células fagocíticas foi obtida com a localização ultraestrutural de dois marcadores enzimáticos bam conhecidos: a enzima 5'-Nucleotidase marcadora de membrana plasmática de células maduras e a enzima peroxidase, presente em grânulos, marcadora de células imaturas. A atividade da enzima 5'-Nucleotidase foi encontrada apenas em alguns macrófagos, presentes no foco inflamatório, em projeções da membrana plasmática e em algumas vesículas citoplasmáticas. Macrófagos peritoneais de camundongo apresentaram a mesma reatividade para este marcador. Contudo a análise da atividade peroxidásica demonstrou a predominância da presença de fagócitos mononucleares imaturos nas lesões crônicas induzidas neste sistema por Leishmania mexicana amazonensis.

  6. Ineficácia in vivo da terbinafina em leishmaniose cutânea causada por Leishmania (Leishmania amazonensis em camundongos C57BL/6 Terbinafine in vivo inefficacy on cutaneous leishmaniasis caused by Leishmania (Leishmania amazonensis in C57BL/6 mice

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    Raimunda Nonata Ribeiro Sampaio

    2003-07-01

    Full Text Available Testou-se, em camundongos C57BL/6 inoculados com a cepa MHOM/BR/PH8 de Leishmania (Leishmania amazonensis, terbinafina via oral 100mg/kg/dia, por 20 dias, solução salina 0,9% via oral como controle e stibogluconato de sódio 400mg SbV/kg/dia via subcutânea como padrão-ouro. A terbinafina mostrou-se ineficaz, clínica e parasitologicamente, e pelo ensaio por diluição limitante, quando comparada aos controles.The efficiency of terbinafine was tested in C57BL/6 mice inoculated with the Leishmania (Leishmania amazonensis strain MHOM/BR/PH8. The mice were administered: terbinafine at a dose of 100mg/kg/d by via oral; 0.9% saline solution orally as the control; and subcutaneous sodium stibogluconate 400mg SbV/kg/d as gold standard, for 20 days. Terbinafine was demonstrated to be ineffective when compared to the controls, using clinical and parasitological parameters and the limiting dilution assay.

  7. Ineficácia in vivo da terbinafina em leishmaniose cutânea causada por Leishmania (Leishmania) amazonensis em camundongos C57BL/6 / Terbinafine in vivo inefficacy on cutaneous leishmaniasis caused by Leishmania (Leishmania) amazonensis in C57BL/6 mice

    Scientific Electronic Library Online (English)

    Raimunda Nonata Ribeiro, Sampaio; Gustavo Henrique Soares, Takano; Ana Cristina Barbieri, Malacarne; Tércio Rodrigues, Pereira; Albino Verçosa de, Magalhães.

    2003-07-01

    Full Text Available Testou-se, em camundongos C57BL/6 inoculados com a cepa MHOM/BR/PH8 de Leishmania (Leishmania) amazonensis, terbinafina via oral 100mg/kg/dia, por 20 dias, solução salina 0,9% via oral como controle e stibogluconato de sódio 400mg SbV/kg/dia via subcutânea como padrão-ouro. A terbinafina mostrou-se [...] ineficaz, clínica e parasitologicamente, e pelo ensaio por diluição limitante, quando comparada aos controles. Abstract in english The efficiency of terbinafine was tested in C57BL/6 mice inoculated with the Leishmania (Leishmania) amazonensis strain MHOM/BR/PH8. The mice were administered: terbinafine at a dose of 100mg/kg/d by via oral; 0.9% saline solution orally as the control; and subcutaneous sodium stibogluconate 400mg S [...] bV/kg/d as gold standard, for 20 days. Terbinafine was demonstrated to be ineffective when compared to the controls, using clinical and parasitological parameters and the limiting dilution assay.

  8. Effects of Brazilian propolis on Leishmania amazonensis

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    Diana Copi Ayres

    2007-03-01

    Full Text Available Leishmaniasis, an endemic parasitosis that leads to chronic cutaneous, mucocutaneous or visceral lesions, is part of those diseases, which still requires improved control tools. Propolis has shown activities against different bacteria, fungi, and parasites. In this study we investigated the effect of four ethanolic extracts of typified propolis collected in different Brazilian states, on Leishmania amazonensis performing assays with promastigote forms, extracellular amastigotes, and on infected peritoneal macrophages. Ethanolic extracts of all propolis samples (BRG, BRPG, BRP-1, and BRV were capable to reduce parasite load as monitored by the percentage of infected macrophages and the number of intracellular parasites. BRV sample called red propolis, collected in the state of Alagoas, and containing high concentration of prenylated and benzophenones compounds, was the most active extract against L. amazonensis. The anti-Leishmania effect of BRV sample was increased in a concentration and time dependent manner. BRV treatment proved to be non-toxic to macrophage cultures. Since BRV extract at the concentration of 25 µg/ml reduced the parasite load of macrophages while presented no direct toxic to promastigotes and extracellular amastigotes, it was suggested that constituents of propolis intensify the mechanism of macrophage activation leading to killing of L. amazonensis. Our results demonstrate, for the first time, that ethanolic extracts of Brazilian propolis reduce L. amazonensis infection in macrophages, and encourage further studies of this natural compound in animal models of leishmaniasis.

  9. Leishmaniose cutânea experimental. III- Aspectos histopatológicos do comportamento evolutivo da lesão cutânea produzida em Cebus apella (Primates: Cebidae) por Leishmania (Viannia) lainsoni, L. (V.) braziliensis e L. (Leishmania) amazonensis / Experimental cutaneous Leishmaniasis: III -Histopathological aspects of the evolution of cutaneous lesions produced in Cebus apella (Primates: Cebidae) by Leishmania (Viannia) lainsoni, L. (V.) braziliensis and L. (Leishmania) amazonensis

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    Fernando T., Silveira; Mário A. P., Moraes; Ralph, Lainson; Jeffrey J., Shaw.

    1990-12-01

    Full Text Available Estudaram-se os aspectos histopatológicos relativos à evolução da infecção experimental produzida em Cebus apella (Primates: Cebidae) por Leishmania (V.) lainsoni, L. (V.) braziliensis e L. (L.) amazonensis. O exame microscópico de biópsias seqüênciais, obtidas dos animais a intervalos definidos de [...] tempo (a primeira, às 48 ou 72 horas após a inoculação, e as seguintes, a cada 30 dias), mostrou que o desenvolvimento das lesões, independentemente da espécie de Leishmania inoculada, passa por uma seqüência de etapas a nível tecidual - 1) infiltrado inespecífico crônico; 2) nódulo macrofágico (com numerosos parasitas); 3) necrose das células parasitadas; 4) granuloma epitelióide; 5) absorção da área necrosada (às vezes formando granuloma de corpo estranho); 6) infiltrado inespecífico crônico residual); e 7) cicatrização - que representaria a formação e a resolução das lesões. Discutiram-se também os prováveis mecanismos imunopatológicos que determinam esta seqüência de eventos e sua possível semelhança com a evolução das lesões na leishmaniose tegumentar humana. Abstract in english We have studied the histopathological aspects related to the evolution of cutaneous lesions experimentally produced in the monkey Cebus apella (Primates: Cebidae) by Leishmania (V.) lainsoni, L. (V.) braziliensis and L. (L.) amazonensis. Microscopical examination of a serie of biopsies obtained from [...] these animals showed the kinetics of the cutaneous lesions regarding three species of Leishmania inoculated, as follows: 1) an initial non-specific chronic inflammatory infiltrate; 2) macrophagic nodules; 3) necrosis of parasitized phagocytic cells; 4) epithelioide granuloma; 5) absorption of the necrotic area (sometimes forming "foreign-body granuloma"); 6) a non-specific residual inflammatory infiltration; and 7) cicatrization. These pathological processes are, of course, responsible for both development and resolution of the leishmaniotic lesion. We also discuss some immunopathological mechanisms probably related with the sequencial events, and that could be also responsible for the different clinical aspects found in man.

  10. Activity evaluation from different native or irradiated with {sup 60} Co gamma rays snake venoms and their inhibitory effect on Leishmania (Leishmania) amazonensis; Avaliacao da atividade de diferentes venenos de serpentes, nativos ou irradiados, com radiacao gama de {sup 60} Co, quanto ao poder inibitorio do crescimento de Leishmania (Leishmania) amazonensis

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    Lourenco, Cecilia de Oliveira

    2000-07-01

    Cutaneous leishmaniasis is a disease, caused by Leishmania parasites, that occurs frequently in tropical and sub-tropical regions of the world. Skin lesions that could results in disfiguring aspect characterize it. The treatment is based on few drugs as antimony salts or pentamidine that are toxic with increasing resistance by the parasite. Alternative forms of disease treatment are in constant search, including natural components as snake venoms. Previous studies demonstrate that some components of snake venoms have an inhibitory effect against those parasites, including Leishmania species. Although snake venoms presented high toxicity, several methods have been described to detoxify most or some of their toxic components, with favorable results by the use of gamma irradiation. In this report we tested several native and irradiated snake venoms for inhibitory effect against Leishmania (Leishmania) amazonensis parasite and LLCMK{sub 2} mammalian cells, with enzymatic tests and electrophoresis. There are significant activity in Acanthophis antarcticus, Agkistrodon bilineatus, Bothrops moojeni, Bothrops jararaca, Hoplocephalus stephensi, Naja melanoleuca, Naja mossambica, Pseudechis australis, Pseudechis colletti, Pseudechis guttatus and Pseudechis porphyriacus, venom being inactive Pseudonaja textilis, Notechis ater niger, Notechis scutatus. Oxyuranus microlepidotus and Oxyuranus scutellatus venoms. After 2 KGy of {sup 60}Co irradiation most venom loses significantly their activity. Venoms with antileishmanial activity presented L-amino acid oxidase (L-AO) activity and showed common protein with a molecular weight about 60kDa in SDS-PAGE. These results indicate that L-AO activity in those venoms are probably related with antileishmanial effect. (author)

  11. Analysis and chromosomal mapping of Leishmania (Leishmania) amazonensis amastigote expressed sequence tags

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    Luciana Girotto, Gentil; Fernanda, Lasakosvitsch; José Franco da, Silveira; Márcia Regina Machado dos, Santos; Clara Lúcia, Barbiéri.

    2007-09-01

    Full Text Available The characterization of expressed sequence tags (ESTs) generated from a cDNA library of Leishmania (Leishmania) amazonensis amastigotes is described. The sequencing of 93 clones generated new L. (L.) amazonensis ESTs from which 32% are not related to any other sequences in database and 68% presented [...] significant similarities to known genes. The chromosome localization of some L. (L.) amazonensis ESTs was also determined in L. (L.) amazonensis and L. (L.) major. The characterization of these ESTs is suitable for the genome physical mapping, as well as for the identification of genes encoding cysteine proteinases implicated with protective immune responses in leishmaniasis.

  12. Analysis and chromosomal mapping of Leishmania (Leishmania amazonensis amastigote expressed sequence tags

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    Luciana Girotto Gentil

    2007-09-01

    Full Text Available The characterization of expressed sequence tags (ESTs generated from a cDNA library of Leishmania (Leishmania amazonensis amastigotes is described. The sequencing of 93 clones generated new L. (L. amazonensis ESTs from which 32% are not related to any other sequences in database and 68% presented significant similarities to known genes. The chromosome localization of some L. (L. amazonensis ESTs was also determined in L. (L. amazonensis and L. (L. major. The characterization of these ESTs is suitable for the genome physical mapping, as well as for the identification of genes encoding cysteine proteinases implicated with protective immune responses in leishmaniasis.

  13. Studies on the effectiveness of diarylheptanoids derivatives against Leishmania amazonensis

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    Catarina AC, Araujo; Leila V, Alegrio; Denise CF, Gomes; Marco Edilson F, Lima; Leonardo, Gomes-Cardoso; Leonor L, Leon.

    1999-11-01

    Full Text Available In a previous work we demonstrated that diarylheptanoids extracted from Centrolobium sclerophyllum are very active against Leishmania amazonensis promastigotes. In order to continue our studies with these class of compounds, we decided to evaluate the activity of several diarylheptanoids derived fro [...] m curcumin (diferuloyl methane) against the extracellular form (promastigotes) of L. amazonensis. Furthermore, an experiment against the intracellular form of the parasite (amastigotes) was carried out, comparing the most active compound among the curcumin derivatives (the methylcurcumin) with des-O-methylcentrolobine, the most active diarylheptanoid derived from C. sclerophyllum.

  14. Leishmania amazonensis in dog with clinical diagnosis of visceral leishmaniasis in Paraná State, Brazil – a case reportLeishmania amazonensis em cão com quadro clínico de leishmaniose visceral no Estado do Paraná, Brasil – relato de caso

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    Patrícia Mendes Pereira

    2012-02-01

    Full Text Available Leishmaniasis are antropozoonoses involving various species of Leishmania and hosts, with variable clinical presentations and considered of great importance to public health. This article describes a case of canine leishmaniasis caused by Leishmania amazonensis, from the town of Cambé, Paraná state, Brazil. This state is considered endemic for American cutaneous leishmaniasis (ACL in humans, but this is the first report in a dog in the region, caused by L. amazonensis. The taxonomic characterization of Leishmania was performed by multilocus enzyme electrophoresis technique (MLEE. Clinical signs were similar to those observed in visceral leishmaniasis as polyarthritis and progressive weight loss. It is concluded that the related polyarthritis when related to leishmaniasis, cannot be considered a clinical sign associated only with L. chagasi in view of the L. amazonensis also has this as one of its presentations. The LTA becomes a differential diagnosis of polyarthritis in dogs.As leishmanioses são antropozoonoses que envolvem diversas espécies de Leishmania e hospedeiros, tendo apresentações clínicas variáveis e consideradas de grande importância para a saúde pública. Este artigo descreve um caso de leishmaniose canina causada por Leishmania amazonensis, proveniente da cidade de Cambé, Paraná – Brasil. Este estado é considerado endêmico para a leishmaniose tegumentar americana (LTA em humanos, porém este é o primeiro relato em cão na região, provocado pela L. amazonensis. A caracterização taxonômica da Leishmania foi realizada através da técnica multilocus enzyme electrophoresis (MLEE. Os sinais clínicos foram similares aos observados na leishmaniose visceral como poliartrite e emagrecimento progressivo. Conclui-se que a poliartrite quando relacionada à leishmaniose não pode ser considerada um sinal clínico associado somente a L. chagasi, tendo em vista que a L. amazonensis também tem esta como uma das suas formas de apresentação. A LTA torna-se um diagnóstico diferencial de poliartrite em cães.

  15. Antigenic differences of Leishmania amazonensis isolates causing diffuse cutaneous leishmaniasis.

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    Leon, L L; Machado, G M; Paes, L E; Grimaldi Júnior, G

    1990-01-01

    Six geographically distinct isolates of Leishmania amazonensis causing diffuse cutaneous leishmaniasis (DCL) (from Bahia and Maranhão, in Brazil and Guarico, in Venezuela) were characterized by immunoblot analysis to see whether any geographical or strain-related differences existed in antigenic composition. Western blots of promastigote homogenates were reacted with polyclonal sera from patients infected with L. amazonensis with the various forms of clinical disease. The pattern of antigenic reactivity of these strains revealed the presence of shared antigenic components between geographically distinct L. amazonensis isolates causing DCL, when tested with the sera of the infected patients. In certain cases, however, some polyclonal sera also detected antigenic fractions unique to the strains examined. Variation was observed between the antigenic components of some isolates of L. amazonensis that were recognized by a single serum, and between the antigenic components of a single isolate of L. amazonensis that were recognized by the different patients' sera. However, no constant association was found between the antigenic components identified in these isolates and the geographical area of isolation. These results indicate that, although these parasites appear to be closely related antigenically, they also possess some strain-related antigenic differences. PMID:2278069

  16. Metaciclogénesis y desarrollo de Leishmania (Leishmania) amazonensis (Kinetoplastida: Trypanosomatidae) en Lutzomyia ovallesi (Diptera: Psychodidae) / Metacyclogenesis and development of Leishmania (Leishmania) amazonensis (Kinetoplastida: Trypanosomatidae) in Lutzomyia ovallesi (Diptera: Psychodidae)

    Scientific Electronic Library Online (English)

    Elsa, Nieves; Maritza, Rondón.

    2010-12-01

    Full Text Available La interacción Leishmania-vector es un proceso complejo y multifactorial. En los últimos años se han logrado avances importantes en la relación Leishmania-vector, pero aún se desconocen algunos aspectos. Este trabajo estudia la interacción de Leishmania (Leishmania) amazonensis (Kinetoplastida: Tryp [...] anosomadidae) en Lutzomyia ovallesi (Diptera: Psychodidae). Hembras de L. ovallesi fueron infectadas artificialmente y se determinó el patrón de desarrollo, densidad parasitaria, secuencia de morfotipos de promastigotes en las regiones del intestino, efecto de la realimentación, y el patrón de carbohidratos de la superficie de las leptomonas usando lectinas fluorescentes. Los resultados mostraron desarrollo suprapilórico de L. (L.) amazonensis. La densidad parasitaria mostró dos ciclos de división separados en tiempo y espacio, se observaron morfotipos caracterizados como paramastigotes en división. La realimentación no alteró la migración de los morfotipos y la mayor proporción de leptomonas en el intestino medio torácico sugiere migración natural que pudiera ser modulada por el gradiente de carbohidratos en el intestino del flebótomo y por la exposición de saliva. Las leptomonas exhibieron diferentes patrones de unión de glicoconjugados, que sugieren presencia de distintos residuos de carbohidratos en la superficie celular del parásito y/o flagelo, que pueden tener funciones importantes en la interacción y estabilidad de los parásitos en el lumen del intestino y su migración a las diferentes regiones del mismo. Se aportan diferentes aspectos del desarrollo de L. (L.) amazonensis en L. ovallesi importantes para entender su potencial vectorial Abstract in english Leishmania-vector interaction is a complex and multifactorial process. During the last few years important advances in the Leishmania-vector interaction have been reached but there still some unknown aspects. This work studies the Leishmania (Leishmania) amazonensis (Kinetoplastide: Trypanosomatidae [...] ) interaction in Lutzomia ovallesi (Diptera: Psychodidae). L. ovallesi females were artificially infected and the development pattern, parasite density, and promastigote morphotype sequence in the intestinal regions were determined, as well as the effect of re-feeding, and the carbohydrate pattern of the leptomonad surface using fluorescent lectins. The results showed suprapiloric development of L. (L.)amazonensis. Parasite density showed two division cycles separated in time and space, morphotypes characterized as dividing paramastigotes were observed. Re-feeding did not alter the migration of morphotypes and the greater proportion of leptomonads in the middle intestine suggests a natural migration which could be modulated by the carbohydrate gradient in the phlebotom intestine and by exposure to saliva. Leptomonads exhibited different glyconjugate union patterns which suggest the presence of various carbohydrate residues in the cell surface of the parasite and/or flagellum, which can have important functions in the interaction and stability of the parasites in the intestinal lumen and their migration to the different intestinal regions. Several aspects of L. (L.)amazonensis development in L. ovallesi important for understanding its vectorial potency are discussed

  17. Comparative two-dimensional gel electrophoresis maps for promastigotes of Leishmania amazonensis and Leishmania major

    Directory of Open Access Journals (Sweden)

    Reynolds K. B. Brobey

    2006-02-01

    Full Text Available The outcome of Leishmania infections is determined by both the parasite species and the host genetic makeup. While much has been learned regarding immune responses to this parasite, our knowledge on parasite-derived factors is limited. The recent completion of the L. major and L. infantum genome sequence projects and concurrent advancement in proteomics technology would greatly accelerate the search for novel Leishmania proteins. Using a proteomics-based approach to study species-specific Leishmania proteins, we developed high-resolution, broad pH (3-10 two-dimensional gel electrophoresis (2-DE separations to determine protein-expression profiles between highly infectious forms of the parasitic species L. amazonensis (New World and L. major (Old World. Approximately 1,650 and 1,530 distinct protein spots were detected in the L. amazonensis and L. major gels, respectively. While a vast majority of the spots had similar distribution and intensity, a few were computationally defined as preferentially expressed in L. amazonensis in comparison to L. major, or vice versa. These data attest to the feasibility of establishing a 2-DE-based protein array for inter-species profiling of Leishmania proteins and provide the framework for future design of proteome studies of Leishmania.

  18. Leishmanicidal activity of Himatanthus sucuuba latex against Leishmania amazonensis.

    Science.gov (United States)

    Soares, Deivid C; Andrade, Alexandre L S; Delorenzi, Jan C; Silva, Jefferson R A; Freire-de-Lima, Leonardo; Falcão, Camila A B; Pinto, Angelo C; Rossi-Bergmann, Bartira; Saraiva, Elvira M

    2010-06-01

    Himatanthus sucuuba (HsL) latex exhibited a potent leishmanicidal activity against intracellular amastigotes of Leishmania amazonensis, a causative agent of cutaneous leishmaniasis. HsL inhibited intracellular amastigote growth in a dose-dependent manner (IC(50)=15.7microg/mL). Moreover, HsL increased nitric oxide (NO) and Tumor Nuclear Factor-alpha (TNF-alpha) and decreased Transforming Growth Factor-beta (TGF-beta) production in macrophages. As assessed by plasma membrane integrity and mitochondrial activity, HsL showed low toxicity for host macrophages. HsL in vivo was active by the oral route, reducing the parasite load in established footpad lesions after only five doses. In summary, these findings support HsL as an interesting candidate for further evaluations regarding its potential application as a therapeutical agent against Leishmania. PMID:20096374

  19. Metacyclic promastigotes of Leishmania amazonensis selection using gamma irradiation

    International Nuclear Information System (INIS)

    Leishmania spp. causes a spectrum of human diseases, ranging from self-healing skin lesions to severe and lethal visceral disease. In previous work we demonstrated that the protein and nucleic acid metabolism and oxidative respiration were severely affected by irradiation, in a dose response way, but a small but representative fractions are relatively radio resistant, surviving after 800 Gy of 60Co irradiation. The best explanation could be a selection of metacyclic promastigotes. In these forms, the G0 state allows the adequate correction of DNA repair after the irradiation insult. In this work, we are looking for the ideal radiation dose to select the higher proportion of metacyclic forms of L.. (L.) amazonensis in culture. Parasites were grown in RPMI 1640 medium, plus 20% fetal calf serum, than they were irradiated with different doses ranging between 25 and 400 Gy. Parasites irradiated at 400 Gy infected, proportionally, more cells than parasites irradiated at other doses. To confirm this metacyclogenesis, a complement lysis assay was performed with 5, 10 and 20% of male guinea pig blood serum at 20 deg C for 3 hours, and parasites counted. Guinea pig serum a 10% promotes more lysis, with 200 Gy irradiated parasites being less affected, probably due to metacyclic selection. These preliminary results suggests that the ionizing radiation, specially between 200 and 400 Gy, could be a alternative tool for the selection of metacyclic forms of Leishmania amazonensis in culture. (author)

  20. In Vitro Metacyclogenesis of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis Clinical Field Isolates, as Evaluated by Morphology, Complement Resistance, and Infectivity to Human Macrophages

    OpenAIRE

    da Silva, Ildefonso Alves; Morato, Camila Imai; Quixabeira, Valéria Bernadete Leite; Pereira, Ledice Inácia de Araújo; Dorta, Miriam Leandro; de Oliveira, Milton Adriano Pelli; Horta, Maria de Fátima; Ribeiro-Dias, Fátima

    2015-01-01

    This study was designed to assess in vitro metacyclogenesis of Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis clinical field isolates obtained from patient lesions (L. braziliensis IMG3 and PPS6m; L. amazonensis MAB6). Metacyclogenesis was evaluated by different criteria, namely, promastigote size (morphometric analysis and flow cytometry), surface modifications (loss of lectin or monoclonal antibody (mAb) binding, complement resistance), and infectivity to human ma...

  1. Heme Uptake Mediated by LHR1 Is Essential for Leishmania amazonensis Virulence

    OpenAIRE

    Miguel, Danilo C; Flannery, Andrew R.; Mittra, Bidyottam; Andrews, Norma W

    2013-01-01

    The protozoan parasite Leishmania amazonensis is a heme auxotroph and must acquire this essential factor from the environment. Previous studies showed that L. amazonensis incorporates heme through the transmembrane protein LHR1 (Leishmania Heme Response 1). LHR1-null promastigotes were not viable, suggesting that the transporter is essential for survival. Here, we compared the growth, differentiation, and infectivity for macrophages and mice of wild-type, LHR1-single-knockout (LHR1/?lhr1), an...

  2. Immune responses induced by a Leishmania (Leishmania) amazonensis recombinant antigen in mice and lymphocytes from vaccinated subjects

    OpenAIRE

    Fernandes, Ana Paula; Elizabeth Cortez HERRERA; Mayrink, Wilson; Gazzinelli, Ricardo T; LIU Wen Yu; COSTA Carlos Alberto da; Tavares, Carlos Alberto Pereira; MELO Maria Norma; MICHALICK Marilene Susan Marques; Gentz, Reiner; Nascimento, Evaldo

    1997-01-01

    In the search for Leishmania recombinant antigens that can be used as a vaccine against American Cutaneous Leishmaniasis, we identified a Leishmania (Leishmania) amazonensis recombinant protein of 33 kD (Larp33) which is recognized by antibodies and peripheral blood leukocytes (PBL) from subjects vaccinated with Leishvacin ®, Larp33 was expressed in Escherichia coli after cloning of a 2,2 kb Sau3A digested genomic fragment of L. (L.) amazonensis into the pDS56-6 His vector. Immunoblotting ana...

  3. Inhibition of growth of Leishmania mexicana mexicana by Leishmania mexicana amazonensis during "in vitro" co-cultivation Inibição do crescimento de Leishmania mexicana mexicana por Leishmania mexicana amazonensis durante o co-cultivo "in vitro"

    OpenAIRE

    Pacheco, Raquel S; Gabriel Grimaldi Júnior; Carlos M Morel

    1987-01-01

    Inhibition of one Leishmania subspecies by exometabolites of another subspecies, a phenomenon not previously reported, is suggested by our recent observations in cell cloning experiments with Leishmania mexicana mexicana and Leishmania mexicana amazonensis. Clones were identified using the technique of schizodeme analysis. The phenomenon observed is clearly relevant to studies of parasite isolation, leishmanial metabolism, cross-immunity and chemotherapy.Inhibição do crescimento de um subespé...

  4. A dhfr-ts- Leishmania major Knockout Mutant Cross-protects against Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    PST Veras

    1999-07-01

    Full Text Available E10-5A3 is a dhfr-ts- Leishmania major double knockout auxotrophic shown previously to induce substantial protection against virulent L. major infection in both genetically susceptible and resistant mice. We investigated the capacity of dhfr-ts- to protect against heterologous infection by L. amazonensis. The degree of protection was evaluated by immunization of BALB/c or C57BL/6 mice with E10-5A3, followed by L. amazonensis challenge. Whether immunized by subcutaneous (SC or intravenous (IV inoculation, susceptible and resistant mice displayed a partial degree of protection against challenge with virulent L. amazonensis. SC-immunized BALB/c mice developed lesions 40 to 65% smaller than non immunized mice, while IV immunization led to protection ranging from 40 to 75% in four out of six experiments compared to non immunized animals. The resistant C57BL/6 mice displayed comparable degrees of protection, 57% by SC and 49% by IV immunization. Results are encouraging as it has been previously difficult to obtain protection by SC vaccination against Leishmania, the preferred route for human immunization.

  5. LaRbp38: A Leishmania amazonensis protein that binds nuclear and kinetoplast DNAs

    International Nuclear Information System (INIS)

    Leishmania amazonensis causes a wide spectrum of leishmaniasis. There are no vaccines or adequate treatment for leishmaniasis, therefore there is considerable interest in the identification of new targets for anti-leishmania drugs. The central role of telomere-binding proteins in cell maintenance makes these proteins potential targets for new drugs. In this work, we used a combination of purification chromatographies to screen L. amazonensis proteins for molecules capable of binding double-stranded telomeric DNA. This approach resulted in the purification of a 38 kDa polypeptide that was identified by mass spectrometry as Rbp38, a trypanosomatid protein previously shown to stabilize mitochondrial RNA and to associate with nuclear and kinetoplast DNAs. Western blotting and supershift assays confirmed the identity of the protein as LaRbp38. Competition and chromatin immunoprecipitation assays confirmed that LaRbp38 interacted with kinetoplast and nuclear DNAs in vivo and suggested that LaRbp38 may have dual cellular localization and more than one function

  6. Study of ionizing radiation as a tool for select promastigotes forms of Leishmania Amazonensis, and the megalomaniac response in experimental models; Estudo do uso da radiacao ionizante como ferramenta de selecao de formas promastigotas metaciclicas de Leishmania amazonensis, e a inducao de resposta imunologica em modelos experimentais

    Energy Technology Data Exchange (ETDEWEB)

    Bonetti, Franco Claudio

    2006-07-01

    Actually, millions of people around the globe are under the risk of infection by a protozoan transmitted by a bit of a sand fly. This parasite is a Leishmania spp. This causes a wide spectrum disease, since a cutaneous disease to a visceral one. The cutaneous form is the major clinical manifestation (above 90%). The ionizing radiation, produced in a {sup 60}Co font, had being successes used to promote physical-chemical transformations on different protozoan, including Leishmania spp. In previous work was determined that promastigotes forms of Leishmania amazonensis, irradiated with different doses of radiation, lost their viability maintaining, however, their immunogenicity. In this work, was studied the use of ionizing radiation as a tool for selection of meta cyclic forms of the parasite in axenic culture, for a possible efficient irradiated immuno gene production. Our results shown that cultures irradiated with 400 Gy of gamma irradiation, has 75% of metacyclic form, which are capable to produce, in vitro, an infection that is similar the natural occurrence. These irradiated parasites have their internal cellular structure modified, maintaining their external structure intact. Susceptible strain of mice immunized with leishmania irradiated with different doses had high immunoglobulin production, and maintained this production after the challenge with naive parasites. In other strains this default was similar, however in lower titles. Immunodeficient mice didn't produce immunoglobulin nor on the immunization or on the challenge. (author)

  7. Reprogramming Neutral Lipid Metabolism in Mouse Dendritic Leucocytes Hosting Live Leishmania amazonensis Amastigotes

    OpenAIRE

    2013-01-01

    Once they have gained entry to mammals, live Leishmania (L) amazonensis amastigotes are known to subvert both macrophages and dendritic leucocytes (DLs) as host cells. These L. amazonensis amastigotes then may or may not proliferate in these two phagocytic leucocyte lineages, but in both cases the otherwise versatile differentiation program of these lineages is known to be rapidly remodeled. Here, we describe the rapid reprogramming of C57BL/6 and DBA/2 mouse bone marrow-derived DLs, with a s...

  8. IL-18 contributes to susceptibility to Leishmania amazonensis infection by macrophage-independent mechanisms

    Science.gov (United States)

    Sousa, Louisa M.A.; Carneiro, Matheus B.H.; dos Santos, Liliane M.; Natale, Caio Cotta; Resende, Magda E.; Mosser, David M.; Vieira, Leda Q.

    2016-01-01

    We evaluated the role of IL-18 during Leishmania amazonensis infection in C57BL/6 mice, using IL-18KO mice. We showed that IL-18 is involved in susceptibility to L. amazonensis, since IL-18KO mice presented reduced lesions and parasite loads. Because macrophages are the host cells of the parasite, we investigated if macrophages were involved in IL-18-mediated susceptibility to L. amazonensis. We showed that macrophages obtained from WT or IL-18KO responded similarly to L. amazonensis infection. Moreover, we showed that C57BL/6 macrophages do not respond to IL-18, since they do not express IL-18R. Therefore, macrophages are not involved in IL-18-mediated susceptibility to L. amazonensis. PMID:26009021

  9. Protection of C57BL/10 mice by vaccination with association of purified proteins from Leishmania (Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    MORA Ana Mariela

    1999-01-01

    Full Text Available In the past few years, induction of protective immunity to cutaneous leishmaniasis has been attempted by many researchers using a variety of antigenic preparations, such as living promastigotes or promastigote extracts, partially purified, or defined proteins. In this study, eleven proteins from Leishmania (Leishmania amazonensis (LLa with estimated molecular mass ranging from 97 to 13.5kDa were isolated by polyacrylamide gel electrophoresis and electro-elution. The proteins were associated as vaccine in different preparations with gp63 and BCG (Bacilli Calmette-Guérin. The antigenicity of these vaccines was measured by their ability to induce the production of IFN-g by lymphocyte from subjects vaccinated with Leishvacinâ . The immunogenicity was evaluated in vaccinated mice. C57BL/10 mice were vaccinated with three doses of each vaccine consisting of 30 mg of each protein at 15 days interval. One hundred mg of live BCG was only used in the first dose. Seven days after the last dose, they received a first challenge infection with 105 infective promastigotes and four months later, a second challenge was done. Two months after the second challenge, 42.86% of protection was obtained in the group of mice vaccinated with association of proteins of gp63+46+22kDa, gp63+13.5+25+42kDa, gp63+46+42kDa, gp63+66kDa, and gp63+97kDa; 57.14% of protection was demonstrated with gp63+46+97+13.5kDa, gp63+46+97kDa, gp63+46+33kDa, and 71.43% protection for gp63 plus all proteins. The vaccine of gp63+46+40kDa that did not protect the mice, despite the good specific stimulation of lymphocytes (LSI = 7.60 and 10.77UI/ml of IFN-g production. When crude extract of L. (L. amazonensis was used with BCG a 57.14% of protection was found after the first challenge and 28.57% after the second, the same result was observed for gp63. The data obtained with the vaccines can suggest that the future vaccine probably have to contain, except the 40kDa, a cocktail of proteins that would protect mice against cutaneous leishmaniasis.

  10. Further observations on clinical, histopathological, and immunological features of borderline disseminated cutaneous leishmaniasis caused by Leishmania (Leishmania) amazonensis

    OpenAIRE

    Fernando T. Silveira; Ralph Lainson; Carlos EP Corbett

    2005-01-01

    Leishmania (Leishmania) amazonensis has for some time been considered as the causative agent of two distinct forms of American cutaneous leishmaniasis (ACL): localized cutaneous leishmaniasis (LCL), and anergic diffuse cutaneous leishmaniasis (ADCL). Recently, a new intermediate form of disease, borderline disseminated cutaneous leishmaniasis (BDCL), was introduced into the clinical spectrum of ACL caused by this parasite, and in this paper we record the clinical, histopathological, and immun...

  11. Sand fly captures with Disney traps in area of occurrence of Leishmania (Leishmania amazonensis in the state of Mato Grosso do Sul, mid-western Brazil Capturas de flebotomíneos com armadilhas de Disney em área de ocorrência de Leishmania (Leishmania amazonensis no estado de Mato Grosso do Sul, região Centro-Oeste do Brasil

    Directory of Open Access Journals (Sweden)

    Maria Elizabeth Cavalheiros Dorval

    2010-10-01

    Full Text Available INTRODUCTION: The work was conducted to study phlebotomine fauna (Diptera: Psychodidae and aspects of American cutaneous leishmaniasis transmission in a forested area where Leishmania (Leishmania amazonensis occurs, situated in the municipality of Bela Vista, State of Mato Grosso do Sul, Brazil. METHODS: The captures were conducted with modified Disney traps, using hamster (Mesocricetus auratus as bait, from May 2004 to January 2006. RESULTS: Ten species of phlebotomine sandflies were captured: Brumptomyia avellari, Brumptomyia brumpti, Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni and Sciopemyia sordellii. The two predominant species were Ev bourrouli (57.3% and Bi flaviscutellata (41.4%, present at all sampling sites. Two of the 36 hamsters used as bait presented natural infection with Leishmania. The parasite was identified as Leishmania (Leishmania amazonensis. CONCLUSIONS: Analysis of the results revealed the efficiency of Disney traps for capturing Bichromomyia flaviscutellata and the simultaneous presence of both vector and the Leishmania species transmitted by the same can be considered a predictive factor of the occurrence of leishmaniasis outbreaks for the human population that occupies the location.INTRODUÇÃO: O estudo foi realizado com o objetivo de estudar a fauna de flebotomíneos (Diptera: Psychodidae e aspectos ligados à transmissão da leishmaniose tegumentar americana em uma área florestal com ocorrência de Leishmania (Leishmania amazonensis, situada no município de Bela Vista, Estado do Mato Grosso do Sul, Brasil. MÉTODOS: As capturas de flebotomíneos foram realizadas utilizando-se armadilhas tipo Disney modificadas, com isca roedor, Mesocricetus auratus, no período de maio de 2004 a janeiro de 2006. RESULTADOS: As coletas resultaram na identificação de 10 espécies de Phlebotominae: Brumptomyia avellari, Brumptomyia brumpti, Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni e Sciopemyia sordellii. As duas espécies predominantes foram Ev bourrouli, com 57,3% dos espécimes coletados, e Bi. flaviscutellata, representada por 41,4% e que esteve presente em todos os locais amostrados. Dois hamsters sentinelas adquiriram a infecção natural, sendo os isolados identificados como Leishmania amazonensis. CONCLUSÕES: Os resultados mostram a eficiência das armadilhas Disney para captura de Bichromomyia flaviscutellata, e a presença simultânea de ambos, o vetor e a espécie de Leishmania por ele transmitida pode ser considerada um fator preditor da ocorrência de leishmaniose para a população humana que permanecer nesse local.

  12. Sand fly captures with Disney traps in area of occurrence of Leishmania (Leishmania) amazonensis in the state of Mato Grosso do Sul, mid-western Brazil / Capturas de flebotomíneos com armadilhas de Disney em área de ocorrência de Leishmania (Leishmania) amazonensis no estado de Mato Grosso do Sul, região Centro-Oeste do Brasil

    Scientific Electronic Library Online (English)

    Maria Elizabeth Cavalheiros, Dorval; Tulia Peixoto, Alves; Geucira, Cristaldo; Hilda Carlos da, Rocha; Murilo Andrade, Alves; Elisa Teruya, Oshiro; Alessandra Gutierrez de, Oliveira; Reginaldo Peçanha, Brazil; Eunice Aparecida Bianchi, Galati; Rivaldo Venancio da, Cunha.

    2010-10-01

    Full Text Available INTRODUÇÃO: O estudo foi realizado com o objetivo de estudar a fauna de flebotomíneos (Diptera: Psychodidae) e aspectos ligados à transmissão da leishmaniose tegumentar americana em uma área florestal com ocorrência de Leishmania (Leishmania) amazonensis, situada no município de Bela Vista, Estado d [...] o Mato Grosso do Sul, Brasil. MÉTODOS: As capturas de flebotomíneos foram realizadas utilizando-se armadilhas tipo Disney modificadas, com isca roedor, Mesocricetus auratus, no período de maio de 2004 a janeiro de 2006. RESULTADOS: As coletas resultaram na identificação de 10 espécies de Phlebotominae: Brumptomyia avellari, Brumptomyia brumpti, Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni e Sciopemyia sordellii. As duas espécies predominantes foram Ev bourrouli, com 57,3% dos espécimes coletados, e Bi. flaviscutellata, representada por 41,4% e que esteve presente em todos os locais amostrados. Dois hamsters sentinelas adquiriram a infecção natural, sendo os isolados identificados como Leishmania amazonensis. CONCLUSÕES: Os resultados mostram a eficiência das armadilhas Disney para captura de Bichromomyia flaviscutellata, e a presença simultânea de ambos, o vetor e a espécie de Leishmania por ele transmitida pode ser considerada um fator preditor da ocorrência de leishmaniose para a população humana que permanecer nesse local. Abstract in english INTRODUCTION: The work was conducted to study phlebotomine fauna (Diptera: Psychodidae) and aspects of American cutaneous leishmaniasis transmission in a forested area where Leishmania (Leishmania) amazonensis occurs, situated in the municipality of Bela Vista, State of Mato Grosso do Sul, Brazil. M [...] ETHODS: The captures were conducted with modified Disney traps, using hamster (Mesocricetus auratus) as bait, from May 2004 to January 2006. RESULTS: Ten species of phlebotomine sandflies were captured: Brumptomyia avellari, Brumptomyia brumpti, Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni and Sciopemyia sordellii. The two predominant species were Ev bourrouli (57.3%) and Bi flaviscutellata (41.4%), present at all sampling sites. Two of the 36 hamsters used as bait presented natural infection with Leishmania. The parasite was identified as Leishmania (Leishmania) amazonensis. CONCLUSIONS: Analysis of the results revealed the efficiency of Disney traps for capturing Bichromomyia flaviscutellata and the simultaneous presence of both vector and the Leishmania species transmitted by the same can be considered a predictive factor of the occurrence of leishmaniasis outbreaks for the human population that occupies the location.

  13. Distinct Macrophage Fates after in vitro Infection with Different Species of Leishmania: Induction of Apoptosis by Leishmania (Leishmania) amazonensis, but Not by Leishmania (Viannia) guyanensis

    Science.gov (United States)

    DaMata, Jarina Pena; Mendes, Bárbara Pinheiro; Maciel-Lima, Kátia; Menezes, Cristiane Alves Silva; Dutra, Walderez Ornelas; Sousa, Lirlândia Pires; Horta, Maria Fátima

    2015-01-01

    Leishmania is an intracellular parasite in vertebrate hosts, including man. During infection, amastigotes replicate inside macrophages and are transmitted to healthy cells, leading to amplification of the infection. Although transfer of amastigotes from infected to healthy cells is a crucial step that may shape the outcome of the infection, it is not fully understood. Here we compare L. amazonensis and L. guyanensis infection in C57BL/6 and BALB/c mice and investigate the fate of macrophages when infected with these species of Leishmania in vitro. As previously shown, infection of mice results in distinct outcomes: L. amazonensis causes a chronic infection in both strains of mice (although milder in C57BL/6), whereas L. guyanensis does not cause them disease. In vitro, infection is persistent in L. amazonensis-infected macrophages whereas L. guyanensis growth is controlled by host cells from both strains of mice. We demonstrate that, in vitro, L. amazonensis induces apoptosis of both C57BL/6 and BALB/c macrophages, characterized by PS exposure, DNA cleavage into nucleosomal size fragments, and consequent hypodiploidy. None of these signs were seen in macrophages infected with L. guyanensis, which seem to die through necrosis, as indicated by increased PI-, but not Annexin V-, positive cells. L. amazonensis-induced macrophage apoptosis was associated to activation of caspases-3, -8 and -9 in both strains of mice. Considering these two species of Leishmania and strains of mice, macrophage apoptosis, induced at the initial moments of infection, correlates with chronic infection, regardless of its severity. We present evidence suggestive that macrophages phagocytize L. amazonensis-infected cells, which has not been verified so far. The ingestion of apoptotic infected macrophages by healthy macrophages could be a way of amastigote spreading, leading to the establishment of infection. PMID:26513474

  14. Leishmania amazonensis exhibits phosphatidylserine-dependent procoagulant activity, a process that is counteracted by sandfly saliva

    Directory of Open Access Journals (Sweden)

    Natalia Cadaxo Rochael

    2013-09-01

    Full Text Available Leishmania parasites expose phosphatidylserine (PS on their surface, a process that has been associated with regulation of host's immune responses. In this study we demonstrate that PS exposure by metacyclic promastigotes of Leishmania amazonensis favours blood coagulation. L. amazonensis accelerates in vitro coagulation of human plasma. In addition, L. amazonensis supports the assembly of the prothrombinase complex, thus promoting thrombin formation. This process was reversed by annexin V which blocks PS binding sites. During blood meal, Lutzomyia longipalpis sandfly inject saliva in the bite site, which has a series of pharmacologically active compounds that inhibit blood coagulation. Since saliva and parasites are co-injected in the host during natural transmission, we evaluated the anticoagulant properties of sandfly saliva in counteracting the procoagulant activity of L. amazonensis . Lu. longipalpis saliva reverses plasma clotting promoted by promastigotes. It also inhibits thrombin formation by the prothrombinase complex assembled either in phosphatidylcholine (PC/PS vesicles or in L. amazonensis . Sandfly saliva inhibits factor X activation by the intrinsic tenase complex assembled on PC/PS vesicles and blocks factor Xa catalytic activity. Altogether our results show that metacyclic promastigotes of L. amazonensis are procoagulant due to PS exposure. Notably, this effect is efficiently counteracted by sandfly saliva.

  15. Leishmania amazonensis exhibits phosphatidylserine-dependent procoagulant activity, a process that is counteracted by sandfly saliva

    Scientific Electronic Library Online (English)

    Natalia Cadaxo, Rochael; Luize Goncalves, Lima; Sandra Maria Pereira de, Oliveira; Marcello Andre, Barcinski; Elvira Maria, Saraiva; Robson Queiroz, Monteiro; Lucia Helena, Pinto-da-Silva.

    2013-09-01

    Full Text Available Leishmania parasites expose phosphatidylserine (PS) on their surface, a process that has been associated with regulation of host's immune responses. In this study we demonstrate that PS exposure by metacyclic promastigotes of Leishmania amazonensis favours blood coagulation. L. amazonensis accelera [...] tes in vitro coagulation of human plasma. In addition, L. amazonensis supports the assembly of the prothrombinase complex, thus promoting thrombin formation. This process was reversed by annexin V which blocks PS binding sites. During blood meal, Lutzomyia longipalpis sandfly inject saliva in the bite site, which has a series of pharmacologically active compounds that inhibit blood coagulation. Since saliva and parasites are co-injected in the host during natural transmission, we evaluated the anticoagulant properties of sandfly saliva in counteracting the procoagulant activity of L. amazonensis . Lu. longipalpis saliva reverses plasma clotting promoted by promastigotes. It also inhibits thrombin formation by the prothrombinase complex assembled either in phosphatidylcholine (PC)/PS vesicles or in L. amazonensis . Sandfly saliva inhibits factor X activation by the intrinsic tenase complex assembled on PC/PS vesicles and blocks factor Xa catalytic activity. Altogether our results show that metacyclic promastigotes of L. amazonensis are procoagulant due to PS exposure. Notably, this effect is efficiently counteracted by sandfly saliva.

  16. Combined effect of the essential oil from Chenopodium ambrosioides and antileishmanial drugs on promastigotes of Leishmania amazonensis Efeito combinado do óleo de essência de Chenopodium ambrosioides e drogas anti-leishmaniose nos promastigotas de Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    Lianet Monzote

    2007-08-01

    Full Text Available To date, there are no vaccines against Leishmania, and chemotherapy remains the mainstay for the control of leishmaniasis. The drugs of choice used for leishmaniasis therapy are significantly toxic, expensive and with a growing frequency of refractory infections. Because of these limitations, a combination therapy is the better hope. This work demonstrates that the essential oil from Chenopodium ambrosioides shows a synergic activity after incubation in conjunction with pentamidine against promastigotes of Leishmania amazonensis. However, an indifferent effect has been found for combinations of meglumine antimoniate or amphotericin B and the essential oil.Até hoje não temos vacina contra a Leishmania e a quimioterapia é a indicação para o controle desta doença. Os remédios que hoje utilizamos são tóxicos e muito caros e além disso o resultado não é sempre o desejado. Por isso, uma terapia de combinação é a melhor opção. Este trabalho mostra que o óleo de essência de C. ambrosioides tem atividade sinérgica junto com a pentamidina sobre os promastigotas de L. amazonensis, diferente do resultado da combinação de antimônio de meglumine e anfotericina B e o óleo de essência.

  17. Combined effect of the essential oil from Chenopodium ambrosioides and antileishmanial drugs on promastigotes of Leishmania amazonensis / Efeito combinado do óleo de essência de Chenopodium ambrosioides e drogas anti-leishmaniose nos promastigotas de Leishmania amazonensis

    Scientific Electronic Library Online (English)

    Lianet, Monzote; Ana Margarita, Montalvo; Ramón, Scull; Migdalia, Miranda; Juan, Abreu.

    2007-08-01

    Full Text Available Até hoje não temos vacina contra a Leishmania e a quimioterapia é a indicação para o controle desta doença. Os remédios que hoje utilizamos são tóxicos e muito caros e além disso o resultado não é sempre o desejado. Por isso, uma terapia de combinação é a melhor opção. Este trabalho mostra que o óle [...] o de essência de C. ambrosioides tem atividade sinérgica junto com a pentamidina sobre os promastigotas de L. amazonensis, diferente do resultado da combinação de antimônio de meglumine e anfotericina B e o óleo de essência. Abstract in english To date, there are no vaccines against Leishmania, and chemotherapy remains the mainstay for the control of leishmaniasis. The drugs of choice used for leishmaniasis therapy are significantly toxic, expensive and with a growing frequency of refractory infections. Because of these limitations, a comb [...] ination therapy is the better hope. This work demonstrates that the essential oil from Chenopodium ambrosioides shows a synergic activity after incubation in conjunction with pentamidine against promastigotes of Leishmania amazonensis. However, an indifferent effect has been found for combinations of meglumine antimoniate or amphotericin B and the essential oil.

  18. Diseminación tisular y efectos histopatológicos producidos por Leishmania mexicana amazonensis en roedores infectados experimentalmente / Tissue dissemination and histopathology effects produced by Leishmania mexicana amazonensis in rodents experimentally infected

    Scientific Electronic Library Online (English)

    Elizabeth, Bruzual; Lucila, Arcay; María Antonia, de la Parte-Pérez.

    2008-12-01

    Full Text Available Al género Leishmania pertenecen diferentes especies responsables de diversas formas clínicas de leishmaniasis. En este trabajo comparamos los efectos histopatológicos producidos en hamsters (Mesocricetus auratus) y ratones (Mus musculus) infectados experimentalmente por vía subcutánea con 20 x 10(5) [...] amastigotes de Leishmania mexicana amazonensis. Nuestros resultados mostraron en hamsters daños histopatológicos en intestino, escroto, testículo, epidídimo, bazo, hígado, riñón, corazón y pulmón y en ratones en testículo, bazo, riñón y corazón. Estos resultados evidencian diferencias en la diseminación tisular, metástasis cutáneas, efectos histopatológicos y mortalidad, siendo más amplios, tempranos y graves en hamsters que en ratones. Abstract in english Leishmania comprises different species responsible of a variety of clinical forms of leishmaniasis. In this investigation we compared the histopathological effects produced in hamsters (Mesocricetus auratus) and mice (Mus musculus), inoculated by the subcutaneous route with 20 x 10(5) amastigotes of [...] Leishmania mexicana amazonensis. Our results in hamsters show tissue damage in gut, scrotum, testicle, epididimus, spleen, liver, kidney, heart and lung and in mice we found histopathological changes in testicle, spleen, kidney and heart. These results show differences on tissue dissemination, cutaneous metastasis, histopathological effects and mortality, being more extensive, early and severe in hamsters then in mice.

  19. The T-cell anergy induced by Leishmania amazonensis antigens is related with defective antigen presentation and apoptosis

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    Roberta O. Pinheiro

    2004-09-01

    Full Text Available Leishmania amazonensis is the main agent of diffuse cutaneous leishmaniasis, a disease associated with anergic immune responses. In this study we show that the crude antigen of Leishmania amazonensis (LaAg but not L. braziliensis promastigotes (LbAg contains substances that suppress mitogenic and spontaneous proliferative responses of T cells. The suppressive substances in LaAg are thermoresistant (100ºC/1h and partially dependent on protease activity. T cell anergy was not due to a decreased production of growth factors as it was not reverted by addition of exogenous IL-2, IL-4, IFN-gamma or IL-12. LaAg did not inhibit anti-CD3-induced T cell activation, suggesting that anergy was due to a defect in antigen presentation. It was also not due to cell necrosis, but was accompanied by expressive DNA fragmentation in lymph node cells, indicative of apoptosis. Although pre-incubation of macrophages with LaAg prevented their capacity to present antigens, this effect was not due to apoptosis of the former. These results suggest that the T cell anergy found in diffuse leishmaniasis may be the result of parasite antigen-driven apoptosis of those cells following defective antigen presentation.A Leishmania amazonensis é o principal agente etiológico da leishmaniose cutânea difusa, uma doença associada a respostas imunes anérgicas. Neste estudo nós mostramos que o extrato bruto de promastigotas de Leishmania amazonensis (LaAg, mas não de L. braziliensis (LbAg, contém substâncias que suprimem respostas proliferativas, espontâneas e mitogênicas, de células T. As substâncias supressoras no LaAg são termo-resistentes (100°C/1h e parcialmente dependentes da atividade de proteases. A anergia de células T não foi devida à diminuição na produção de fatores de crescimento, uma vez que não foi revertida pela adição de: IL-2, IL-4, IFN-gama ou IL-12. O LaAg não inibiu a ativação de células T induzida por anti-CD3, sugerindo que a anergia é devida a um defeito na apresentação de antígenos. A anergia não foi devida à necrose celular, mas foi acompanhada de uma expressiva fragmentação de DNA nas células de linfonodos, indicativo de apoptose. Apesar da pré-incubação de macrófagos com LaAg ter inibido sua capacidade de apresentação de antigenos, este efeito não foi devido à apoptose dos primeiros. Estes resultados sugerem que a anergia de células T encontrada na leishmaniose difusa deve ser devida à apoptose dessas células que se segue à apresentação defeituosa de antígenos pelo antígeno do parasito.

  20. Correlation of meta 1 expression with culture stage, cell morphology and infectivity in Leishmania (Leishmania amazonensis promastigotes

    Directory of Open Access Journals (Sweden)

    Marcos Gonzaga dos Santos

    2011-03-01

    Full Text Available The parasitic protozoan Leishmania (Leishmania amazonensis alternates between mammalian and insect hosts. In the insect host, the parasites proliferate as procyclic promastigotes andthen differentiate into metacyclic infective forms. The meta 1 gene is preferentially expressed during metacyclogenesis. Meta 1 expression profile determination along parasite growth curves revealed that the meta 1 mRNA level peaked at the early stationary phase then decreased to an intermediate level. No correlation was observed between meta 1 expression and infectivity. Conversely, infectivity correlated with the increase of apoptotic cells in the late stationary phase.

  1. Subversion of Immunity by Leishmania amazonensis Parasites: Possible Role of Phosphatidylserine as a Main Regulator

    OpenAIRE

    Joao Luiz Mendes Wanderley; Jaqueline França Costa; Valéria Matos Borges; Marcello Barcinski

    2012-01-01

    Leishmania amazonensis parasites cause progressive disease in most inbred mouse strains and are associated with the development of diffuse cutaneous leishmaniasis in humans. The poor activation of an effective cellular response is correlated with the ability of these parasites to infect mononuclear phagocytic cells without triggering their activation or actively suppressing innate responses of these cells. Here we discuss the possible role of phosphatidylserine exposure by these parasites as ...

  2. Selective Effect of 2?,6?-Dihydroxy-4?-Methoxychalcone Isolated from Piper aduncum on Leishmania amazonensis

    OpenAIRE

    Torres-Santos, Eduardo Caio; Moreira, Davyson Lima; Maria Auxiliadora C Kaplan; Meirelles, Maria Nazareth; Rossi-Bergmann, Bartira

    1999-01-01

    2?,6?-Dihydroxy-4?-methoxychalcone (DMC) was purified from the dichloromethane extract of Piper aduncum inflorescences. DMC showed significant activity in vitro against promastigotes and intracellular amastigotes of Leishmania amazonensis, with 50% effective doses of 0.5 and 24 ?g/ml, respectively. Its inhibitory effect on amastigotes is apparently a direct effect on the parasites and is not due to activation of the nitrogen oxidative metabolism of macrophages, since the production of nitric ...

  3. Activity evaluation from different native or irradiated with 60 Co gamma rays snake venoms and their inhibitory effect on Leishmania (Leishmania) amazonensis

    International Nuclear Information System (INIS)

    Cutaneous leishmaniasis is a disease, caused by Leishmania parasites, that occurs frequently in tropical and sub-tropical regions of the world. Skin lesions that could results in disfiguring aspect characterize it. The treatment is based on few drugs as antimony salts or pentamidine that are toxic with increasing resistance by the parasite. Alternative forms of disease treatment are in constant search, including natural components as snake venoms. Previous studies demonstrate that some components of snake venoms have an inhibitory effect against those parasites, including Leishmania species. Although snake venoms presented high toxicity, several methods have been described to detoxify most or some of their toxic components, with favorable results by the use of gamma irradiation. In this report we tested several native and irradiated snake venoms for inhibitory effect against Leishmania (Leishmania) amazonensis parasite and LLCMK2 mammalian cells, with enzymatic tests and electrophoresis. There are significant activity in Acanthophis antarcticus, Agkistrodon bilineatus, Bothrops moojeni, Bothrops jararaca, Hoplocephalus stephensi, Naja melanoleuca, Naja mossambica, Pseudechis australis, Pseudechis colletti, Pseudechis guttatus and Pseudechis porphyriacus, venom being inactive Pseudonaja textilis, Notechis ater niger, Notechis scutatus. Oxyuranus microlepidotus and Oxyuranus scutellatus venoms. After 2 KGy of 60Co irradiation most venom loses significantly their activity. Venoms with antileishmanial activity presented L-amino acid oxidase (L-AO) activity and showed common protein with a molecular weight about 60kDa in SDS-PAGE. These results indicate that L-AO activity in those venoms are probably related with antileishmanial effect. (author)

  4. INTRACELLULAR Leishmania amazonensis KILLING INDUCED BY THE GUANINE NUCLEOSIDE 8-BROMOGUANOSINE

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    GIORGIO Selma

    1998-01-01

    Full Text Available In this study we investigated the effect of 8-Bromoguanosine, an immunostimulatory compound, on the cytotoxicity of macrophages against Leishmania amazonensis in an in vitro system. The results showed that macrophages treated with 8-Bromoguanosine before or after infection are capable to reduce parasite load, as monitored by the number of amastigotes per macrophage and the percentage of infected cells (i.e. phagocytic index. Since 8-Bromoguanosine was not directly toxic to the promastigotes, it was concluded that the ribonucleoside induced macrophage activation. Presumably, 8-Bromoguanosine primed macrophages by inducing interferon alpha and beta which ultimately led to L. amazonensis amastigote killing. The results suggest that guanine ribonucleosides may be useful to treat infections with intracellular pathogens.

  5. Identificação de espécies de Leishmania isoladas de casos humanos em Mato Grosso do Sul por meio da reação em cadeia da polimerase / Identification of Leishmania species isolated in human cases in Mato Grosso do Sul, by means of the polymerase chain reaction

    Scientific Electronic Library Online (English)

    Manoel Sebastião da Costa, Lima Junior; Renato, Andreotti; Maria Elizabeth Moraes Cavalheiros, Dorval; Elisa Teruya, Oshiro; Alessandra Gutierrez de, Oliveira; Maria de Fatima Cepa, Matos.

    2009-06-01

    Full Text Available As leishmanioses são zoonoses endêmicas em Mato Grosso do Sul e têm por agentes etiológicos nessa região Leishmania (Leishmania) chagasi, Leishmania (Leishmania) amazonensis e Leishmania (Viannia) braziliensis. Como método para identificação de espécies de Leishmania, a reação em cadeia da polimeras [...] e é uma ferramenta com elevada especificidade e sensibilidade. Analisaram-se 39 isolados de Leishmania criopreservados, obtidos por meio de aspirado medular e/ou biópsia de lesão, conforme a suspeita clínica. Os isolados foram submetidos à extração de DNA e à reação em cadeia da polimerase com os iniciadores: RV1/RV2 para Leishmania (Leishmania) chagasi, a1/a2 para a identificação de Leishmania (Leishmania) amazonensis e b1/b2 para Leishmania (Viannia) braziliensis. Leishmania (Leishmania) chagasi foi a única espécie identificada em 37 casos de leishmaniose visceral. Leishmania (Leishmania) amazonensis foi identificada em dois isolados de pacientes com diagnóstico de leishmaniose tegumentar. Os resultados obtidos confirmam a possibilidade do uso dos três pares de iniciadores como uma ferramenta na caracterização de isolados de Leishmania. Abstract in english Leishmaniases are endemic zoonoses in the State of Mato Grosso do Sul. Their etiological agents in this region of Brazil are Leishmania (Leishmania) chagasi, Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis. The polymerase chain reaction (PCR) is a tool with high specificity [...] and sensitivity for identifying Leishmania species. This study examined 39 cryopreserved isolates of Leishmania that had been collected by bone marrow aspiration and/or lesion biopsy, depending on the clinical suspicion. The isolates were subjected to DNA extraction and PCR using the following primers: RV1/RV2 for identifying Leishmania (Leishmania) chagasi, a1/a2 for Leishmania (Leishmania) amazonensis and b1/b2 for Leishmania (Viannia) braziliensis.Leishmania (Leishmania) chagasi was the only species identified in the 37 cases of visceral leishmaniasis.Leishmania (Leishmania) amazonensis was identified in two isolates from patients with a diagnosis of cutaneous leishmaniasis. The results obtained confirm that it is possible to use these three pairs of primers as a tool for characterizing Leishmania isolates.

  6. Evaluation of Antileishmanial Activity of Albaha Medicinal Plants against Leishmania amazonensis

    Science.gov (United States)

    Al-Sokari, Saeed S.; Ali, Nasser A. Awadh; Monzote, Lianet; Al-Fatimi, Mohamed A.

    2015-01-01

    Sixteen methanolic extracts obtained from thirteen plant species, selected either from ethnobotanical or chemotaxonomical data, were screened for their antileishmanial activity against Leishmania amazonensis. The cytotoxic activity against normal peritoneal macrophages from normal BALB/c mice was also determined. Eight extracts had IC50 values ranging from Euphorbia helioscopia, and Solanum incanum leaf extracts showed antileishmanial activities with IC50 between <12.5–26.9?µg/mL and acceptable selectivity indices of 8–5. The other leishmanicidal plant extracts, with IC50 ranging from 18.0 to 29.5?µg/mL, exhibited low selectivity indices. PMID:26357662

  7. Experimental infection with Leishmania (Viannia braziliensis, and Leishmania (Leishmania amazonensis in the marmoset, Callithrix penicillata (Primates: Callithricidae

    Directory of Open Access Journals (Sweden)

    César A. Cuba Cuba

    1990-12-01

    Full Text Available Foureen marmosets (Callithrix penicillata were inoculated intradermally with promastigotes and/or amastigotes of Leishmania (Viannia brazilensis (L. (V b. strains MHOM/BR/83/LTB-300MHOM/BR/85/LTB-12 MHOM/BR/81/LTB-179 and MHOM/BR/82/LTB-250. The evolution of subsequent lesions was studied for 15 to 75 weeks post-inoculation (PI. All but of the L. (V b. injected marmosets developed a cutaneous lesion at the point of inoculation after 3 to 9 weeks, characterized by the appearance of subcutaneous nodules containing parasites. parasites were isolated by culture (Difco Blood Agar from all 11 positive animals. The maximum size of the lesions was variable and ranged between 37 mm² to 107 mm². Ulceration of primary nodules became evident after 3 to 12 weeks in all infected marmosets, but was faster and larger in 5 of the 11 animals. The active lesions persisted in 9 out of 11 Callithrix until the en of the observation period, which varied from 15-75 weeks. In 3 animals spontaneous healing of their lesions (13 to 25 weeks, PI was observed buth with cryptic parasitism. In another 2 infected animals there was regression followed by reactivation of the cutaneous lesions. The appearance of smaller satellite lesions adjacent to primary ones, as well as metastatic lesions to the ear lobes, were documented in 2 animals. Promastigotes of L. (Leishmania amazonensis (L.(La. MHOM/BR/77/LTB-16 were inoculated in 1 marmoset. This animal remained chronically infected for 6 months and the lesions developed in a similar manner to L.(Vb. infected marmosets. No significant differences in clinical and parasitological behaviour were observed between promastigote or amastigote derived infections of the 2 species. Both produced chronic, long lasting lesions which eventually healed. The same was true for parameters of size and ulceration. Skin tests converted to parasite in 11 of 15 inected masmosets and in 10 of 12 parasite positive animnals. Moderate levels of circulating antibodies were also observed by IFAT /IgG assays. In spite of the failure to reproduce the mucosal form of the disease, an important aspect of the Callithrix model in experimental cutaneous leishmaniasis lies in the reproduction of 2 clinical events that are common in humans, namely, the chronic ulceration and spontaneous healing of the lesions.

  8. Experimental infection with Leishmania (Viannia) braziliensis, and Leishmania (Leishmania) amazonensis in the marmoset, Callithrix penicillata (Primates: Callithricidae)

    Scientific Electronic Library Online (English)

    César A. Cuba, Cuba; Vera, Ferreira; Maria, Bampi; Albino, Magalhães; Philip D., Marsden; Alejandro, Vexenat; Milton Thiago de, Mello.

    1990-12-01

    Full Text Available Foureen marmosets (Callithrix penicillata) were inoculated intradermally with promastigotes and/or amastigotes of Leishmania (Viannia) brazilensis (L. (V) b.) strains MHOM/BR/83/LTB-300MHOM/BR/85/LTB-12 MHOM/BR/81/LTB-179 and MHOM/BR/82/LTB-250. The evolution of subsequent lesions was studied for 15 [...] to 75 weeks post-inoculation (PI). All but of the L. (V) b. injected marmosets developed a cutaneous lesion at the point of inoculation after 3 to 9 weeks, characterized by the appearance of subcutaneous nodules containing parasites. parasites were isolated by culture (Difco Blood Agar) from all 11 positive animals. The maximum size of the lesions was variable and ranged between 37 mm² to 107 mm². Ulceration of primary nodules became evident after 3 to 12 weeks in all infected marmosets, but was faster and larger in 5 of the 11 animals. The active lesions persisted in 9 out of 11 Callithrix until the en of the observation period, which varied from 15-75 weeks. In 3 animals spontaneous healing of their lesions (13 to 25 weeks, PI) was observed buth with cryptic parasitism. In another 2 infected animals there was regression followed by reactivation of the cutaneous lesions. The appearance of smaller satellite lesions adjacent to primary ones, as well as metastatic lesions to the ear lobes, were documented in 2 animals. Promastigotes of L. (Leishmania) amazonensis (L.(L)a.) MHOM/BR/77/LTB-16 were inoculated in 1 marmoset. This animal remained chronically infected for 6 months and the lesions developed in a similar manner to L.(V)b. infected marmosets. No significant differences in clinical and parasitological behaviour were observed between promastigote or amastigote derived infections of the 2 species. Both produced chronic, long lasting lesions which eventually healed. The same was true for parameters of size and ulceration. Skin tests converted to parasite in 11 of 15 inected masmosets and in 10 of 12 parasite positive animnals. Moderate levels of circulating antibodies were also observed by IFAT /IgG assays. In spite of the failure to reproduce the mucosal form of the disease, an important aspect of the Callithrix model in experimental cutaneous leishmaniasis lies in the reproduction of 2 clinical events that are common in humans, namely, the chronic ulceration and spontaneous healing of the lesions.

  9. Benzaldehyde Thiosemicarbazone Derived from Limonene Complexed with Copper Induced Mitochondrial Dysfunction in Leishmania amazonensis

    Science.gov (United States)

    Britta, Elizandra Aparecida; Barbosa Silva, Ana Paula; Ueda-Nakamura, Tânia; Dias-Filho, Benedito Prado; Silva, Cleuza Conceição; Sernaglia, Rosana Lázara; Nakamura, Celso Vataru

    2012-01-01

    Background Leishmaniasis is a major health problem that affects more than 12 million people. Treatment presents several problems, including high toxicity and many adverse effects, leading to the discontinuation of treatment and emergence of resistant strains. Methodology/Principal Findings We evaluated the in vitro antileishmanial activity of benzaldehyde thiosemicarbazone derived from limonene complexed with copper, termed BenzCo, against Leishmania amazonensis. BenzCo inhibited the growth of the promastigote and axenic amastigote forms, with IC50 concentrations of 3.8 and 9.5 µM, respectively, with 72 h of incubation. Intracellular amastigotes were inhibited by the compound, with an IC50 of 10.7 µM. BenzCo altered the shape, size, and ultrastructure of the parasites. Mitochondrial membrane depolarization was observed in protozoa treated with BenzCo but caused no alterations in the plasma membrane. Additionally, BenzCo induced lipoperoxidation and the production of mitochondrial superoxide anion radicals in promastigotes and axenic amastigotes of Leishmania amazonensis. Conclusion/Significance Our studies indicated that the antileishmania activity of BenzCo might be associated with mitochondrial dysfunction and oxidative damage, leading to parasite death. PMID:22870222

  10. A novel alkyl phosphocholine-dinitroaniline hybrid molecule exhibits biological activity in vitro against Leishmania amazonensis.

    Science.gov (United States)

    Godinho, Joseane Lima Prado; Georgikopoulou, Kalliopi; Calogeropoulou, Theodora; de Souza, Wanderley; Rodrigues, Juliany Cola Fernandes

    2013-09-01

    Parasitic protozoa of the Leishmania genus cause leishmaniasis, an important complex of tropical diseases that affect about 12 million people around the world. The drugs used to treat leishmaniasis are pentavalent antimonials, miltefosine, amphotericin B and pentamidine. In the present study, we evaluated the effect of a novel alkyl phosphocholine-dinitroaniline hybrid molecule, TC95, against Leishmania amazonensis promastigotes and intracellular amastigotes. Antiproliferative assays indicated that TC95 is a potent inhibitor of promastigotes and intracellular amastigotes with IC50 values of 2.6 and 1.2 μM, respectively. Fluorescence microscopy with anti-α-tubulin antibody revealed changes in the cytoskeleton, whilst scanning electron microscopy showed alterations in the shape, plasma membrane, length of the flagellum, and cell cycle. Flow cytometry confirmed the cell cycle arrest mainly in G1 phase, however a significant population appeared in sub G0/G1 and super-G2. The alterations in the plasma membrane integrity were confirmed by fluorometric analysis using Sytox Blue. Transmission electron microscopy also revealed an accumulation of lipid bodies, confirmed by fluorescence microscopy and fluorometric analysis using Nile Red. Important lesions were also observed in organelles such as mitochondrion, endoplasmic reticulum and Golgi complex. In summary, our study suggests that TC95, an alkyl phosphocholine-trifluralin hybrid molecule, is a promising novel compound against L. amazonensis. PMID:23845259

  11. A Trypanosomatid Iron Transporter that Regulates Mitochondrial Function Is Required for Leishmania amazonensis Virulence

    Science.gov (United States)

    Mittra, Bidyottam; Laranjeira-Silva, Maria Fernanda; Perrone Bezerra de Menezes, Juliana; Jensen, Jennifer; Michailowsky, Vladimir; Andrews, Norma W.

    2016-01-01

    Iron, an essential co-factor of respiratory chain proteins, is critical for mitochondrial function and maintenance of its redox balance. We previously reported a role for iron uptake in differentiation of Leishmania amazonensis into virulent amastigotes, by a mechanism that involves reactive oxygen species (ROS) production and is independent of the classical pH and temperature cues. Iron import into mitochondria was proposed to be essential for this process, but evidence supporting this hypothesis was lacking because the Leishmania mitochondrial iron transporter was unknown. Here we describe MIT1, a homolog of the mitochondrial iron importer genes mrs3 (yeast) and mitoferrin-1 (human) that is highly conserved among trypanosomatids. MIT1 expression was essential for the survival of Trypanosoma brucei procyclic but not bloodstream forms, which lack functional respiratory complexes. L. amazonensis LMIT1 null mutants could not be generated, suggesting that this mitochondrial iron importer is essential for promastigote viability. Promastigotes lacking one LMIT1 allele (LMIT1/?lmit1) showed growth defects and were more susceptible to ROS toxicity, consistent with the role of iron as the essential co-factor of trypanosomatid mitochondrial superoxide dismutases. LMIT1/?lmit1 metacyclic promastigotes were unable to replicate as intracellular amastigotes after infecting macrophages or cause cutaneous lesions in mice. When induced to differentiate axenically into amastigotes, LMIT1/?lmit1 showed strong defects in iron content and function of mitochondria, were unable to upregulate the ROS-regulatory enzyme FeSOD, and showed mitochondrial changes suggestive of redox imbalance. Our results demonstrate the importance of mitochondrial iron uptake in trypanosomatid parasites, and highlight the role of LMIT1 in the iron-regulated process that orchestrates differentiation of L. amazonensis into infective amastigotes. PMID:26741360

  12. In vitro activity of the hydroethanolic extract and biflavonoids isolated from Selaginella sellowii on Leishmania (Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    Yasmin Silva Rizk

    2014-12-01

    Full Text Available This study is the first phytochemical investigation of Selaginella sellowii and demonstrates the antileishmanial activity of the hydroethanolic extract from this plant (SSHE, as well as of the biflavonoids amentoflavone and robustaflavone, isolated from this species. The effects of these substances were evaluated on intracellular amastigotes of Leishmania (Leishmania amazonensis, an aetiological agent of American cutaneous leishmaniasis. SSHE was highly active against intracellular amastigotes [the half maximum inhibitory concentration (IC50 = 20.2 µg/mL]. Fractionation of the extract led to the isolation of the two bioflavonoids with the highest activity: amentoflavone, which was about 200 times more active (IC50 = 0.1 ?g/mL and less cytotoxic than SSHE (IC50 = 2.2 and 3 ?g/mL, respectively on NIH/3T3 and J774.A1 cells, with a high selectivity index (SI (22 and 30, robustaflavone, which was also active against L. amazonensis (IC50 = 2.8 µg/mL, but more cytotoxic, with IC50 = 25.5 µg/mL (SI = 9.1 on NIH/3T3 cells and IC50 = 3.1 µg/mL (SI = 1.1 on J774.A1 cells. The production of nitric oxide (NO was lower in cells treated with amentoflavone (suggesting that NO does not contribute to the leishmanicidal mechanism in this case, while NO release was higher after treatment with robustaflavone. S. sellowii may be a potential source of biflavonoids that could provide promising compounds for the treatment of cutaneous leishmaniasis.

  13. In vitro activity of the hydroethanolic extract and biflavonoids isolated from Selaginella sellowii on Leishmania (Leishmania) amazonensis

    Scientific Electronic Library Online (English)

    Yasmin Silva, Rizk; Alice, Fischer; Marillin de Castro, Cunha; Patrik Oening, Rodrigues; Maria Carolina Silva, Marques; Maria de Fátima Cepa, Matos; Mônica Cristina Toffoli, Kadri; Carlos Alexandre, Carollo; Carla Cardozo Pinto de, Arruda.

    2014-12-01

    Full Text Available This study is the first phytochemical investigation of Selaginella sellowii and demonstrates the antileishmanial activity of the hydroethanolic extract from this plant (SSHE), as well as of the biflavonoids amentoflavone and robustaflavone, isolated from this species. The effects of these substances [...] were evaluated on intracellular amastigotes of Leishmania (Leishmania) amazonensis, an aetiological agent of American cutaneous leishmaniasis. SSHE was highly active against intracellular amastigotes [the half maximum inhibitory concentration (IC50) = 20.2 µg/mL]. Fractionation of the extract led to the isolation of the two bioflavonoids with the highest activity: amentoflavone, which was about 200 times more active (IC50 = 0.1 ?g/mL) and less cytotoxic than SSHE (IC50 = 2.2 and 3 ?g/mL, respectively on NIH/3T3 and J774.A1 cells), with a high selectivity index (SI) (22 and 30), robustaflavone, which was also active against L. amazonensis (IC50 = 2.8 µg/mL), but more cytotoxic, with IC50 = 25.5 µg/mL (SI = 9.1) on NIH/3T3 cells and IC50 = 3.1 µg/mL (SI = 1.1) on J774.A1 cells. The production of nitric oxide (NO) was lower in cells treated with amentoflavone (suggesting that NO does not contribute to the leishmanicidal mechanism in this case), while NO release was higher after treatment with robustaflavone. S. sellowii may be a potential source of biflavonoids that could provide promising compounds for the treatment of cutaneous leishmaniasis.

  14. In vitro activity of Surfactant Protein A against Leishmania amazonensis / Actividad in vitro de la proteína A del surfactante pulmonar frente a Leishmania amazonensis

    Scientific Electronic Library Online (English)

    Yuliannis, Lugones; Odalys, Blanco; Roberto, Faure; Lianet, Monzote.

    2012-03-01

    Full Text Available La proteína surfactante A (SP-A), es una proteína pulmonar hidrofílica, de la familia de las colectinas, que contribuye al mantenimiento de la homeostasis en el pulmón y la regulación de los mecanismos de defensa e inflamación. En este estudio se reporta la actividad de la SP-A frente a Leishmania a [...] mazonensis en su forma promastigote y amastigote. La SP-A muestra actividad similar contra ambas formas del parásito, con un valor de CI50 de 34.0 ± 3.1 y 33.6 ± 1.1 µg/mL frente a promastigotes y amastigotes, respectivamente. La proteína causó una citotoxicidad moderada en macrófagos peritoneales procedentes de ratones BALB/c, con un valor de CI50 de 172.0 ± 6.1 µg/mL. Este es el primer reporte de la actividad anti-leishmanial de la SP-A. Abstract in english Surfactant protein A (SP-A) is a hydrophilic pulmonary protein belonging to the collectin family, involved in lung homeostasis, the regulation of host defenses and inflammation. The present study addresses the activity of SP-A against promastigotes and amastigotes of Leishmania amazonensis and its c [...] ytotoxicity for host cells. SP-A exhibited similar activity against both parasite forms, with an IC50 of 34.0 ± 3.1 and 33.6 ± 1.1 µg/mL against promastigotes and amastigotes, respectively. It was moderately cytotoxic to peritoneal macrophages from BALB/c mice, exhibiting an IC50 value of 172.0 ± 6.1 µg/mL. This is the first report on the anti-leishmanial activity of SP-A.

  15. In Vitro and In Vivo Miltefosine Susceptibility of a Leishmania amazonensis Isolate from a Patient with Diffuse Cutaneous Leishmaniasis

    Science.gov (United States)

    Coelho, Adriano C.; Trinconi, Cristiana T.; Costa, Carlos H. N.; Uliana, Silvia R. B.

    2014-01-01

    Miltefosine was the first oral compound approved for visceral leishmaniasis chemotherapy, and its efficacy against Leishmania donovani has been well documented. Leishmania amazonensis is the second most prevalent species causing cutaneous leishmaniasis and the main etiological agent of diffuse cutaneous leishmaniasis in Brazil. Driven by the necessity of finding alternative therapeutic strategies for a chronic diffuse cutaneous leishmaniasis patient, we evaluated the susceptibility to miltefosine of the Leishmania amazonensis line isolated from this patient, who had not been previously treated with miltefosine. In vitro tests against promastigotes and intracellular amastigotes showed that this parasite isolate was less susceptible to miltefosine than L. amazonensis type strains. Due to this difference in susceptibility, we evaluated whether genes previously associated with miltefosine resistance were involved. No mutations were found in the miltefosine transporter gene or in the Ros3 or pyridoxal kinase genes. These analyses were conducted in parallel with the characterization of L. amazonensis mutant lines selected for miltefosine resistance using a conventional protocol to select resistance in vitro, i.e., exposure of promastigotes to increasing drug concentrations. In these mutant lines, a single nucleotide mutation G852E was found in the miltefosine transporter gene. In vivo studies were also performed to evaluate the correlation between in vitro susceptibility and in vivo efficacy. Miltefosine was effective in the treatment of BALB/c mice infected with the L. amazonensis type strain and with the diffuse cutaneous leishmaniasis isolate. On the other hand, animals infected with the resistant line bearing the mutated miltefosine transporter gene were completely refractory to miltefosine chemotherapy. These data highlight the difficulties in establishing correlations between in vitro susceptibility determinations and response to chemotherapy in vivo. This study contributed to establish that the miltefosine transporter is essential for drug activity in L. amazonensis and a potential molecular marker of miltefosine unresponsiveness in leishmaniasis patients. PMID:25033218

  16. The action of ionizing radiation on the morphology, physiology and growth of Leishmania Amazonensis, with evaluation of their immunogenic power in experimental models; Acao da radiacao ionizante sobre a morfologia, fisiologia e crescimento da Leishmania amazonensis, com avaliacao de seu poder imunogenico em modelos experimentais

    Energy Technology Data Exchange (ETDEWEB)

    Bonetti, Franco Claudio

    2002-07-01

    Leishmaniasis is a disease which affects thousands of people in the Tropical regions around the world, is caused by a protozoan of the genus Leishmania spp., with urban and wild mammals acting as reservoirs. In the mammal host, the amastigote form of the parasite infects and multiplies into macrophages. Treatments for leishmaniasis have a high cost and are long lasting, frequently resulting in therapy interruption. This procedure culminates with a selection of resistant parasite strains, inducing tolerance to the therapy. Either the control of vectors or the mammal host are difficult due the social and economic implications. Thus, the search for alternatives treatments against these protozoans have been stimulated. The gamma radiation ({sup 60}CO) shown to be an efficient toll to kill these parasites maintaining their immunogenicity. Cellular viability, Electronically microscopy and Multiplex-PCR techniques showed that, after irradiation, the parasites had their growth inhibited by cytoplasmatic and nucleic material disorganisation, appointing the gamma radiation as important in terms of immunogens improvement. (author)

  17. Transcriptional signatures of BALB/c mouse macrophages housing multiplying Leishmania amazonensis amastigotes

    Directory of Open Access Journals (Sweden)

    Lang Thierry

    2009-03-01

    Full Text Available Abstract Background Mammal macrophages (M? display a wide range of functions which contribute to surveying and maintaining tissue integrity. One such function is phagocytosis, a process known to be subverted by parasites like Leishmania (L. Indeed, the intracellular development of L. amazonensis amastigote relies on the biogenesis and dynamic remodelling of a phagolysosome, termed the parasitophorous vacuole, primarily within dermal M?. Results Using BALB/c mouse bone marrow-derived M? loaded or not with amastigotes, we analyzed the transcriptional signatures of M? 24 h later, when the amastigote population was growing. Total RNA from M? cultures were processed and hybridized onto Affymetrix Mouse430_2 GeneChips®, and some transcripts were also analyzed by Real-Time quantitative PCR (RTQPCR. A total of 1,248 probe-sets showed significant differential expression. Comparable fold-change values were obtained between the Affymetrix technology and the RTQPCR method. Ingenuity Pathway Analysis software® pinpointed the up-regulation of the sterol biosynthesis pathway (p-value = 1.31e-02 involving several genes (1.95 to 4.30 fold change values, and the modulation of various genes involved in polyamine synthesis and in pro/counter-inflammatory signalling. Conclusion Our findings suggest that the amastigote growth relies on early coordinated gene expression of the M? lipid and polyamine pathways. Moreover, these M? hosting multiplying L. amazonensis amastigotes display a transcriptional profile biased towards parasite-and host tissue-protective processes.

  18. Cell populations in lesions of cutaneous leishmaniasis of leishmania (L.) amazonensis- infected rhesus macaques, Macaca mulatta

    Scientific Electronic Library Online (English)

    VF, Amaral; C, Pirmez; AJS, Gonçalves; V, Ferreira; G, Grimaldi Jr.

    2000-04-01

    Full Text Available The cellular nature of the infiltrate in cutaneous lesion of rhesus monkeys experimentally infected with Leishmania (L.) amazonensis was characterized by immunohistochemistry. Skin biopsies from infected animals with active or healing lesions were compared to non-infected controls (three of each typ [...] e) to quantitate inflammatory cell types. Inflammatory cells (composed of a mixture of T lymphocyte subpopulations, macrophages and a small number of natural killer cells and granulocytes) were more numerous in active lesions than in healing ones. T-cells accounted for 44.7 ± 13.1% of the infiltrate in active lesions (versus CD2+= 40.3 ± 5.7% in healing lesions) and T-cell ratios favor CD8+ cells in both lesion types. The percentage of cells expressing class II antigen (HLA-DR+) in active lesions (95 ± 7.1%) was significantly higher (P

  19. Brazilian brown propolis elicits antileishmanial effect against promastigote and amastigote forms of Leishmania amazonensis.

    Science.gov (United States)

    Santana, Lorena C L R; Carneiro, Sabrina Maria P; Caland-Neto, Laurentino B; Arcanjo, Daniel D R; Moita-Neto, José M; Citó, Antônia M G L; Carvalho, Fernando Aécio A

    2014-01-01

    Propolis is a complex matrix of chemical constituents extracted from plants and produced by bees which is used in folk medicine due to its several pharmacological properties. Its chemical composition varies according to the region where it is produced. This work has studied the antileishmanial activity and cytotoxicity of brown propolis (BP) originating from the semi-arid region of Piauí, Brazil. The BP showed significant inhibition of the Leishmania amazonensis promastigotes growth as well as being effective in reducing infection of murine macrophages and the number of internalised amastigotes in these cells. The dichloromethane fraction was the most active and showed the best selectivity index. The studied samples presented good activity and the fractioning improved the antileishmanial activity without an increase in the cytotoxicity against mammalian cells. Therefore, BP is a potential source for development of apitherapeutic products for the treatment of leishmaniasis. PMID:24261482

  20. Impacto de Leishmania amazonensis y la Sangre de Ave en el Potencial Biológico y Fecundidad de Lutzomyia migonei y Lutzomyia ovallesi (Diptera: Psychodidae

    Directory of Open Access Journals (Sweden)

    Elsa Nieves

    2011-03-01

    Resumo. Nos flebotomíneos (Diptera: Psychodidae o hábito pela hematofagia é responsável pela indução de vários processos fisiológicos também na transmissão de Leishmania Ross. O presente estudo compara o sangue de ave, de mamífero e com infecção por Leishmania amazonensis Lainson & Shaw sobre o potencial biológico de Lutzomyia migonei (França e de Lutzomyia ovallesi Ortiz. Foram utilizadas fêmeas das duas espécies alimentadas artificialmente com sangue de hamster (Mesocricetus auratus Waterhouse e frango (Gallus gallus Linnaeus, infectados com L. amazonensis. Os grupos controle foram alimentados somente com sangue, sem parasitas. Foram determinados o grau de repasto sanguíneo, o tempo de digestão, o padrão de diurese, o tempo de oviposição, a sobrevivencia a oviposição e a fecundidade. A espécie L. migonei quando alimentada com sangue de hamster e frango apresentaram maior fecundidade do que as fêmeas de L. ovallesi, a maior fecundidade foi com sangue de frango. A presença de Leishmania no sangue de frango ou sangue de hamster diminuiu significativamente o seu consumo, o que resultou na diminuição da sobrevida das fêmeas após a oviposição em L. migonei alimentados com sangue de frango e não com sangue de hamsters. Entretanto, não afetar a quantidade de sangue e a sobrevivência de oviposição de L. ovallesi. A infecção com L. amazonensis causo um aumento no número de ovos retidos e diminuiu o número de ovos postos por L. migonei e L. ovallesi, especialmente com sangue de frango e também reduz o tempo de digestão do sangue em ambas as espécies com sangue de frango, mas não com sangue de hamster. Embora o sangue de frango foi menos eficaz do que o sangue de hamster sobre o potencial biológico de L. migonei e L. ovallesi, não exclui o sangue de frango como uma fonte de sangue para a manutenção das populações de ambas as espécies nas casas.

  1. The effect of the phytol-rich fraction from Lacistema pubescens against Leishmania amazonensis is mediated by mitochondrial dysfunction.

    Science.gov (United States)

    da Silva, Josiane Mello; Antinarelli, Luciana Maria Ribeiro; Ribeiro, Antônia; Coimbra, Elaine Soares; Scio, Elita

    2015-12-01

    Leishmaniasis is a complex disease caused by protozoan parasite Leishmania and the treatment remains a serious problem since the available drugs exhibited high toxicity and side effects. Plant-derived natural products are promising leads for the development of novel chemotherapeutics. In this work the phytol-rich hexane fraction (PRF) from the leaves of Lacistema pubescens was obtained and identified by GC-MS analysis. When assayed for antileishmanial effects, PRF was active against promastigote and amastigote forms of Leishmania amazonensis (IC50 values of 44.0 and 25.8 ?g/mL respectively). Furthermore, PRF did not show significant cytotoxicity on peritoneal macrophages being more destructive to the intracellular parasite than to mammalian cells. In addition, possible targets of PRF were investigated against L. amazonensis promastigotes. The results showed that PRF exerted its antipromastigote activity by marked depolarization of the mitochondrial membrane potential followed by the increase of ROS levels in L. amazonensis promastigotes. During these events, no rupture of the cell membrane integrity was observed. Our results indicated that PRF was effective and selective against L. amazonensis, and that this effect was mainly mediated by mitochondrial dysfunction associated to ROS production. PMID:26424529

  2. Degranulating Neutrophils Promote Leukotriene B4 Production by Infected Macrophages To Kill Leishmania amazonensis Parasites.

    Science.gov (United States)

    Tavares, Natália; Afonso, Lilian; Suarez, Martha; Ampuero, Mariana; Prates, Deboraci Brito; Araújo-Santos, Théo; Barral-Netto, Manoel; DosReis, George A; Borges, Valéria Matos; Brodskyn, Cláudia

    2016-02-15

    Neutrophils mediate early responses against pathogens, and they become activated during endothelial transmigration toward the inflammatory site. In the current study, human neutrophils were activated in vitro with immobilized extracellular matrix proteins, such as fibronectin (FN), collagen, and laminin. Neutrophil activation by FN, but not other extracellular matrix proteins, induces the release of the granules' contents, measured as matrix metalloproteinase 9 and neutrophil elastase activity in culture supernatant, as well as reactive oxygen species production. Upon contact with Leishmania amazonensis-infected macrophages, these FN-activated neutrophils reduce the parasite burden through a mechanism independent of cell contact. The release of granule proteases, such as myeloperoxidase, neutrophil elastase, and matrix metalloproteinase 9, activates macrophages through TLRs, leading to the production of inflammatory mediators, TNF-? and leukotriene B4 (LTB4), which are involved in parasite killing by infected macrophages. The pharmacological inhibition of degranulation reverted this effect, abolishing LTB4 and TNF production. Together, these results suggest that FN-driven degranulation of neutrophils induces the production of LTB4 and TNF by infected macrophages, leading to the control of Leishmania infection. PMID:26800873

  3. Further observations on clinical, histopathological, and immunological features of borderline disseminated cutaneous leishmaniasis caused by Leishmania (Leishmania) amazonensis

    Scientific Electronic Library Online (English)

    Fernando T, Silveira; Ralph, Lainson; Carlos EP, Corbett.

    2005-08-01

    Full Text Available Leishmania (Leishmania) amazonensis has for some time been considered as the causative agent of two distinct forms of American cutaneous leishmaniasis (ACL): localized cutaneous leishmaniasis (LCL), and anergic diffuse cutaneous leishmaniasis (ADCL). Recently, a new intermediate form of disease, bor [...] derline disseminated cutaneous leishmaniasis (BDCL), was introduced into the clinical spectrum of ACL caused by this parasite, and in this paper we record the clinical, histopathological, and immunological features of eight more BDCL patients from Brazilian Amazonia, who acquired the disease in the Pará state, North Brazil. Seven of them had infections of one to two years' evolution and presented with primary skin lesions and the occurrence of metastases at periods varying from six to 12 months following appearance of the first lesion. Primary skin lesions ranged from 1-3 in number, and all had the aspect of an erythematous, infiltrated plaque, variously located on the head, arms or legs. There was lymphatic dissemination of infection, with lymph node enlargement in seven of the cases, and the delayed hypersensitivity skin-test (DTH) was negative in all eight patients prior to their treatment. After that, there was a conversion of DTH to positive in five cases re-examined. The major histopathological feature was a dermal mononuclear infiltration, with a predominance of heavily parasitized and vacuolated macrophages, together with lymphocytes and plasma cells. In one case, with similar histopathology, the patient had acquired his infection seven years previously and he presented with the largest number of disseminated cutaneous lesions. BDCL shows clinical and histopathological features which are different from those of both LCL and ADCL, and there is a good prognosis of cure which is generally not so in the case of frank ADCL.

  4. In vitro evaluation of (-)?-bisabolol as a promising agent against Leishmania amazonensis.

    Science.gov (United States)

    Rottini, Mariana Margatto; Amaral, Ana Claudia Fernandes; Ferreira, Jose Luiz Pinto; Silva, Jefferson Rocha de Andrade; Taniwaki, Noemi Nosomi; Souza, Celeste da Silva Freitas de; d'Escoffier, Luiz Ney; Almeida-Souza, Fernando; Hardoim, Daiana de Jesus; Gonçalves da Costa, Sylvio Celso; Calabrese, Kátia da Silva

    2015-01-01

    Current treatments for leishmaniasis present some difficulties due to their toxicity, the use of the intravenous route for administration and therapy duration, which may lead to treatment discontinuation. The aim of this study is to investigate new treatment alternatives to improve patients well being. Therefore, we evaluated the inhibitory effect of (-)?-bisabolol, a sesquiterpene alcohol found in various essential oils of different plant species, against the promastigotes and intracellular amastigotes forms of Leishmania amazonensis, as well as the cytotoxic, morphological and ultrastructural alterations of treated cells. Promastigotes forms of L.?amazonensis were incubated with (-)?-bisabolol to determine the antileishmanial activity of this compound. The cytotoxicity effect was evaluated by testing against J774.G8 cells. After these tests, the infected and uninfected cells with L.?amazonensis were used to determine if the (-)?-bisabolol was able to kill intracellular parasites and to cause some morphological changes in the cells. The (-)?-bisabolol compound showed significant antileishmanial activity against promastigotes with a 50% effective concentration of 8.07?µg/ml (24?h) and 4.26?µg/ml (48 h). Against intracellular amastigotes the IC50 (inhibitory concentration) of (-)?-bisabolol (24?h) was 4.15?µg/ml. The (-)?-bisabolol also showed a cytotoxic effect against the macrophage strain J774.G8. The value of 50% cytotoxic concentration was 14.82?µg/ml showing that (-)?-bisabolol is less toxic to macrophages than to the parasite. Ultrastructural studies of treated promastigotes and amastigotes showed several alterations, such as loss of cytoplasmic organelles, including the nucleus, and the presence of lipid inclusions. This study showed that (-)?-bisabolol has promising antileishmanial properties, as it can act against the promastigote forms and is able to penetrate the cell, and is also active against the amastigote forms. About 69% of the promastigotes forms suffered mitochondrial membrane damage after treatment with IC50 of (-)?-bisabolol, suggesting inhibition of the metabolic activity of parasites. These results open new prospects for research that can contribute to the development of products based on essential oils or isolated compounds from plants for the treatment of cutaneous leishmaniasis. PMID:25448354

  5. In vitro and in vivo activity of major constituents from Pluchea carolinensis against Leishmania amazonensis.

    Science.gov (United States)

    Montrieux, Elly; Perera, Wilmer H; García, Marley; Maes, Louis; Cos, Paul; Monzote, Lianet

    2014-08-01

    The search for new therapeutic agents from natural sources has been a constant for the treatment of diseases such as leishmaniasis. Herein, in vitro and in vivo pharmacological activities of pure major phenolic constituents (caffeic acid, chlorogenic acid, ferulic acid, quercetin, and rosmarinic acid) from Pluchea carolinensis against Leishmania amazonensis are presented. Pure compounds showed inhibitory activity against promastigotes (IC50?=?0.2-0.9 ?g/mL) and intracellular amastigotes (IC50?=?1.3-2.9 ?g/mL). Four of them were selected after testing against macrophages of BALB/c mice: caffeic acid, ferulic acid, quercetin, and rosmarinic acid, with selective indices of 11, 17, 10, and 20, respectively. Ferulic acid, rosmarinic acid, and caffeic acid controlled lesion size development and parasite burden in footpads from BALB/c experimentally infected mice, after five injections of compounds by intralesional route at 30 mg/kg every 4 days. Pure compounds from P. carolinensis demonstrated antileishmanial properties. PMID:24906989

  6. The in Vitro Biological Activity of the Brazilian Brown Seaweed Dictyota mertensii against Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    Amanda Silva dos Santos Aliança

    2014-09-01

    Full Text Available Seaweeds present a wide variety of interesting bioactive molecules. In the present work we evaluated the biological activity of the dichloromethane/methanol (2:1 extract (DME from the brown seaweed Dictyota mertensii against Leishmania amazonensis and its cytotoxic potential on mammalian cells. The extract showed significant inhibitory effect on the growth of promastigote forms (IC50 = 71.60 ?g/mL and low toxicity against mammalian cells (CC50 = 233.10 ?g/mL. The DME was also efficient in inhibiting the infection in macrophages, with CC50 of 81.4 ?g/mL and significantly decreased the survival of amastigote forms within these cells. The selectivity index showed that DME was more toxic to both promastigote (SI = 3.25 and amastigote (SI = 2.86 forms than to macrophages. Increased NO production was observed in treated macrophages suggesting that besides acting directly on the parasites, the DME also shows an immunomodulatory effect on macrophages. Drastic ultrastructural alterations consistent with loss of viability and cell death were observed in treated parasites. Confocal microscopy and cytometry analyzes showed no significant impairment of plasma membrane integrity, whereas an intense depolarization of mitochondrial membrane could be observed by using propidium iodide and rhodamine 123 staining, respectively. The low toxicity to mammalian cells and the effective activity against promastigotes and amastigotes, point to the use of DME as a promising agent for the treatment of cutaneous leishmaniasis.

  7. Benzophenone derivatives as cysteine protease inhibitors and biological activity against Leishmania(L.) amazonensis amastigotes.

    Science.gov (United States)

    de Almeida, Letícia; Alves, Karina Ferreira; Maciel-Rezende, Claudia Mara; Jesus, Larissa de Oliveira Passos; Pires, Francieli Ribeiro; Junior, Claudio Viegas; Izidoro, Mario Augusto; Júdice, Wagner Alves de Souza; Dos Santos, Marcelo Henrique; Marques, Marcos José

    2015-10-01

    The leishmanicidal potential of benzophenones has been described, some of them highlighting their potential as cysteine protease inhibitors. Therefore, this work described leishmanicidal activity of nine benzophenone derivatives (1a-c;2a-c;3a-c) against intramacrophage amastigote forms of Leishmania(L.)amazonensis (IC50) and the cytotoxic effect on murine peritoneal macrophages (CC50). The derivative 1c exhibited a selectivity index SI (CC50/IC50) of 6.7, besides cytotoxicity lower than Amphotericin B (pactivity against papain (42.8±0.3, p<0.05), and when tested on trypanosomatids cysteine proteases 1c also proved to be a potent inhibitor of rCPB2.8, rCPB3.0 and cruzain, showing non-competitive inhibition mechanism by enzymatic assays in vitro.So, benzophenone 1c is interesting drug candidate prototype, with a multi-target directed mode of action, inhibiting rCPB2.8, rCPB3.0 and cruzain. PMID:26463637

  8. Leishmaniose cutânea na Amazônia: registro do primeiro caso humano de infecção mista, determinado por duas espécies distintas de Leishmnias: Leishmania brasiliensis e Leishmania mexicana amazonensis

    Directory of Open Access Journals (Sweden)

    F. T. Silveira

    1984-10-01

    Full Text Available Fez-se o registro, na Amazônia, do primeiro caso humano de infecção cutânea mista determinada por duas espécies distintas de Leishmania: a Leishmania braziliensis braziliensis e a Leishmania mexicana amazonensis. As duas amostras, em questão, foram isoladas de lesões distintas de um mesmo paciente, e a caracterização das espécies foi feita com base em observações de infecção experimental em hamsters, comportamento em meios artificiais de cultura, desenvolvimento de infecção experimental em Lutzomyia longipalpis, e eletroforese de isoenzimas em gel de amido. Conclui-se ser de interesse o achado que, combinado com o fato já conhecido de ausência de imunidade cruzada entre a maioria das leishmânias, sugere a necessidade do emprego de uma vacina polivalente para a região.

  9. Dynamic of the Cellular Immune Response at the Dermal Site of Leishmania (L.) amazonensis and Leishmania (V.) braziliensis Infection in Sapajus apella Primate

    Science.gov (United States)

    Laurenti, Márcia Dalastra; Passero, Luiz Felipe Domingues; Tomokane, Thaise Yumie; Francesquini, Fernanda de Camargo; Rocha, Mussya Cisotto; Gomes, Claudia Maria de Castro; Corbett, Carlos Eduardo Pereira; Silveira, Fernando Tobias

    2014-01-01

    The purpose of this study was to characterize the immunopathological response in the skin of S. apella infected with Leishmania (L.) amazonensis and L. (V.) braziliensis parasites, the main causative agents of localized cutaneous leishmaniasis in South America. In infected animals, amastigote forms of L. (L.) amazonensis could be detected till 120 days postinfection (PI), while, in L. (V.) braziliensis infection, parasites could be detected until 180 days PI in the skin sections. CD20+ cells were detected throughout the experimental time in both groups as well as in CD3+ cells, which appeared to be activated because high densities of inducible nitric oxide synthase (iNOS+) cells were detected at 60 and 90 days PI in both studied groups. After 60 and 120 days PI, decrease in iNOS+ cells was observed in L. (L.) amazonensis and L. (V.) braziliensis, respectively, which was associated with parasite clearance. Increase in lysozyme+ cells was observed during the experimental infections, which also can be associated with parasite killing. PMID:25309902

  10. Effect of Apigenin on Leishmania amazonensis Is Associated with Reactive Oxygen Species Production Followed by Mitochondrial Dysfunction.

    Science.gov (United States)

    Fonseca-Silva, Fernanda; Canto-Cavalheiro, Marilene M; Menna-Barreto, Rubem F S; Almeida-Amaral, Elmo E

    2015-04-24

    Leishmaniasis is an important neglected disease caused by protozoa of the genus Leishmania that affects more than 12 million people worldwide. Leishmaniasis treatment requires the administration of toxic and poorly tolerated drugs, and parasite resistance greatly reduces the efficacy of conventional medications. Apigenin (1), a naturally occurring plant flavone, has a wide range of reported biological effects. In this study, antileishmanial activity of 1 in vitro was investigated, and its mechanism of action against Leishmania amazonensis promastigotes was described. Treatment with 1 for 24 h resulted in concentration-dependent inhibition of cellular proliferation (IC50 = 23.7 ?M) and increased reactive oxygen species (ROS) generation. Glutathione and N-acetyl-l-cysteine protected L. amazonensis from the effects of 1 and reduced ROS levels after the treatment. By contrast, oxidized glutathione did not reduce the levels of ROS caused by 1 by not preventing the proliferation inhibition. Apigenin 1 also induced an extensive swelling in parasite mitochondria, leading to an alteration of the mitochondrial membrane potential, rupture of the trans-Golgi network, and cytoplasmic vacuolization. These results demonstrate the leishmanicidal effect of 1 and suggest the involvement of ROS leading to mitochondrial collapse as part of the mechanism of action. PMID:25768915

  11. Glycosphingolipid antigens from Leishmania (L.) amazonensis amastigotes: Binding of anti-glycosphingolipid monoclonal antibodies in vitro and in vivo

    Scientific Electronic Library Online (English)

    A.H., Straus; V.B., Valero; C.M., Takizawa; S.B., Levery; M.S., Toledo; E., Suzuki; M.E.K., Salyan; S., Hakomori; C.L., Barbieri; H.K., Takahashi.

    1997-03-01

    Full Text Available Specific glycosphingolipid antigens of Leishmania (L.) amazonensis amastigotes reactive with the monoclonal antibodies (MoAbs) ST-3, ST-4 and ST-5 were isolated, and their structure was partially elucidated by negative ion fast atom bombardment mass spectrometry. The glycan moieties of five antigens [...] presented linear sequences of hexoses and N-acetylhexosamines ranging from four to six sugar residues, and the ceramide moieties were found to be composed by a sphingosine d18:1 and fatty acids 24:1 or 16:0. Affinities of the three monoclonal antibodies to amastigote glycosphingolipid antigens were also analyzed by ELISA. MoAb ST-3 reacted equally well with all glycosphingolipid antigens tested, whereas ST-4 and ST-5 presented higher affinities to glycosphingolipids with longer carbohydrate chains, with five or more sugar units (slow migrating bands on HPTLC). Macrophages isolated from footpad lesions of BALB/c mice infected with Leishmania (L.) amazonensis were incubated with MoAb ST-3 and, by indirect immunofluorescence, labeling was only detected on the parasite, whereas no fluorescence was observed on the surface of the infected macrophages, indicating that these glycosphingolipid antigens are not acquired from the host cell but synthesized by the amastigote. Intravenous administration of 125I-labeled ST-3 antibody to infected BALB/c mice showed that MoAb ST-3 accumulated significantly in the footpad lesions in comparison to blood and other tissues

  12. Glycosphingolipid antigens from Leishmania (L. amazonensis amastigotes: Binding of anti-glycosphingolipid monoclonal antibodies in vitro and in vivo

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    A.H. Straus

    1997-03-01

    Full Text Available Specific glycosphingolipid antigens of Leishmania (L. amazonensis amastigotes reactive with the monoclonal antibodies (MoAbs ST-3, ST-4 and ST-5 were isolated, and their structure was partially elucidated by negative ion fast atom bombardment mass spectrometry. The glycan moieties of five antigens presented linear sequences of hexoses and N-acetylhexosamines ranging from four to six sugar residues, and the ceramide moieties were found to be composed by a sphingosine d18:1 and fatty acids 24:1 or 16:0. Affinities of the three monoclonal antibodies to amastigote glycosphingolipid antigens were also analyzed by ELISA. MoAb ST-3 reacted equally well with all glycosphingolipid antigens tested, whereas ST-4 and ST-5 presented higher affinities to glycosphingolipids with longer carbohydrate chains, with five or more sugar units (slow migrating bands on HPTLC. Macrophages isolated from footpad lesions of BALB/c mice infected with Leishmania (L. amazonensis were incubated with MoAb ST-3 and, by indirect immunofluorescence, labeling was only detected on the parasite, whereas no fluorescence was observed on the surface of the infected macrophages, indicating that these glycosphingolipid antigens are not acquired from the host cell but synthesized by the amastigote. Intravenous administration of 125I-labeled ST-3 antibody to infected BALB/c mice showed that MoAb ST-3 accumulated significantly in the footpad lesions in comparison to blood and other tissues

  13. Leishmania amazonensis promastigotes in 3D Collagen I culture: an in vitro physiological environment for the study of extracellular matrix and host cell interactions

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    Debora B. Petropolis

    2014-04-01

    Full Text Available Leishmania amazonensis is the causative agent of American cutaneous leishmaniasis, an important neglected tropical disease. Once Leishmania amazonensis is inoculated into the human host, promastigotes are exposed to the extracellular matrix (ECM of the dermis. However, little is known about the interaction between the ECM and Leishmania promastigotes. In this study we established L. amazonensis promastigote culture in a three-dimensional (3D environment mainly composed of Collagen I (COL I. This 3D culture recreates in vitro some aspects of the human host infection site, enabling the study of the interaction mechanisms of L. amazonensis with the host ECM. Promastigotes exhibited “freeze and run” migration in the 3D COL I matrix, which is completely different from the conventional in vitro swimming mode of migration. Moreover, L. amazonensis promastigotes were able to invade, migrate inside, and remodel the 3D COL I matrix. Promastigote trans-matrix invasion and the freeze and run migration mode were also observed when macrophages were present in the matrix. At least two classes of proteases, metallo- and cysteine proteases, are involved in the 3D COL I matrix degradation caused by Leishmania. Treatment with a mixture of protease inhibitors significantly reduced promastigote invasion and migration through this matrix. Together our results demonstrate that L. amazonensis promastigotes release proteases and actively remodel their 3D environment, facilitating their migration. This raises the possibility that promastigotes actively interact with their 3D environment during the search for their cellular “home”—macrophages. Supporting this hypothesis, promastigotes migrated faster than macrophages in a novel 3D co-culture model.

  14. Effect of Elatol, Isolated from Red Seaweed Laurencia dendroidea, on Leishmania amazonensis

    OpenAIRE

    Celso Vataru Nakamura; Renato Crespo Pereira; Daniela Bueno Sudatti; Éverson Miguel Bianco; Benedito Prado Dias Filho; Tânia Ueda-Nakamura; Adriana Oliveira dos Santos; Phercyles Veiga-Santos

    2010-01-01

    In the present study, we investigated the antileishmanial activity of sesquiterpene elatol, the major constituent of the Brazilian red seaweed Laurencia dendroidea (Hudson) J.V. Lamouroux, against L. amazonensis. Elatol after 72 h of treatment, showed an IC50 of 4.0 µM and 0.45 µM for promastigote and intracellular amastigote forms of L. amazonensis, respectively. By scanning and transmission electron microscopy, parasites treated with elatol revealed notable changes compared with control cel...

  15. Kinetoplastid membrane protein-11 is present in promastigotes and amastigotes of Leishmania amazonensis and its surface expression increases during metacyclogenesis

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    Denise CS Matos

    2010-05-01

    Full Text Available Kinetoplastid membrane protein-11 (KMP-11, a protein present in all kinetoplastid protozoa, is considered a potential candidate for a leishmaniasis vaccine. A suitable leishmaniasis vaccine candidate molecule must be expressed in amastigotes, the infective stage for mammals. However, the expression of KMP-11 in Leishmania amastigotes has been a subject of controversy. We evaluated the expression of this molecule in logarithmic and stationary growth phase promastigotes, as well as in amastigotes, of Leishmania amazonensis by immunoblotting, flow cytometry and immunocytochemistry, using a monoclonal antibody against KMP-11. We found that KMP-11 is present in promastigotes and amastigotes. In both stages, the protein was found in association with membrane structures (at the cell surface, flagellar pocket and intracellular vesicles. More importantly, its surface expression is higher in amastigotes than in promastigotes and increases during metacyclogenesis. The increased expression of KMP-11 in metacyclic promastigotes, and especially in amastigotes, indicates a role for this molecule in the parasite relationship with the mammalian host. The presence of this molecule in amastigotes is consistent with the previously demonstrated immunoprotective capacity of vaccine prototypes based on the KMP-11-coding gene and the presence of humoral and cellular immune responses to KMP-11 in Leishmania-infected humans and animals.

  16. Construcción de una biblioteca genómica de Leishmania amazonensis y su expresión en músculo de ratones BALB/c

    Scientific Electronic Library Online (English)

    Ana Margarita, Montalvo Álvarez; Esteban, Alberti Amador; Marta M., González Elías; Rocío, García Miniet; María E., Sarmiento García; Armando, Acosta Domínguez.

    2001-12-01

    Full Text Available Se construyó una biblioteca genómica de Leishmania amazonensis mediante el vector pcDNA3, con promotor de expresión en células eucariotas, con el objetivo de contribuir a la aplicación de la tecnología de inmunización con ácidos nucleicos en la leishmaniosis. Para demostrar la expresión de la genote [...] ca en el músculo de ratones inmunizados con esta, se realizó la técnica de inmunofluorescencia indirecta. Como anticuerpo primario se utilizó una mezcla de sueros con alto título antileishmania, de una zona donde predomina la infección con L. braziliensis. Se obtuvo una biblioteca con 80 % de clones recombinantes. Se demostró la expresión de determinantes antigénicos en el músculo de ratones BALB/c inmunizados, según resultados de la inmunofluorescencia. Abstract in english A genomic library of Leishaminia amazonensis was built through a pcDNA3 vector, with expression promoter in eukaryot cells, to contribute to the application of immunization technology with nucleic acids in leishmaniasis. To show the expression genomic library in the muscles of mice immunized with it [...] , the indirect immunofluoresce technique was used. A mix of sera with high antileishmania titers from an area where L.braziliensis infection is predominant was used as primary antibody. A library of 80% recombinant clones was obtained. Antigen determinant expression was confirmed in immunized BALB/c mice´s muscles, according to the results of immunofluorescence testing.

  17. In vitro and in vivo activity of meglumine antimoniate produced at Farmanguinhos-Fiocruz, Brazil, against Leishmania (Leishmania amazonensis, L (L. chagasi and L (Viannia braziliensis

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    Eliane de Morais-Teixeira

    2008-06-01

    Full Text Available The leishmanicidal activity of four batches of meglumine antimoniate, produced in Farmanguinhos-Fiocruz, Brazil (TAMs, was assessed and compared to Glucantime®-Aventis Pharma Ltda. Using the amastigote-like in vitro model, the active concentrations of Sb v varied from 10µg/ml to 300 µg/ml for L. (L. chagasi and from 50µg/ml to 300µg/ml for L. (L. amazonensis, with no statistically significant differences among the four batches of TAMs and Glucantime®. The inhibitory concentrations (IC50 determined by the amastigote-infected macrophage model for TAM01/03 and Glucantime® were, respectively: 26.3µg/ml and 127.6µg/ml for L. chagasi, 15.4µg /ml and 22.9µg/ml for L. amazonensis, and 12.1µg/ml and 24.2µg/ml for L. (V. braziliensis. The activities of the four batches of TAMs were confirmed in an in vivo model by assessing, during eight weeks skin lesions caused by L. braziliensis in hamster that were treated with 20mg Sb v/Kg/day for 30 consecutive days. The meglumine antimoniate produced by Farmanguinhos was as effective as the reference drug, Glucantime®-Aventis, against three species of Leishmania that are of medical importance in Brazil.

  18. Oral Tolerance promotes reduction of inflammatory lesions in mice infected with Leishmania amazonensis.

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    Daniel Tavares

    2013-06-01

    Full Text Available Introduction: The Oral Tolerance is a physiological mechanism of systemic hyporeactivity to an immunogen previously ingested and bystander suppression is an inhibited response to a second immunogen when it is presented along with the immunogen for which it was established oral tolerance. We use for bystander suppression study, two strains of mice genetically selected for extreme phenotypes of susceptibility (TS and resistance (TR to oral tolerance (Silva, AC et al. 1998. TR strain presents good inflammatory responses and a non-tolerogenic profile while TS strain presents non-inflammatory but high-tolerogenic profile, with high percentages of regulatory T cells (Treg cells, CD4+CD25+Foxp3+. Furthermore, TS strain is able to produce high levels of inhibitory citokynes such as IL-10 (Silva, MF et al. 2010. Previous studies in mice strains with extreme phenotypes to susceptibility (TS strain and resistance (TR strain to Oral Tolerance showed that when infected with Leishmania amazonensis, TR strain develops an exacerbate lesion while a regulatory activity of the TS strain depresses the inflammation and avoid acute lethal response (Tavares, D et al. 2006. Material and Methods: TR and TS strains of mice from the F25 generation, obtained by two-way genetic selection according to susceptibility (TS or resistance (TR to ovalbumin oral tolerance (Silva, AC et al. 1998. Bystander suppression was done by modification of Miller et al. (1991. Female mice were gavaged with 2 mg OVA (Sigma Chemical Co., St Louis, MO in PBS during a two-consecutive-day protocol (total dose of 4 mg. Seven days afterwards, OVA-gavaged mice were challenged with 50 µg of soluble OVA in PBS 8 h after subcutaneously infected with 10 million viable L. amazonensis stationary promastigotes in a volume of 25 µl in the left hind footpad. Evolution of the lesion was monitored by weekly measuring of footpad thickness with a Mitutoyo caliper (MTI Co USA and expressed as the difference in thickness between the infected and the uninfected counterlateral footpad. The Committee for the Care and Use of Laboratory Animals of the Universidade do Estado do Rio de Janeiro, approved the protocols of the experiments described in this work (CEUA/013/2012/UERJ. Treg cells from bystander draining lymph nodes were evaluated by flow cytometry according Silva, MF et al. (2010. Briefly, 106 cells were fixed and permeabilized using the mouse regulatory T-cell staining kit (eBioscience, following the manufacturer’s instructions. Data were acquired on a FACSCalibur (BD Biosciences, San Jose, CA and analyzed using CELLQUEST (BD Biosciences software. Results: The bystander suppression reduced the inflammatory lesion in TR strain similarly to the swelling of infected TS strain (Figure 1A. Non infected (normal group TS strain presents a higher proportion of Treg cells than TR strain (Figure 2B. In both strains, infected mice present a higher proportion of Treg cells than non infected mice (Figure 2B. Mice previously tolerized and immunized (bystander suppression group showed an increase of CD4+ CD25+ FoxP3+ cells (Figure 1B. Discussion: The suppression of immune reactivity is currently attributed to the regulatory T cells, population dedicated to maintaining peripheral tolerance to self-antigens or prevent harmful immunopathological responses, by inhibiting the release of cytokines such as IL-10 and TGF-? or by mechanisms contact-dependent. The TS strain presents intrinsically a higher proportion of Treg cells that the TR strain (Figure 2B, and initially, the difference of the inflammatory profiles of these strains could be attributed to a greater or lesser amount of Treg cells. Bystander suppression by oral tolerance has been described as a regulatory T cell mechanism producing IL-10 and TGF-b that reduces inflammation (Miller A et al. 1991. It has been reported that T CD4+ cells induced by oral antigen, use IL-4 and IL-10 to “educate” Dendritic Cells (DCs, which in turn induce naive T cells to produce the same cytokines as those produced by the orally tolerized

  19. Ecological Niche Modelling Predicts Southward Expansion of Lutzomyia (Nyssomyia) flaviscutellata (Diptera: Psychodidae: Phlebotominae), Vector of Leishmania (Leishmania) amazonensis in South America, under Climate Change

    Science.gov (United States)

    Carvalho, Bruno M.; Ready, Paul D.

    2015-01-01

    Vector borne diseases are susceptible to climate change because distributions and densities of many vectors are climate driven. The Amazon region is endemic for cutaneous leishmaniasis and is predicted to be severely impacted by climate change. Recent records suggest that the distributions of Lutzomyia (Nyssomyia) flaviscutellata and the parasite it transmits, Leishmania (Leishmania) amazonensis, are expanding southward, possibly due to climate change, and sometimes associated with new human infection cases. We define the vector’s climatic niche and explore future projections under climate change scenarios. Vector occurrence records were compiled from the literature, museum collections and Brazilian Health Departments. Six bioclimatic variables were used as predictors in six ecological niche model algorithms (BIOCLIM, DOMAIN, MaxEnt, GARP, logistic regression and Random Forest). Projections for 2050 used 17 general circulation models in two greenhouse gas representative concentration pathways: “stabilization” and “high increase”. Ensemble models and consensus maps were produced by overlapping binary predictions. Final model outputs showed good performance and significance. The use of species absence data substantially improved model performance. Currently, L. flaviscutellata is widely distributed in the Amazon region, with records in the Atlantic Forest and savannah regions of Central Brazil. Future projections indicate expansion of the climatically suitable area for the vector in both scenarios, towards higher latitudes and elevations. L. flaviscutellata is likely to find increasingly suitable conditions for its expansion into areas where human population size and density are much larger than they are in its current locations. If environmental conditions change as predicted, the range of the vector is likely to expand to southeastern and central-southern Brazil, eastern Paraguay and further into the Amazonian areas of Bolivia, Peru, Ecuador, Colombia and Venezuela. These areas will only become endemic for L. amazonensis, however, if they have competent reservoir hosts and transmission dynamics matching those in the Amazon region. PMID:26619186

  20. Leishmania amazonensis DNA in wild females ofLutzomyia cruzi (Diptera: Psychodidae) in the state of Mato Grosso do Sul, Brazil.

    Science.gov (United States)

    Oliveira, Everton Falcão de; Casaril, Aline Etelvina; Mateus, Nathália Lopes Fontoura; Murat, Paula Guerra; Fernandes, Wagner Souza; Oshiro, Elisa Teruya; Oliveira, Alessandra Gutierrez de; Galati, Eunice Aparecida Bianchi

    2015-12-01

    Studies on natural infection by Leishmania spp of sandflies collected in endemic and nonendemic areas can provide important information on the distribution and intensity of the transmission of these parasites. This study sought to investigate the natural infection by Leishmaniain wild female sandflies. The specimens were caught in the city of Corumbá, state of Mato Grosso do Sul (Brazil) between October 2012-March 2014, and dissected to investigate flagellates and/or submitted to molecular analysis to detect Leishmania DNA. A total of 1,164 females (77.56% of which were Lutzomyia cruzi) representing 11 species were investigated using molecular analysis; 126 specimens of Lu. cruziwere dissected and also submitted to molecular analysis. The infection rate based on the presence of Leishmania DNA considering all the sandfly species analysed was 0.69%; only Leishmania (Leishmania) amazonensis was identified in Lu. cruzi by the molecular analysis. The dissections were negative for flagellates. This is the first record of the presence of L. (L.) amazonensis DNA in Lu. cruzi, and the first record of this parasite in this area. These findings point to the need for further investigation into the possible role of this sandfly as vector of this parasite. PMID:26602870

  1. Lutzomyia reducta Feliciangeli et al., 1988, a host of Leishmania amazonensis, sympatric with two other members of the Flaviscutellata complex in southern Amazonas and Rondônia, Brazil (Diptera: Psychodidae Lutzomyia reducta Feliciangeli et al., 1988 um hospedeiro de Leishmania amazonensis, simpátrico com duas outras espécies do complexo flaviscutellata no sul do Amazonas e Rondônica, Brasil (Diptera: Psychodidae

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    R. A. Freitas

    1989-09-01

    Full Text Available A member of the Lutzomyia flaviscutellata complex from Rondônia and southern Amazonas States, Brazil, is so close to the Venezuelan Lutzomyia olmeca recuta Feliciangeli et al., 1988, that it is regarded as belonging to the same species. Since this phlebotomine co-extis with L. olmeca nociva in Brazil, the subspecific status of the former is untenable and is rased to specific rank, as Lutzomyia reducta. The Brazilian material is described and illustrated, and compared with specimens of L. o. nociva and L. flaviscutellata from the same area. Keys to the known taxa of the flaviscutellata complex are presented. Leishmania amazonensis was isolated from one heavily infected specimen of L. reducta, making this the third species of the flaviscutellata complex to be implicated as a vector of this parasite in Brazil. The relative abundance of the three sympatric flaviscutellata complex species varies locally and appears to be related to soil drainage. L. reducta constituted about 25% if all phlebotomines captured in Disney traps at poorly drained and well drained site, but appears not to coloniza areas subject to periodic flooding. L. olmeca nociva was restricted to poorly drained areas not subject to flooding, whereas L. flaviscutellata was ubiquitous L. reducta has never been detected north of the Amazon river in Brazil, but absence of recosrds from western and northwestern Amazonas State may reflect lack of collecting in these areas.Um flebotomíneo do complexo Lutzomyia flaviscutellata, de Rondônia e sul do Amazonas, Brasil é tão parecido com Lutzomyia olmeca reducta, que é considerado como sendo da mesma espécie. Este flebotomíneo ocorre junto com L. olmeca nociva, portanto o nome é emendado para o nível de espécie, como Lutzomyia reducta. O material do Brasil é descrito e ilustrado, e comparado com exemplares de L. o. nociva e L. flaviscutellata da mesma área. Chaves para as espécies e subespécies do complexo flaviscutellata são incluídas. Leishmania amazonensis foi isolada em um exemplar de L. reducta altamente infectado, tornando esta espécie a terceira a ser implicada como vetor desta leishmania no Brasil. A abundância relativa das três espécies simpátricas do complexo flaviscutellata varia em escala local e aparenta ter relação com a drenagem do solo. L. reducta constituiu cerca de 25% dos flebotomíneos capturados em armadilhas Disney em locais mal e bem drenados, porém não foi encontrada em locais sujeitos a inundações. L. olmeca nociva era restrita às áreas mal drenadas não sujeitas a inundações, enquanto L. flaviscutellata foi capturada neste dois ambientes e também numa área periodicamente inundada. L. reducta não tem sido assinalada ao norte do Rio Amazonas no Brasil, porém a ausência de registros do oeste e noroeste do Estado do Amazonas possa refletri a falta de levantamento nestas áreas.

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

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    Silva-Jr FP

    2002-01-01

    Full Text Available 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.

  3. Papel de la vacuola parasitófora de macrófagos de ratón infectados por Leishmania amazonensis en la adquisición de moléculas / Role of the parasitophorous vacuole of murine macrophages infected with Leishmania amazonensis in molecule acquisition

    Scientific Electronic Library Online (English)

    Tania M, Cortázar; Joselín, Hernández; María Clara, Echeverry; Marcela, Camacho.

    2006-10-01

    Full Text Available Introducción. Leishmania son parásitos intracelulares de macrófagos, confinados en compartimentos denominados vacuolas parasitóforas. La permeabilidad de este compartimento depende de su interacción con el tráfico vesicular y transportadores presentes en su membrana. Objetivo. En este trabajo se est [...] udió la permeabilidad de la membrana de la vacuola parasitófora en la línea celular J774.A1 infectada con Leishmania amazonensis, in situ y en compartimentos aislados. Materiales y métodos. El aislamiento de vacuolas parasitóforas se hizo por gradiente de densidad. La permeabilidad de la membrana de estas se valoró por distribución de sondas fluorescentes y electrofisiología. Para establecer indirectamente el transporte de protones se usó naranja de acridina. La presencia de transportadores ABC sensibles a probenecid se estableció con amarillo lucifer y calceína. Por primera vez con la técnica de patch-clamp se registraron corrientes en la membrana de este compartimento aislado. Resultados. La vacuola parasitófora colorea de rojo con naranja de acridina indicando un pH ácido. Concentra amarillo lucifer a través de un transportador sensible a probenecid, pero excluye la sonda calceína. Vacuolas aisladas se marcan de rojo con naranja de acridina y concentran amarillo lucifer a través de un transportador sensible a probenecid. Estas vacuolas excluyeron calceína y presentaron en su membrana una corriente iónica que se activa a diferencias de potencial cercanas a 60 mV, con una conductancia de 46 ± 3 pS. Conclusiones. Se pueden aislar vacuolas parasitóforas con propiedades de permeabilidad que preservan mecanismos de transporte similares a los encontrados in situ. Se registra por primera vez la presencia de una corriente iónica poco selectiva en la membrana de este compartimiento. Abstract in english Introduction. Leishmania are intracellular parasites of macrophages, confined into compartments known as parasitophorous vacuoles. The permeability of this compartment depends on its interaction with the endocytic pathway and transport proteins present on its membrane. Objective. The membrane permea [...] bility of the parasitophorous vacuole was studied in J774.A1- macrophage like cells infected with Leishmania amazonensis, in situ and on isolated compartments. Materials and methods. The parasitophorous vacuoles were isolated by density gradients. Fluorescent probe distribution and electrophysiological recordings were used to determine parasitophorous vacuole membrane permeability. Proton transport was evaluated indirectly by acridine orange staining. Probenecid sensitive ABC transporters were detected using the fluorescent probes lucifer yellow and calcein. For the first time ion currents were recorded on the membrane of isolated parasitophorous vacuoles using the patch clamp technique. Results. The parasitophorous vacuole stains red with acridine orange indicating an acidic compartment. It concentrates lucifer yellow by means of a probenecid sensitive transporter but excludes calcein. Isolated vacuoles stained red with acridine orange and concentrated lucifer yellow by means of a probenecid sensitive transporter. These vacuoles excluded calcein and showed an ion current in their membrane which is activated at potentials close to 60 mV with a mean conductance of 46 ± 3 pS. Conclusions. Isolated parasitophorous vacuoles with permeability properties preserving transport mechanisms similar to those found in situ can be purified. A poorly selective ion current on the parasitophorous vacuole membrane is reported for the first time.

  4. Identification and characterization of new Leishmania promastigote surface antigens, LaPSA-38S and LiPSA-50S, as major immunodominant excreted/secreted components of L. amazonensis and L. infantum.

    Science.gov (United States)

    Bras-Gonçalves, Rachel; Petitdidier, Elodie; Pagniez, Julie; Veyrier, Renaud; Cibrelus, Prisca; Cavaleyra, Mireille; Maquaire, Sarah; Moreaux, Jérôme; Lemesre, Jean-Loup

    2014-06-01

    We have previously demonstrated that sera from dogs vaccinated with excreted/secreted antigens (ESA) of Leishmania infantum promastigotes (LiESAp) mainly recognized an immunodominant antigen of 54 kDa. An anti-LiESAp-specific IgG2 humoral response was observed and associated to Th1-type response in vaccinated dogs. This response was highly correlated with a long-lasting and strong LiESAp-vaccine protection toward L. infantum experimental infection. In addition, it was also shown that dogs from the vaccinated group developed a selective IgG2 response against an immunodominant antigen of 45 kDa of Leishmania amazonensis ESA promastigotes (LaESAp). In order to identify and characterize these immunodominant antigens, a mouse monoclonal antibody (mAb F5) was produced by immunization against LaESAp. It was found to recognize the major antigenic targets of both LaESAp and LiESAp. Analysis with mAb F5 of L. amazonensis amastigote and promastigote cDNA expression libraries enabled the identification of clones encoding proteins with significant structural homology to the promastigote surface antigens named PSA-2/gp-46. Among them, one clone presented a full-length cDNA and encoded a novel L. amazonensis protein of 38.6 kDa calculated molecular mass (LaPSA-38S) sharing an amino acid sequence consistent with that of the PSA polymorphic family and a N-terminal signal peptide, characteristic of a secreted protein. We then screened a L. infantum promastigote DNA cosmid library using a cDNA probe derived from the LaPSA-38S gene and identified a full-length clone of a novel excreted/secreted protein of L. infantum with a calculated molecular mass of 49.2 kDa and named LiPSA-50S. The fact that a significant immunological reactivity was observed against PSA, suggests that these newly identified proteins could have an important immunoregulatory influence on the immune response. This hypothesis is supported by the fact that (i) these proteins were naturally excreted/secreted by viable Leishmania promastigotes and amastigotes, and (ii) they are selectively recognized by vaccinated and protected dogs. PMID:24614507

  5. Effect of Elatol, Isolated from Red Seaweed Laurencia dendroidea, on Leishmania amazonensis

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    Celso Vataru Nakamura

    2010-10-01

    Full Text Available In the present study, we investigated the antileishmanial activity of sesquiterpene elatol, the major constituent of the Brazilian red seaweed Laurencia dendroidea (Hudson J.V. Lamouroux, against L. amazonensis. Elatol after 72 h of treatment, showed an IC50 of 4.0 µM and 0.45 µM for promastigote and intracellular amastigote forms of L. amazonensis, respectively. By scanning and transmission electron microscopy, parasites treated with elatol revealed notable changes compared with control cells, including: pronounced swelling of the mitochondrion; appearance of concentric membrane structures inside the organelle; destabilization of the plasma membrane; and formation of membrane structures, apparently an extension of the endoplasmic reticulum, which is suggestive of an autophagic process. A cytotoxicity assay showed that the action of the isolated compound is more specific for protozoa, and it is not toxic to macrophages. Our studies indicated that elatol is a potent antiproliferative agent against promastigote and intracellular amastigote forms, and may have important advantages for the development of new anti-leishamanial chemotherapies.

  6. Novel 3,4-methylenedioxyde-6-X-benzaldehyde-thiosemicarbazones: Synthesis and antileishmanial effects against Leishmania amazonensis.

    Science.gov (United States)

    de Melos, Jorge Luiz R; Torres-Santos, Eduardo Caio; Faiões, Viviane Dos S; de Nigris Del Cistia, Catarina; Sant'Anna, Carlos Maurício R; Rodrigues-Santos, Cláudio Eduardo; Echevarria, Aurea

    2015-10-20

    A series of eleven 3,4-methylenedioxyde-6-X-benzaldehyde-thiosemicarbazones (16-27) was synthesised as part of a study to search for potential new drugs with a leishmanicidal effect. The thiosemicarbazones, ten of which are new compounds, were prepared in good yields (85-98%) by the reaction of 3,4-methylenedioxyde-6-benzaldehydes (6-X-piperonal), previously synthesised for this work by several methodologies, and thiosemicarbazide in ethanol with a few drops of H2SO4. These compounds were evaluated against Leishmania amazonensis promastigotes, and derivatives where X = I (22) and X = CN (23) moieties showed impressive results, having IC50 = 20.74 ?M and 16.40 ?M, respectively. The intracellular amastigotes assays showed IC50 = 22.00 ?M (22) and 17.00 ?M (23), and selectivity index >5.7 and >7.4, respectively, with a lower toxicity compared to pentamidine (positive control, SI = 4.5). The results obtained from the preliminary QSAR study indicated the hydrophobicity (log P) as a fundamental parameter for the 2D-QSAR linear model. A molecular docking study demonstrated that both compounds interact with flavin mononucleotide (FMN), important binding site of NO synthase. PMID:26375353

  7. Sobre a sensibilidade da cultura de leucócitos circulantes na detecção de Leishmania no sangue periférico de pacientes com leishmaniose tegumentar

    Directory of Open Access Journals (Sweden)

    Fernando T. Silveira

    1989-09-01

    Full Text Available Foi investigada a presença de Leishmania, através da cultura de leucócitos circulantes, no sangue periférico de 60 pacientes portadores de leishmaniose tegumentar americana, nas suas diferentes formas clínicas, assim como nas principais fases evolutivas da doença. Biópsias de lesões cutâneas e/ou de mucosa desses pacientes foram obtidas com a finalidade de isolar e caracterizar os parasitas, através da técnica de anticorpos monoclonais. Dos 60 pacientes examinados, foram isoladas 40 amostras de Leishmania das lesões biopsiadas, sendo 5 de Leishmania (V. brasiliensis, 3 de L. (V. guyanensis, 1 de L. (V. lainsoni, 13 de L. (L. amazonensis e 18 não puderam ser caracterizados a nível específico, porém, reagiram com anticorpos monoclonais do grupo braziliensis. Quanto àpesquisa através das culturas de leucócitos circulantes, esta revelou resultados completamente negativos. Com base nesses achados, os autores concluíram ser pouco consistente atribuir valor à cultura de leucócitos para o diagnóstico da leishmaniose tegumentar.

  8. Sobre a sensibilidade da cultura de leucócitos circulantes na detecção de Leishmania no sangue periférico de pacientes com leishmaniose tegumentar

    Directory of Open Access Journals (Sweden)

    Fernando T. Silveira

    1989-09-01

    Full Text Available Foi investigada a presença de Leishmania, através da cultura de leucócitos circulantes, no sangue periférico de 60 pacientes portadores de leishmaniose tegumentar americana, nas suas diferentes formas clínicas, assim como nas principais fases evolutivas da doença. Biópsias de lesões cutâneas e/ou de mucosa desses pacientes foram obtidas com a finalidade de isolar e caracterizar os parasitas, através da técnica de anticorpos monoclonais. Dos 60 pacientes examinados, foram isoladas 40 amostras de Leishmania das lesões biopsiadas, sendo 5 de Leishmania (V. brasiliensis, 3 de L. (V. guyanensis, 1 de L. (V. lainsoni, 13 de L. (L. amazonensis e 18 não puderam ser caracterizados a nível específico, porém, reagiram com anticorpos monoclonais do grupo braziliensis. Quanto àpesquisa através das culturas de leucócitos circulantes, esta revelou resultados completamente negativos. Com base nesses achados, os autores concluíram ser pouco consistente atribuir valor à cultura de leucócitos para o diagnóstico da leishmaniose tegumentar.The possible presence of Leishmania in the peripheral blood of 60 patients with American cutaneous leishmaniasis was investigated by the culture of circulating leucocytes. Patients were selected with a variety ofclinical forms ofthe disease and in different evolutionary stages of infection. Biopsies of skin and/or mucosal lesions were made in order to isolate the parasites, which were identified using monoclonal antibodies. 40 isolations were obtained, including 5 of Leishmania (Viannia braziliensis, 3 L. (V. guyanensis, 1 L. (V. lainsoni, 13 L. (Leishmania amazonensis and 18 which could only be identified as parasites of the braziliensis complex. Cultures of circulanting leucocytes were consistently negative, and the authors conclude that this method is of little use in diagnosis of cutaneous or mucocutaneous leishmaniasis.

  9. The human parasite Leishmania amazonensis downregulates iNOS expression via NF-?B p50/p50 homodimer: role of the PI3K/Akt pathway

    Science.gov (United States)

    Calegari-Silva, Teresa C.; Vivarini, Áislan C.; Miqueline, Marina; Dos Santos, Guilherme R. R. M.; Teixeira, Karina Luiza; Saliba, Alessandra Mattos; Nunes de Carvalho, Simone; de Carvalho, Laís; Lopes, Ulisses G.

    2015-01-01

    Leishmania amazonensis activates the NF-?B transcriptional repressor homodimer (p50/p50) and promotes nitric oxide synthase (iNOS) downregulation. We investigated the role of PI3K/Akt in p50/p50 NF-?B activation and the effect on iNOS expression in L. amazonensis infection. The increased occupancy of p50/p50 on the iNOS promoter of infected macrophages was observed and we demonstrated that both p50/p50 NF-?B induction and iNOS downregulation in infected macrophages depended on PI3K/Akt activation. Importantly, the intracellular growth of the parasite was also impaired during PI3K/Akt signalling inhibition and in macrophages knocked-down for Akt 1 expression. It was also observed that the increased nuclear levels of p50/p50 in L. amazonensis-infected macrophages were associated with reduced phosphorylation of 907 Ser p105, the precursor of p50. Corroborating these data, we demonstrated the increased levels of phospho-9 Ser GSK3? in infected macrophages, which is associated with GSK3? inhibition and, consequently, its inability to phosphorylate p105. Remarkably, we found that the levels of pPTEN 370 Ser, a negative regulator of PI3K, increased due to L. amazonensis infection. Our data support the notion that PI3K/Akt activity is sustained during the parasite infection, leading to NF-?B 105 phosphorylation and further processing to originate p50/p50 homodimers and the consequent downregulation of iNOS expression. PMID:26400473

  10. Lutzomyia reducta Feliciangeli et al., 1988, a host of Leishmania amazonensis, sympatric with two other members of the Flaviscutellata complex in southern Amazonas and Rondônia, Brazil (Diptera: Psychodidae) / Lutzomyia reducta Feliciangeli et al., 1988 um hospedeiro de Leishmania amazonensis, simpátrico com duas outras espécies do complexo flaviscutellata no sul do Amazonas e Rondônica, Brasil (Diptera: Psychodidae)

    Scientific Electronic Library Online (English)

    R. A., Freitas; T. V., Barrett; R. D., Naiff.

    1989-09-01

    Full Text Available Um flebotomíneo do complexo Lutzomyia flaviscutellata, de Rondônia e sul do Amazonas, Brasil é tão parecido com Lutzomyia olmeca reducta, que é considerado como sendo da mesma espécie. Este flebotomíneo ocorre junto com L. olmeca nociva, portanto o nome é emendado para o nível de espécie, como Lutzo [...] myia reducta. O material do Brasil é descrito e ilustrado, e comparado com exemplares de L. o. nociva e L. flaviscutellata da mesma área. Chaves para as espécies e subespécies do complexo flaviscutellata são incluídas. Leishmania amazonensis foi isolada em um exemplar de L. reducta altamente infectado, tornando esta espécie a terceira a ser implicada como vetor desta leishmania no Brasil. A abundância relativa das três espécies simpátricas do complexo flaviscutellata varia em escala local e aparenta ter relação com a drenagem do solo. L. reducta constituiu cerca de 25% dos flebotomíneos capturados em armadilhas Disney em locais mal e bem drenados, porém não foi encontrada em locais sujeitos a inundações. L. olmeca nociva era restrita às áreas mal drenadas não sujeitas a inundações, enquanto L. flaviscutellata foi capturada neste dois ambientes e também numa área periodicamente inundada. L. reducta não tem sido assinalada ao norte do Rio Amazonas no Brasil, porém a ausência de registros do oeste e noroeste do Estado do Amazonas possa refletri a falta de levantamento nestas áreas. Abstract in english A member of the Lutzomyia flaviscutellata complex from Rondônia and southern Amazonas States, Brazil, is so close to the Venezuelan Lutzomyia olmeca recuta Feliciangeli et al., 1988, that it is regarded as belonging to the same species. Since this phlebotomine co-extis with L. olmeca nociva in Brazi [...] l, the subspecific status of the former is untenable and is rased to specific rank, as Lutzomyia reducta. The Brazilian material is described and illustrated, and compared with specimens of L. o. nociva and L. flaviscutellata from the same area. Keys to the known taxa of the flaviscutellata complex are presented. Leishmania amazonensis was isolated from one heavily infected specimen of L. reducta, making this the third species of the flaviscutellata complex to be implicated as a vector of this parasite in Brazil. The relative abundance of the three sympatric flaviscutellata complex species varies locally and appears to be related to soil drainage. L. reducta constituted about 25% if all phlebotomines captured in Disney traps at poorly drained and well drained site, but appears not to coloniza areas subject to periodic flooding. L. olmeca nociva was restricted to poorly drained areas not subject to flooding, whereas L. flaviscutellata was ubiquitous L. reducta has never been detected north of the Amazon river in Brazil, but absence of recosrds from western and northwestern Amazonas State may reflect lack of collecting in these areas.

  11. Study of ionizing radiation as a tool for select promastigotes forms of Leishmania Amazonensis, and the megalomaniac response in experimental models

    International Nuclear Information System (INIS)

    Actually, millions of people around the globe are under the risk of infection by a protozoan transmitted by a bit of a sand fly. This parasite is a Leishmania spp. This causes a wide spectrum disease, since a cutaneous disease to a visceral one. The cutaneous form is the major clinical manifestation (above 90%). The ionizing radiation, produced in a 60Co font, had being successes used to promote physical-chemical transformations on different protozoan, including Leishmania spp. In previous work was determined that promastigotes forms of Leishmania amazonensis, irradiated with different doses of radiation, lost their viability maintaining, however, their immunogenicity. In this work, was studied the use of ionizing radiation as a tool for selection of meta cyclic forms of the parasite in axenic culture, for a possible efficient irradiated immuno gene production. Our results shown that cultures irradiated with 400 Gy of gamma irradiation, has 75% of metacyclic form, which are capable to produce, in vitro, an infection that is similar the natural occurrence. These irradiated parasites have their internal cellular structure modified, maintaining their external structure intact. Susceptible strain of mice immunized with leishmania irradiated with different doses had high immunoglobulin production, and maintained this production after the challenge with naive parasites. In other strains this default was similar, however in lower titles. Immunodeficient mice didn't produce immunoglobulin nor on the immunization or on the challenge. (author)

  12. Short-term protection conferred by Leishvacin® against experimental Leishmania amazonensis infection in C57BL/6 mice.

    Science.gov (United States)

    Carneiro, Matheus Batista Heitor; de Andrade e Sousa, Louisa Maria; Vaz, Leonardo Gomes; Dos Santos, Liliane Martins; Vilela, Luciano; de Souza, Carolina Carvalho; Gonçalves, Ricardo; Tafuri, Wagner Luis; Afonso, Luís Carlos Crocco; Côrtes, Denise Fonseca; Vieira, Leda Quercia

    2014-12-01

    To date, there is no vaccine available against human leishmaniasis. Although some vaccination protocols can induce immunity in murine models, they fail to induce protection in humans. The reasons for that remain unclear. The aim of the present study was to characterize the changes in the pattern of the immune response during subcutaneous vaccination with Leishvacin® in mice. We also investigated whether IFN-? and nitric oxide synthase are indispensable for the protection elicited by the vaccine. C57BL/6 WT vaccinated mice showed smaller lesions and fewer numbers of parasites in footpads until 8 weeks post-infection. Up to this time, they produced higher levels of IFN-?, IL-2, IL-4, IL-17A and IL-10 and higher specific antibody response than control non-vaccinated mice. Moreover, we showed that IFN-?, most likely by induction of iNOS expression, is essential for immunity. However, after 12 weeks of infection, we observed loss of difference in lesion size and parasite burden between the groups. Loss of resistance was associated with the disappearance of differences in cytokine patterns between vaccinated and control mice, but not of antibody response, which remained different until a later time of infection. The reversal of resistance to L. amazonensis could not be explained by upregulation of regulatory cytokines. Our data point to a subversion of the host immune response by L. amazonensis even when a protective response was previously induced. PMID:25102355

  13. DFT/PCM, QTAIM, 1H NMR conformational studies and QSAR modeling of thirty-two anti-Leishmania amazonensis Morita-Baylis-Hillman Adducts

    Science.gov (United States)

    Filho, Edilson B. A.; Moraes, Ingrid A.; Weber, Karen C.; Rocha, Gerd B.; Vasconcellos, Mário L. A. A.

    2012-08-01

    Morita-Baylis-Hillman Adducts (MBHA) has been recently synthesized and bio-evaluated by our research group against Leishmania amazonensis, parasite that causes cutaneous and mucocutaneous leishmaniasis. We present here a theoretical conformational study of thirty-two leismanicidal MBHA by B3LYP/6-31+g(d) calculations with Polarized Continuum Model (PCM) to simulate water influence. Intramolecular Hydrogen Bonds (IHBs) indicated to control the most conformational preferences of MBHA. Quantum Theory Atoms in Molecules (QTAIM) calculations were able to characterize these interactions at Bond Critical Point level. Compounds presenting an unusual seven member IHB between NO2 group and hydroxyl moiety, supported by experimental spectroscopic data, showed a considerable improvement of biological activity (lower IC50 values). These results are in accordance to redox NO2 mechanism of action. Based on structural observations, some molecular descriptors were calculated and submitted to Quantitative Structure-Activity Relationship (QSAR) studies through the PLS Regression Method. These studies provided a model with good validation parameters values (R2 = 0.71, Q2 = 0.61 and Qext2 = 0.92).

  14. LFR1 Ferric Iron Reductase of Leishmania amazonensis Is Essential for the Generation of Infective Parasite Forms*

    OpenAIRE

    Flannery, Andrew R.; Huynh, Chau; Mittra, Bidyottam; Mortara, Renato A.; Andrews, Norma W

    2011-01-01

    The protozoan parasite Leishmania is the causative agent of serious human infections worldwide. The parasites alternate between insect and vertebrate hosts and cause disease by invading macrophages, where they replicate. Parasites lacking the ferrous iron transporter LIT1 cannot grow intracellularly, indicating that a plasma membrane-associated mechanism for iron uptake is essential for the establishment of infections. Here, we identify and functionally characterize a second member of the Lei...

  15. El rol de tres pruebas de ELISA con antígenos de promastigotes de Leishmania braziliensis, L. amazonensis y L. guyanensis en el diagnóstico de leishmaniasis tegumentaria / Role of three ELISA tests using promastigote homogenates of Leishmania braziliensis, L. amazonensis and L. guyanensis in the diagnosis of tegumentary leishmaniasis

    Scientific Electronic Library Online (English)

    José F., Gil; Carlos L., Hoyos; Rubén O., Cimino; Alejandro J., Krolewiecki; Inés, Lopéz Quiroga; Silvana P., Cajal; Marisa, Juárez; María F., García Bustos; María C., Mora; Jorge D., Marco; Julio R., Nasser.

    2011-10-01

    Full Text Available Es importante conocer si la variabilidad de especies de Leishmania circulantes en una región afecta la performance de las pruebas de ELISA estandarizadas para el diagnostico de la leishmaniasis. El objetivo de este trabajo fue analizar la reactividad de la prueba de ELISA utilizando homogenados de p [...] romastigotes de Leishmania (V.) braziliensis (ELISAb), L (L) amazonensis (ELISAa) y L (V.) guyanensis (ELISAg) frente a distintos grupos de sueros. Se estudiaron muestras de personas con leishmaniasis cutánea (n = 37), leishmaniasis mucocutánea (n = 8), no infectados (n = 52), infectadas por Trypanosoma cruzi (n = 11) e infecciones mixtas (n = 14). Se calcularon las sensibilidades, especificidades, cut off, valores predictivos, y se compararon las tres pruebas usando ANOVA, índice de concordancia kappa, comparación de curvas ROC e intervalos de confianza construidos por el método de bootstrap. Se encontraron diferencias significativas al comparar los niveles de DO de los sueros de pacientes con leishmaniasis cutánea respecto a los controles negativos, pero no se encontraron diferencias entre pruebas. Las sensibilidades calculadas fueron de 84.6% para ELISAb y ELISAa y de 88.5 para ELISAg, mientras que el valor de especificidad para las tres pruebas fue de 96.2. El índice de concordancia kappa y la comparación de curvas ROC mostraron performances similares para las tres pruebas (p = 0.225). La elevada reactividad obtenida para estas ELISAs frente a sueros de pacientes con leishmaniasis mucocutánea indica un importante potencial de esta técnica como complemento en el diagnóstico de la enfermedad. Abstract in english It is important to know whether the variability of species of Leishmania parasites circulating in a region affects the performance of the ELISA test for the diagnosis of leishmaniasis. Therefore, the aim of this study was to analyze the reactivity of the ELISA using homogenates of promastigotes of L [...] eishmania (V.) braziliensis (ELISAb), Leishmania (L) amazonensis (ELISAa) and Leishmania (V.) guyanensis (ELISAg) against different sera groups. Samples from individuals with cutaneous leishmaniasis (n = 37), mucocutaneous leishmaniasis (n = 8), healthy controls (n = 52), persons infected with Trypanosoma cruzi (n = 11) and mixed infections (n = 14) were included in the study. We calculated sensitivities, specificities, cut offs, and predictive values for the three tests and compared them using ANOVA, kappa index, ROC curves comparison, and confidence intervals calculated by the bootstrap method. Significant differences were found when comparing the OD levels of sera from patients with cutaneous leishmaniasis against healthy controls, but there were no differences when comparing the different ELISAs. The sensitivities calculated for ELISAb and ELISAa were 84.6 and of 88.5% for ELISAg, while the value of specificity for the three tests was 96.2. The kappa index (0.87) and comparison of ROC curves showed similar performance for the three ELISAs (p = 0.225). The high reactivity obtained for these ELISAs in sera of patients with mucocutaneous leishmaniasis indicates this test as an important complement in the diagnosis of the disease.

  16. Determinación de la producción de Öxido Nítrico en Macrófagos Activados J774.1 como respuesta al tamaño de la carga fagocítica de partículas de látex y Leishmania amazonensis Determination of Nitric Oxide Production in Activate Macrophages J774.1 as effect of Phagocitic Load Volume of Latex Beads and Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    Camargo Jiménez Maria Helena

    2000-06-01

    Full Text Available La leishmaniosis es una enfermedad parasitaria causada por el protozoario Leishmania. Cercade 12 millones de personas padecen esta enfermedad y 350 millones de personas están enriesgo de contraerla. Existe evidencia de que la infección por Leishmania amazonensisdisminuyela producción de óxido nítrico (NO de macrófagos; se ha interpretado que este deterioroes inducido por el parásito. Esta investigación corrobora esta idea, pero contradice que seaexclusivo de la infección. En este trabajo, cultivos celulares de macrófagos fueron expuestosa partículas de látex en diferentes proporciones buscando cargas fagocíticas y volúmenes defagosoma semejantes a los de la infección. Las concentraciones de nitrito y parámetros mor-fológicos se midieron a las 48 horas post fagocitosis e infección con 24 horas de activaciónmediante IFN-?y LPS. Se determinó que volúmenes similares de fagosomas generados porpartículas de látex o amastigotes de L. amazonensis, deterioran en la misma proporción laproducción de NO. Esto sugiere que este deterioro no depende de la naturaleza de la partículafagocitada. El incremento en el volumen de fagosoma se correlaciona con la disminución en laproducción de nitrito, por tanto, la expansión del fagosoma puede ser uno de los mecanismosimplicados en la disminución de la producción de NO. Los resultados apoyan la evidencia deque Leishmaniadisminuye la producción de NO pero contradicen la interpretación,comúnmente aceptada, de que este fenómeno es específico de la infección.Leishmaniasis is a parasitic disease caused by a protozoa of the genera Leishmania. Around 12million people are infected and 350 millions of people are risk to contract it. There is evidencethat infection by Leishmania amazonensisdecreases nitric oxide production; it has beeninterpreted that this impairment is induced by the parasite. This study confirmed this idea, butcontradicts that this impairment is exclusive of infection. In this work, cultured macrophageswere exposed to latex beads at diferents proportions, to simulate phagocitic loads andphagosome volumes resembling those of the infection. Nitrite concentration and morphologicalparameters were measured at 48 hours post phagocytosis and infection with 24 hours ofactivation induced by IFN-?and LPS. Our results determined that macrophages with similarvolume of phagosomes generated by latex beads or L. amazonensisamastigotes have the samedecrease in inducible NO production. This suggest that this partial inhibition does not depenton the nature of phagocited particle. Phagosome volume increase correlates with the decreasein inducible NO production. Therefore, phagosome expansion may be one of the mechanismimplicated in the disminution of inducible NO production. The results support the evidencethat Leishmaniareduces inducible NO production, but contradicts the common interpretationthat this fenomena is specific of infection.

  17. Acercamiento al estudio de la interacción y salida de Leishmania amazonensis en un modelo in vitro con macrófagos murinos de la línea celular J774a.1

    Directory of Open Access Journals (Sweden)

    León Cabrera Sonia Andrea

    2006-06-01

    Full Text Available Los miembros del género Leishmania son parásitos intracelulares obligados, responsables de numerosas enfermedades humanas. Cumplen una parte muy importante de su ciclo de vida dentro del mamífero hospedero, en donde después de la fagocitosis por los macrófagos, los parásitos son confinados dentro de un compartimiento endolisosomal denominado vacuola parasitófora (VP, en el cual se replican siendo finalmente liberados infectando otros macrófagos y de esta forma ampliando la infección. Poco es el conocimiento que se tiene acerca de cómo ocurre el proceso de liberación de amastigotes de Leishmania que infectan macrófagos. Se sospecha que este mecanismo puede estar ocurriendo por un proceso de fusión de membranas. Mediciones de capacitancia
    de la membrana del macrófago y el uso de inhibidores de fusión de membranas soportan esta idea. El objetivo de este trabajo fue realizar seguimientos del ciclo infectivo de Leishmania amazonensis, para confirmar los hallazgos
    previos en cuanto a los tiempos en que probablemente puede estar ocurriendo la salida del amastigote. Además, se buscó determinar la viabilidad del parásito a lo largo del ciclo infectivo con el fin de comprender mejor la interacción hospedero-patógeno en el modelo in vitro; para ello se midió: viabilidad del parásito con tinción de diacetato de fluoresceína (DAF y ioduro de propidio (IP, porcentaje de infección y número de parásitos por célula (p/c. Los resultados sugieren que la salida de los parásitos puede presentarse entre las 72 y 78 horas post infección (hpi y entre las 96 y 120 hpi. Con los resultados de trabajos previos, y los datos presentados en este estudio, se ha propuesto que L. amazonensis puede presentar dos ciclos infectivos que se desarrollan durante cinco días en nuestras condiciones de cultivo in vitro. En las primeras 36-48 hpi el parásito se diferencia a amastigote. Después de su diferenciación comienza su división celular. Luego de las 72 hpi ocurre una disminución en el número de parásitos por célula (p/c que ha sido relacionada con el momento en el cual podría salir el parásito de su célula hospedera. La recuperación del número de p/c a las 96 hpi y la disminución presentada a las 120 hpi sugieren la ocurrencia de un nuevo ciclo infectivo. La viabilidad del amastigote se vio afectada a medida que transcurrió la infección. Durante las primeras 24 hpi prácticamente todos los parásitos fueron viables (93,85%
    y se observaron de color verde intenso dentro de las VP por marcaje con la sonda DAF. Entre el tercer y cuarto
    día se presentó una disminución significativa en la viabilidad de los parásitos p = 0,017 y p = 0,0097 respectivamente.
    Entre el tercer y quinto día post infección el cultivo en general se observó más deteriorado y se encontró
    una cantidad considerable de macrófagos no viables, pero aún con parásitos viables en el interior de la VP. Estas
    observaciones se han interpretado como competencia en el cultivo, lo que generaría déficit alimenticio,
    explicando la drástica disminución en la viabilidad general del cultivo. El descenso diario de un grupo de células infectadas podría ser la causa de la disminución en los porcentajes de infección. En este trabajo se desarrolló un método eficiente para marcar la membrana de macrófagos infectados con los análogos fluorescentes de fosfolípidos NBD-PE y RHO-PE con el fin de implementar la técnica FRET, y así evidenciar la fusión de una membrana no marcada como la de la VP, con una membrana previamente marcada como la del macrófago. Se estipuló que la concentración de 5 ?g/mL y 10 ?g/mL para las pruebas RHO-PE y NBD-PE respectivamente, puestas en contacto con macrófagos infectados en nuestras condiciones, fueron capaces de marcar clara y continuamente la membrana celular del 95,9% y 97,0% de los macrófagos. Asi mismo, con la menor formación de vesículas de la sonda comparada con otras concentraciones y con una

  18. Métodos subsidiários para o diagnóstico da Leishmaniose tegumentar americana (LTA: comparação dos resultados do seqüenciamento de DNA e da PCR-RFLP para determinação da espécie de leishmania em amostras cutâneo-mucosas Subsidiary methods for the diagnosis of American tegumentar leishmaniasis (ATL: comparison of sequencing of DNA and PCR-RFLP for identification of leishmania species in skin samples

    Directory of Open Access Journals (Sweden)

    Flávio C. Barbosa Garcia

    2005-12-01

    Full Text Available FUNDAMENTOS: Métodos moleculares têm-se mostrados mais eficazes para o diagnóstico da LTA. OBJETIVOS: Comparar os resultados da intradermorreação de Montenegro (IRM, presença de leishmania em biópsia (Bc, reação de imunofluorescência indireta (Rifi, seqüenciamento de DNA e PCR-RFLP (-restriction fragment lenght polymorphism para o diagnóstico da LTA. MÉTODOS: Foram estudados 152 pacientes com LTA. Para a PCR em Bc, utilizaram-se primers específicos para seqüência de 120bp do kDNA do minicírculo comum a todas as espécies de leishmanias. O produto da PCR, utilizado para seqüenciamento e para restrição enzimática com Hae III, foi comparado às culturas L. (L. amazonensis e L. (V. braziliensis. RESULTADOS: Houve predomínio do sexo masculino (75%, da cor branca (80% e da profissão urbana (48%. A idade variou de três a 77 anos, com 56,5% entre 21 e 50 anos. 65,8% eram do Estado de São Paulo, prevalecendo a forma cutânea (79,6%. A IRM foi positiva em 73,4%, e a Rifi em 59,7%, enquanto a Bc evidenciou presença de leishmania em 30,6%. A PCR foi positiva em 81,6%, e a PCR-RFLP identificou L. (V. braziliensis como espécie predominante (66%, o que também ocorreu com o seqüenciamento. Comparando PCR-RFLP e seqüenciamento, houve 61% de concordância entre os resultados, mostrando significância da PCR-RFLP para L. (V. braziliensis. CONCLUSÕES: A IRM e a PCR foram estatisticamente equivalentes como métodos subsidiários para o diagnóstico da LTA, a PCR-RFLP e o seqüenciamento também o foram na identificação das espécies de leishmania, o primeiro apresentando menores custo e tempo de execução comparado ao seqüenciamento de DNA.BACKGROUND: ATL is endemic in Brazil, and molecular methods have been shown more effective for its diagnosis. OBJECTIVES: Our purpose was to compare the results of Montenegro’s skin reaction (MR, presence of leishmania in skin biopsy (Bx, indirect immunofluorescence (IIF for leishmania in sera samples, PCR, sequencing of DNA and PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism as subsidiary exams for ATL diagnose. METHODS: 152 patients were studied, with accomplishment of MR, Bx, IIF and PCR for leishmania in skin samples. For PCR, a specific pair of primers for a sequence of 120bp from kDNA minicircle, common to all leishmanias species, was used. The product of PCR was used for DNA sequencing and for RFLP with the enzyme Hae III. The analysis of the restriction pattern was compared to the cultures of L. (L. amazonensis and L. (V. braziliensis. RESULTS: The predominant sex was male (75%, the white color (80% and urban professional occupation (48%. The age varied from 3 to 77 years, with prevalence from 21 to 50 years (56.5%. In relation to the origin, 65.8% was from the state of São Paulo, being the cutaneous form the more prevalent (79.6%. MR was positive in 73.4% and there was presence of leishmania in 30.6% of the samples, while IIF presented 59.7% of positivity. PCR was positive in 81.6% of skin samples, and PCR-RFLP identified L. (V. braziliensis (66% as predominant species, fact that also happened with DNA sequencing, with 64.4% of the positive results for L. (V. braziliensis. Comparing PCR-RFLP and DNA sequencing there was 61% of agreement amongst the results, being significant in PCR-RFLP for L. (V. braziliensis. CONCLUSION: MR and PCR were equivalent as subsidiary methods for the diagnosis of ATL, such as PCR-RFLP and DNA sequencing in the identification of the leishmania species. On the other hand, the method PCR-RFLP presented smaller cost and smaller time of execution compared to the sequencing of DNA.

  19. Métodos subsidiários para o diagnóstico da Leishmaniose tegumentar americana (LTA): comparação dos resultados do seqüenciamento de DNA e da PCR-RFLP para determinação da espécie de leishmania em amostras cutâneo-mucosas / Subsidiary methods for the diagnosis of American tegumentar leishmaniasis (ATL): comparison of sequencing of DNA and PCR-RFLP for identification of leishmania species in skin samples

    Scientific Electronic Library Online (English)

    Flávio C. Barbosa, Garcia; Sandra Silva Rodrigues dos, Santos; Maria Fernanda, Chociay; Ângela C. Rapela, Medeiros; Ana Maria F., Roselino.

    2005-12-01

    Full Text Available FUNDAMENTOS: Métodos moleculares têm-se mostrados mais eficazes para o diagnóstico da LTA. OBJETIVOS: Comparar os resultados da intradermorreação de Montenegro (IRM), presença de leishmania em biópsia (Bc), reação de imunofluorescência indireta (Rifi), seqüenciamento de DNA e PCR-RFLP (-restriction [...] fragment lenght polymorphism) para o diagnóstico da LTA. MÉTODOS: Foram estudados 152 pacientes com LTA. Para a PCR em Bc, utilizaram-se primers específicos para seqüência de 120bp do kDNA do minicírculo comum a todas as espécies de leishmanias. O produto da PCR, utilizado para seqüenciamento e para restrição enzimática com Hae III, foi comparado às culturas L. (L.) amazonensis e L. (V.) braziliensis. RESULTADOS: Houve predomínio do sexo masculino (75%), da cor branca (80%) e da profissão urbana (48%). A idade variou de três a 77 anos, com 56,5% entre 21 e 50 anos. 65,8% eram do Estado de São Paulo, prevalecendo a forma cutânea (79,6%). A IRM foi positiva em 73,4%, e a Rifi em 59,7%, enquanto a Bc evidenciou presença de leishmania em 30,6%. A PCR foi positiva em 81,6%, e a PCR-RFLP identificou L. (V.) braziliensis como espécie predominante (66%), o que também ocorreu com o seqüenciamento. Comparando PCR-RFLP e seqüenciamento, houve 61% de concordância entre os resultados, mostrando significância da PCR-RFLP para L. (V.) braziliensis. CONCLUSÕES: A IRM e a PCR foram estatisticamente equivalentes como métodos subsidiários para o diagnóstico da LTA, a PCR-RFLP e o seqüenciamento também o foram na identificação das espécies de leishmania, o primeiro apresentando menores custo e tempo de execução comparado ao seqüenciamento de DNA. Abstract in english BACKGROUND: ATL is endemic in Brazil, and molecular methods have been shown more effective for its diagnosis. OBJECTIVES: Our purpose was to compare the results of Montenegro’s skin reaction (MR), presence of leishmania in skin biopsy (Bx), indirect immunofluorescence (IIF) for leishmania in sera sa [...] mples, PCR, sequencing of DNA and PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) as subsidiary exams for ATL diagnose. METHODS: 152 patients were studied, with accomplishment of MR, Bx, IIF and PCR for leishmania in skin samples. For PCR, a specific pair of primers for a sequence of 120bp from kDNA minicircle, common to all leishmanias species, was used. The product of PCR was used for DNA sequencing and for RFLP with the enzyme Hae III. The analysis of the restriction pattern was compared to the cultures of L. (L.) amazonensis and L. (V.) braziliensis. RESULTS: The predominant sex was male (75%), the white color (80%) and urban professional occupation (48%). The age varied from 3 to 77 years, with prevalence from 21 to 50 years (56.5%). In relation to the origin, 65.8% was from the state of São Paulo, being the cutaneous form the more prevalent (79.6%). MR was positive in 73.4% and there was presence of leishmania in 30.6% of the samples, while IIF presented 59.7% of positivity. PCR was positive in 81.6% of skin samples, and PCR-RFLP identified L. (V.) braziliensis (66%) as predominant species, fact that also happened with DNA sequencing, with 64.4% of the positive results for L. (V.) braziliensis. Comparing PCR-RFLP and DNA sequencing there was 61% of agreement amongst the results, being significant in PCR-RFLP for L. (V.) braziliensis. CONCLUSION: MR and PCR were equivalent as subsidiary methods for the diagnosis of ATL, such as PCR-RFLP and DNA sequencing in the identification of the leishmania species. On the other hand, the method PCR-RFLP presented smaller cost and smaller time of execution compared to the sequencing of DNA.

  20. Vaccination of C57BL/10 mice against cutaneous leishmaniasis using killed promastigotes of different strains and species of Leishmania Vacinação de camundongos C57BL/10 contra leishmaniose com promastigotas mortas de diferentes cepas e espécies de Leishmania

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    Wilson Mayrink

    2002-04-01

    Full Text Available Antigenic extracts from five Leishmania stocks were used to vaccinate C57BL/10 mice. The Leishvacin® and PH8 monovalent vaccine yielded the highest IFN-gamma levels in the supernatants of spleen cell culture from vaccinated animals. Each single strain immunized group showed evidence of protective immunity six months after the challenge with promastigotes of Leishmania (Leishmania amazonensis. No differences were detected between the vaccinated groups. It can be concluded that vaccines composed of single Leishmania stocks can provide protection to C57BL/10 mice against L. (L. amazonensis infection.Estudos anteriores revelaram que uma vacina preparada com promastigotas mortas de cinco cepas de Leishmania pode induzir uma imunidade protetora para a leishmaniose tegumentar americana no homem e em modelos experimentais. Um dos problemas do uso desta vacina é a complexidade de sua composição e a necessidade de se incorporar diferentes cepas de Leishmania. Por esta razão, extratos antigênicos de cada uma das cinco cepas constituintes da vacina foram preparados e usados individualmente em estudos imunológicos com camundongos C57BL/10. A Leishvacin® e a vacina monovalente PH8 induziram os maiores níveis de Interferon-g (IFN-gama detectado no sobrenadante de células esplênicas dos animais vacinados. Cada grupo imunizado com vacinas monovalentes desenvolveram uma imunidade protetora seis meses após a infecção desafio com promastigotas de Leishmania (Leishmania amazonensis e nenhuma diferença estatística foi observada entre os grupos vacinados. Pode-se concluir que vacinas compostas por cepas isoladas de Leishmania protegem camundongos C57BL/10 contra, pelo menos, da infecção por L. (L. amazonensis.

  1. Leishmania mexicana in Proechimys iheringi denigratus Moojen (Rodentia, Echimyidae in a region endemic for American cutaneous leishmaniasis

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    Air C. Barretto

    1985-12-01

    Full Text Available Three isolates of Leishmania were recovered from five of 27 specimens of the rodent Proechimys iheringi denigratus Moojen captured near Três Braços in the Atlantic Forest region of Bahia, Brazil. Two of these isolates were recovered from hamsters inoculated with a pooled triturate of liver, spleen and skin tissue from apparently healthy P. i. denigratus. The third isolate was recovered from a triturate of only skin tissue from another. Metastasis was observed in the inoculated hamsters, the parasites grew abundantly in artificial media and a typical suprapylarial pattern of infection in Lutzomyia longipalpis was produced indicating that the parasites belong to the Leishmania mexicana complex. All isolates reacted with Leishmania mexicana mexicana and Leishmania mexicana amazonensis monoclonal antibodies. The isoenzyme analysis differentiated these isolates from standard isolates of L. m. mexicana, L. m. amazonensis, L. m. aristedesi, L. m. pifanoi, L. m. garnhami and L. m. ssp.(Goiás-W. Barbosa. These isolates seem to be a subspecies of L. mexicana very closely related to L. m. amazonensis from which they differ by decreased electrophoretic mobility of GPI, PEP and ALAT. This is the first record of the isolation of a parasite of thegenus Leishmania in a rodent captured in the State of Bahia.Três isolados de Leishmania foram obtidos de cinco entre 27 exemplares do roedor Proechimys iheringi denigratus, capturados na região de Três Braços, na mata atlântica do Estado da Bahia, Brasil. O isolamento desse parasito foi feito através de inoculação de triturado de pele, baço e fígado em patas de hamsters. Em pelo menos um dos casos, (MTB-574, o parasito foi isolado da pele. Metas- tase foi observada nos hamsters inoculados, os parasitos cresceram abundantemente em meios artificiais de cultura e um padrão suprapapilario típico foi obtido em Lutzomyia longipalpis, indicando que o parasito pertence ao complexo L. mexicana. Todos os isolados reagiram positivamente com anticorpos monoclonais de L. m. mexicana e L. m. amazonensis. A análise isoenzimática diferenciou o parasito de isolados padrões de L. m. mexicana. L. m. amazonensis. L. m. aristedesi. L. m. pifanoi. L. m. garnhami e L. m. ssp(Goiás-W. Barbosa. O parasito parece ser uma subespécie de L. mexicana muito próxima à L. m. amazonensis, da qual difere pela menor mobilidade eletroforêtica de GPI, PEP e ALAT. Este e o primeiro registro do isolamento de um parasito do gênero Leishmania em um roedor capturado no Estado da Bahia.

  2. Polyclonal B cell activation in hamsters infected with parasites of the genus Leishmania.

    OpenAIRE

    Campos-Neto, A.; Bunn-Moreno, M M

    1982-01-01

    Mesocricetus auratus (golden hamsters) infected with leishmania developed characteristic B cell immune responses that depended on the infecting species of leishmania. Thus, hamsters infected with viscerotropic leishmania (Leishmania donovani) developed antileishmania antibodies and hypergammaglobulinemia due to polyclonal activation of B cells as measured by reverse hemolytic plaque assay. In contrast, dermotropic leishmanias (L. braziliensis braziliensis and L. mexicana amazonensis) stimulat...

  3. Molecular and parasitological detection of Leishmania spp. in a dipteran of the species Tabanus importunus / Detecção molecular e parasitológica de Leishmania spp. em díptero da espécie Tabanus importunus

    Scientific Electronic Library Online (English)

    Willian Marinho Dourado, Coelho; Katia Denise Saraiva, Bresciani.

    2013-12-01

    Full Text Available A leishmaniose é uma importante zoonose, de caráter crônico, causada por protozoários do gênero Leishmania spp. Esta protozoose tem como principal vetor os flebotomíneos, sendo que, no Brasil, o Lutzomyia longipalpis é a principal espécie incriminada na transmissão da leishmaniose Visceral Americana [...] . A presença do ácido desoxirribonucleico (DNA) do parasito em ectoparasitos, como carrapatos e pulgas, tem gerado especulações quanto a existência de novos vetores no ciclo da leishmaniose. Foi objetivo deste estudo relatar a detecção molecular de Leishmania spp. em uma mutuca da espécie Tabanus importunus que parasitava um cão oligossintomático infectado por Leishmania spp. A análise molecular amplificou o DNA do protozoário na cabeça, na região torácica e no abdomen do tabanídeo, resultando como positivo para complexo Leishmania. Este é o primeiro relato da presença de DNA de Leishmania spp. em insetos dipteros da espécie T. importunus. Abstract in english Leishmaniasis is an important chronic zoonosis caused by protozoa of the genus Leishmania spp. The major vectors of this protozoosis are sand flies, and Lutzomyia longipalpis is considered the main species implicated in the transmission of American Visceral Leishmaniasis in Brazil. The presence of t [...] he parasite's deoxyribonucleic acid (DNA) in ectoparasites such as ticks and fleas has prompted speculations about the existence of new vectors in the cycle of leishmaniasis. The aim of this paper is to report the molecular detection of Leishmania spp. in a horse fly of the species Tabanus importunus which parasitized an oligosymptomatic dog infected with Leishmania spp. Molecular amplification of the protozoan's DNA in the head, thoracic region and abdomen of the tabanid tested positive for Leishmania complex. This is the first report of the presence of DNA from Leishmania spp. in dipterous insects of the species T. importunus.

  4. The action of ionizing radiation on the morphology, physiology and growth of Leishmania Amazonensis, with evaluation of their immunogenic power in experimental models

    International Nuclear Information System (INIS)

    Leishmaniasis is a disease which affects thousands of people in the Tropical regions around the world, is caused by a protozoan of the genus Leishmania spp., with urban and wild mammals acting as reservoirs. In the mammal host, the amastigote form of the parasite infects and multiplies into macrophages. Treatments for leishmaniasis have a high cost and are long lasting, frequently resulting in therapy interruption. This procedure culminates with a selection of resistant parasite strains, inducing tolerance to the therapy. Either the control of vectors or the mammal host are difficult due the social and economic implications. Thus, the search for alternatives treatments against these protozoans have been stimulated. The gamma radiation (60CO) shown to be an efficient toll to kill these parasites maintaining their immunogenicity. Cellular viability, Electronically microscopy and Multiplex-PCR techniques showed that, after irradiation, the parasites had their growth inhibited by cytoplasmatic and nucleic material disorganisation, appointing the gamma radiation as important in terms of immunogens improvement. (author)

  5. Infective stages of Leishmania in the sandfly vector and some observations on the mechanism of transmission Formas infectante de Leishmania no vetor flebotomíneo e algumas observações sobre o mecanismo de transmissão

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    Ralph Lainson

    1987-09-01

    Full Text Available Infective stages of Leishmania (Leishmania amazonensis, capable of producing amastigote infections in hamster skin, were shown to be present in the experimentally infected sandfly vector Lutzomyia flaviscutellata 15, 25, 40, 49, 70, 96 and 120 hours after the flies had received their infective blood-meal. Similarly, infective stages of Leishmania (L. chagasi were demonstrated in the experimentally infected vector Lu. longipalpis examined 38, 50, 63, 87, 110, 135, 171 and 221 hours following the infective blood-meal, by the intraperitoneal inoculation of the flagellates into hamsters. The question of whether or not transmission by the bite of the sandfly is dependent on the presence of [quot ]metacyclic[quot ] promastigotes in the mouthparts of the vector is discussed.Foi demonstrado através de infecção experimental, que estágios infectivos de Leishmania (L. amazonensis, capazes de produzir infecção na pele do hamster, encontram-se presentes no vetor flebotomíneo Lutzomyia flaviscutellata 15, 25, 40, 49, 70, 96 e 120 horas após o inseto ter recebido sua refeição sangüínea infectiva. Da mesma maneira, foi comprovada a presença de estágios infectivos de L. (L. chagasi em exemplares do vetor Lu. longipalpis, examinados 38, 50, 63, 87, 110, 135, 171 e 221 horas após o repasto sangüíneo infectivo - através da inoculação em hamster por via intraperitoneal dos flagelados obtidos desses fle botomíneos. A questão sobre a transmissão do gênero Leishmania pelo flebotomíneo ser ou não dependente da presença de promastigotos "metacíclios" na proboscis do vetor, é discutida.

  6. ALTERAÇÕES DA MATRIZ EXTRACELULAR ESPLÊNICA EM CÃES NATURALMENTE INFECTADOS COM Leishmania (Leishmania) infantum chagasi / CHANGES IN THE SPLENIC EXTRACELLULAR MATRIX IN DOGS NATURALLY INFECTED WITH Leishmania (Leishmania) infantum chagasi

    Scientific Electronic Library Online (English)

    Nathálya dos Santos, Martins; Glaucia Barbosa, Coelho; Larissa Sarmento dos, Santos; Rudson Almeida de, Oliveira; Ana Lúcia Abreu, Silva; Ferdinan Almeida, Melo.

    2015-03-01

    Full Text Available Objetivou-se estudar as alterações da matriz extracelular no baço de cães naturalmente infectados com Leishmania (Leishmania) infantum chagasi correlacionando-as com aspectos clínicos, histopatológicos e parasitológicos. Foram utilizados 18 cães, divididos em três grupos: seis animais não infectados [...] (grupo controle) e doze animais infectados. Todos sem raça e idade definidas, provenientes da região do Município de São Luis-MA. Cortes parafinados do baço foram corados pela Hematoxilina e Eosina (H&E); Prata amoniacal de Gomori, para marcação das fibras reticulares e pela técnica Imuno-histoquímica da estreptoavidina-peroxidase para detecção de formas amastigotas de Leishmania. As análises morfométricas foram feitas utilizando-se o programa KS300 e o sistema de análise de imagens Kontron Elektronic/Carl Zeiss, Germany. Os resultados mostram que há um aumento significativo da deposição de fibras colágenas no baço quando comparadas aos animais controles, revelando diferenças significativas entre os animais sintomáticos e assintomáticos. Encontraram-se correlações positivas entre a presença do parasitismo tecidual e a deposição de colágeno. Os animais sintomáticos apresentaram uma maior deposição de colágeno no baço, que pode estar associada ao maior parasitismo tecidual encontrado. Os resultados demonstram que na leishmaniose visceral canina há uma fibrogênese intensa no baço, sendo esta associada ao parasitismo tecidual e a processos degenerativos decorrentes da doença. Abstract in english The aim of this study was to study the changes in the splenic extracellular matrix of dogs naturally infected with Leishmania (Leishmania) infantum chagasi and its correlation to clinical, histopathological, and parasitological aspects. Eighteen dogs were used, separated into three groups: six non-i [...] nfected animals (control group) and twelve infected animals. The dogs had undefined breed and age, from the township region of São Luís-MA. Paraffined slices of the spleen were stained with Hematoxilin and Eosin (H&E); Gomori’s ammoniacal Silver, to mark reticular fibers; and the Immunohistochemistry technique of streptavidin peroxidase to detect amastigote forms of Leishmania. The morphometrical analyses were done using the KS300 program and the images analysis system Kontron Elektronic/Carl Zeiss, Germany. The results showed that there is a significant increase in the deposition of collagen fibers in the spleen, compared to control animals, showing significant differences for symptomatic and asymptomatic animals. Positive correlations were found between the presence of the parasite in the tissue and collagen deposition. Symptomatic animals showed higher collagen deposition in the spleen, which can be associated to the high parasitism found in the tissue. The results showed that there is an intense fibrogenesis in the spleen in the canine visceral leishmaniasis, thus being associated to the parasitism of the tissue and the degenerative processes of the disease.

  7. Kinetics of growth of Leishmania (Leishmania chagasi cycle in McCoy cell culture Cinéticas de crescimento do ciclo da Leishmania (Leishmania chagasi em cultura de células McCoy

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    Yeda L. Nogueira

    2006-12-01

    Full Text Available The kinetics of growth of Leishmania performed in vitro after internalization of the promastigote form in the cell and the occurrence of the transformation of the parasite into the amastigote form have been described by several authors. They used explants of macrophages in hamster spleen cell culture or in a human macrophage lineage cell, the U937. Using microscopy, the description of morphologic inter-relationship and the analysis of the production of specific molecules, it has been possible to define some of the peculiarities of the biology of the parasite. The present study shows the growth cycle of Leishmania chagasi during the observation of kinetic analysis undertaken with a McCoy cell lineage that lasted for a period of 144 hours. During the process, the morphologic transformation was revealed by indirect immunofluorescence (IF and the molecules liberated in the extra cellular medium were observed by SDS-PAGE at 24-hour intervals during the whole 144-hour period. It was observed that in the first 72 hours the promastigote form of L. chagasi adhered to the cell membranes and assumed a rounded (amastigote-like form. At 96 hours the infected cells showed morphologic alterations; at 120 hours the cells had liberated soluble fluorescent antigens into the extra cellular medium. At 144 hours, new elongated forms of the parasites, similar to promastigotes, were observed. In the SDS-PAGE, specific molecular weight proteins were observed at each point of the kinetic analysis showing that the McCoy cell imitates the macrophage and may be considered a useful model for the study of the infection of the Leishmania/cell binomial.Cinéticas de crescimento de Leishmania realizadas in vitro após a internalização da forma promastigota na célula e a ocorrência da transformação do parasito na forma amastigota foram descritas por vários autores, seja com a utilização de explantes de macrófagos em células de baço de hamster ou atualmente da célula de linhagem de macrófago humano U937. Aliando a microscopia à descrição das inter-relações morfológicas e à síntese de moléculas específicas foi possível esclarecer pontos sobre a biologia do parasito. O presente estudo mostra o acompanhamento do ciclo de crescimento da Leishmania chagasi em uma cinética realizada com células de linhagem McCoy, no período de 144 horas. Durante o processo, as transformações morfológicas foram reveladas pela reação de imunofluorescência indireta (RIFI e as moléculas liberadas no meio extracelular foram observadas pelo método de SDS-PAGE, em intervalos de 24 horas no período de 144 horas. Observou-se que nas primeiras 72 horas, a forma promastigota da L. chagasi fica aderida à membrana das células com aspecto arredondado (amastigota-like. Em 96 horas as células infectadas apresentaram alterações morfológicas; em 120 horas, as células liberaram, para o meio extracelular, antígenos fluorescentes solúveis; e em 144 horas foram observadas novas formas alongadas dos parasitos como se fossem promastigotas. No SDS-PAGE, proteínas com pesos moleculares específicos são observadas em cada ponto da cinética, mostrando que a célula McCoy parece mimetizar o macrófago e que pode ser um modelo útil para o estudo da infecção do binômio leishmânia/célula.

  8. Diagnosis of Leishmania infantum infection by Polymerase Chain Reaction in wild mammals / Diagnóstico de infecção por Leishmania infantum pela reação em cadeia da polimerase em mamíferos silvestres

    Scientific Electronic Library Online (English)

    Mayara C., Lombardi; Andréia P., Turchetti; Herlandes P., Tinoco; Angela T., Pessanha; Semiramis A., Soave; Marcelo C.C., Malta; Tatiane A., Paixão; Renato L., Santos.

    2014-12-01

    Full Text Available A leishmaniose visceral é uma doença infecciosa crônica de mamíferos causada, no Brasil, pelo protozoário Leishmania infantum (sinonímia: Leishmania chagasi) e transmitida pelo flebótomo Lutzomyia longipalpis. Trata-se de uma zoonose endêmica em muitas regiões do Brasil, inclusive em Belo Horizonte, [...] Minas Gerais. Em centros urbanos, leishmaniose visceral acomete principalmente o cão doméstico. Entretanto, L. infantum já foi diagnosticada em outras espécies, incluindo canídeos e primatas de cativeiro em zoológicos. Este estudo buscou avaliar a presença do DNA deste agente em animais de cativeiro e de vida livre da Fundação Zoobotânica de Belo Horizonte através da reação em cadeia da polimerase. Foram analisadas oitenta e uma amostras de sangue oriundas de primatas, carnívoros, ruminantes, edentatos, marsupial e herbívoro de estômago simples. Três primatas Alouatta guariba (bugio marrom) e dois canídeos Speothos venaticus (cachorro-do-mato-vinagre), foram positivos, demonstrando a importância do controle da leishmaniose em áreas endêmicas com a finalidade de conservar a fauna silvestre mantida em cativeiro. Abstract in english Visceral leishmaniasis is a chronic infectious disease caused by Leishmania infantum (synonym: Leishmania chagasi) and transmitted by the sandfly Lutzomyia longipalpis in Brazil. It is an endemic zoonosis in several regions of the country, including Belo Horizonte (State of Minas Gerais). In urban a [...] reas, the domestic dog is susceptible and considered the most important animal reservoir. However, L. infantum has been previously diagnosed in other species, including captive primates and canids. This study aimed to evaluate the presence of the agent DNA in captive animals as well as some free ranging animals from the Zoo-Botanical Foundation of Belo Horizonte by Polymerase Chain Reaction. Eighty one blood samples from primates, carnivores, ruminants, edentates, marsupial, and a monogastric herbivore were analyzed. Three primates Alouatta guariba (brown howler monkey), and two canids Speothos venaticus (bush dog) were positive, demonstrating the importance of leishmaniasis control in endemic areas for preservation of wildlife species in captivity.

  9. Evaluation of HIV-Leishmania co-infection in patients from the northwestern Paraná State, Brazil = Avaliação da co-infecção HIV-Leishmania em pacientes da região noroeste do Estado do Paraná, Brasil

    Directory of Open Access Journals (Sweden)

    Élide Aparecida Oliveira

    2011-01-01

    Full Text Available Leishmaniasis occurs throughout the world and is one of the opportunistic infections that attack HIV-infected individuals. Few data are available on American cutaneous leishmaniasis (ACL in HIV-infected patients. Current research investigates the occurrence ofHIV-Leishmania co-infection in HIV-infected individuals in an endemic region in Southern of Brazil. A non-randomized transversal investigation, molecular and serum epidemiologic type, on the occurrence of ACL in 169 HIV-infected patients was undertaken. The patients were followed up at the Integrated Nucleus of Health of the city Maringá, Southern of Brazil. Results showed that 13 (7.7% of the HIV-infected patients also presented Leishmania (Viannia DNA, detectable in blood by PCR. Serology, direct research, culture and PCR in skin material produced negative results. PCR positiveness for Leishmania was not associated with CD4 T lymphocytes count, opportunistic disease, treatment, use of proteases inhibitors, tattooing/piercing or use of injectable drugs, residential environment or previous ACL history. Results show that HIVinfected patients who live in endemic areas may reveal Leishmania DNA in the blood without any ACL symptoms. Above findings may be attributed to anti-retrovirus medicine that controls viral replication and maintains the functionality of the immune system and to a possible anti- Leishmania activity of these drugs.As leishmanioses ocorrem em todo o mundo e são infecções oportunistas que afetam indivíduos portadores do vírus HIV. Este estudo investigou a ocorrência da co-infecção HIV-Leishmania em portadores do HIV numa região endêmica para LTA do Sul do Brasil. Foi realizado estudo transversal, não randomizado, utilizando metodologia molecular e sorológica, sobre a ocorrência de LTA em 169 portadores do HIV. Foram estudados pacientes atendidos no Núcleo Integrado de Saúde de Maringá, Paraná, Sul do Brasil. Observou-se que 13 (7,7% dos pacientes infectados pelo HIV também apresentavam o DNA de Leishmania (Viannia, detectável no sangue por PCR. A sorologia, pesquisa direta de Leishmania, cultura e PCR de lesões de pele foram negativas. A positividade da PCR não estava associada à contagem de linfócitos T CD4+, doença oportunista, tratamento, uso de inibidores de protease, tatuagem, uso de drogas injetáveis, ambiente da residência ou história prévia de LTA. Os resultados mostraram que indivíduos portadores do vírus HIV que residem em área endêmica podem apresentar o DNA de Leishmania sem manifestar sintomas de LTA. Estes resultados podem ser atribuídos a ação dos medicamentos anti retrovirais que controlam a replicação viral mantendo a integridade do sistema imunológico ou a uma possível atividade anti-Leishmania destas drogas.

  10. Conjunctival swab PCR to detect Leishmania spp. in cats / Uso da PCR de suabe conjuntival para detecção de Leishmania spp. em gatos

    Scientific Electronic Library Online (English)

    Trícia Maria Ferreira de Sousa, Oliveira; Vanessa Figueredo, Pereira; Graziella Ulbricht, Benvenga; Maria Fernanda Alves, Martin; Julia Cristina, Benassi; Diogo Tiago da, Silva; Wilma Aparecida, Starke-Buzetti.

    2015-06-01

    Full Text Available A importância do cão como fonte de infecção da leishmaniose visceral já é conhecida, mas o papel dos gatos como reservatórios das leishmanioses ainda não está totalmente esclarecido. O presente estudo avaliou a eficácia da PCR de suabe conjuntival (PCR-SC) na detecção de gatos infectados por Leishma [...] nia spp. Foram encontrados sete (13,5%) gatos positivos para Leishmania spp. na PCR de suabe conjuntival, dentre 52 animais de Pirassununga - SP e Ilha Solteira - SP testados. Sendo positivos 28,6% (02/07) dos gatos do município de Pirassununga e 11,1% (5/45) dos gatos do município de Ilha Solteira. Os resultados demonstraram que o suabe de conjuntiva ocular foi capaz de detectar gatos infectados por esse protozoário. A coleta de amostras da conjuntiva mostrou ser um método simples, menos invasivo e pouco estressante para os gatos e seus proprietários, o que pode facilitar estudos sobre a frequência e distribuição da leishmaniose felina. Abstract in english The relevance of the dog as a source of visceral leishmaniasis infection is known, but the role of cats as reservoir hosts for leishmaniasis is not yet fully clear. This study assessed the efficacy of conjunctival swab PCR (CS-PCR) in the detection of cats infected by Leishmania spp. The results wer [...] e seven (13.5%) cats positive for Leishmania spp. in the PCR, in 52 cats tested from Pirassunuga-SP and Ilha Solteira-SP. From the city of Pirassununga – SP 28.6% (2/7) were positive and from the city of Ilha Solteira – SP 11.1% (5/45) were positive. The results showed that CS-PCR was capable of detecting cats infected by this protozoan. Conjunctival swab samples proved easier to perform in cats, which might facilitate studies on the frequency and distribution of feline leishmaniasis.

  11. Effects of seco-steroids purified from Physalis angulata L., Solanaceae, on the viability of Leishmania sp Efeitos de seco-esteróides purificados de Physalis angulata L., Solanaceae na viabilidade de Leishmania sp

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    Elisalva T. Guimarães

    2010-12-01

    Full Text Available Physalis angulata L., Solanaceae, is an annual herb commonly used in popular medicine in many tropical and subtropical countries. P. angulata extracts contain a variety of substances, but little is known about their pharmacological activities. In this work we investigated the in vitro antileishmanial activity of seco-steroids (physalins purified from P. angulata. Addition of physalins B, F, and G caused a concentration-dependent inhibition in the growth of L. amazonensis promastigotes, being the IC50 values were 6.8, 1.4, and 9.2 ?M, respectively. Physalin D was less active and had an IC50 value of 30.5 ?M. Physalins were also active in cultures of other Leishmania species (L. major, L. braziliensis, and L. chagasi. Our results demonstrate the potent antileishmanial activity of physalins in cultures of Leishmania species of the New and Old Worlds and suggest the therapeutic potential of these seco-steroids in leishmaniasis.Physalis angulata L., Solanaceae, é uma erva anual utilizada na medicina popular em muitos países tropicais e subtropicais. Apesar dos extratos da P. angulata apresentarem uma grande variedade de substâncias, pouco é conhecido sobre a sua atividade farmacológica. Neste trabalho foi investigado a atividade antileishmania in vitro de seco-esteroides (fisalinas purificados da P. angulata. O tratamento com as fisalinas B, F e G causou uma inibição concentração-dependente do crescimento de promastigotas de Leishmania amazonensis em cultura axênica, com valores de IC50 de 6,8, 1,4, e 9,2 ?M respectivamente. A fisalina D foi menos ativa, com valores de IC50 de 30,5 ?M. Foi também observada uma atividade leishmanicida em culturas de outras espécies de Leishmania (L. major, L. braziliensis e L. chagasi. Nossos resultados demonstram que as fisalinas inibem o crescimento dos promastigotas com o tratamento de espécies de Leishmania do Velho e do Novo Mundos e sugerem o potencial terapêutico destas moléculas na leishmaniose.

  12. Cura espontânea da leishmaniose causada por Leishmania Viannia Braziliensis em lesões cutâneas

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    Jackson Maurício Lopes Costa

    1990-12-01

    Full Text Available Os autores relatam que durante 14 anos de trabalho clínico em campo, realizado nas comunidades de Três Braços e Corte de Pedra, Bahia, acompanharam 1.416 pacientes portadores de Leishmaniose Tegumentar Americana, cuja espécie envolvida na transmissão, é predominantemente a Leishmania Viannia brasilienses. A terapêutica utilizada rotineiramente nos casos é o antimoniato-N-metilglucamina (Glucantime. Contudo, 16 pacientes do sexo masculino recusaram-se a utilizar a medicação e 6 do sexo feminino encontravam-se em período gestacional, portanto não utilizaram o medicamento. Estes pacientes foram acompanhados por um período entre 4 a 12 anos, a partir do diagnóstico. Observou-se que em 9 pacientes (40,9% desta casuística, o tempo de cicatrizaçâo após o aparecimento da lesão, pode ser calculado em 6 meses de evolução. Quando se eleva a observação para 12 meses, temos que 19 pacientes (86,3% cicatrizaram suas lesões neste período. Em 3 casos (13,6% as lesões permaneceram ativas por mais de 12 meses. Conclui-se que os determinantes da cicatrizaçâo natural das lesões produzidas por Leishmania Viannia Braziliensis permanecem desconhecidos, dificultando para nós entendermos e compararmos aos efeitos das drogas utilizadas no tratamento da leishmaniose tegumentar.In field clinics in the comunities of Três Braços and Corte de Pedra, Bahia, we have attended 1.416 patients with tegumentary leishmaniasis in fourteen years, the predomi nant species in transmission is Leishmania Viannia brasiliensis (LVB. Because of the danger of metastasis with this infection treatment was routinely recomended with Glucantime. However sixteen patients refused injection therapy and six women were pregnant when seen and not treated. All patients were followed up in our clinic. All these patients closed their skin ulcers although one subsequently relapsed. Patients were followed up for variable periods (four to twelve years, after the diagnosis. In nine patients (40,9% of the cohort, the time to healing after initiation of the lesion was calculated as six months of evolution. At twelve months, nineteen patients (86,3% had complete healing of their lesions. In three patients an active lesion was present for longer than one year. The determinants of this variable natural evolution of human LVB lesion remains completely unknown. It is difficult for us to understand and compare the effects of therapeutic agents in mucocutaneous leishmaniasis.

  13. Análise histomorfométrica da matriz extracelular do linfonodo poplíteo de cães naturalmente infectados por Leishmania (L. chagasi

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    Kris Régia J. Kondo

    2009-08-01

    Full Text Available Nas Américas, a leishmaniose visceral canina é causada por Leishmania (Leishmania chagasi, um protozoário intracelular obrigatório do sistema fagocítico mononuclear; as principais alterações histológicas associadas a essa doença ocorrem nos em órgãos linfóides. Apesar de o cão ser considerado o principal mantenedor e disseminador da leishmaniose no ambiente urbano, são escassos estudos dos aspectos histopatológicos e histomorfométricos, em cães naturalmente infectados com L. chagasi, que investiguem a interação entre o parasito e a matriz extracelular. Este estudo visou caracterizar e quantificar as alterações dos componentes celulares e da matriz extracelular (colágenos I e III do linfonodo poplíteo de 22 cães com infecção natural por L. chagasi detectada através da reação de imunofluorescência indireta (RIF e compará-las com as alterações encontradas no linfonodo poplíteo de 10 cães não-infectados, negativos na RIF e clinicamente saudáveis. Fragmentos dos linfonodos foram seccionados longitudinalmente, processados rotineiramente para exame histológico e corados por hematoxilina-eosina. Cortes adicionais do mesmo linfonodo incluídos em glicol metacrilato foram corados pelo azul de toluidina para histomorfometria. Linfonodos de cães infectados apresentaram linfadenopatia generalizada, aumento do tamanho e do número dos folículos linfóides, hipertrofia da cápsula e hiperplasia linfóide significativa. Nos linfonodos de cães do grupo infectado, a análise quantitativa de fibras colágenas mostrou significativo predomínio do colágeno I sobre o colágeno III. Esses resultados demonstram que cães infectados por L. chagasi apresentam degradação dos constituintes da matriz extracelular e conseqüente destruição do arcabouço linfóide, alterando a morfologia do órgão.In the Americas, canine visceral leishmaniasis is caused by Leishmania (Leishmania chagasi, an obligatory intracellular parasite of the phagocytic-monocytic system; the main histological changes associated with this disease occur in the lymphoid organs. Although dogs are considered to be the main carriers and disseminators of leishmaniasis in urban areas, there are few studies on the histopathologic and histomorphometric aspects in dogs naturally infected by L.chagasi analyzing the interaction between parasite and extracellular matrix. The current study characterize and quantify changes in the cellular and extracellular matrix (collagens type I and III components of the popliteal lymph node from of 22 dogs with the natural infection by L. chagasi confirmed by indirect immuno-fluorescence assay (IFA and compare theses findings with those fund in the popliteal lymph node from 10 non-infected dogs, that reacted negative in the IFA, and were clinically healthy. Lymph node fragments were longitudinally sliced and sections were processed for routine histopathology and stained by hematoxylin and eosin. For histomorphometry, additional sections from the same lymph node were fixed in glycol methacrylate and stained with toluidine blue. Lymph nodes from affected dogs were systemically enlarged, had increased numbers of lymphoid follicles, capsule hyperplasia and hypertrophy, and significant hyperplasia of lymphoid cells. In the lymph nodes from infected dogs, quantitative analyses of collagen fibers revealed predominance of type I collagen over type III fibers. These results demonstrate that dogs infected by L.chagasi experience degradation of the extracellular matrix components and consequently destruction of the lymphoid framework, thus altering nodal morphology.

  14. Análise histomorfométrica da matriz extracelular do linfonodo poplíteo de cães naturalmente infectados por Leishmania (L.) chagasi

    Scientific Electronic Library Online (English)

    Kris Régia J., Kondo; Cláudio César, Fonseca; Sérgio Luis P. da, Matta; Marlene Isabel V., Viloria.

    2009-08-01

    Full Text Available Nas Américas, a leishmaniose visceral canina é causada por Leishmania (Leishmania) chagasi, um protozoário intracelular obrigatório do sistema fagocítico mononuclear; as principais alterações histológicas associadas a essa doença ocorrem nos em órgãos linfóides. Apesar de o cão ser considerado o pri [...] ncipal mantenedor e disseminador da leishmaniose no ambiente urbano, são escassos estudos dos aspectos histopatológicos e histomorfométricos, em cães naturalmente infectados com L. chagasi, que investiguem a interação entre o parasito e a matriz extracelular. Este estudo visou caracterizar e quantificar as alterações dos componentes celulares e da matriz extracelular (colágenos I e III) do linfonodo poplíteo de 22 cães com infecção natural por L. chagasi detectada através da reação de imunofluorescência indireta (RIF) e compará-las com as alterações encontradas no linfonodo poplíteo de 10 cães não-infectados, negativos na RIF e clinicamente saudáveis. Fragmentos dos linfonodos foram seccionados longitudinalmente, processados rotineiramente para exame histológico e corados por hematoxilina-eosina. Cortes adicionais do mesmo linfonodo incluídos em glicol metacrilato foram corados pelo azul de toluidina para histomorfometria. Linfonodos de cães infectados apresentaram linfadenopatia generalizada, aumento do tamanho e do número dos folículos linfóides, hipertrofia da cápsula e hiperplasia linfóide significativa. Nos linfonodos de cães do grupo infectado, a análise quantitativa de fibras colágenas mostrou significativo predomínio do colágeno I sobre o colágeno III. Esses resultados demonstram que cães infectados por L. chagasi apresentam degradação dos constituintes da matriz extracelular e conseqüente destruição do arcabouço linfóide, alterando a morfologia do órgão. Abstract in english In the Americas, canine visceral leishmaniasis is caused by Leishmania (Leishmania) chagasi, an obligatory intracellular parasite of the phagocytic-monocytic system; the main histological changes associated with this disease occur in the lymphoid organs. Although dogs are considered to be the main c [...] arriers and disseminators of leishmaniasis in urban areas, there are few studies on the histopathologic and histomorphometric aspects in dogs naturally infected by L.chagasi analyzing the interaction between parasite and extracellular matrix. The current study characterize and quantify changes in the cellular and extracellular matrix (collagens type I and III) components of the popliteal lymph node from of 22 dogs with the natural infection by L. chagasi confirmed by indirect immuno-fluorescence assay (IFA) and compare theses findings with those fund in the popliteal lymph node from 10 non-infected dogs, that reacted negative in the IFA, and were clinically healthy. Lymph node fragments were longitudinally sliced and sections were processed for routine histopathology and stained by hematoxylin and eosin. For histomorphometry, additional sections from the same lymph node were fixed in glycol methacrylate and stained with toluidine blue. Lymph nodes from affected dogs were systemically enlarged, had increased numbers of lymphoid follicles, capsule hyperplasia and hypertrophy, and significant hyperplasia of lymphoid cells. In the lymph nodes from infected dogs, quantitative analyses of collagen fibers revealed predominance of type I collagen over type III fibers. These results demonstrate that dogs infected by L.chagasi experience degradation of the extracellular matrix components and consequently destruction of the lymphoid framework, thus altering nodal morphology.

  15. Assessment of PCR in the detection of Leishmania spp in experimentally infected individual phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae)

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    MICHALSKY Érika M.; FORTES-DIAS Consuelo L.; Pimenta, Paulo F. P.; SECUNDINO Nágila F.C.; Edelberto S. DIAS

    2002-01-01

    DNA amplification by the polymerase chain reaction (PCR) was applied in the investigation of the presence of Leishmania (Kinetoplastida: Trypanosomatidae) parasites in single phlebotomine sandflies. Three phlebotomine/parasite pairs were used: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis and Lutzomyia migonei/Leishmania braziliensis, all of them incriminated in the transmission of visceral or cutaneous leishmaniasis. DNA extraction was performed with whol...

  16. Modulation of Na+/K+ ATPase Activity by Hydrogen Peroxide Generated through Heme in L. amazonensis

    OpenAIRE

    Rocco-Machado, Nathália; Cosentino-Gomes, Daniela; Meyer-Fernandes, José Roberto

    2015-01-01

    Leishmania amazonensis is a protozoan parasite that occurs in many areas of Brazil and causes skin lesions. Using this parasite, our group showed the activation of Na+/K+ ATPase through a signaling cascade that involves the presence of heme and protein kinase C (PKC) activity. Heme is an important biomolecule that has pro-oxidant activity and signaling capacity. Reactive oxygen species (ROS) can act as second messengers, which are required in various signaling cascades. Our goal in this work ...

  17. Diagnóstico molecular da taxa de infecção natural de flebotomíneos (Psychodidae, Lutzomyia por Leishmania sp na Amazônia maranhense Molecular diagnosis of the natural infection rate due to Leishmania sp in sandflies (Psychodidae, Lutzomyia in the Amazon region of Maranhão, Brazil

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    Yrla Nívea Oliveira-Pereira

    2006-12-01

    Full Text Available A taxa de infecção natural de três diferentes espécies de flebotomíneos por Leishmania foi estudada usando a técnica de reação em cadeia da polimerase. Primers específicos para Leishmania foram designados para examinar se os pools de flebotomíneos estavam infectadas. Um total de 1.100 fêmeas separadas em pools de 10 indivíduos foram examinados, consistindo de 50 Lutzomyia whitmani, 43 Lutzomyia triacantha e 17 Lutzomyia choti. De todos os pools analisados, 4 de Lutzomyia whitmani estavam positivos, mas nenhum pool das duas espécies restantes estava infectado. Deste modo, uma taxa de infecção de 0,4% foi verificada neste estudo. Esta taxa de infecção associada a estudos anteriores sugere que Lutzomyia whitmani transmite Leishmania aos mamíferos em Buriticupu, Maranhão.The natural infection rate due to Leishmania was studied in three different sandfly species using the polymerase chain reaction technique. Leishmania specific primers were designed to examine whether sandfly pools were infected. In total 1,100 female sandflies separated into pools of 10 individuals, consisting of 50 pools of Lutzomyia whitmani, 43 of Lutzomyia triacantha and 17 of Lutzomyia choti, were analyzed. Among all the pools examined, four pools of Lutzomyia whitmani were positive, but none of the pools of the other two species were infected. Thus, a total infection rate of 0.4% was established in this study. A similar infection rate was found in previous studies, suggesting that Lutzomyia whitmani transmits Leishmania to mammals in Buriticupu, Maranhão.

  18. Effect of ionizing radiation on the morphology, physiology and growth of Leishmania ssp; Acao da radiacao ionizante sobre a morfologia, fisiologia e crescimento da Leishmania spp

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    Bonetti, Franco C.; Spencer, Patrick J.; Nascimento, Nanci do [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil); Junior A, Heitor F. [Sao Paulo Univ., SP (Brazil). Faculdade de Medicina. Instituto de Medicina Tropical

    2000-07-01

    The Leishmania spp is a pathogenic protozoan, which cause different diseases in man. The human diseases, in America, caused by this group of protozoa are divided in cutaneous or tegumentar and visceral, known as kala-azar. In this work, our principal study object was the specie that causes tegumentar leishmaniasis, in Brazil. Metabolic studies of cellular respiration and proteins and nucleic acids synthesis were accomplished using radiation as a form of sterilizing the parasites without however affecting their immunogenic capacity The promastigotes forms of irradiated Leishmania spp were totally sterilized with the dose of 1500 Gy, with their reproductive and nucleic acids, as well as protein synthesis capacity blocked. (author)

  19. Estado atual da leishmaniose cutânea difusa (LCD no Estado do Maranhão: II. aspectos epidemiológicos, clínico-evolutivos

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    Jackson Maurício Lopes Costa

    1992-06-01

    Full Text Available Os Autores fazem um estudo retrospectivo e prospectivo de 6 pacientes portadores de leishmaniose cutânea difusa, observados no Estado do Maranhão a partir de 1974. Os casos abordados são oriundos de diversas regiões do estado, observando-se em todos eles o envolvimento da leishmania (Leishmania amazonensis, sendo que 5 (84% dos pacientes apresentaram início de doença na 1ª década de vida. Em todos os pacientes envolvidos no estudo, houve relato de lesão inicial nodular única, que, posteriormente, em período variável de tempo, disseminou-se adquirindo outros aspectos. Evolutivamente apresentaram múltiplas lesões nodulares e ulceradas, intradermorreação de Montenegro(- e refratariedade aos esquemas terapêuticos utilizados até ao presente momento.The authors describe a retrospective and prospective study of 6 patients with diffuse cutaneous leishmaniasis observed in the State of Maranhão, since 1974. The patients comefromdifferentruralregions of the state and in all of them Leishmania (Leishmania amazonensis was the cause five of the patients initiated their disease in the first decade of life. All the patients first had a solitary, nodular lesion, that after a variable period of time, disseminated and acquired other aspects. Sequentially the patients presented multiple nodular and ulcerative lesions, negative leishmania skin-lests and a refractory response to the therapeutic schedules used up to the present.

  20. DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL / Detecção de Leishmania (Viannia) em Nyssomyia neivai e Nyssomyia whitmani por Multiplex Reação em Cadeia da Polimerase, no sul do Brasil

    Scientific Electronic Library Online (English)

    Herintha Coeto, Neitzke-Abreu; Kárin Rosi, Reinhold-Castro; Mateus Sabaini, Venazzi; Regiane Bertin de Lima, Scodro; Alessandra de Cassia, Dias; Thaís Gomes Verzignassi, Silveira; Ueslei, Teodoro; Maria Valdrinez Campana, Lonardoni.

    2014-09-01

    Full Text Available Flebotomíneos transmitem os patógenos das leishmanioses. Foi avaliada a infecção natural de flebotomíneos por Leishmania (Viannia) em municípios do Estado do Paraná, sul do Brasil. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon. Após dissecação para pesquisa de flagelados no tub [...] o digestório e identificação das espécies, as fêmeas de flebotomíneos foram submetidas a Multiplex Reação em Cadeia da Polimerase (multiplex PCR) para a detecção do fragmento do kDNA de Leishmania (Viannia) e do fragmento do gene IVS6 da cacofonia de flebotomíneos. A análise foi realizada em pools contendo sete a 12 tubos digestórios de fêmeas da mesma espécie. Um total de 510 fêmeas foram analisadas, incluindo nove Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai e 268 Nyssomyia whitmani. Embora nenhuma fêmea tenha sido encontrada naturalmente infectada com flagelados pela dissecação, o fragmento de DNA de Leishmania (Viannia) foi mostrado por multiplex PCR em uma amostra de Ny. neivai (0,46%) e três amostras de Ny. whitmani (1,12%). Conclui-se que Ny. neivai e Ny. whitmani são suscetíveis à infecção por Leishmania, e que multiplex PCR, devido à sua sensibilidade, especificidade e viabilidade, pode ser utilizada em estudos epidemiológicos para a detecção da infecção natural do inseto vetor. Abstract in english Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes a [...] nd identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

  1. Kinetics of growth of Leishmania (Leishmania) chagasi cycle in McCoy cell culture Cinéticas de crescimento do ciclo da Leishmania (Leishmania) chagasi em cultura de células McCoy

    OpenAIRE

    Yeda L. NOGUEIRA; Paulo M Nakamura; Eunice A. B. Galati

    2006-01-01

    The kinetics of growth of Leishmania performed in vitro after internalization of the promastigote form in the cell and the occurrence of the transformation of the parasite into the amastigote form have been described by several authors. They used explants of macrophages in hamster spleen cell culture or in a human macrophage lineage cell, the U937. Using microscopy, the description of morphologic inter-relationship and the analysis of the production of specific molecules, it has been possible...

  2. Gamma radiation affects the anti-Leishmania activity of Bothrops moojeni venom and correlates with L-amino acid oxidase activity

    Energy Technology Data Exchange (ETDEWEB)

    Tempone, A.G.; Lourenco, C.O.; Spencer, P.J.; Rogero, J.R.; Nascimento, N. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Div. de Radiobiologia; Andrade Junior, H.F. [Sao Paulo Univ., SP (Brazil). Faculdade de Medicina. Inst. de Medicina Tropical

    1999-11-01

    Leishmania causes human disfiguring skin disease in endemic areas of Amazon and North Eastern Brazil. Those parasites present a remarkable resistance to most treatments, except those using toxic antimonial salts. We detected a specific anti-Leishmania activity in snake venoms, using an in vitro promastigote assay. In this report, we analyzed the activity of Bothrops moojeni venom against L. Amazonensis, using whole venom or fractions of L-amino acid oxidase (L-AO). Crude venom of B.moojeni, was fractionated by molecular exclusion chromatography. Activity against promastigotes was detected by respiratory oxidative conversion of MTT in a colorimetric assay and L-AO activity was detected by a colorimetric assay with peroxidase and OPD as revealing reagents. Crude venom was irradiated with 500, 1000, and 2000 Gy in a {sup 60} Co gamma radiation source. The venom had an anti-Leishmania activity of 33 pg/promastigote and the active fraction migrates as 100-150 kDa, close to the size described for L-AOs, and also presented L-AO activity. The radiation reduces both the L-AO and anti-Leishmania activity in a dose dependent effect. Those data suggests the anti-Leishmania activity in this venom is closely related to the L-amino acid oxidase activity and also that radiation could be used as a tool to detect specific activities reduction in water solutions, similarly to observed in dry preparations. (author) 13 refs., 3 figs.

  3. Gamma radiation affects the anti-Leishmania activity of Bothrops moojeni venom and correlates with L-amino acid oxidase activity

    International Nuclear Information System (INIS)

    Leishmania causes human disfiguring skin disease in endemic areas of Amazon and North Eastern Brazil. Those parasites present a remarkable resistance to most treatments, except those using toxic antimonial salts. We detected a specific anti-Leishmania activity in snake venoms, using an in vitro promastigote assay. In this report, we analyzed the activity of Bothrops moojeni venom against L. Amazonensis, using whole venom or fractions of L-amino acid oxidase (L-AO). Crude venom of B.moojeni, was fractionated by molecular exclusion chromatography. Activity against promastigotes was detected by respiratory oxidative conversion of MTT in a colorimetric assay and L-AO activity was detected by a colorimetric assay with peroxidase and OPD as revealing reagents. Crude venom was irradiated with 500, 1000, and 2000 Gy in a 60 Co gamma radiation source. The venom had an anti-Leishmania activity of 33 pg/promastigote and the active fraction migrates as 100-150 kDa, close to the size described for L-AOs, and also presented L-AO activity. The radiation reduces both the L-AO and anti-Leishmania activity in a dose dependent effect. Those data suggests the anti-Leishmania activity in this venom is closely related to the L-amino acid oxidase activity and also that radiation could be used as a tool to detect specific activities reduction in water solutions, similarly to observed in dry preparations. (author)

  4. Characterization of Leishmania infantum species in dogs from the urban area of Cuiabá, State of Mato Grosso, Brazil / Caracterização da espécie Leishmania infantum em cães de área urbana de Cuiabá, Estado do Mato Grosso

    Scientific Electronic Library Online (English)

    Bianca De, Santis; Elizabeth Gloria Oliveira Barbosa, Santos; Elisa, Cupolillo; Renato, Porrozzi; Amanda dos Santos, Cavalcanti; Bárbara Neves dos, Santos; Saulo Teixeira De, Moura; Kellen, Malhado; Sergio Augusto Miranda, Chaves.

    2011-12-01

    Full Text Available INTRODUÇÃO: A leishmaniose visceral apresenta comportamento urbano em algumas cidades brasileiras, sendo os cães domésticos as principais fontes de infecção. Em Cuiabá-MT, a leishmaniose visceral canina foi diagnosticada e caracterizada, como recomendação do Ministério da Saúde. MÉTODOS: Amostras bi [...] ológicas de cães suspeitos foram analisadas por eletroforese de isoenzimas. Foram utilizadas a enzima 6PGDH e a cepa de referência IOC/L0566 (MHOM/BR/1975/M2903) de Leishmania (Leishmania) infantum, como um dos controles. RESULTADOS: A análise eletroforética revelou que os isolados pertenciam à espécie L. (L.) infantum. CONCLUSÕES: Os autores ressaltam a importância da caracterização da espécie, principalmente em cidades com infecção mista, como Cuiabá. Abstract in english INTRODUCTION: Visceral leishmaniasis presents urban behavior in some Brazilian cities, with domestic dogs as the main infection source. In Cuiabá, MT, canine visceral leishmaniasis was diagnosed and characterized as recommended by the Ministry of Health. METHODS: Biological samples from suspected ca [...] nine carriers were analyzed by the isoenzyme electrophoresis technique. The 6PGDH enzyme and reference strain IOC/L0566 (MHOM/BR/1975/M2903) of Leishmania (Leishmania) infantum was used as one of the controls. RESULTS: Electrophoresis analysis revealed that the canine isolates belonged to the species L. (L.) infantum. CONCLUSIONS: The authors emphasize the importance of species characterization, particularly in areas of mixed infection like Cuiabá.

  5. Padronização de condições para detecção de DNA de Leishmania spp. em flebotomíneos (Diptera, Psychodidae pela reação em cadeia da polimerase Standardization of conditions for PCR detection of Leishmania spp. DNA in sand flies (Diptera, Psychodidae

    Directory of Open Access Journals (Sweden)

    Byanca Regina de Paiva

    2007-01-01

    Full Text Available A correta identificação dos agentes etiológicos em insetos vetores é de crucial importância aos estudos epidemiológicos. A pesquisa de flagelado nesses vetores, pela dissecção de seu trato digestivo, observação microscópica do seu conteúdo ou por isolamento dos parasitas provenientes de insetos em meios de cultura, tem-se mostrado operacionalmente inadequada e com baixa especificidade do diagnóstico, pois fêmeas de flebotomíneos também podem albergar outros flagelados como Trypanosoma e Endotrypanum. Acreditamos que por sua eficiência e especificidade, a amplificação de seqüências-alvo do DNA da Leishmania, por meio da reação em cadeia de polimerase, pode ser aplicada na investigação de sua presença em flebotomíneos, desde que estes estejam devidamente acondicionados e o DNA do parasita extraído a partir de metodologia adequada. Este trabalho descreve metodologias utilizadas na padronização da conservação dos espécimes de flebotomíneos e extração do DNA da Leishmania como uma alternativa mais prática que os métodos tradicionais.The correct identification of etiological agents in vector insects is crucial for epidemiological studies. Identification of flagellates in such vectors, usually by dissection of the digestive tract and microscopic observation of the contents as well as attempts at parasite isolation from insects in culture media, have proven operationally inadequate and with poor diagnostic specificity, since female sand flies are also hosts for other flagellates like Trypanosoma and Endotrypanum. Due to the efficiency and specificity of DNA target sequence amplification by polymerase chain reaction (PCR, the latter could be used to investigate the presence of Leishmania in sand flies, although the insects need to be properly stored and the Leishmania DNA extracted using appropriate methodology. This paper describes methodologies to standardize sand fly storage and Leishmania DNA extraction in such specimens as a more practical method in field studies.

  6. Histological evaluation of the lesion induced by inoculation of Leishmania mexicana in the cheek pouch of the hamster / Avaliação histológica da lesão induzida pela inoculação de Leishmania mexicana na bolsa jugal do hamster

    Scientific Electronic Library Online (English)

    Maria Sueli Parreira de, Arruda; Maria Esther Salles, Nogueira; Ana Paula, Bordon.

    2002-08-01

    Full Text Available No presente estudo, investigamos o papel da resposta imune na morfologia do granuloma leishmaniótico induzido na bolsa jugal do hamster, um local imunologicamente privilegiado, após inoculação de 3x10(5) Leishmania mexicana. Os animais foram avaliados histológica e imunologicamente até os 120 dias d [...] a inoculação. Independente da época do sacrifício, os animais foram sempre não reatores ao teste do coxim plantar. Histologicamente, a inoculação de Leishmania mexicana na bolsa jugal resultou na formação de abcesso que evoluiu para reação granulomatosa rica em formas amastigotas e, posteriormente, para resolução. Esses resultados sugerem que o desenvolvimento da resposta imune não é preponderante no controle da infecção induzida pela Leishmania mexicana inoculada subcutaneamente na bolsa jugal do hamster. Sugerem ainda que os macrófagos que compõe os granulomas leishmanióticos são capazes de eliminar esse parasita, independente da presença de resposta imune avaliável pelo teste do coxim plantar. Abstract in english We have studied the role of the immune response in the morphology of the leishmaniotic granuloma induced in the cheek pouch of hamsters, an immunologically privileged site, after inoculation of 3 x 10(5) Leishmania mexicana. Animals were histologically and immunologically evaluated until 120 days af [...] ter inoculation. Independent of the time of sacrifice, the animals were always non-reactors to the footpad test (FPT). At histology, the introduction of L. mexicana in the cheek pouch leads to an abscess that evolves to a granulomatous reaction rich in amastigote forms, and later it leads to resolution, even in the absence of immune response detectable by FPT. Our results demonstrate that the development of immune response is not preponderant for the control of infection induced by L. mexicana inoculated subcutaneously in the cheek pouch of the hamster. It also suggests that the macrophages present in the leishmaniotic granuloma are capable of eliminating this parasite, even in the absence of immune response evaluated by FPT.

  7. Assessment of PCR in the detection of Leishmania spp in experimentally infected individual phlebotomine sandflies (Diptera: Psychodidae: Phlebotominae

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    MICHALSKY Érika M.

    2002-01-01

    Full Text Available DNA amplification by the polymerase chain reaction (PCR was applied in the investigation of the presence of Leishmania (Kinetoplastida: Trypanosomatidae parasites in single phlebotomine sandflies. Three phlebotomine/parasite pairs were used: Lutzomyia longipalpis/Leishmania chagasi, Lutzomyia migonei/Leishmania amazonensis and Lutzomyia migonei/Leishmania braziliensis, all of them incriminated in the transmission of visceral or cutaneous leishmaniasis. DNA extraction was performed with whole insects, with no need of previous digestive tract dissection or pooling specimens. The presence of either mouse blood in the digestive tract of the sandflies or the digestive tract itself did not interfere in the PCR. Infection by as few as 10 Leishmania sp. per individual were sufficient for DNA amplification with genus-specific primers. Using primers for L. braziliensis and L. mexicana complexes, respectively, it was possible to discriminate between L. braziliensis and L. amazonensis in experimentally infected vectors (L. migonei.

  8. An agent-based model for Leishmania major infection

    Science.gov (United States)

    Dancik, Garrett M.; Jones, Douglas E.; Dorman, Karin S.

    Leishmania are protozoan parasites transmitted by bites of infected sandflies. Over 20 species of Leishmania, endemic in 88 countries, are capable of causing human disease. Disease is either cutaneous, where skin ulcers occur on exposed surfaces of the body, or visceral, with near certain mortality if untreated. C3HeB/FeJ mice are resistant to L. major, but develop chronic cutaneous lesions when infected with another species L. amazonensis. The well-characterized mechanism of resistance to L. major depends on a CD4+ Thl immune response, macrophage activation, and elimination of the parasite [Sacks 2002]. The factors that account for host susceptibility to L. Amazonensis, however, are not completely understood, despite being generally attributed to a weakened Th1 response [Vanloubbeck 2004].

  9. Avaliação da atividade anti-Trypanosoma e anti-Leishmania de Mentha arvensis e Turnera ulmifolia

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    Karla K.A. SANTOS

    2012-01-01

    Full Text Available Tripanosomiasis or "Chagas disease", caused by Trypanosoma cruzi, affect 10 million people in Latin America. Today, the chemotherapy is the only specific treatment against this disease, being the most used drugs the nifurtimox and benznidazole. Leishmaniasis is a disease caused by parasites of the genus Leishmania, mainly founded in regions with forests, as the Amazonia. Recent reports about the Leishmaniasis indicate a deficit of therapeutical drugs available against this disease and reinforce the necessity of the discovering of new drugs. An interesting approach against these diseases is the use of natural products, as the extracts of plants as Mentha arvensis and Turnera ulmifolia. For the in vitro assays against T. cruzi and Leishmania, was used the clone CL-B5 and promastigote forms, respectively. The cytotoxic assay was performed using fibroblasts. Our results indicated that M. arvensis was active against all strains assayed, inhibiting 65 e 47% of the assayed strains (IC50 = 192.3 and 531.9 ¿g/mL respectively, representing an interesting and alternative source of natural products with anti-kinetoplastida activity.

  10. Analysis of the specificity of human antibodies to antigens of Leishmania braziliensis braziliensis Análise da especificidade de anticorpos humanos a antígenos de Leishmania braziliensis braziliensis

    OpenAIRE

    Aoi Masuda; Sueli Fátima do Nascimento; Carmem Silvia Guerra; Gláucia da Silva Paranhos; Antonio Walter Ferreira

    1989-01-01

    The antigenicity of promastigotes of Leishmania braziliensis braziliensis (L. b.braziliensis) treated with 1% sodium desoxycholate in 10 mM Tris-Hcl pH 8.2 was analysed by immunoblot using as probes sera from American cutaneous leishmaniasis (ACL), American visceral leishmaniasis (AVL), schistosomiasis, malaria and Chagas' disease. The ACL sera reacted constantly with a 60 kD band. No reactivity to this protein was observed with sera from the other diseases above mentioned indicating that the...

  11. BODY WEIGHT AS A DETERMINANT OF CLINICAL EVOLUTION IN HAMSTERS (Mesocricetus auratus) INFECTED WITH Leishmania (Viannia) panamensis / Peso como determinante da evolução clínica em hamsters (Mesocricetus auratus) infectados com Leishmania (Viannia) panamensis

    Scientific Electronic Library Online (English)

    Angela Maria, Gomez-Galindo; Lucy Gabriela, Delgado-Murcia.

    2013-09-01

    Full Text Available RESUMO O resultado clínico da infecção por espécies de Leishmania do subgênero Viannia no modelo de hamster (Mesocricetus auratus) tem se mostrado diferente, dependendo do protocolo experimental. O peso corporal tem sido um importante determinante da evolução clínica da infecção em hamsters com lei [...] shmaniose visceral, mas sua importância como parâmetro clínico em hamsters com leishmaniose cutânea não é conhecido. Neste estudo, a evolução clínica da infecção com L. (V) panamensis foi avaliada em jovens e adultos hamsters machos durante 11 semanas, comparando os parâmetros clínicos tais como a atitude, a temperatura, a frequência respiratória, a aparência das fezes, e o peso corporal entre infectado e grupos não infectados. Os resultados mostraram que o peso corporal diminuiu em hamsters adultos após infecção por L. (V) panamensis. Esta observação suporta a utilização do peso corporal, como um parâmetro adicional para definir a administração ou o tratamento de leishmaniose cutânea em hamsters adultos infectados usados como modelo animal experimental para a leishmaniose. Abstract in english SUMMARY The clinical outcome of infection with Leishmania species of the subgenus Viannia in hamster model (Mesocricetus auratus) has shown to be different depending on experimental protocol. Body weight has been a relevant determinant of the clinical outcome of the infection in hamsters with visce [...] ral leishmaniasis but its importance as a clinical parameter in hamsters with cutaneous leishmaniasis is not known. In this study, the clinical evolution of infection with L. (V) panamensis was evaluated in juvenile and adult male hamsters during 11 weeks by comparing clinical parameters such as attitude, temperature, respiratory rate, appearance of the stool, and body weight between infected and non-infected groups. Results showed that body weight decreased in adult hamsters after infection by L. (V) panamensis; this observation supports the use of body weight as an additional parameter to define the management or treatment of cutaneous leishmaniasis in infected adult hamsters used as an animal experimental model for leishmaniasis.

  12. Potential utility of hyperbaric oxygen therapy and propolis in enhancing the leishmanicidal activity of glucantime / A utilidade da terapia de oxigenação hiperbárica e própolis em potencializar a atividade leishmanicida do glucantime

    Scientific Electronic Library Online (English)

    Diana Copi, Ayres; Thiago Antonio, Fedele; Maria Cristina, Marcucci; Selma, Giorgio.

    2011-12-01

    Full Text Available Nesse trabalho foi avaliada a eficácia da terapia da oxigenação hiperbárica (HBO), aplicada em combinação ou não com o tratamento com glucantime, durante a infecção com Leishmania amazonensis. O efeito de gel da própolis vermelha de origem brasileira (propaina) aplicado em combinação ou não com o tr [...] atamento com glucantime, também foi avaliado durante infecção com esse parasita. A inibição da infecção de macrófagos tratados com glucantime em combinação com HBO foi maior que a de macrófagos tratados apenas com glucantime ou HBO. A linhagem murina susceptível, BALB/c, infectada no dorso com L. amazonensis, tratada com glucantime e exposta a HBO, mostrou durante o curso da doença, fases em que as lesões eram menores do que a de camundongos apenas tratados com glucantime; observou-se revascularização da pele da lesão e baixa produção de interferon-gama em células de linfonodos desses animais. O tratamento com propaina não foi eficiente na cura das lesões, apesar de lesões menos exsudativas serem observadas em animais tratados com propaina ou propaina combinada ao tratamento com glucantime. Os resultados demonstram que tanto HBO como a própolis vermelha em combinação com glucantime, são promissoras no tratamento da leishmaniose cutânea. Novos estudos devem ser realizados para avaliar tratamentos e outros protocolos em diferentes modelos murinos da leishmaniose Abstract in english In this study we investigated the efficacy of hyperbaric oxygen (HBO) therapy, alone or combined with the pentavalent antimonial glucantime on Leishmania amazonensis infection. In parallel, the effect of Brazilian red propolis gel (propain) alone or combined with glucantime on L. amazonensis infecti [...] on was evaluated. The inhibition of the infection in macrophages treated with glucantime in combination with HBO exposition was greater than that of macrophages treated with glucantime alone or HBO alone. The susceptible mouse strain BALB/c infected in the shaved rump with L. amazonensis treated with glucantime and exposed to HBO showed: time points in the course of the disease in which lesions were smaller than those of mice treated with glucantime alone and revascularization of the skin in the lesion site; interferon-gamma (IFN-g) levels were not elevated in lymph node cells from these animals. Propain alone was not efficient against lesions, although less exudative lesions were observed in animals treated with propain alone or combined with glucantime. These results reveal the potential value of HBO and red propolis in combination with glucantime for treating cutaneous leishmaniasis and encourage further studies on the effect of more aggressive HBO, propolis and glucantime therapies on different mouse models of leishmaniasis.

  13. Occurrence of anti-Leishmania spp., Neospora caninum, and Toxoplasma gondii antibodies in dog sera from Veterinary Hospital from Universidade Estadual de LondrinaOcorrência de anticorpos contra Leishmania spp., Neospora caninum E Toxoplasma gondii em soros de cães atendidos no Hospital Veterinário da Universidade Estadual de Londrina-Pr

    Directory of Open Access Journals (Sweden)

    Dauton Luiz Zulpo

    2012-10-01

    Full Text Available The aim of this study was to detect the presence of IgG antibodies anti-Leishmania spp., Toxoplasma gondii and Neospora caninum in dogs from a Veterinary Hospital from Universidade Estadual de Londrina. Blood samples from 112 animals were obtained by jugular venipuncture to obtain sera. The samples were tested by indirect immunofluorescence to detect antibodies anti-Leishmania spp., anti-N. caninum and anti-T. gondii. Thirteen (11.61%, 25 (22.32%, and 57 (50.89% samples were positive for Leishmania spp., N. caninum, and T. gondii, respectively. The co-presence of anti-Leishmania spp. and N. caninum was observed in 6 (5.36%, anti-Leishmania spp. and anti-T. gondii in 8 (14.7%, and anti-N. caninum and anti-T. gondii in 18 (16.07% samples. The co-presence of anti-Leishmania spp., anti-N. caninum and anti-T. gondii was observed in 5 (4.46% dogs. There was a higher prevalence of Leishmania in Toxoplasma and Neospora positive animals, however, these results were not statistically significant (range p = 0.052 p = 0.06. The dogs have an important role in the epidemiological cycle of these diseases, which are important in animal and public health. The northern state of Paraná is an endemic area for human cutaneous leishmaniasis, therefore, studies should be conducted to uncover the real role of dogs as reservoirs of Leishmania to humans in the state. O objetivo do presente trabalho foi detectar a presença de anticorpos contra Leishmania spp., Neospora caninum e Toxoplasma gondii em cães atendidos no Hospital Veterinário, da Universidade Estadual de Londrina. Amostras de 112 animais foram obtidas por venopunção jugular ou cefálica com posterior obtenção dos soros. Estas foram submetidas à técnica de imunofluorescência indireta para detecção de anticorpos da classe IgG anti-Leishmania spp, anti-N. caninum e anti-T. gondii. Dos 112 soros examinados, 13 (11,61%, 25 (22,32% e 57 (50,89% foram positivos para Leishmania spp., N. caninum e T. gondii, respectivamente. A co-presença de anticorpos anti-Leishmania spp. e N. caninum foi observada em 6 (5,36% amostras, anticorpos anti-Leishmania spp. e anti-T. gondii em 8 (7,14%, e anticorpos anti-N. caninum e anti-T. gondii em 18 (16,07% amostras. A co-presença de anticorpos anti- Leishmania spp., anti-N. caninum e anti-T. gondii foi observada em 5 (4,46% cães. Verificou-se uma maior prevalência de Leishmania nos animais positivos para Toxoplasma e Neospora, embora estes resultados não tenham sido estatisticamente significativos (p?0.06. Os cães são importantes no ciclo epidemiológico das enfermidades em estudo, sendo estas doenças importantes do ponto de vista de saúde animal, ou de saúde pública. A região norte do estado do Paraná é uma área endêmica para leishmaniose tegumentar humana, portanto, estudos devem ser realizados para desvendar o real papel dos cães como reservatórios da Leishmania para seres humanos no estado.

  14. Histopatologia da leishmaniose tegumentar por Leishmania braziliensis braziliensis: 1. Padrões histopatológicos e estudo evolutivo das lesões / Histopathology of mucocutaneous leishmaniasis caused by Leishmania braziliensis braziliensis: 1. Histological patterns and evolutive study of the lesions

    Scientific Electronic Library Online (English)

    Albino Verbosa de, Magalhães; Mario A. P., Moraes; Alberto N., Raick; Alejandro, Llanos-Cuentas; Jackson M. L., Costa; Cesar C., Cuba; Philip D., Marsden.

    1986-08-01

    Full Text Available Os autores analisam material de biópsias de 378 casos de Leishmaniose Tegumentar, causada por Leishmania braziliensis braziliensis, da localidade endêmica de Três Braços (Estado da Bahia). O parásitos, embora escassos, foram encontrados em 63,7% dos casos da forma cutânea e em 37,5% dos casos da for [...] ma mucosa. As alterações dérmicas ou do córion da mucosa permitiram identificar cinco padrões histopatológicos: 1) Reação Exsudativa Celular, constituída por um infiltrado histiolinfoplasmocitário; 2) Reação Exsudativa e Necrótica, na qual ocorre uma necrose no seio do infiltrado inflamatório; 3) Reação Exsudativa e Necrótico-Granulomatosa, que corresponde ao quadro descrito como inflamação crônica granulomatosa com necrose; 4) Reação Exsudativa e Granulomatosa, onde se observa uma reação granulomatosa desorganizada, sem que esteja presente necrose tecidual; 5) Reação Exsudativa e Tuberculóide, caracterizada pelo granuloma tuberculóide. O estudo evolutivo realizado em 49 casos, mostrou que houve uma mudança de padrão histopatológico observada, em biópsias sucessivas, em 63,2% dos casos da forma cutânea e em 45,4% dos casos da forma mucosa. Através desse estudo, é possível afirmar-se que o padrão de Reação Exsudativa Celular constitui o quadro inicial e final da lesão, com os demais padrões aparecendo interposto durante a evolução da doença. Abstract in english The Authors describe the histopathological findings observed in biopsies from 378 cases of Mucocutaneous Leishmaniasis, caused by Leishmania braziliensis braziliensis. All the patients lived in the locality of Três Braços, State of Bahia, an endemic area of Leishmaniasis in Brazil. Parasites were fo [...] und in 63.7% of patients with cutaneous form and in 35.7% of cases with mucous lesions, although always in reduced number. Histopathologic findings allowed the caracterization of five different patterns: 1. Exsudative-cellular reaction, due to infiltraction of histiocytes, lymphocytes and plasm cells; 2. Exsudative-necrotic reaction, characterized by the association of necrosis with cellular infiltration; 3. Exsudative and necrotic-granulomatous reaction, corresponding to pattern described as chronic-granulomatous inflammation with necrosis; 4. Exsudativegranulomatous reaction, characterized a disorganized granulomatous reaction, without tissue necrosis; 5. Exsudative-tuberculoid reaction — in which a typical tuberculoid granuloma is formed. The follow-up study in 49 patients showed, in sucessive biopsies, changes in the histopathologic patterns in 63.2% of cutaneous forms and in 45.4% of the mucous forms. It was concluded that the exsudative-cellular reaction represents both the inicial and final pattern of Mucocutaneous Leishmaniasis lesion. The other four histopathologic patterns characterize intermediate forms found during the evolution of the disease.

  15. Targeting host syntaxin-5 preferentially blocks Leishmania parasitophorous vacuole development in infected cells and limits experimental Leishmania infections.

    Science.gov (United States)

    Canton, Johnathan; Kima, Peter E

    2012-10-01

    Our previous observations established a role for syntaxin-5 in the development of Leishmania parasitophorous vacuoles (LPVs). In this study, we took advantage of the recent identification of Retro-2, a small organic molecule that can cause the redistribution of syntaxin-5; we show herein that Retro-2 blocks LPV development within 2 hours of adding it to cells infected with Leishmania amazonensis. In infected cells incubated for 48 hours with Retro-2, LPV development was significantly limited; furthermore, infected cells harbored four to five times fewer parasites than infected cells incubated in vehicle alone. In vivo studies revealed that Retro-2 curbed experimental L. amazonensis infections in a dose-dependent manner. Retro-2 did not have any appreciable effect on the host cell physiological characteristics; furthermore, it had no apparent toxicity in experimental animals. An unexpected, but welcome, finding was that Retro-2 inhibited the replication of Leishmania parasites in axenic cultures. This study is significant because it identifies an endoplasmic reticulum/Golgi SNARE as a potential target for the control of Leishmania infections; moreover, it suggests that small organic molecules can be identified that can selectively disrupt the vesicle fusion machinery that promotes the development of pathogen-containing compartments without exerting toxic effects on the host. PMID:22885104

  16. Estado atual da leishmaniose cutânea difusa (LCD) no Estado do Maranhão: II. aspectos epidemiológicos, clínico-evolutivos

    Scientific Electronic Library Online (English)

    Jackson Maurício Lopes, Costa; Ana Cristina Rodrigues, Saldanha; Ana Carla de Melo e, Silva; Artur, Serra Neto; Clovis Eduardo Santos, Galvão; Conceição de Maria Pedroso e, Silva; Antonio Rafael da, Silva.

    1992-06-01

    Full Text Available Os Autores fazem um estudo retrospectivo e prospectivo de 6 pacientes portadores de leishmaniose cutânea difusa, observados no Estado do Maranhão a partir de 1974. Os casos abordados são oriundos de diversas regiões do estado, observando-se em todos eles o envolvimento da leishmania (Leishmania) ama [...] zonensis, sendo que 5 (84%) dos pacientes apresentaram início de doença na 1ª década de vida. Em todos os pacientes envolvidos no estudo, houve relato de lesão inicial nodular única, que, posteriormente, em período variável de tempo, disseminou-se adquirindo outros aspectos. Evolutivamente apresentaram múltiplas lesões nodulares e ulceradas, intradermorreação de Montenegro(-) e refratariedade aos esquemas terapêuticos utilizados até ao presente momento. Abstract in english The authors describe a retrospective and prospective study of 6 patients with diffuse cutaneous leishmaniasis observed in the State of Maranhão, since 1974. The patients comefromdifferentruralregions of the state and in all of them Leishmania (Leishmania) amazonensis was the cause five of the patien [...] ts initiated their disease in the first decade of life. All the patients first had a solitary, nodular lesion, that after a variable period of time, disseminated and acquired other aspects. Sequentially the patients presented multiple nodular and ulcerative lesions, negative leishmania skin-lests and a refractory response to the therapeutic schedules used up to the present.

  17. Leishmania infection in a population of dogs: an epidemiological investigation relating to visceral leishmaniasis control / Infecção por Leishmania em uma população de cães: uma investigação epidemiológica relacionada ao controle da leishmaniose visceral

    Scientific Electronic Library Online (English)

    Klauber Menezes, Penaforte; Vinicius Silva, Belo; Rafael Goncalves, Teixeira-Neto; Renata Aparecida Nascimento, Ribeiro; Robson Bruniera de, Oliveira; Dante Alighieri, Schettini; Eduardo Sergio da, Silva.

    2013-12-01

    Full Text Available A identificação dos fatores associados à infecção por Leishmania em cães é essencial para o direcionamento das ações de controle da leishmaniose visceral. Desta forma, o presente estudo analisa alguns destes fatores em uma população de cães, bem como as limitações das estratégias de controle executa [...] das em um município brasileiro. Por meio de modelos de regressão logística, analisou-se a associação entre as variáveis de exposição e a ocorrência de infecção. As ações de controle da doença foram analisadas qualitativamente. Dos 755 animais avaliados por meio de sorologia, 13,6% (103/755) foram soro-reagentes. Destes, 23,3% (24/103) eram assintomáticos e 76,7% (79/103) apresentaram pelo menos um sinal clínico possivelmente associado com a leishmaniose visceral. Com associações fracas, cães de raça pura, de pelo curto, com mais de cinco anos, machos e de grande porte foram mais propensos à infecção. As duas últimas variáveis compuseram o modelo final de regressão, sendo significativa a associação com o tamanho do cão. As estratégias de controle desenvolvidas apresentaram limitações e um número elevado de cães sorologicamente negativos foi submetido a eutanásia. Os dados apresentados contribuem para um melhor entendimento da dinâmica da infecção na leishmaniose visceral canina e indicam que ações visando a adequada execução do programa de controle da doença em áreas endêmicas brasileiras devem ser priorizadas. Abstract in english Identification of factors associated with Leishmania infection in dogs is essential for targeting visceral leishmaniasis control actions. Thus, the present study analyzed some of these factors in a population of dogs in a Brazilian municipality, along with the limitations of control strategies imple [...] mented there. The association between the exposure variables and occurrences of infection was analyzed through logistic regression models. The disease control interventions were treated qualitatively. Out of the 755 animals examined, 13.6% (103/755) were seropositive. Of these, 23.3% (24/103) were asymptomatic and 76.7% (79/103) presented at least one clinical sign possibly associated with visceral leishmaniasis. With weak associations, purebred, shorthaired, over 5 years of age, male and large dogs were more prone to infection. The latter two variables formed the final regression model and the association with dog size was statistically significant. The control strategies developed presented limitations and a great number of seronegative dogs was culled. The data presented contribute towards better understanding of the dynamics of infection in canine visceral leishmaniasis and indicate that actions aimed towards adequate implementation of Visceral Leishmaniasis control program in Brazilian endemic areas should be prioritized.

  18. Behavior of Leishmania major metacyclic promastigotes during the course of infection and immune response development in resistant versus susceptible hosts Comportamento de promastigoteas metacíclicos de Leishmania major durante o curso da infecção e da resposta imune em hospedeiros resistentes versus suscetíveis

    Directory of Open Access Journals (Sweden)

    Regina Coeli Cunha Dórea

    2003-11-01

    Full Text Available Little is known on the epitopes derived from metacyclic promastigotes of Leishmania that are important on the regulation or destruction of the parasite, as targets of immune attack in the vertebrate host. In this study we investigated an alternative method to obtain metacyclic promasigotes of Leishmania major, as evaluated by the course of infection and delayed-type hipersensitivity (DTH in resistant versus susceptible inbred mice. Non-infective (procyclic promastigotes of L. major recently transformed from tissue amastigotes were attached to a negatively charged glass-wool column, whereas metacyclic promastigotes were not bound to columns and could be easily recovered. Optimal chromatography conditions were validated through statistical analyses. Parasite average yield from glass wool columns and promastigote viability were estimated by light microscopy. Metacyclic promastigotes yielded 43.5% to 57.5%. Different patterns of cutaneous lesions were obtained in BALB/c (susceptible and C57BL/6 (resistant mice, the former with highly infective lesions induced by metacyclic promastigotes. DTH responses proved to be higher in groups of C57BL/6 mice which were infected with metacyclic promastigotes. These results indicate that the new method could be integrated with the investigation of metacyclogenesis of Leishmania in vivo.Pouco se conhece sobre os epítopos derivados de promastigotas metacíclicos de Leishmania que são importantes para a regulação ou destruição do parasita, como alvos de ação imunológica no hospedeiro vertebrado. Neste estudo, nós investigamos um método alternativo para obter promastigotas metacíclicos de Leishmania major, pela avaliação do curso da infecção e reação de hipersensibilidade do tipo retardado (HTR em hospedeiros resistentes e susceptíveis. Promastigotas não-infectantes (procíclicos de L. major, recentemente isolados de amastigotas, foram selecionados pela adesão a colunas de lã de vidro negativamente carregadas, enquanto que promastigotas metacíclicos não se aderem à coluna e podem ser recuperados com facilidade. Condições ótimas de cromatografia foram validadas por análise estatística. O rendimento médio de parasitas obtidos após separação em colunas de lã de vidro e a viabilidade dos promastigotas foram estimados por microscopia óptica. Os promastigotas metacíclicos tiveram um rendimento médio de 43,5% a 57,5%. Camundongos BALB/c (susceptíveis e camundongos C57BL/6 (resistentes apresentaram padrões distintos de lesões cutâneas, os primeiros com lesões mais agressivas, induzidas por promastigotas metacíclicos. As respostas à reação de HTR foram maiores nos grupos de camundongos C57BL/6, submetidos à infecção com promastigotas metacíclicos. Estes resultados indicam que o novo método poderia ser integrado aos protocolos existentes para estudar a metaciclogênese de parasitas do gênero Leishmania in vivo.

  19. Comparative analysis of the geographic distribution of the histopathological spectrum and Leishmania species of American cutaneous leishmaniasis in Brazil Análise comparativa da distribuição geográfica do espectro histopatológico e espécies de Leishmania da leishmaniose tegumentar americana no Brasil

    OpenAIRE

    Linton Wallis Figueiredo Souza; Simone Vilas Trancoso Souza; Ana Cristina de Carvalho Botelho

    2012-01-01

    BACKGROUND: Knowledge of the relationship between specific geographical differences and histopathology of the American cutaneous leishmaniasis is limited because host-parasite interactions in space and time are complex. OBJECTIVE: To describe the geographic analysis of the histopathological pattern of localized American cutaneous leishmaniasis and the relationship with Leishmania species. METHODS: A cross-sectional, descriptive and analytical study of a sample consisting of 55 patients of mun...

  20. Action of Bothrops moojeni venom and its L-amino acid oxidase fraction, treated with 60Co gamma rays, in Leishmania spp

    International Nuclear Information System (INIS)

    Bothrops moojeni venom showed an anti leishmania activity in vitro, as determined by a cell viability assay using the reduction of MTT. After venom purification, by chromatography techniques, the fractions with anti leishmania and L-amino acid oxidase activities, eluted in the same positions. The molecular weight of the enzyme was estimated to be 140 kDa by molecular exclusion chromatography, and 69 kDa, by SDS-PAGE, migrating as a single band, with an isoelectric point of 4.8 as determined by isoelectric focusing. The purified LAO from B. moojeni venom, 135-fold more active than crude venom, showed homo dimeric constitution, and was active against Leishmania spp from the New World, with an effective concentration against L(L). amazonensis of 1.80 μg/ml (EC50), L.(V.) panamensis (0.78 |μg/ml) and L.(L.) chagasi (0.63 (μg/ml). Ultrastructural studies of promastigotes affected by LAO demonstrated cell death, with edema in several organelles such as mitochondria and nuclear membrane, before cell disruption and necrosis. The action of LAO was demonstrated to be hydrogen peroxide-dependent. Studies with LLCMK-2 cells, treated with LAO, showed a toxic effect, with an EC50 of 11|μg/ml. Irradiation of LAO with 60Co gamma rays, did not affect its whole oxidative activity, neither detoxified the enzyme. Amastigotes treated with LAO were not affected by its hydrogen peroxide, otherwise, the exogenous product, killed amastigotes with an EC50 of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  1. ITS1 PCR-RFLP Diagnosis and Characterization of Leishmania in Clinical Samples and Strains from Cases of Human Cutaneous Leishmaniasis in States of the Mexican Southeast

    Science.gov (United States)

    Monroy-Ostria, Amalia; Nasereddin, Abedelmajeed; Monteon, Victor M.; Guzmán-Bracho, Carmen; Jaffe, Charles L.

    2014-01-01

    American cutaneous leishmaniasis includes a spectrum of clinical forms localized cutaneous, diffuse cutaneous, and mucocutaneous leishmaniasis which can be caused by different strains of Leishmania belonging to the L. mexicana or L. braziliensis complexes which may coexist in the same endemic area. We evaluated the PCR-RFLP assay of the ITS1 genes for direct identification of Leishmania species in 163 clinical samples and 21 Mexican isolates of Leishmania. In relation to the Mexican isolates of Leishmania 52% displayed a pattern similar to the L. (L.) mexicana, 5% showed a mixed pattern compatible with L. (L.) mexicana and L. (V.) braziliensis, eight with L. (L.) amazonensis and L. (L.) mexicana, and one to L. (V.) braziliensis. Most of the clinical samples, 109/116 (94%), gave a pattern similar to that of the L. mexicana, two clinical samples gave similar patterns to that of Leishmania braziliensis, and 5 samples gave patterns that suggest a coinfection of L. (L.) mexicana and L. (V.) braziliensis or L. (L.) mexicana and L. (L.) amazonensis. The ITS1 PCR-RFLP assay is a multipurpose tool for diagnosis of Leishmania from clinical samples and enables determination of the infecting species of New World Leishmania in the field in relatively short time and low cost. PMID:25104958

  2. Efficacy of the photodynamic antimicrobial therapy (PACT) with the use of methylene blue associated with the λ660nm laser in Leishmania (Leishmania) amazonesis: in vitro study

    Science.gov (United States)

    Pires-Santos, Gustavo M.; Marques, Aparecida M. C.; Alves, Eliomara S. S.; Oliveira, Susana C. P. S.; Monteiro, Juliana S. C.; Rosa, Cristiane B.; Colombo, Fabio; Pinheiro, Antônio L. B.; Vannier-Santos, Marcos A.

    2012-03-01

    The present studied evaluated the in vitro effects of PDT on Leishmania (Leishmania) amazonensis promastigotes. For this examination L. amazonensis promastigotes, stain Josefa, were used and maintained in Warren media supplement with fetal bovine serum at 26°C for 96 hours. A viability curve was accomplished using different concentrations of methylene blue photosensitizer associated to red laser light in order to obtain the most effective interaction to inhibit the parasite's growth. Two pre-irradiation periods, 5 and 30 minutes, were evaluated and the promastigotes were counted by colorimetry. On fluorescence microscopy the autophagic processes and reactive oxygen species were detected. Promastigotes treated with Photodynamic Therapy (PDT) by concentrations of 5 and 0,315ug/mL, presented cellular proliferation inhibition when compared to the control. In the first condition, the cells had structural alterations such as truncated cells, cells with two flagella, bleb formation and cells body deformation, while none of these modifications could be visualized in the control group. When analyzed through fluorescence microscopy, the promastigotes treated were positives for free radicals immediately after light application and also 1 hour after treatment presenting signs of autophagia. PDT on L. (L.) amazonensis is effective causing alterations that can help elucidate the mechanisms of the parasite's death when treated with methilene

  3. Combining diagnostic procedures for the management of leishmaniasis in areas with high prevalence of Leishmania guyanensis Procedimentos diagnósticos combinados no manejo da leishmaniose em áreas com alta prevalência de Leishmania guyanensis

    OpenAIRE

    Ednelza de Almeida Benicio; Ellen Pricilla Nunes Gadelha; Anette Talhari; Roberto Moreira da Silva Jr; Luis Carlos Ferreira; Mayara Cristina Cordeiro dos Santos; Marcelo Távora Mira; Cintia Mara Costa de Oliveira; Carolina Talhari; Sinésio Talhari; Paulo Roberto Machado; Albert Schriefer

    2011-01-01

    BACKGROUND: The Amazon region corresponds to approximately 40% of the cases of leishmaniasis in Brazil. We report a prospective study with 180 patients conducted in a health care unit that diagnoses 10% of the cases of leishmaniasis in the Brazilian Amazon. The study addresses how a combination of procedures improves diagnosis in areas with high prevalence of Leishmania guyanensis. OBJECTIVES: to evaluate diagnostic methods in areas with high prevalence of Leishmania guyanensis. METHODS: All ...

  4. Monoclonal antibody affinity purification of a 78 kDa membrane protein of Leishmania donovani of Indian origin and its role in host–parasite interaction

    Indian Academy of Sciences (India)

    Mandira Mukherjee; Anindita Bhattacharyya; Swadesh Duttagupta

    2002-12-01

    Monoclonal antibodies were raised against pathogenic promastigotes of Leishmania donovani of Indian origin. Among these, one was used for immuno-affinity purification of a 78 kDa membrane protein present in both the amastigote and promastigote forms of the parasite. Results of immunoblot experiments with the anti-78 kDa antibody revealed that the protein was present only in parasites belonging to the L. donovani complex. The expression of the protein was observed to be the same during different phases of growth of the promastigotes. Therefore, the 78 kDa protein is neither stage-specific nor differentially regulated. Surface iodination and subcellular fractionation of the promastigotes indicated that the protein was localized on the cell surface. The 78 kDa protein was found to inhibit the binding of promastigotes to macrophages significantly, suggesting that it may play a role in the process of infection. Thus, here we report the purification of a surface protein of L. donovani of Indian origin, which may play an important role in the process of infection.

  5. Identification of Leishmania chagasi from skin in Leishmania/HIV co-infection: a case report / Identificação de Leishmania chagasi na pele em co-infecção Leishmania/HIV: relato de caso

    Scientific Electronic Library Online (English)

    Marcela, Orsini; Marcelo, Silva; Zélia Maria Profeta da, Luz; Jolandt, Disch; Octávio, Fernandes; Dalton, Moreira; Antonio Carlos Martins, Guedes; Ana, Rabello.

    2002-06-01

    Full Text Available É descrito um caso de co-infecção leishmania/HIV com manifestações cutâneas e visceral. Infecção pela leishmania foi confirmada através de métodos convencionais (parasitológicos e sorológicos) e através da PCR. A espécie Leishmania chagasi isolada da pele foi caracterizada por eletroforese enzimátic [...] a e por polimorfismo de fragmento obtido por enzima de restrição. Abstract in english A case of HIV/Leishmania co-infection presenting both visceral and cutaneous manifestations is reported. Leishmania infection was confirmed by conventional methods (parasitological approach and serology) and by PCR. Leishmania chagasi isolated from the skin lesion was characterized by enzyme electro [...] phoresis and by restriction fragment length polymorphism of the internal transcribed spacer of the ribosomal gene.

  6. Action of Bothrops moojeni venom and its L-amino acid oxidase fraction, treated with {sup 60}Co gamma rays, in Leishmania spp; Acao do veneno de Bothrops moojeni e sua fracao L-aminoacido oxidase, submetida ao tratamento com raios gama de {sup 60}Co, em Leishmania spp

    Energy Technology Data Exchange (ETDEWEB)

    Cardoso, Andre Gustavo Tempone

    1999-07-01

    Bothrops moojeni venom showed an anti leishmania activity in vitro, as determined by a cell viability assay using the reduction of MTT. After venom purification, by chromatography techniques, the fractions with anti leishmania and L-amino acid oxidase activities, eluted in the same positions. The molecular weight of the enzyme was estimated to be 140 kDa by molecular exclusion chromatography, and 69 kDa, by SDS-PAGE, migrating as a single band, with an isoelectric point of 4.8 as determined by isoelectric focusing. The purified LAO from B. moojeni venom, 135-fold more active than crude venom, showed homo dimeric constitution, and was active against Leishmania spp from the New World, with an effective concentration against L(L). amazonensis of 1.80 {mu}g/ml (EC{sub 50}), L.(V.) panamensis (0.78 |{mu}g/ml) and L.(L.) chagasi (0.63 ({mu}g/ml). Ultrastructural studies of promastigotes affected by LAO demonstrated cell death, with edema in several organelles such as mitochondria and nuclear membrane, before cell disruption and necrosis. The action of LAO was demonstrated to be hydrogen peroxide-dependent. Studies with LLCMK-2 cells, treated with LAO, showed a toxic effect, with an EC{sub 50} of 11|{mu}g/ml. Irradiation of LAO with 6{sup 0C}o gamma rays, did not affect its whole oxidative activity, neither detoxified the enzyme. Amastigotes treated with LAO were not affected by its hydrogen peroxide, otherwise, the exogenous product, killed amastigotes with an EC{sub 50} of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  7. The efficacy of 2-nitrovinylfuran derivatives against Leishmania in vitro and in vivo

    Scientific Electronic Library Online (English)

    Sergio, Sifontes-Rodríguez; Lianet, Monzote-Fidalgo; Nilo, Castañedo-Cancio; Ana Margarita, Montalvo-Álvarez; Yamilé, López-Hernández; Niurka Mollineda, Diogo; Juan Francisco, Infante-Bourzac; Oliver, Pérez-Martín; Alfredo, Meneses-Marcel; José Antonio Escario, García-Trevijano; Miguel Ángel, Cabrera-Pérez.

    Full Text Available Despite recent advances in the treatment of some forms of leishmaniasis, the available drugs are still far from ideal due to inefficacy, parasite resistance, toxicity and cost. The wide-spectrum antimicrobial activity of 2-nitrovinylfuran compounds has been described, as has their activity against T [...] richomonas vaginalis and other protozoa. Thus, the aim of this study was to test the antileishmanial activities of six 2-nitrovinylfurans in vitro and in a murine model of leishmaniasis. Minimum parasiticide concentration (MPC) and 50% inhibitory concentration (IC50) values for these compounds against the promastigotes of Leishmania amazonensis, Leishmania infantum and Leishmania braziliensis were determined, as were the efficacies of two selected compounds in an experimental model of cutaneous leishmaniasis (CL) caused by L. amazonensis in BALB/c mice. All of the compounds were active against the promastigotes of the three Leishmania species tested. IC50 and MPC values were in the ranges of 0.8-4.7 µM and 1.7-32 µM, respectively. The compounds 2-bromo-5-(2-bromo-2-nitrovinyl)-furan (furvina) and 2-bromo-5-(2-methyl-2-nitrovinyl)-furan (UC245) also reduced lesion growth in vivo at a magnitude comparable to or higher than that achieved by amphotericin B treatment. The results demonstrate the potential of this class of compounds as antileishmanial agents and support the clinical testing of Dermofural(r) (a furvina-containing antifungal ointment) for the treatment of CL.

  8. The efficacy of 2-nitrovinylfuran derivatives against Leishmania in vitro and in vivo

    Scientific Electronic Library Online (English)

    Sergio, Sifontes-Rodríguez; Lianet, Monzote-Fidalgo; Nilo, Castañedo-Cancio; Ana Margarita, Montalvo-Álvarez; Yamilé, López-Hernández; Niurka Mollineda, Diogo; Juan Francisco, Infante-Bourzac; Oliver, Pérez-Martín; Alfredo, Meneses-Marcel; José Antonio Escario, García-Trevijano; Miguel Ángel, Cabrera-Pérez.

    2015-04-01

    Full Text Available Despite recent advances in the treatment of some forms of leishmaniasis, the available drugs are still far from ideal due to inefficacy, parasite resistance, toxicity and cost. The wide-spectrum antimicrobial activity of 2-nitrovinylfuran compounds has been described, as has their activity against T [...] richomonas vaginalis and other protozoa. Thus, the aim of this study was to test the antileishmanial activities of six 2-nitrovinylfurans in vitro and in a murine model of leishmaniasis. Minimum parasiticide concentration (MPC) and 50% inhibitory concentration (IC50) values for these compounds against the promastigotes of Leishmania amazonensis, Leishmania infantum and Leishmania braziliensis were determined, as were the efficacies of two selected compounds in an experimental model of cutaneous leishmaniasis (CL) caused by L. amazonensis in BALB/c mice. All of the compounds were active against the promastigotes of the three Leishmania species tested. IC50 and MPC values were in the ranges of 0.8-4.7 µM and 1.7-32 µM, respectively. The compounds 2-bromo-5-(2-bromo-2-nitrovinyl)-furan (furvina) and 2-bromo-5-(2-methyl-2-nitrovinyl)-furan (UC245) also reduced lesion growth in vivo at a magnitude comparable to or higher than that achieved by amphotericin B treatment. The results demonstrate the potential of this class of compounds as antileishmanial agents and support the clinical testing of Dermofural(r) (a furvina-containing antifungal ointment) for the treatment of CL.

  9. Terpenes Increase the Lipid Dynamics in the Leishmania Plasma Membrane at Concentrations Similar to Their IC50 Values

    OpenAIRE

    Camargos, Heverton Silva; Moreira, Rodrigo Alves; Mendanha, Sebastião Antonio; Fernandes, Kelly Souza; Dorta, Miriam Leandro; Alonso, Antonio

    2014-01-01

    Although many terpenes have shown antitumor, antibacterial, antifungal, and antiparasitic activity, the mechanism of action is not well established. Electron paramagnetic resonance (EPR) spectroscopy of the spin-labeled 5-doxyl stearic acid revealed remarkable fluidity increases in the plasma membrane of terpene-treated Leishmania amazonensis promastigotes. For an antiproliferative activity assay using 5×106 parasites/mL, the sesquiterpene nerolidol and the monoterpenes (+)-limonene, ?-terpin...

  10. Modulation of Na+/K+ ATPase Activity by Hydrogen Peroxide Generated through Heme in L. amazonensis.

    Science.gov (United States)

    Rocco-Machado, Nathália; Cosentino-Gomes, Daniela; Meyer-Fernandes, José Roberto

    2015-01-01

    Leishmania amazonensis is a protozoan parasite that occurs in many areas of Brazil and causes skin lesions. Using this parasite, our group showed the activation of Na+/K+ ATPase through a signaling cascade that involves the presence of heme and protein kinase C (PKC) activity. Heme is an important biomolecule that has pro-oxidant activity and signaling capacity. Reactive oxygen species (ROS) can act as second messengers, which are required in various signaling cascades. Our goal in this work is to investigate the role of hydrogen peroxide (H2O2) generated in the presence of heme in the Na+/K+ ATPase activity of L. amazonensis. Our results show that increasing concentrations of heme stimulates the production of H2O2 in a dose-dependent manner until a concentration of 2.5 ?M heme. To confirm that the effect of heme on the Na+/K+ ATPase is through the generation of H2O2, we measured enzyme activity using increasing concentrations of H2O2 and, as expected, the activity increased in a dose-dependent manner until a concentration of 0.1 ?M H2O2. To investigate the role of PKC in this signaling pathway, we observed the production of H2O2 in the presence of its activator phorbol 12-myristate 13-acetate (PMA) and its inhibitor calphostin C. Both showed no effect on the generation of H2O2. Furthermore, we found that PKC activity is increased in the presence of H2O2, and that in the presence of calphostin C, H2O2 is unable to activate the Na+/K+ ATPase. 100 ?M of Mito-TEMPO was capable of abolishing the stimulatory effect of heme on Na+/K+ ATPase activity, indicating that mitochondria might be the source of the hydrogen peroxide production induced by heme. The modulation of L. amazonensis Na+/K+ ATPase by H2O2 opens new possibilities for understanding the signaling pathways of this parasite. PMID:26070143

  11. Activity of Brazilian and Bulgarian propolis against different species of Leishmania

    Directory of Open Access Journals (Sweden)

    Gérzia Maria de Carvalho Machado

    2007-02-01

    Full Text Available Extracts of propolis samples collected in Brazil and Bulgaria were assayed against four Leishmania species - Leishmania amazonensis, L. braziliensis, L. chagasi from the New World, and L. major from the Old World - associated to different clinical forms of leishmaniasis. The composition of the extracts has been previously characterized by high temperature high resolution gas chromatography coupled to mass spectrometry. Considering the chemical differences among the extracts and the behavior of the parasites, it was observed significant differences in the leishmanicidal activities with IC50/1 day values in the range of 2.8 to 229.3 µg/ml . An overall analysis showed that for all the species evaluated, Bulgarian extracts were more active than the ethanol Brazilian extract. As the assayed propolis extracts have their chemical composition determined it merits further investigation the effect of individual components or their combinations on each Leishmania species.

  12. Activity of Brazilian and Bulgarian propolis against different species of Leishmania

    Scientific Electronic Library Online (English)

    Gérzia Maria de Carvalho, Machado; Leonor Laura, Leon; Solange Lisboa de, Castro.

    2007-02-01

    Full Text Available Extracts of propolis samples collected in Brazil and Bulgaria were assayed against four Leishmania species - Leishmania amazonensis, L. braziliensis, L. chagasi from the New World, and L. major from the Old World - associated to different clinical forms of leishmaniasis. The composition of the extra [...] cts has been previously characterized by high temperature high resolution gas chromatography coupled to mass spectrometry. Considering the chemical differences among the extracts and the behavior of the parasites, it was observed significant differences in the leishmanicidal activities with IC50/1 day values in the range of 2.8 to 229.3 µg/ml . An overall analysis showed that for all the species evaluated, Bulgarian extracts were more active than the ethanol Brazilian extract. As the assayed propolis extracts have their chemical composition determined it merits further investigation the effect of individual components or their combinations on each Leishmania species.

  13. Study of cross-reactivity in serum samples from dogs positive for Leishmania sp., Babesia canis and Ehrlichia canis in enzyme-linked immunosorbent assay and indirect fluorescent antibody test Estudo da reatividade cruzada em amostras de soro de cães positivos para Leishmania sp., Babesia canis e Ehrlichia canis, pelo ensaio imunoenzimático indireto e pela reação de imunofluorescência indireta

    Directory of Open Access Journals (Sweden)

    Trícia Maria F. de Sousa Oliveira

    2008-03-01

    Full Text Available To verify the presence of cross-reaction among leishmaniosis, ehrlichiosis and babesiosis in serological diagnostics used in human visceral leishmaniasis control programs, serum samples from leishmaniasis endemic and non-endemic areas were collected and tested by Indirect Fluorescent Antibody (IFAT and Enzyme-linked immunosorbent assay (ELISA. All serum samples from endemic areas were positive for Leishmania sp., by ELISA and IFAT, 51% positive for Babesia canis and 43% for Ehrlichia canis by IFAT. None of the serum samples from non-endemic areas were positive for Leishmania sp., by IFAT, but 67% were positive for B. canis and 78% for E. canis using the same test. When tested by ELISA for Leishmania sp., four samples from non-endemic area were positive. These dogs were then located and no clinical signs, parasites or antibody was detected in new tests for a six month period. Only one of these 4 samples was positive for B. canis by IFAT and ELISA and three for E. canis by IFAT. The results of the work suggest a co-infection in the endemic area and no serological cross-reaction among these parasites by IFAT and ELISA.Para verificar a existência de reação cruzada entre leishmaniose visceral, erliquiose e babesiose, nos testes sorológicos utilizados em programas de controle da leishmaniose visceral humana, amostras de soro canino provenientes de áreas endêmicas e não endêmicas para essa enfermidade, foram testadas pela Reação de Imunofluorescência (RIFI e Ensaio imunoenzimático (ELISA. Todos os soros provenientes de área endêmica foram positivos para Leishmania sp pelo ELISA e RIFI, 51% para Babesia canis e 43% para Ehrlichia canis pela RIFI. Pela RIFI, nenhum dos soros provenientes de área não endêmica foi positivo para Leishmania sp, sendo 67% positivos para B. canis e 78% para E. canis pelo mesmo teste. Quando testados pelo ELISA para Leishmania sp., quatro soros da área não endêmica foram positivos. Os cães foram localizados e nenhum sinal clínico, parasito ou anticorpo foi detectado em novos exames realizados ao longo de seis meses. Os resultados desse trabalho sugerem portanto, a presença de uma co-infecção entre os três parasitos citados nas áreas endêmicas e não a reação cruzada entre eles, nos testes sorológicos de RIFI e ELISA descritos.

  14. Mucosal Leishmaniasis Caused by Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in the Brazilian Amazon

    Science.gov (United States)

    de Oliveira Guerra, Jorge Augusto; Prestes, Suzane Ribeiro; Silveira, Henrique; Coelho, Leila Inês de Aguiar Raposo Câmara; Gama, Pricila; Moura, Aristoteles; Amato, Valdir; Barbosa, Maria das Graças Vale; de Lima Ferreira, Luiz Carlos

    2011-01-01

    Background Leishmania (Viannia) braziliensis is a parasite recognized as the most important etiologic agent of mucosal leishmaniasis (ML) in the New World. In Amazonia, seven different species of Leishmania, etiologic agents of human Cutaneous Leishmaniasis, have been described. Isolated cases of ML have been described for several different species of Leishmania: L. (V.) panamensis, L. (V.) guyanensis and L. (L.) amazonensis. Methodology Leishmania species were characterized by polymerase chain reaction (PCR) of tissues taken from mucosal biopsies of Amazonian patients who were diagnosed with ML and treated at the Tropical Medicine Foundation of Amazonas (FMTAM) in Manaus, Amazonas state, Brazil. Samples were obtained retrospectively from the pathology laboratory and prospectively from patients attending the aforementioned tertiary care unit. Results This study reports 46 cases of ML along with their geographical origin, 30 cases caused by L. (V.) braziliensis and 16 cases by L. (V.) guyanensis. This is the first record of ML cases in 16 different municipalities in the state of Amazonas and of simultaneous detection of both species in 4 municipalities of this state. It is also the first record of ML caused by L. (V.) guyanensis in the states of Pará, Acre, and Rondônia and cases of ML caused by L. (V.) braziliensis in the state of Rondônia. Conclusions/Significance L. (V.) braziliensis is the predominant species that causes ML in the Amazon region. However, contrary to previous studies, L. (V.) guyanensis is also a significant causative agent of ML within the region. The clinical and epidemiological expression of ML in the Manaus region is similar to the rest of the country, although the majority of ML cases are found south of the Amazon River. PMID:21408116

  15. The transmission of suprapylarian Leishmania by bite of experimentally infected sand flies (Diptera: Psychodidae A trasnmissão de Leishmania suprapilária pela picada do flebotomíneo infectado experimentalmente

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    L. Ryan

    1987-09-01

    Full Text Available Lutzomyia furcata transmitted Leishmania chagasi to a hamster 10 days after being experimentally fed on an infected spleen. An individual female Psychodopygus carrerai carrerai that had fed on a hamster lesion caused by Leishmania mexicana amazonensis transmitted this parasite 6 days later to another hamster. Transmission electron microscopy of this fly's head revealed a small number of degenerate promastigotes in the foregut, but only a few were attached.O protozoário Leishmania (L. chagasi foi transmitido experimentalmente a um hamster pela picada do flebotomíneo Lutzomyia furcata. Os insetos foram infectados através de uma membrana (pele de pinto, utilizando-se formas amastigotas provenientes do baço de um hamster infectado. O baço foi triturado em sangue de coelho. A L. (L. amazonensis foi transmitida a um hamster pela picada do flebotomíneo Psychodopygus c. carrerai, previamente alimentado em lesão de pele de um outro hamster infectado com o parasita. O exame desse flebotomíneo, através de microscópio eletrônico, revelou um número pequeno de flagelados degenerados, livres no lumen do intestino anterior.

  16. Differentiation of Leishmania species by FT-IR spectroscopy

    Science.gov (United States)

    Aguiar, Josafá C.; Mittmann, Josane; Ferreira, Isabelle; Ferreira-Strixino, Juliana; Raniero, Leandro

    2015-05-01

    Leishmaniasis is a parasitic infectious disease caused by protozoa that belong to the genus Leishmania. It is transmitted by the bite of an infected female Sand fly. The disease is endemic in 88 countries Desjeux (2001) [1] (16 developed countries and 72 developing countries) on four continents. In Brazil, epidemiological data show the disease is present in all Brazilian regions, with the highest incidences in the North and Northeast. There are several methods used to diagnose leishmaniasis, but these procedures have many limitations, are time consuming, have low sensitivity, and are expensive. In this context, Fourier Transform Infrared Spectroscopy (FT-IR) analysis has the potential to provide rapid results and may be adapted for a clinical test with high sensitivity and specificity. In this work, FT-IR was used as a tool to investigate the promastigotes of Leishmaniaamazonensis, Leishmaniachagasi, and Leishmaniamajor species. The spectra were analyzed by cluster analysis and deconvolution procedure base on spectra second derivatives. Results: cluster analysis found four specific regions that are able to identify the Leishmania species. The dendrogram representation clearly indicates the heterogeneity among Leishmania species. The band deconvolution done by the curve fitting in these regions quantitatively differentiated the polysaccharides, amide III, phospholipids, proteins, and nucleic acids. L. chagasi and L. major showed a greater biochemistry similarity and have three bands that were not registered in L. amazonensis. The L. amazonensis presented three specific bands that were not recorded in the other two species. It is evident that the FT-IR method is an indispensable tool to discriminate these parasites. The high sensitivity and specificity of this technique opens up the possibilities for further studies about characterization of other microorganisms.

  17. Heterorhabditis amazonensis RSC5 (Rhabditida: Heterorhabditidae movement and host recognition

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    VANESSA ANDALÓ

    2014-06-01

    Full Text Available Response of Heterorhabditis amazonensis RSC5 to compounds released by different host insects and its virulence level to several insect hosts like Galleria mellonella, Mycotretus apicalis and Tenebrio molitor were evaluated in this study, and compared with other entomopathogenic nematode species like Steinernema carpocapsae All and Steinernema riobrave 355. Tests were performed in Petri dishes with agar-water 2% to determine nematode movement toward the insect with and without opportunity of choosing different insect hosts. Evaluations were made quantifying the proximity of infective juveniles (IJs to the insect as a source of allurement. In order to determine the displacement of IJs in a closed soil condition, a test was carried out in an arena with sand. The nematode was virulent to the target insects. When nematode and insect were released on agar-water, IJs moved toward the stimulus, with H. amazonensis howing preference for certain insects. In the arena with sand S. carpocapsae caused lower insect mortality (70% ± 8.9 for G. mellonella than H. amazonensis and S. riobrave (80% ± 6.5 and 99% ± 0.0. Heterorhabditis amazonensis was able to find and choose its hosts (G. mellonella and T. molitor, similarly to S. riobrave behavior, and located them more effectively than S. carpocapsae. The virulence of H. amazonensis was thus similar to S. riobrave, and this characteristic could be promising to introduce this native species in integrated pest management programs.

  18. Evaluation of Leishmania (Leishmania chagasi strains isolated from dogs originating from two visceral leishmaniasis-endemic areas in Brazil using multilocus enzyme electrophoresis Avaliação de amostras de Leishmania (Leishmania chagasi isoladas de cães oriundos de duas áreas endêmicas de leishmaniose visceral no Brasil através da eletroforese de isoenzimas

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    Carlos Eduardo Ribeiro Coutinho

    2011-10-01

    Full Text Available INTRODUCTION: Domestic dogs are the most important reservoir in the peridomestic transmission cycle of Leishmania (Leishmania chagasi. The genetic variability of subpopulations of this parasite circulating in dogs has not been thoroughly analyzed in Brazil, even though this knowledge has important implications in the clinical-epidemiological context. METHODS: The objective of this study was to evaluate and compare the phenotypic variability of 153 L. chagasi strains isolated from dogs originating from the municipalities of Rio de Janeiro (n = 57 and Belo Horizonte (n = 96, where the disease is endemic. Strains isolated only from intact skin were selected and analyzed by multilocus enzyme electrophoresis using nine enzyme systems (6PG, GPI, NH1 and NH2, G6P, PGM, MDH, ME, and IDHNADP. RESULTS: The electrophoretic profile was identical for all isolates analyzed and was the same as that of the L. chagasi reference strain (MHOM/BR/74/PP75. Phenetic analysis showed a similarity index of one for all strains, with the isolates sharing 100% of the characteristics analyzed. CONCLUSIONS: The results demonstrate that the L. chagasi populations circulating in dogs from Rio de Janeiro and Belo Horizonte belong to a single zymodeme.INTRODUÇÃO: Cães domésticos são considerados os reservatórios mais importantes no ciclo peridoméstico de transmissão de Leishmania (Leishmania chagasi. No entanto, a variabilidade genética de sub-populações que circulam neste hospedeiro é ainda pouco explorada no Brasil, sendo tal conhecimento de grande importância no contexto clínico-epidemiológico. MÉTODOS: O objetivo deste estudo foi avaliar e comparar a variabilidade fenotípica de 153 amostras de L. chagasi isoladas de cães oriundos dos municípios do Rio de Janeiro (n = 57 e Belo Horizonte (n = 96, onde a doença é endêmica. Foram selecionadas somente amostras isoladas de pele íntegra e analisadas por eletroforese de isoenzimas (MLEE empregando nove sistemas enzimáticos (6PG, GPI, NH1 e NH2, G6P, PGM, MDH, ME, IDHNADP. RESULTADOS: Todas as amostras analisadas apresentaram perfil eletroforético idêntico entre si e com a amostra de L. chagasi utilizada como referência neste estudo (MHOM/BR/74/PP75. A análise fenética demonstrou índice de similaridade igual a um para todas as amostras, revelando um compartilhamento de 100% dos caracteres avaliados. CONCLUSÕES: A partir desses resultados, podemos inferir que as populações de L. chagasi que estão circulando nos cães do Rio de Janeiro e Belo Horizonte podem ser agrupadas em um único zimodema.

  19. A novel A2 allele found in Leishmania (Leishmania) infantum chagasi / Novo alelo do gene A2 descrito em Leishmania (Leishmania) infantum chagasi

    Scientific Electronic Library Online (English)

    Trícia Maria Ferreira de Sousa, Oliveira; Elton José Rosas de, Vasconcelos; Andréa Cristina Higa, Nakaghi; Tânia Paula Aquino, Defina; Márcia Mariza Gomes, Jusi; Cristiane Divan, Baldani; Ângela Kaysel, Cruz; Rosangela Zacarias, Machado.

    2011-03-01

    Full Text Available A leishmaniose visceral (LV) é uma zoonose amplamente disseminada, causada no Brasil pela Leishmania (Leishmania) infantum chagasi. Flebotomíneos vetores adquirem o agente etiológico, alimentando-se do sangue de animais contaminados, como cachorros ou animais selvagens. A doença é endêmica no Brasil [...] , e focos de epidemia são relatados em cidades densamente povoadas por todo o país. Muitas manifestações clínicas relacionadas à infecção por Leishmania estão ligadas à relação parasito-hospedeiro, e vários possíveis fatores de virulência dos parasitas, que causam a LV, são alvos de estudo, tais como os genes A2. O gene A2 foi isolado pela primeira vez em 1994 e, em seguida, em 2005, três novos alelos foram descritos em Leishmania (Leishmania) infantum. No presente estudo, um fragmento do gene A2 de uma população clonal de L.(L.) infantum chagasi foi amplificado por PCR e sua sequência de nucleotídeos determinada. O fragmento mostrou 90% de similaridade com alelos do gene A2 de Leishmania (Leishmania) donovani e de L. (L.) infantum, descritos na literatura. Entretanto, a tradução da sequência de nucleotídeos mostra diferenças na sequência de aminoácidos da proteína, que podem ser essenciais em determinar a variabilidade do gene A2 em espécies do complexo L. (L.) donovani e representa uma ferramenta adicional na compreenssão do papel dessa família de genes na virulência e imunidade da leishmaniose visceral. O conhecimento dessa variação é importante para o desenvolvimento de testes diagnósticos mais precisos e ferramentas mais eficazes no controle da doença. Abstract in english Visceral leishmaniasis (VL) is a widely spread zoonotic disease. In Brazil the disease is caused by Leishmania (Leishmania) infantum chagasi. Peridomestic sandflies acquire the etiological agent by feeding on blood of infected reservoir animals, such as dogs or wildlife. The disease is endemic in Br [...] azil and epidemic foci have been reported in densely populated cities all over the country. Many clinical features of Leishmania infection are related to the host-parasite relationship, and many candidate virulence factors in parasites that cause VL have been studied such as A2 genes. The A2 gene was first isolated in 1994 and then in 2005 three new alleles were described in Leishmania (Leishmania) infantum. In the present study we amplified by polymerase chain reaction (PCR) and sequenced the A2 gene from the genome of a clonal population of L. (L.) infantum chagasi VL parasites. The L. (L.) infantum chagasi A2 gene was amplified, cloned, and sequenced in. The amplified fragment showed approximately 90% similarity with another A2 allele amplified in Leishmania (Leishmania) donovani and in L.(L.) infantum described in literature. However, nucleotide translation shows differences in protein amino acid sequence, which may be essential to determine the variability of A2 genes in the species of the L. (L.) donovani complex and represents an additional tool to help understanding the role this gene family may have in establishing virulence and immunity in visceral leishmaniasis. This knowledge is important for the development of more accurate diagnostic tests and effective tools for disease control.

  20. A novel A2 allele found in Leishmania (Leishmania infantum chagasi Novo alelo do gene A2 descrito em Leishmania (Leishmania infantum chagasi

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    Trícia Maria Ferreira de Sousa Oliveira

    2011-03-01

    Full Text Available Visceral leishmaniasis (VL is a widely spread zoonotic disease. In Brazil the disease is caused by Leishmania (Leishmania infantum chagasi. Peridomestic sandflies acquire the etiological agent by feeding on blood of infected reservoir animals, such as dogs or wildlife. The disease is endemic in Brazil and epidemic foci have been reported in densely populated cities all over the country. Many clinical features of Leishmania infection are related to the host-parasite relationship, and many candidate virulence factors in parasites that cause VL have been studied such as A2 genes. The A2 gene was first isolated in 1994 and then in 2005 three new alleles were described in Leishmania (Leishmania infantum. In the present study we amplified by polymerase chain reaction (PCR and sequenced the A2 gene from the genome of a clonal population of L. (L. infantum chagasi VL parasites. The L. (L. infantum chagasi A2 gene was amplified, cloned, and sequenced in. The amplified fragment showed approximately 90% similarity with another A2 allele amplified in Leishmania (Leishmania donovani and in L.(L. infantum described in literature. However, nucleotide translation shows differences in protein amino acid sequence, which may be essential to determine the variability of A2 genes in the species of the L. (L. donovani complex and represents an additional tool to help understanding the role this gene family may have in establishing virulence and immunity in visceral leishmaniasis. This knowledge is important for the development of more accurate diagnostic tests and effective tools for disease control.A leishmaniose visceral (LV é uma zoonose amplamente disseminada, causada no Brasil pela Leishmania (Leishmania infantum chagasi. Flebotomíneos vetores adquirem o agente etiológico, alimentando-se do sangue de animais contaminados, como cachorros ou animais selvagens. A doença é endêmica no Brasil, e focos de epidemia são relatados em cidades densamente povoadas por todo o país. Muitas manifestações clínicas relacionadas à infecção por Leishmania estão ligadas à relação parasito-hospedeiro, e vários possíveis fatores de virulência dos parasitas, que causam a LV, são alvos de estudo, tais como os genes A2. O gene A2 foi isolado pela primeira vez em 1994 e, em seguida, em 2005, três novos alelos foram descritos em Leishmania (Leishmania infantum. No presente estudo, um fragmento do gene A2 de uma população clonal de L.(L. infantum chagasi foi amplificado por PCR e sua sequência de nucleotídeos determinada. O fragmento mostrou 90% de similaridade com alelos do gene A2 de Leishmania (Leishmania donovani e de L. (L. infantum, descritos na literatura. Entretanto, a tradução da sequência de nucleotídeos mostra diferenças na sequência de aminoácidos da proteína, que podem ser essenciais em determinar a variabilidade do gene A2 em espécies do complexo L. (L. donovani e representa uma ferramenta adicional na compreenssão do papel dessa família de genes na virulência e imunidade da leishmaniose visceral. O conhecimento dessa variação é importante para o desenvolvimento de testes diagnósticos mais precisos e ferramentas mais eficazes no controle da doença.

  1. Reproduction of Leishmania (Leishmania infantum chagasi in conditioned cell culture growth medium Isolamento e crescimento de Leishmania (Leishmania infantum chagasi em meio de crescimento condicionado de cultura celular

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    Yeda L. Nogueira

    2006-06-01

    Full Text Available Leishmanias can be produced by inoculation in conditioned McCoy cell culture growth medium (CGM. Leishmania (Leishmania infantum chagasi (100 parasites grown in NNN medium was inoculated in 2.5 mL CGM, kept in plates (24 wells and its multiplication was observed for five days (120 hours. After day 5, the medium was saturated with the flagellate forms of the parasite (promastigotes. The reproduction of the leishmanias was observed every 24 hours and the number of parasites was calculated by counting the parasites in a drop of 10 µ L and photomicrographied. So the number of Leishmanias was adjusted to 1 mL volume. The advantage of the technique by isolation of Leishmania in CGM demonstrated in this study is its low cost and high efficacy even with a small quantity of parasites (10² promastigotes used as inoculum. Additionally, isolation of the leishmania can be obtained together with an increase in their density (180 times as observed by growth kinetics, within a shorter time. These results justify the use of this low-cost technique for the isolation and investigation of the behavior and multiplication of Leishmania both in vertebrates and invertebrates, besides offering means of obtaining antigens, whether whole antigens (leishmanias or the soluble antigens produced by the parasites which may be useful for the production of new diagnostic kits.Leishmânias podem ser produzidas em meio de crescimento condicionado, após o cultivo de células McCoy (MCC. Promastigotas crescidas em meio semi-sólido NNN foram inoculadas em MCC, inicialmente, 100 parasitos por poço com 2,5 mL de McCoyMCC, em placas com 24 poços, sua multiplicação foi observada por uma cinética de 120 horas. Após este tempo, o meio estava saturado de promastigotas. A reprodução das leishmânias foi acompanhada a cada 24 horas, com contagem do número de parasitos por campo fotomicrografado. Como vantagem da técnica do crescimento da leishmânia em MCC tem-se o seu baixo custo, com pequena quantidade de parasitos pode-se obter o aumento da densidade de promastigotas em tempo reduzido. Com o emprego dessa técnica pode-se estudar o comportamento e a multiplicação das leishmânias nos vertebrados e invertebrados, assim como, obter antígenos, tanto brutos (leishmânia como solúveis, produzidos pelos parasitos, que poderão ser úteis para se desenvolver kits de diagnósticos.

  2. Características clínicas da leishmaniose tegumentar em crianças de 0 a 5 anos em uma área endêmica de Leishmania (Viannia braziliensis

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    Ampuero Julia

    2006-01-01

    Full Text Available Para caracterizar a clínica da leishmaniose tegumentar em crianças de 0 a 5 anos de idade, foram avaliadas, retrospectivamente, 4.464 fichas clínicas do Centro de Saúde de Corte de Pedra, Presidente Tancredo Neves, Bahia, Brasil, área endêmica de leishmaniose tegumentar americana, entre maio de 1987 e dezembro de 1995. Foram registrados neste período 4.275 casos novos de leishmaniose, dos quais, 491 (11,5% correspondiam a crianças de 0 a 5 anos. A razão entre gênero masculino e feminino nas crianças foi 1,1:1. A forma clínica predominante foi a cutânea (98% e as lesões ulceradas foram as mais freqüentes (99%. A localização das lesões ocorreu, principalmente, acima da cintura (p<0,05, e 35,5% apresentaram lesões múltiplas. A magnitude da doença em crianças, a freqüência semelhante observada em ambos os gêneros e a localização das lesões sugere a possibilidade de transmissão vetorial no domicílio ou peridomicílio.

  3. Peripheral blood fibrocytes: new information to explain the dynamics of Leishmania infection

    Science.gov (United States)

    Macedo-Silva, Roger Magno; dos Santos, Carina de Lima Pereira; Diniz, Vanessa Alvaro; de Carvalho, Jorge José; Guerra, Camila; Côrte-Real, Suzana

    2013-01-01

    Fibrocytes are important for understanding the progression of many diseases because they are present in areas where pathogenic lesions are generated. However, the morphology of fibrocytes and their interactions with parasites are poorly understood. In this study, we examined the morphology of peripheral blood fibrocytes and their interactions with Leishmania (L.) amazonensis . Through ultrastructural analysis, we describe the details of fibrocyte morphology and how fibrocytes rapidly internalise Leishmania promastigotes. The parasites differentiated into amastigotes after 2 h in phagolysosomes and the infection was completely resolved after 72 h. Early in the infection, we found increased nitric oxide production and large lysosomes with electron-dense material. These factors may regulate the proliferation and death of the parasites. Because fibrocytes are present at the infection site and are directly involved in developing cutaneous leishmaniasis, they are targets for effective, non-toxic cell-based therapies that control and treat leishmaniasis. PMID:24626303

  4. Peripheral blood fibrocytes: new information to explain the dynamics of Leishmania infection

    Scientific Electronic Library Online (English)

    Roger Magno, Macedo-Silva; Carina de Lima Pereira dos, Santos; Vanessa Alvaro, Diniz; Jorge Jose de, Carvalho; Camila, Guerra; Suzana, Corte-Real.

    2014-02-01

    Full Text Available Fibrocytes are important for understanding the progression of many diseases because they are present in areas where pathogenic lesions are generated. However, the morphology of fibrocytes and their interactions with parasites are poorly understood. In this study, we examined the morphology of periph [...] eral blood fibrocytes and their interactions with Leishmania (L.) amazonensis . Through ultrastructural analysis, we describe the details of fibrocyte morphology and how fibrocytes rapidly internalise Leishmania promastigotes. The parasites differentiated into amastigotes after 2 h in phagolysosomes and the infection was completely resolved after 72 h. Early in the infection, we found increased nitric oxide production and large lysosomes with electron-dense material. These factors may regulate the proliferation and death of the parasites. Because fibrocytes are present at the infection site and are directly involved in developing cutaneous leishmaniasis, they are targets for effective, non-toxic cell-based therapies that control and treat leishmaniasis.

  5. Peripheral blood fibrocytes: new information to explain the dynamics of Leishmania infection

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    Roger Magno Macedo-Silva

    2014-02-01

    Full Text Available Fibrocytes are important for understanding the progression of many diseases because they are present in areas where pathogenic lesions are generated. However, the morphology of fibrocytes and their interactions with parasites are poorly understood. In this study, we examined the morphology of peripheral blood fibrocytes and their interactions with Leishmania (L. amazonensis . Through ultrastructural analysis, we describe the details of fibrocyte morphology and how fibrocytes rapidly internalise Leishmania promastigotes. The parasites differentiated into amastigotes after 2 h in phagolysosomes and the infection was completely resolved after 72 h. Early in the infection, we found increased nitric oxide production and large lysosomes with electron-dense material. These factors may regulate the proliferation and death of the parasites. Because fibrocytes are present at the infection site and are directly involved in developing cutaneous leishmaniasis, they are targets for effective, non-toxic cell-based therapies that control and treat leishmaniasis.

  6. Peripheral blood fibrocytes: new information to explain the dynamics of Leishmania infection.

    Science.gov (United States)

    Macedo-Silva, Roger Magno; Santos, Carina de Lima Pereira dos; Diniz, Vanessa Alvaro; Carvalho, Jorge José de; Guerra, Camila; Côrte-Real, Suzana

    2014-02-01

    Fibrocytes are important for understanding the progression of many diseases because they are present in areas where pathogenic lesions are generated. However, the morphology of fibrocytes and their interactions with parasites are poorly understood. In this study, we examined the morphology of peripheral blood fibrocytes and their interactions with Leishmania (L.) amazonensis . Through ultrastructural analysis, we describe the details of fibrocyte morphology and how fibrocytes rapidly internalise Leishmania promastigotes. The parasites differentiated into amastigotes after 2 h in phagolysosomes and the infection was completely resolved after 72 h. Early in the infection, we found increased nitric oxide production and large lysosomes with electron-dense material. These factors may regulate the proliferation and death of the parasites. Because fibrocytes are present at the infection site and are directly involved in developing cutaneous leishmaniasis, they are targets for effective, non-toxic cell-based therapies that control and treat leishmaniasis. PMID:24626303

  7. Expresión heteróloga de ARN mensajeros de Leishmania en ovocitos de anfibios

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    Arroyo Olarte Rubén Darío

    2006-06-01

    Full Text Available La técnica de expresión heteróloga en ovocitos de Xenopus laevis ha sido ampliamente utilizada para la caracterización funcional de canales iónicos. En este estudio se reporta el uso del modelo nativo de ovocitos de Bufo marinus para la expresión heteróloga de ARN poliadenilado (ARNm poli(A+ de Leishmania amazonensis y la posterior identificación de canales iónicos mediante registros electrofisiológicos utilizando la técnica de voltaje controlado (voltage-clamp. Se logró realizar la maduración, extracción y cultivo de ovocitos de B. marinus bajo condiciones similares a las estandarizadas para X. laevis. El potencial de reposo promedio así como las corrientes endógenas mostraron ser definidas por canales de cloruro dependientes de calcio y rectificadores de salida, de manera similar a lo reportado en X. laevis. La inyección de ARNm poli(A+ de L. amazonensis generó la expresión de corrientes de cloruro de  amplitud, cinética y características farmacológicas diferentes a las corrientes endógenas, reportadas en estudios previos con X. laevis como posible resultado de la expresión heteróloga de canales iónicos de L. amazonensis.

  8. Intermediary metabolism of Leishmania.

    Science.gov (United States)

    Blum, J J

    1993-04-01

    In the course of their existence, parasites develop several metabolic pathways that differ significantly from those of their hosts. Despite the fairly close evolutionary kinship between Leishmania donovani and Trypanosoma brucei, the forms that live in the insect vectors have evolved different strategies for the disposition of available food resources. In this brief review, Joseph Blum will focus on the data available from studies on Leishmania spp and will largely ignore the information available from Trypanosoma spp. PMID:15463727

  9. Palaearctic origin of Leishmania

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    Sara F Kerr

    2000-01-01

    Full Text Available The hypothesis of a Palaearctic origin of Leishmania in the early Cenozoic, dispersal to the Nearctic in the late Eocene and to the Neotropical in the Pliocene is presented. It is further hypothesized that murid rodents and their immediate ancestors have been important mammalian reservoirs since the origination of Leishmania. Biochemical, molecular, biogeographical, entomological, mammalalogical and ecological support for these hypotheses are reviewed.

  10. PCR-RFLP y RAPD para la tipificación de Leishmania neotropical PCR-RFLP and RAPD for typing neotropical Leishmania

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    Iván Darío Vélez

    2008-12-01

    Full Text Available Introducción. El análisis de la longitud de los fragmentos de restricción del producto amplificado y el estudio del ADN polimórfico amplificado al azar han demostrado ser herramientas útiles para la tipificación de Leishmania.
    Objetivos. Estudiar la utilidad de las técnicas moleculares para la identificación y tipificación de cepas de referencia de Leishmania spp. del Nuevo Mundo y valorar su aplicabilidad a muestras clínicas.
    Materiales y métodos. Se aplicó PCR para amplificar el gen que codifica la cisteíno-proteinasa B, y el análisis de la longitud de los fragmentos de restricción del producto amplificado utilizando ácido desoxirribonucleico de 16 cepas de referencia de Latinoamérica y de muestras clínicas de pacientes colombianos con leishmaniasis, y la técnica del ácido desoxirribonucleico polimórfico amplificado al azar utilizando ocho cepas de referencia. Se establecieron los patrones de bandas en cada caso.
    Resultados. Se obtuvo producto de amplificación en la PCR para Leishmania braziliensis, L. peruviana, L. panamensis y L. guyanensis. Para el resto, no fue posible amplificar el gen con los cebadores utilizados. La restricción mostró un patrón de bandas común para L. peruviana, L. guyanensis y L. panamensis, mientras L. braziliensis, presentaba un perfil individual único. El análisis de restricción del producto amplificado generó un patrón de bandas similar en los cinco pacientes estudiados, que se correspondía con el patrón generado por L. peruviana, L. guyanensis o L. panamensis. Mediante la amplificación al azar se obtuvieron patrones de bandas reproducibles con todas las cepas estudiadas, que posibilitaron la diferenciación. Se discuten las ventajas y limitaciones de ambos procederes. Conclusiones. El combinar ambas metodologías resultaría útil para identificar especies de importancia médica, tomando en cuenta sus ventajas y desventajas.Introduction. The analysis of the PCR-restriction fragment length polymorphism and random amplified polymorphic DNA have been useful tools for Leishmania identification.
    Objectives. Molecular procedures were demonstrated for identification and typing of reference strains of New World Leishmania and their applicability was validated for clinical samples.
    Materials and methods. DNA was extracted from 16 reference strains of Latin American Leishmania as well as from clinical samples of leishmaniasis patients. A sequence coding for cysteine proteinase B was amplified by PCR and subjected to restriction fragment length polymorphism analysis. The enzyme used was Taq1. For eight of the reference strains, the random amplified polymorphic desoxyribonucleic acid technique (RAPD was applied. Band patterns for Leishmania species differentiation were established each each method. The sample size of the clinical sample was of 5.
    Results. PCR products of the cysteine proteinase B gene were obtained for L. braziliensis, L. peruviana, L. panamensis and L. guyanensis. For the other species, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, no amplification occurred. The patterns of restriction fragments revealed band patterns in common for L. peruviana, L. guyanensis and L. panamensis, whereas L. braziliensis had a distinctive pattern. When human samples were examined, amplification occurred for all cases, and the profiles corresponded to the common profile of L. peruviana, L. guyanensis and L. panamensis. The RAPD technique demonstrated reproducible and distinctive patterns for each of the 8 reference strains, L. mexicana, L. amazonensis, L. garnhami, L. lainsoni, L. chagasi, L. naiffi, making possible to differentiate all them. The advantages and limitations of each procedure are discussed.
    Co

  11. Leishmania replication protein A-1 binds in vivo single-stranded telomeric DNA

    International Nuclear Information System (INIS)

    Replication protein A (RPA) is a highly conserved heterotrimeric single-stranded DNA-binding protein involved in different events of DNA metabolism. In yeast, subunits 1 (RPA-1) and 2 (RPA-2) work also as telomerase recruiters and, in humans, the complex unfolds G-quartet structures formed by the 3' G-rich telomeric strand. In most eukaryotes, RPA-1 and RPA-2 bind DNA using multiple OB fold domains. In trypanosomatids, including Leishmania, RPA-1 has a canonical OB fold and a truncated RFA-1 structural domain. In Leishmania amazonensis, RPA-1 alone can form a complex in vitro with the telomeric G-rich strand. In this work, we show that LaRPA-1 is a nuclear protein that associates in vivo with Leishmania telomeres. We mapped the boundaries of the OB fold DNA-binding domain using deletion mutants. Since Leishmania and other trypanosomatids lack homologues of known telomere end binding proteins, our results raise questions about the function of RPA-1 in parasite telomeres

  12. Leishmania is not prone to develop resistance to tamoxifen

    Directory of Open Access Journals (Sweden)

    Adriano C. Coelho

    2015-12-01

    Full Text Available Tamoxifen, an antineoplastic agent, is active in vitro and in vivo against the parasitic protozoa Leishmania. As part of our efforts to unravel this drug's mechanisms of action against the parasite and understand how resistance could arise, we tried to select tamoxifen-resistant Leishmania amazonensis. Three different strategies to generate tamoxifen resistant mutants were used: stepwise increase in drug concentration applied to promastigote cultures, chemical mutagenesis followed by drug selection and treatment of infected mice followed by selection of amastigotes. For amastigote selection, we employed a method with direct plating of parasites recovered from lesions into semi-solid media. Tamoxifen resistant parasites were not rescued by any of these methods. Miltefosine was used as a control in selection experiments and both stepwise selection and chemical mutagenesis allowed successful isolation of miltefosine resistant mutants. These findings are consistent with a multi-target mode of action to explain tamoxifen's leishmanicidal properties. Considering that drug resistance is a major concern in anti-parasitic chemotherapy, these findings support the proposition of using tamoxifen as a partner in drug combination schemes for the treatment of leishmaniasis.

  13. Leishmania is not prone to develop resistance to tamoxifen.

    Science.gov (United States)

    Coelho, Adriano C; Trinconi, Cristiana T; Senra, Luisa; Yokoyama-Yasunaka, Jenicer K U; Uliana, Silvia R B

    2015-12-01

    Tamoxifen, an antineoplastic agent, is active in vitro and in vivo against the parasitic protozoa Leishmania. As part of our efforts to unravel this drug's mechanisms of action against the parasite and understand how resistance could arise, we tried to select tamoxifen-resistant Leishmania amazonensis. Three different strategies to generate tamoxifen resistant mutants were used: stepwise increase in drug concentration applied to promastigote cultures, chemical mutagenesis followed by drug selection and treatment of infected mice followed by selection of amastigotes. For amastigote selection, we employed a method with direct plating of parasites recovered from lesions into semi-solid media. Tamoxifen resistant parasites were not rescued by any of these methods. Miltefosine was used as a control in selection experiments and both stepwise selection and chemical mutagenesis allowed successful isolation of miltefosine resistant mutants. These findings are consistent with a multi-target mode of action to explain tamoxifen's leishmanicidal properties. Considering that drug resistance is a major concern in anti-parasitic chemotherapy, these findings support the proposition of using tamoxifen as a partner in drug combination schemes for the treatment of leishmaniasis. PMID:26150922

  14. In vitro and in vivo antileishmanial efficacy of a combination therapy of diminazene and artesunate against Leishmania donovani in BALB/c mice / Estudo in vitro e in vivo da eficácia anti leishmaniótica de terapêutica combinada de Diminazene e Artesunate contra Leishmania donovani em camundongos Balb/c

    Scientific Electronic Library Online (English)

    Joshua Muli, Mutiso; John Chege, Macharia; Mustafa, Barasa; Evans, Taracha; Alain J., Bourdichon; Michael M., Gicheru.

    2011-06-01

    Full Text Available A atividade in vitro e in vivo de Diminazene (Dim), Artezunate (Art) e a combinação Dim e Art (Dim-Art) contra Leishmania donovani foi comparada com a droga de referência Anfotericina B. IC50 da Dim-Art foi 2,28 ± 0,24 µg/mL enquanto aquelas de Dim e Art foram 9,16 ± 0,3 µg/mL e 4,64 ± 0,48 µg/mL re [...] spectivamente. O IC50 da Anfotericina B foi 0,16 ± 0,32 µg/mL contra a fase estacionária de promastigotas. A avaliação in vivo do modelo de L. donovani em camundongos Balb/c indicou que os tratamentos com a terapêutica de drogas combinadas em doses de 12,5 mg/kg por 28 dias consecutivos significantemente (p Abstract in english The in vitro and in vivo activity of diminazene (Dim), artesunate (Art) and combination of Dim and Art (Dim-Art) against Leishmania donovani was compared to reference drug; amphotericin B. IC50 of Dim-Art was found to be 2.28 ± 0.24 µg/mL while those of Dim and Art were 9.16 ± 0.3 µg/mL and 4.64 ± 0 [...] .48 µg/mL respectively. The IC50 for Amphot B was 0.16 ± 0.32 µg/mL against stationary-phase promastigotes. In vivo evaluation in the L. donovani BALB/c mice model indicated that treatments with the combined drug therapy at doses of 12.5 mg/kg for 28 consecutive days significantly (p

  15. Estudo clínico randomizado comparando antimoniato de meglumina, pentamidina e anfotericina B para o tratamento da leishmaniose cutânea ocasionada por Leishmania guyanensis / A randomized clinical trial comparing meglumine antimoniate, pentamidine and amphotericin B for the treatment of cutaneous leishmaniasis by Leishmania guyanensis

    Scientific Electronic Library Online (English)

    Leandro Ourives, Neves; Anette Chrusciak, Talhari; Ellen Priscilla Nunes, Gadelha; Roberto Moreira da, Silva Júnior; Jorge Augusto de Oliveira, Guerra; Luiz Carlos de Lima, Ferreira; Sinésio, Talhari.

    2011-12-01

    Full Text Available FUNDAMENTOS: O tratamento da leishmaniose tegumentar americana (LTA) ainda constitui desafio, pois a maioria dos medicamentos é injetável e têm-se poucos ensaios clínicos randomizados comparando a eficácia das drogas. Além disso, é provável que as espécies de Leishmania tenham influência nas respost [...] as terapêuticas. OBJETIVOS: Avaliar e comparar a eficácia e a segurança dos esquemas de tratamento na LTA, ocasionada por Leishmania (Viannia) guyanensis. MÉTODOS: 185 pacientes foram selecionados, conforme critérios de elegibilidade, e distribuídos, aleatoriamente, em 3 grupos - 2 com 74 enfermos e outro com 37 - que receberam, respectivamente, antimoniato de meglumina, isotionato de pentamidina e anfotericina B em doses, períodos e vias de administração padronizados. Os enfermos foram reexaminados um, dois e seis meses após o final dos tratamentos. RESULTADOS: Não houve diferença entre os grupos terapêuticos em relação ao sexo, idade, número ou local das lesões. A análise por intenção de tratar (ITT) mostrou eficácias de 58,1% para a pentamidina e 55,5% para o antimoniato (p=0,857). O grupo da anfotericina B foi analisado separadamente, pois 28 (75,7%) pacientes negaram-se a continuar no estudo após a randomização. Eventos adversos leves ou moderados foram relatados por 74 (40%) pacientes, principalmente artralgia (20,3%), para o grupo do antimoniato, e dor (35,1%) ou enduração (10,8%) no local das injeções para a pentamidina. CONCLUSÕES: A pentamidina tem eficácia similar ao antimonial pentavalente para o tratamento da LTA ocasionada por L. guyanensis. Face aos baixos resultados de eficácia apresentados por ambas as drogas, necessita-se, com urgência, investigar novas opções terapêuticas para esta enfermidade. Abstract in english FUNDAMENTALS: American tegumentary leishmaniasis (ATL) treatment remains a challenge, since most available drugs are injectable and only a small number of comparative, randomized clinical trials have been performed to support their use. Moreover, treatment outcome may depend on the causative species [...] of Leishmania. OBJECTIVES: To evaluate and compare the efficacy and tolerability of meglumine antimoniate, pentamidine isethionate, and amphotericin B in the treatment of ATL caused by Leishmania (Viannia) guyanensis. METHODS: 185 patients were selected according to the eligibility criteria and randomly allocated into three groups - two groups with 74 patients each, and one group with 37 patients, which underwent meglumine, pentamidine and amphotericin B treatment, respectively. Doses, mode of administration and time periods of treatment followed the current recommendations for each drug. Patients were re-examined one, two and six months after completion of treatment. RESULTS: No differences were observed among the therapeutic groups in relation to gender, age, number or site of lesions. Intention-to-treat (ITT) analysis showed efficacy of 58.1% for pentamidine and 55.5% for meglumine (p=0.857). The amphotericin B group was analyzed separately, since 28 patients (75.7%) in this group refused to continue participating in the study. Mild or moderate adverse effects were reported by 74 (40%) patients, especially arthralgia (20.3%) in the meglumine group, and pain (35.1%) or induration (10.8%) at the site of injection in the pentamidine group. CONCLUSION: Pentamidine and meglumine show similar efficacy in the treatment of ATL caused by L. guyanensis. Given the low efficacy of both drugs, there is an urgent need for new therapeutical approaches.

  16. Leishmania isoenzyme polymorphisms in Ecuador: Relationships with geographic distribution and clinical presentation

    Directory of Open Access Journals (Sweden)

    Mimori Tatsuyuki

    2006-09-01

    Full Text Available Abstract Background Determinants of the clinical presentation of the leishmaniases are poorly understood but Leishmania species and strain differences are important. To examine the relationship between clinical presentation, species and isoenzyme polymorphisms, 56 Leishmania isolates from distinct presentations of American tegumentary leishmaniasis (ATL from Ecuador were analyzed. Methods Isolates were characterized by multilocus enzyme electrophoresis for polymorphisms of 11 isoenzymes. Patients were infected in four different ecologic regions: highland and lowland jungle of the Pacific coast, Amazonian lowlands and Andean highlands. Results Six Leishmania species constituting 21 zymodemes were identified: L. (Viannia panamensis (21 isolates, 7 zymodemes, L. (V. guyanensis (7 isolates, 4 zymodemes, L. (V. braziliensis (5 isolates, 3 zymodemes, L. (Leishmania mexicana (11 isolates, 4 zymodemes, L. (L. amazonensis (10 isolates, 2 zymodemes and L. (L. major (2 isolates, 1 zymodeme. L. panamensis was the species most frequently identified in the Pacific region and was associated with several clinical variants of cutaneous disease (CL; eight cases of leishmaniasis recidiva cutis (LRC found in the Pacific highlands were associated with 3 zymodemes of this species. Mucocutaneous leishmaniasis found only in the Amazonian focus was associated with 3 zymodemes of L. braziliensis. The papular variant of CL, Uta, found in the Andean highlands was related predominantly with a single zymodeme of L. mexicana. Conclusion Our data show a high degree of phenotypic variation within species, and some evidence for associations between specific variants of ATL (i.e. Uta and LRC and specific Leishmania zymodemes. This study further defines the geographic distribution of Leishmania species and clinical variants of ATL in Ecuador.

  17. DISEÑO Y PRODUCCIÓN DE UNA HERRAMIENTA MOLECULAR PARA EL ESTUDIO DEL N-TERMINAL DE LA NICOTINAMIDA MONONUCLEÓTIDO ADENILIL TRANSFERASA (NMNAT) EN Leishmania braziliensis / DESIGN AND PRODUCTION OF A MOLECULAR TOOL TO STUDY OF N-TERMINAL NICOTINAMIDE MONONUCLEOTIDE ADENYLYL TRANSFERASE (NMNAT) IN Leishmania braziliensis / DESIGN E PRODUÇÃO DE UMA FERRAMENTA MOLECULAR PARA O ESTUDO DO TERMINALN DA NICOTINAMIDA CICLASE MONONUCLEÓTIDO TRANSFERASE (NMNAT) EM Leishmania braziliensis

    Scientific Electronic Library Online (English)

    Jesús D, Castaño; Zuly J, Rivera; María H, Ramírez.

    2012-08-01

    Full Text Available Leishmania braziliensis é um parasita protozoário que causa a maioria dos casos de leishmaniose cutânea em pelo menos 15 países das Américas. A Organização Mundial de Saúde (OMS) informou que cerca de 12 milhões de pessoas estão infectadas em todo o mundo e esse número aumenta a cada ano. Devido ao [...] delicado problema de saúde pública decorrentes da prevalência desta doença é necessário estudar o metabolismo do parasita. A este respeito temos estudado a proteína NMNAT deste parasita, que é uma enzima central no metabolismo de todos os organismos de estar envolvido na produção de NAD+, um importante cofator em reações redox de processos centrais de celulares metabolismo. No L. braziliensis NMNAT encontrou uma seqüencia de 43 aminoácidos no terminal N homologia com a proteína faltando host. Este estudo produziu anticorpos IgG específicos para esta seqüência, usando como peptídeos de antígeno contendo a seqüência mencionada. Os anticorpos obtidos mostraram um reconhecimento da NMNAT L. braziliensis recombinantes por meio de julgamento por western blot. Abstract in spanish Leishmania braziliensis es un parásito protozoario causante de la mayor parte de casos de leishmaniasis cutánea en al menos quince países del continente americano. La Organización Mundial de la Salud (OMS) ha reportado que cerca de doce millones de personas están infectadas en el mundo y que este nú [...] mero aumenta cada año. Debido al delicado problema de salud pública derivado de la prevalencia de esta enfermedad se hace necesario el estudio del metabolismo de este parásito. En tal sentido se ha estudiado la proteína NMNAT de este parásito, la cual es una enzima central del metabolismo de todos los organismos al estar encargada de la síntesis del NAD+, un importante cofactor en reacciones redox de procesos centrales del metabolismo celular. En la NMNAT de L. braziliensis se ha encontrado una secuencia de 44 aminoácidos en el extremo N-terminal carente de homología con la proteína del hospedero. En este estudio se produjeron anticuerpos IgG específicos contra esta secuencia, utilizando como antígenos péptidos que contuvieran la secuencia mencionada. Los anticuerpos obtenidos mostraron un reconocimiento de la NMNAT recombinante de L. braziliensis mediante ensayo por western blot. Abstract in english Leishmania braziliensis is a protozoan which is cause of the most of the cutaneous leishmaniasis cases in at least 15 countries from America. World Health Organization (WHO) has reported that around 12 millions of people are infected in the world and this number increase every year. Because of the d [...] elicate problem of public health due to the prevalence of this disease, it is necessary the metabolism study in this parasite. In this way has been studied NMNAT protein of the parasite, which is a central enzyme of the metabolism of all organisms, since it is in charge of synthesizing NAD+, an important cofactor in oxidation-reduction reactions of central processes in the cellular metabolism. In The NMNAT of L. has been found a 43 amino acids sequence in the N terminal, which does not have homology with the protein in the human host. In this study were produced IgG antibodies against this sequence, using like antigens peptides that had the mentioned sequence. The produced antibodies recognized the recombinant NMNAT of L. braziliensis through western blot assay.

  18. In vitro effects of triterpenic acids from olive leaf extracts on the mitochondrial membrane potential of promastigote stage of Leishmania spp.

    Science.gov (United States)

    Sifaoui, Ines; López-Arencibia, Atteneri; Martín-Navarro, Carmen Maria; Ticona, Juan Carlos; Reyes-Batlle, María; Mejri, Mondher; Jiménez, Antonio Ignacio; Lopez-Bazzocchi, Isabel; Valladares, Basilio; Lorenzo-Morales, Jacob; Abderabba, Manef; Piñero, José E

    2014-10-15

    Protozoan diseases, such as leishmaniasis, are a cause of considerable morbidity throughout the world, affecting millions every year. In this study, two triterpenic acids (maslinic and oleanolic acids) were isolated from Tunisian olive leaf extracts and their in vitro activity against the promastigotes stage of Leishmania (L.) infantum and Leishmania (L.) amazonensis was investigated. Maslinic acid showed the highest activity with an IC50 of 9.32 ± 1.654 and 12.460 ± 1.25 ?g/ml against L. infantum and L. amazonensis, respectively. The mechanism of action of these drugs was investigated by detecting changes in the phosphatidylserine (PS) exposure, the plasma membrane permeability, the mitochondrial membrane potential and the ATP level production in the treated parasites. By using the fluorescent probe SYTOX® Green, both triterpenic acids showed that they produce a time-dependent plasma membrane permeabilization in the treated Leishmania species. In addition, spectrofluorimeteric data revealed the surface exposure of PS in promastigotes. Both molecules reduced the mitochondrial membrane potential and decreased the ATP levels to 15% in parasites treated with IC90 for 24h. We conclude that the triterpenic acids tested in this study, show potential as future therapeutic alternative against leishmaniasis. Further studies are needed to confirm this. PMID:25442278

  19. Leishmania (Leishmania) hertigi in a porcupine (Coendou sp.) found in Brasília, Federal District, Brazil / Leishmania (Leishmania) hertigi em porco-espinho (Coendou sp.) encontrado em Brasília, Distrito Federal, Brasil

    Scientific Electronic Library Online (English)

    Denise Amaro da, Silva; Maria de Fatima, Madeira; Carlos Jose Lima, Barbosa Filho; Edvar Yuri Paheco, Schubach; Juliana Helena da Silva, Barros; Fabiano Borges, Figueiredo.

    2013-06-01

    Full Text Available Alguns estudos relatam o encontro de Leishmania (Leishmania) hertigi em estados da região Norte do Brasil. Na presente investigação, os autores descrevem o isolamento de Leishmania (Leishmania) hertigi em um porco-espinho (Coendou sp.) encontrado em Brasília, Distrito Federal, região Centro-oeste do [...] Brasil. Durante um estudo de leishmaniose visceral canina conduzido na cidade de Brasília, Distrito Federal, um porco-espinho foi encontrado morto em via pública. O animal foi identificado e fragmentos de pele íntegra e baço foram coletados para isolamento do parasito em cultura. Esse relato da ocorrência de L. hertigi em outra região do Brasil pode contribuir para evidenciar a distribuição desse parasita no país. Vale ressaltar a necessidade de estudos para compreensão do papel L. hertigi na patologia e patogênese da leishmaniose e a sua sobrevivência em mamíferos e possíveis vetores. Abstract in english Studies report the occurrence of Leishmania (Leishmania) hertigi in northern states of Brazil. In the present investigation, we describe the isolation of L. (L.) hertigi from a porcupine (Coendou sp.) found in Brasília, Federal District, center-west region of Brazil. During a study on canine viscera [...] l leishmaniasis conducted in the city of Brasília, Federal District, a porcupine was found dead on a public road. The animal was identified and fragments of intact skin and spleen were collected for isolation of parasite in the culture. This report of the occurrence of L. hertigi in another part of Brazil may help establish the distribution of this parasite in the country. Further studies are needed to better understand the role of L. hertigi in the pathology and pathogenesis of leishmaniasis and its survival in mammals and possible vectors.

  20. Genomic Organization of Leishmania Species

    Directory of Open Access Journals (Sweden)

    B Kazemi

    2011-09-01

    Full Text Available Leishmania is a protozoan parasite belonging to the family Trypanosomatidae, which is found among 88 different countries. The parasite lives as an amastigote in vertebrate macro­phages and as a promastigote in the digestive tract of sand fly. It can be cultured in the laboratory us­ing appropriate culture media. Although the sexual cycle of Leishmania has not been observed during the promastigote and amastigote stages, it has been reported by some researchers. Leishma­nia has eukaryotic cell organization. Cell culture is convenient and cost effective, and because posttranslational modifications are common processes in the cultured cells, the cells are used as hosts for preparing eukaryotic recombinant proteins for research. Several transcripts of rDNA in the Leishmania genome are suitable regions for conducting gene transfer. Old World Leishmania spp. has 36 chromosomes, while New World Leishmania spp. has 34 or 35 chromo­somes. The genomic organization and parasitic characteristics have been investigated. Leishmania spp. has a unique genomic organization among eukaryotes; the genes do not have introns, and the chromosomes are smaller with larger numbers of genes confined to a smaller space within the nucleus. Leishmania spp. genes are organized on one or both DNA strands and are transcribed as polycistronic (prokaryotic-like transcripts from undefined promoters. Regulation of gene expres­sion in the members of Trypanosomatidae differs from that in other eukaryotes. The trans-splic­ing phenomenon is a necessary step for mRNA processing in lower eukaryotes and is observed in Leishmania spp. Another particular feature of RNA editing in Leishmania spp. is that mitochon­drial genes encoding respiratory enzymes are edited and transcribed. This review will discuss the chromosomal and mitochondrial (kinetoplast genomes of Leishmania spp. as well as the phenome­non of RNA editing in the kinetoplast genome.

  1. Occurrence of Leishmania (Leishmania chagasi in a domestic cat (Felis catus in Andradina, São Paulo, Brazil: case report Ocorrência de Leishmania (Leishmania chagasi em gato doméstico (Felis catus em Andradina, São Paulo, Brasil: relato de caso

    Directory of Open Access Journals (Sweden)

    Willian Marinho Dourado Coelho

    2010-12-01

    Full Text Available This work describes natural infection by Leishmania in a domestic cat where amastigote forms of the parasite were observed in the popliteal lymph node imprint. Positive and negative serological reactions were observed by enzyme-linked immunosorbent assay (ELISA and indirect immunofluorescence assay (IFA, respectively. Polymerase chain reaction (PCR revealed that the nucleotide sequence of the sample was identical to Leishmania (L. chagasi. This is the first report of the disease in felines of the city of Andradina, SP, an area considered endemic for canine and human visceral leishmaniasis.Neste trabalho, é relatada a infecção natural por Leishmania em um gato doméstico no qual, formas amastigotas do parasito foram observadas em imprint de linfonodo poplíteo. Reações sorológicas positivas e negativas foram observadas pelo teste de imunoadsorção enzimática (ELISA e reação de imunofluorescência indireta (RIFI, respectivamente. A reação em cadeia da polimerase (PCR revelou que a sequência de nucleotídeos foi idêntica à Leishmania (L. chagasi. Este é o primeiro relato da doença em felino da cidade de Andradina, Estado de São Paulo, Brasil, área considerada endêmica para leishmaniose visceral canina e humana.

  2. Miocardite crônica em um cão naturalmente infectado com Leishmania (Leishmania) infantum chagasi: aspectos clínicos e patológicos / Chronic myocarditis in a dog naturally infected by Leishmania (Leishmania) infantum chagasi: clinical and pathological aspects

    Scientific Electronic Library Online (English)

    R.S., Mendes; T.A., Gurjão; L.M., Oliveira; V.L., Santana; W.L., Tafuri; J.R.S., Santos; A.F.M., Dantas; A.P., Souza.

    2014-02-01

    Full Text Available A leishmaniose visceral (LV) é uma doença infecciosa crônica frequentemente fatal causada pela Leishmania infantum chagasi nas Américas. A enfermidade pode acometer vários órgãos, determinando diferentes manifestações clínicas. Contudo o envolvimento do coração raramente tem sido reportado em cães i [...] nfectados por Leishmania sp. Dessa forma, descreve-se um caso de miocardite crônica com repercussões clínicas e patológicas em um cão naturalmente infectado por Leishmania infantum chagasi. A positividade para Leishmaniose Visceral foi determinada pela presença de anticorpos antiLeishmania sp. nos testes sorológicos (RIFI, ELISA e DPP) e confirmada por visualização de formas amastigotas de Leishmania sp. em punção aspirativa do linfonodo poplíteo. O exame cardiovascular revelou alterações radiográficas, eletrocardiográficas, na pressão arterial e nos biomarcardores cardíacos. Após eutanásia, amostras de tecido cardíaco foram avaliadas histologicamente e submetidas à imunomarcação, onde foi observado infiltrado mononuclear (plasmo-histiolinfocitário), com presença de estruturas arredondadas de coloracão marrom-amareladas (imunomarcadas), indicando formas amastigotas de Leishmania infantum chagasi no miocárdio. Os aspectos etiopatogênicos da leishmaniose visceral sobre o miocárdio neste caso podem estar relacionados tanto à presença do parasita quanto à resposta "reacional inespecífica" do tecido à agressão do parasita no organismo. Todavia ainda não se sabem se as cepas de Leishmania infantum chagasi da região semiárida paraibana apresentam algum tropismo por tecido cardíaco ou se induzem a reação imunológica cruzada, com implicações clínicas. Abstract in english Visceral leishmaniasis (VL) is a chronic, often fatal infectious disease caused by Leishmania infantum chagasi in the Americas. The disease can affect many organs and may express different clinical forms. However, the involvement of the heart has rarely been reported in dogs infected by Leishmania s [...] p. Thus, we describe a case of chronic myocarditis with clinical and pathological effects in a dog naturally infected by Leishmania infantum chagasi. Positivity for Visceral Leishmaniasis was determined by the presence of anti-Leishmania sp. in serological tests (IFAT, ELISA and DPP) and confirmed by visualization of amastigote forms of the parasite in the popliteal lymph node aspiration samples. The cardiovascular clinical examination showed changes in the radiographic, ECG, blood pressure and heart biomarkers. After euthanasia, heart tissue samples were histologically examined and underwent our immunohistochemistry assessment, where mononuclear infiltrate was observed (plasma cells, lymphocytes and macrophages) with the presence of rounded brownish-yellow (immunomarked) cells, indicating amastigotes of Leishmania infantum chagasi within the myocardium. The etiopathogenic aspects of visceral leishmaniasis in the myocardium in this case may be related either to the presence of the parasite or the "nonspecific reactive" response of the tissue attributable to the aggression of the parasite in the body. However, it is not known if the strains of Leishmania infantum chagasi found in the semi-arid of Paraíba have some tropism for cardiac tissue or if they induce immunological cross-reaction with clinical implications.

  3. Leishmania(Leishmania) chagasi in captive wild felids in Brazil.

    Science.gov (United States)

    Dahroug, Magyda A A; Almeida, Arleana B P F; Sousa, Valéria R F; Dutra, Valéria; Turbino, Nívea C M R; Nakazato, Luciano; de Souza, Roberto L

    2010-01-01

    This study used a PCR-RFLP test to determine the presence of Leishmania (Leishmania) chagasi in 16 captive wild felids [seven Puma concolor (Linnaeus, 1771); five Panthera onca (Linnaeus, 1758) and four Leopardus pardalis (Linnaeus, 1758)] at the zoological park of the Federal University of Mato Grosso, Brazil. Amplification of Leishmania spp. DNA was seen in samples from five pumas and one jaguar, and the species was characterized as L. chagasi using restriction enzymes. It is already known that domestic felids can act as a reservoir of L. chagasi in endemic areas, and further studies are necessary to investigate their participation in the epidemiological chain of leishmaniasis. PMID:19740501

  4. Heterorhabditis amazonensis RSC5 (Rhabditida: Heterorhabditidae) movement and host recognition / Desplazamiento y capacidad de búsqueda del nematodo entomopatógeno nativo Heterorhabditis amazonensis RSC5 (Rhabditida: Heterorhabditida)

    Scientific Electronic Library Online (English)

    VANESSA, ANDALÓ; GRAZIELLE, FURTADO MOREIRA; ALCIDES, MOINO JUNIOR.

    2014-06-01

    Full Text Available La respuesta de Heterorhabditis amazonensis RSC5 en comparación con otras especies de nematodos entomopatógenos como Steinernema carpocapsae All y Steinernema riobrave 355, a los compuestos liberados por diferentes hospederos (Galleria mellonella, Mycotretus apicalis y Tenebrio molitor) y su nivel d [...] e virulencia a estos insectos fue evaluada. Las pruebas se realizaron en placas de Petri con agar-agua 2% para determinar el movimiento de los nematodos con y sin posibilidad de escogencia por diferentes hospederos. Se cuantificó la proximidad de juveniles infectivos (JIs) al hospedero como una fuente de atracción. Con el fin de determinar el desplazamiento de JIs en una condición similar al suelo, un ensayo se llevó a cabo en un área con arena. Los nematodos fueron virulentos para los hospederos. Cuando los nematodos e insectos fueron puestos en agar-agua, JIs se movieron hacia el estímulo, con preferencia de H. amazonensis a ciertos insectos. En la arena, S. carpocapsae causó menor mortalidad (70% ± 8,9 para G. mellonella) que H. amazonensis y S. riobrave (80% ± 6,5 y 99% ± 0,0). Heterorhabditis amazonensis fue capaz de encontrar y elegir a sus hospederos (G. mellonella y T. molitor) similar al comportamiento de S. riobrave y localizar sus hospederos con más eficacia que S. carpocapsae. De este modo, la virulencia de H. amazonensis fue similar a S. riobrave y esta característica podría ser promisoria para introducir esta especie nativa en programas de manejo integrado de plagas. Abstract in english Response of Heterorhabditis amazonensis RSC5 to compounds released by different host insects and its virulence level to several insect hosts like Galleria mellonella, Mycotretus apicalis and Tenebrio molitor were evaluated in this study, and compared with other entomopathogenic nematode species like [...] Steinernema carpocapsae All and Steinernema riobrave 355. Tests were performed in Petri dishes with agar-water 2% to determine nematode movement toward the insect with and without opportunity of choosing different insect hosts. Evaluations were made quantifying the proximity of infective juveniles (IJs) to the insect as a source of allurement. In order to determine the displacement of IJs in a closed soil condition, a test was carried out in an arena with sand. The nematode was virulent to the target insects. When nematode and insect were released on agar-water, IJs moved toward the stimulus, with H. amazonensis howing preference for certain insects. In the arena with sand S. carpocapsae caused lower insect mortality (70% ± 8.9 for G. mellonella) than H. amazonensis and S. riobrave (80% ± 6.5 and 99% ± 0.0). Heterorhabditis amazonensis was able to find and choose its hosts (G. mellonella and T. molitor), similarly to S. riobrave behavior, and located them more effectively than S. carpocapsae. The virulence of H. amazonensis was thus similar to S. riobrave, and this characteristic could be promising to introduce this native species in integrated pest management programs.

  5. Presença de formas amastigotas de Leishmania chagasi e perfil leucocitário no aparelho reprodutivo de cães

    OpenAIRE

    Ariane P. Benites; Fernandes, Carlos E.; Karine B. Brum; Maria Auxiliadora G.S. Abdo

    2011-01-01

    A leishmaniose visceral (LV) é uma zoonose causada pelo protozoário Leishmania (Leishmania) chagasi. A leishmaniose visceral canina (LVC) é a doença de maior relevância zoonótica. Usualmente, a infecção ocorre entre um hospedeiro invertebrado para um hospedeiro vertebrado, entretanto, a transmissão na ausência do vetor já é conhecida. O objetivo principal deste estudo foi identificar a presença de formas amastigotas, quantificar as células leucocitárias, estimar o risco relativo da presença d...

  6. RNA polymerase I promoter and splice acceptor site recognition affect gene expression in non-pathogenic Leishmania species

    Directory of Open Access Journals (Sweden)

    Tereza Cristina Orlando

    2007-11-01

    Full Text Available Leishmania (Sauroleishmania tarentolae has biotechnological potential for use as live vaccine against visceral leishmaniasis and as a system for the over expression of eukaryotic proteins that possess accurate post-translational modifications. For both purposes, new systems for protein expression in this non-pathogenic protozoan are necessary. The ribosomal RNA promoter proved to be a stronger transcription driver since its use yielded increased levels of recombinant protein in organisms of both genera Trypanosoma or Leishmania. We have evaluated heterologous expression systems using vectors with two different polypyrimidine tracts in the splice acceptor site by measuring a reporter gene transcribed from L. tarentolae RNA polymerase I promoter. Our data indicate that the efficiency of chloramphenicol acetyl transferase expression changed drastically with homologous or heterologous sequences, depending on the polypyrimidine tract used in the construct and differences in size and/or distance from the AG dinucleotide. In relation to the promoter sequence the reporter expression was higher in heterologous lizard-infecting species than in the homologous L. tarentolae or in the mammalian-infecting L. (Leishmania amazonensis.

  7. Histopathological and immunohistochemical study of the gastrointestinal tract from a dog naturally infected with Leishmania (Leishmania) chagasi: a case report / Histopatologia e imunoistoquímica do trato gastrintestinal em cão naturalmente infectado com Leishmania (Leishmania) chagasi: relato de caso

    Scientific Electronic Library Online (English)

    F.L., Silva; W.L., Tafuri; M.R., Oliveira; Wg. L., Tafuri.

    2002-08-01

    Full Text Available Um cão assintomático e naturalmente infectado com Leishmania (L.) chagasi foi sacrificado e fragmentos do estômago, duodeno, jejuno, íleo, ceco e cólon foram coletados para confecção de esfregaços por aposição corados pelo Giemsa ("imprints"). Outros fragmentos foram obtidos e fixados em formol tamp [...] onado a 10% para estudos histopatológicos e imunoistoquímicos. Empregou-se a técnica imunoistoquímica de estreptavidina-peroxidase a qual possibilitou detecção de formas amastigotas de Leishmaniachagasi em todos os segmentos do trato gastrintestinal (TGI). A principal lesão observada foi a reação inflamatória crônica de intensidade variável, localizada principalmente na mucosa e submucosa de todos os segmentos do TGI. O exsudato celular era composto de monócitos, plasmócitos e linfócitos. O estudo imunoistoquímico mostrou a presença de amastigotas de Leishmania em todos os fragmentos do TGI. A pesquisa de parasitas não pôde ser observada de forma satisfatória pela técnica da HE. Concluiu-se que a técnica de imunoistoquímica como diagnóstico da doença no caso de envolvimento gastrintestinal é eficaz. Abstract in english Samples of stomach, duodenum, jejunun, ileum, cecum and colon were collected for Giemsa-smears ("imprints") from one asymptomatic mongrel dog, naturally infected with Leishmania (L) chagasi. Other fragments were obtained and fixed in formalin (10% and buffered) for histopathological and immunohistoc [...] hemical studies. The immunohistochemistry was carried out by a streptavidin-peroxidase technique and it allowed to detect amastigote forms of Leishmania chagasi in the different paraffin gut sections. The principal lesion observed was a discrete to moderate chronic inflammatory reaction in the mucosa and submucosa in all fragments of the gastrointestinal tract (GIT). A chronic cellular exsudate was observed in all GIT tissues and it was composed by mononuclear cells (monocytes, plasmocytes and lymphocytes). A comparison between the two techniques showed that the immunohistochemistry study is the best method to detect amastigote forms of Leishmania.

  8. Amastigota forms resembling Leishmania sp. on corneal ulceration in a dog: case report Formas amastigotas compatíveis com Leishmania sp. em ulceração corneal de cão: relato de caso

    Directory of Open Access Journals (Sweden)

    F.L.C. Brito

    2007-02-01

    Full Text Available One dog with visceral leishmaniasis is reported presenting cutaneous lesions, cachexia, and corneal ulceration. Parasitological and serological diagnoses were performed by exam of contents of the aspirative punction of the bone marrow and by immunofluorecence, respectively. Amastigota forms resembling Leishmania sp. were visualized by citology.Relata-se o caso de um cão com leishmaniose visceral apresentando lesões cutâneas, caquexia e úlcera de córnea. Realizou-se o diagnóstico parasitológico e sorológico por meio de exame do material da medula óssea e por imunofluorescência indireta, respectivamente. À citologia da úlcera corneana, visibilizaram-se formas amastigotas compatíveis com Leishmania sp.

  9. Amastigota forms resembling Leishmania sp. on corneal ulceration in a dog: case report / Formas amastigotas compatíveis com Leishmania sp. em ulceração corneal de cão: relato de caso

    Scientific Electronic Library Online (English)

    F.L.C., Brito; L.C., Alves; F.C.L., Maia; A.R., Albuquerque; J.P.D., Ortiz; J.L., Laus.

    2007-02-01

    Full Text Available Relata-se o caso de um cão com leishmaniose visceral apresentando lesões cutâneas, caquexia e úlcera de córnea. Realizou-se o diagnóstico parasitológico e sorológico por meio de exame do material da medula óssea e por imunofluorescência indireta, respectivamente. À citologia da úlcera corneana, visi [...] bilizaram-se formas amastigotas compatíveis com Leishmania sp. Abstract in english One dog with visceral leishmaniasis is reported presenting cutaneous lesions, cachexia, and corneal ulceration. Parasitological and serological diagnoses were performed by exam of contents of the aspirative punction of the bone marrow and by immunofluorecence, respectively. Amastigota forms resembli [...] ng Leishmania sp. were visualized by citology.

  10. Ocorrência de Leishmania spp. em felinos do município de Araçatuba, SP Occurrence de Leishmania spp. in domestic cats from Araçatuba, SP

    Directory of Open Access Journals (Sweden)

    Katia Denise Saraiva Bresciani

    2010-06-01

    Full Text Available Este trabalho teve como objetivo comparar a ocorrência de Leishmania spp. em gatos por dois métodos (citológico e sorológico, bem como associar a ocorrência deste protozoário com as variáveis sexo, idade e raça. Amostras séricas de 283 felinos domésticos foram testadas pela Reação de Imunofluorescência Indireta (RIFI, e o exame parasitológico direto de linfonodos também foi realizado para a verificação da positividade para Leishmania spp. Ocorrência de 0,7% (2/283 foi observada nos felinos examinados, por meio de imprint de linfonodos e nenhum animal apresentou títulos de anticorpos para Leishmania spp. As duas fêmeas positivas eram sem raça definida, sendo uma jovem e outra adulta. Por meio dos resultados obtidos, não foi constatada diferença estatisticamente significante em relação às variáveis sexo, raça e idade nos gatos desta pesquisa (p > 0,05. Ocorrência de Leishmania spp. nos gatos deste estudo foi baixa. Devido a esta baixa incidência sugere-se que estes não assumem importância epidemiológica na área do estudo.This study had the purpose to compare the occurrence of Leishmania spp. in felines through two methods (cytological and serological, as well as to associate the occurrence of this protozoan with the sex, age and breed variables. Serum samples from 283 domestic felines were processed by means of Indirect Immunofluorescence Reaction (IIR, and the direct parasitological test for linfonodes was also carried out in order to verify positivity for Leishmania spp. Occurrence of 0.7% (2/283 was observed in the tested felines by means of linfonode imprinting and no animal showed title of antibodies for Leishmania spp. The two positive females were mongrel, a young female and an adult female feline. From the obtained results, no statistically significant difference was observed as regards the sex, breed and age variables in this research (p > 0.05. Occurrence of Leishmania spp. in the cats of this study was low. Such low incidence suggests that these hosts has no epidemiological relevance in the study area.

  11. Xiphidorus amazonensis n. sp. (Nematoda: Longidoridae) from the Brazilian Amazon Basin

    OpenAIRE

    Uesugi, C. H.; Huang, C.S.; Cares, J. E.

    1985-01-01

    Xiphidorus amazonensis n. sp. was found in the rhizospheres of Jatropha curcas, Musa sp., Anona muricata, Cassia tora, Panicum laxum, Paspalum fasciculatum, Aeschynomene sensitiva, Saccharum officinarum, Manihot esculenta, Abelmoschus esculentus, Tamarindus indica, Mangifera indica, Vigna unguiculata, Zea mays, Commelina sp., Cyperus rotundus, Fimbristylis miliacea, Citrus sinensis, and Eichhornia crassipes on the Amazon River island of Xiborena, approximately 40 km southeast of Manaus, capit...

  12. Prevalence and Distribution of Leishmania RNA Virus 1 in Leishmania Parasites from French Guiana.

    Science.gov (United States)

    Ginouvès, Marine; Simon, Stéphane; Bourreau, Eliane; Lacoste, Vincent; Ronet, Catherine; Couppié, Pierre; Nacher, Mathieu; Demar, Magalie; Prévot, Ghislaine

    2016-01-01

    In South America, the presence of the Leishmania RNA virus type 1 (LRV1) was described in Leishmania guyanensis and Leishmania braziliensis strains. The aim of this study was to determine the prevalence distribution of LRV1 in Leishmania isolates in French Guiana given that, in this French overseas department, most Leishmania infections are due to these parasite species. The presence of the virus was observed in 74% of Leishmania spp. isolates, with a highest presence in the internal areas of the country. PMID:26598572

  13. Leishmania major, the Predominant Leishmania Species Responsible for Cutaneous Leishmaniasis in Mali

    OpenAIRE

    Paz, Carlos; Samake, Sibiry; Anderson, Jennifer M.; Faye, Ousmane; Traore, Pierre; Tall, Koureishi; Cisse, Moumine; Keita, Somita; Valenzuela, Jesus G.; DOUMBIA, SEYDOU

    2013-01-01

    Leishmania major is the only species of Leishmania known to cause cutaneous leishmanisis (CL) in Mali. We amplified Leishmania DNA stored on archived Giemsa-stained dermal scraping slides obtained from self-referral patients with clinically suspected CL seen in the Center National d'Appui A La Lutte Contre La Maladie (CNAM) in Bamako, Mali, to determine if any other Leishmania species were responsible for CL in Mali and evaluate its geographic distribution. Polymerase chain reaction (PCR) amp...

  14. Xiphidorus amazonensis n. sp. (Nematoda: Longidoridae) from the Brazilian Amazon Basin.

    Science.gov (United States)

    Uesugi, C H; Huang, C S; Cares, J E

    1985-07-01

    Xiphidorus amazonensis n. sp. was found in the rhizospheres of Jatropha curcas, Musa sp., Anona muricata, Cassia tora, Panicum laxum, Paspalum fasciculatum, Aeschynomene sensitiva, Saccharum officinarum, Manihot esculenta, Abelmoschus esculentus, Tamarindus indica, Mangifera indica, Vigna unguiculata, Zea mays, Commelina sp., Cyperus rotundus, Fimbristylis miliacea, Citrus sinensis, and Eichhornia crassipes on the Amazon River island of Xiborena, approximately 40 km southeast of Manaus, capital of the State of Amazonas. The type habitat is flooded annually for about 6 months by the Amazon River. Xiphidorus amazonensis n. sp. differs from the closely related species Xiphidorus yepesara Monteiro, 1976 by the larger size, by a, b, and c values, and by the rounded tail terminus. It also resembles Xiphidorus tucumanensis Chaves and Coomans, 1984, but can be distinguished by its larger size, larger a, b, and c values, more conical female tail, bilobed amphidial pouch, and the presence of a spermatheca full of sperm. PMID:19294098

  15. Vaccine to protect animals against leishmania

    OpenAIRE

    Aseginolaza González, Carlos; Ibáñez, Ramiro; Hernández Castillo, J. A.; Curdi Lucientes, J.

    2009-01-01

    The present invention is generally related to the prevention of leishmaniasis in animals, particularly infection caused by Leishmania sp., based on the use of Leishmania infantum P36 protein or an immunogenic fragment of the latter, or involving an expression system for the mentioned protein or fragment - optionally in combination with a compound stimulating the production of a Th1-type cellular immune response - and comprising various vaccination protocols in application to Leishmania sp. ba...

  16. Infecção natural de Lutzomyia longipalpis por Leishmania sp. em Teresina, Piauí, Brasil Natural infection of Lutzomyia longipalpis by Leishmania sp. in Teresina, Piauí State, Brazil

    Directory of Open Access Journals (Sweden)

    Jackellyne Geórgia Dutra e Silva

    2007-07-01

    Full Text Available No Brasil, o controle do vetor Lutzomyia longipalpis é uma das principais estratégias utilizadas para limitar a expansão da leishmaniose visceral americana. Entretanto, poucos são os estudos sobre a infecção natural destes flebotomíneos por espécies de Leishmania. Um estudo sobre a infecção natural de Lu. longipalpis por Leishmania sp. foi realizado no bairro Bela Vista, Teresina, Piauí, um dos principais focos urbanos da leishmaniose visceral americana no Brasil. Entre fevereiro de 2004 e janeiro de 2005 realizaram-se 180 capturas usando-se armadilhas luminosas do tipo CDC. Foram dissecadas e examinadas, em média, dez fêmeas por captura para detecção de formas evolutivas de Leishmania sp., identificando-se 1.832 exemplares de Lu. longipalpis e seis de Lu. whitmani. Vinte (1,1% espécimes, todas de Lu. longipalpis, estavam infectadas com as formas procíclica e nectomonada de Leishmania sp., localizadas, principalmente, na porção posterior do trato digestivo. O maior percentual de insetos infectados foi encontrado quatro meses após o período de maior precipitação pluviométrica, sugerindo que variáveis climáticas podem contribuir para a predição não apenas da abundância destes dípteros, mas também do seu grau de infecção natural.In Brazil, control of the vector Lutzomyia longipalpis is one of the main strategies used to limit the expansion of American visceral leishmaniasis. However, studies on the ecology of this sand fly are rare, especially regarding its natural infection with species of Leishmania. A study of the natural infection of Lu. longipalpis by Leishmania sp. was carried out in the Bela Vista neighborhood in the city of Teresina, Piauí State, Brazil, an important area of American visceral leishmaniasis transmission. From February 2004 to January 2005, sand flies were captured with CDC light traps. Approximately 10 female sand flies in each capture were dissected and examined for the presence of evolutionary forms of Leishmania sp. Two sand fly species were identified: 1,832 were Lu. longipalpis and six Lu. whitmani. Twenty female sand flies (1.1%, all Lu. longipalpis, were infected with procyclic and nectomonad forms of Leishmania sp., found mostly in the hindgut. Higher proportions of infected sand flies were found four months after the rainy season, suggesting that environmental factors may predict not only vector abundance (as already known but also their level of infection.

  17. Activity of Fabaceae species extracts against fungi and Leishmania: vatacarpan as a novel potent anti-Candida agent

    Directory of Open Access Journals (Sweden)

    Dandara Braga Santana

    2015-08-01

    Full Text Available AbstractLeishmaniasis and fungal infection treatment efficacy is limited by toxicity and ever increasing resistance to available drugs, requiring development of alternative compounds. The richness of Cerrado plant antimicrobial secondary metabolites justifies screening of Fabaceae species extracts: Enterolobium ellipticum Benth., Sclerolobium aureum (Tul. Baill. and Vatairea macrocarpa(Benth. Ducke, against Leishmania(Leishmania amazonensis, yeasts and dermatophytes. Among the 26 extracts tested, more than 50% of the total demonstrated significant antifungal activity in comparison to the drug controls (minimal inhibitory concentration 0.12 to ?31.25 µg/ml. Six extracts capable of complete parasitic growth inhibition had the inhibitory concentration index for 50% values from 9.23 to 78.65 µg/ml. The results led to the selection of the V. macrocarpa ethyl acetate root bark extract for chemical fractionation. This plant, traditionally referred to as angelim-do-cerrado or maleiteira, is used to treat superficial mycoses in Amazonia. A previously unreported pterocarpan vatacarpan together with the known compound musizin was isolated. Vatacarpan demonstrated a minimal inhibitory concentration value of 0.98 µg/ml against Candida albicans ATCC 10231, and thus comparable or superior to fluconazole and amphotericin B. The results add to literature's information the ability of pterocarpans to act as antimicrobial agents.

  18. Evaluacion por Western Blot, Inmunofluorescencia Indirecta y ELISA de Perros Infectados con Leishmania (Leishmania infantum Western blot, ELISA and indirect immunofluorescence test evaluation of Leishmania (Leishmania infantum-infected dogs

    Directory of Open Access Journals (Sweden)

    Jimmy J Vargas-Duarte

    2009-08-01

    Full Text Available Objetivo Evaluar el desempeño de las pruebas empleadas en Colombia para el diagnóstico de la leishmaniasis visceral canina y adaptar una técnica de Western blot empleando animales experimental y naturalmente infectados. Metodología Se obtuvieron sueros de 10 perros infectados experimentalmente con L. infantum, 5 perros infectados naturalmente, 16 perros sanos, 26 de reacción cruzada (infectados con Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi, Leishmania (Viannia spp., 40 de zonas no endémicas y 150 de zona endémica. Todos fueron evaluados mediante las pruebas de inmunofluorescencia indirecta (IFI, ELISA y Western blot (WB. Resultados Se encontró que IFI tuvo el mayor porcentaje de positividad en los perros infectados (73 % mientras que el menor porcentaje de falsos positivos se obtuvo por WB (2,5 %. La prueba de ELISA fue la menos eficiente. Fueron reconocidas 24 fracciones antigénicas, las bandas de 29, 34, 50, 69, 75, 86, 99 y 123 kDa fueron responsables de reacciones inespecíficas en los sueros de perros sanos, de zona no endémica y de reacción cruzada. Las bandas por debajo de 29 kDa mostraron ser potencialmente diagnósticas, especialmente la fracción de 13 kDa. Conclusiones Los métodos directos y serológicos pueden subdiagnosticar la infección por Leishmania, solamente un constructo que combine tanto pruebas directas como indirectas sería la forma más eficiente de diagnóstico.Objective Evaluating canine visceral leishmaniasis diagnostic test performance in Colombia and adapting the Western blot test in naturally and experimentally infected dogs. Methods Sera were obtained from 10 experimentally L. Infantum-infected dogs, 5 naturally infected dogs, 16 healthy dogs, 26 Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi and Leishmania (Viannia spp infected dogs, 40 dogs from non-endemic areas and 150 from endemic areas. Sera were tested for L. infantum infection using immunofluorescent antibody (IFAT, ELISA and Western blot (WB tests. Results Positives results were obtained for 73 % of known infected dogs by the IFAT test and false positives were obtained for 2.5 % of non-infected dogs using WB. ELISA was not efficient for diagnosis. 24 antigenic fractions were recognised in tested sera using WB; however, 29, 34, 50, 69, 75, 86, 99 and 123 kDa bands were recognised in sera from dogs from non-endemic areas, healthy dogs and Trypanosoma cruzi, Erhlichia canis, Dirofilaria immitis and Babesia canis infected dogs. The 13 kDa fraction proved potentially useful for diagnosing canine visceral leishmaniasis. ConclusionsThe separate use of parasitological and serological test could lead to misdiagnosis of Leishmania infection; using both kinds of technique simultaneously is thus highly recommended.

  19. Estudo, ao microscópio óptico e eletrônico, do rim de caes natural e experimentalmente infectados com Leishmania (Leishmania) chagasi / Optical and electron microscopical studies of canine kidney naturally and experimentally infected with Leishmania (Leishmania) chagasi

    Scientific Electronic Library Online (English)

    Washington Luiz, Tafuri; Marilena Suzan Marques, Michalick; Magno, Dias; Odair, Genaro; Virginia Hora Rios, Leite; Alfredo José Afonso, Barbosa; Eduardo Alves, Bambirra; Carlos Alberto Da, Costa; Maria Norma, Melo; Wilson, Mayrink.

    1989-06-01

    Full Text Available Os autores estudam os rins de 4 cães infectados com Leishmania (Leishmania) chagasi. Dois animais (um macho e uma fêmea) naturalmente infectados foram sacrificados 18 meses após sua permanência no laboratório. Dois machos foram inoculados por via endovenosa, com lxlO6 promastigotas da cepa MHO/BR/70 [...] /BH46 e sacrificados após 18 meses e 2 anos, respectivamente. Em todos os animais os rins estavam lesados. As alterações encontradas foram: (1) glomerulonefrite mesangioproliferativa focal ou difusa, com pronunciada hipertrofia e hiperplasia das células mesangiais e com alargamento da matriz; (2) espessamento da membrana basal com depósitos eletrondensos; (3) nefrite intersticial intertubular crônica com exsudação plasmocitária intensa. (4) degeneração albuminosa dos túbulos renais. Baseados nos achados os autores discutem os prováveis mecanismos patogenéticos. Abstract in english Two naturally infected dogs (male and fema lei from Teófilo Otoni (MG Brazili were maintained for 18 months in our laboratory. Two other dogs, two months old males were infected with 1 x 10(6) promastigotes of MHO BR 70 BH46 Leishinania (Leishmanial chagasi strain, endo venous route, and autopsied a [...] fter 10 months and two years. The main findings concerning the kidney were: (1) focal or diffuse mesangial glomerulo nephritis with proliferative and enlargement of mesangial cells; (2) increase in thickness of basement membrane with electron dense deposits: (3) chronic interstitial nephritis with intense exudation of plasmocytes: (4) cloud swelling of renal tubules. The authors discuss the probable pathogenetic mechanisms.

  20. Anticorpos antipromastigotas vivas de Leishmania (Viannia braziliensis, detectados pela citometria de fluxo, para identificação da infecção ativa na leishmaniose tegumentar americana Anti-live Leishmania (Viannia braziliensis promastigote antibodies, detected by flow cytometry, to identify active infection in american cutaneous leishmaniasis

    Directory of Open Access Journals (Sweden)

    Roberta Dias Rodrigues Rocha

    2002-12-01

    Full Text Available Neste estudo, descrevemos etapas iniciais de padronização de uma nova metodologia para detecção de anticorpos antipromastigotas vivas de Leishmania (Viannia braziliensis, pela citometria de fluxo e a análise de sua aplicabilidade para estudos clínicos. Foram avaliados 39 indivíduos com sorologia convencional (RIFI positiva para leishmaniose, classificados quanto à ausência/presença de lesão (L- e L+. Os resultados foram expressos sob a forma de percentual de parasitas fluorescentes positivos (PPFP. A análise dos dados, na diluição 1:1.024, permitiu distinguir 95% dos pacientes L+ como um grupo de alta reatividade (PPFP>50% e 72% dos indivíduos L- como um grupo de baixa reatividade (PPFPIn the current study we described initial standardization steps of a new methodology to detect anti-live Leishmania (Viannia braziliensis promastigote antibodies by flow cytometry, followed by analysis of its applicability to clinical studies. We have studied 39 individuals with positive conventional serology to leishmaniasis, classified according to the absence/presence of cutaneous lesions (L- and L+. The results were expressed as percentage of positive fluorescent parasites (PPFP. Data analysis at dilution of 1:1,024, allowed the distinction of 95% of L+ patients as a group of high reactivity (PPFP>50% and 72% of L- individuals as a group of low reactivity (PPFP<50%. The analysis of immunofluorescence assay titers did not show any relationship with the absence/presence of lesion. Together, our data support the applicability of flow cytometry to identify cases of active infection, which has not been possible through conventional serological reactions.

  1. The genetics of Leishmania virulence.

    Science.gov (United States)

    Bifeld, Eugenia; Clos, Joachim

    2015-12-01

    The ability of Leishmania parasites to infect and persist in the antigen-presenting cell population of their mammalian hosts is dependent on their ability to gain entry to their host and host cells, to survive the mammalian cell environment, and to suppress or evade the protective immune response mechanisms of their hosts. A multitude of genes and their products have been implicated in each of these virulence-enhancing strategies to date, and we present an overview of the nature and known function of such virulence genes. PMID:26047933

  2. Miocardite crônica em um cão naturalmente infectado com Leishmania (Leishmania) infantum chagasi: aspectos clínicos e patológicos

    OpenAIRE

    Mendes, R. S.; T.A. Gurjão; L. M. Oliveira; V.L. Santana; Tafuri, W. L.; J.R.S. Santos; A.F.M. Dantas; A.P. Souza

    2014-01-01

    A leishmaniose visceral (LV) é uma doença infecciosa crônica frequentemente fatal causada pela Leishmania infantum chagasi nas Américas. A enfermidade pode acometer vários órgãos, determinando diferentes manifestações clínicas. Contudo o envolvimento do coração raramente tem sido reportado em cães infectados por Leishmania sp. Dessa forma, descreve-se um caso de miocardite crônica com repercussões clínicas e patológicas em um cão naturalmente infectado por Leishmania infantum chagasi. A posit...

  3. Quantiferon-Leishmania as an Epidemiological Tool for Evaluating the Exposure to Leishmania Infection

    OpenAIRE

    Turgay, Nevin; Balcioglu, I. Cuneyt; Toz, Seray Ozensoy; Ozbel, Yusuf; Jones, Stephen L.

    2010-01-01

    The aim of the present preliminary study was to investigate the potential of measurement of IFN-? secretion by T cells into blood plasma using QuantiFERON assay with leishmanial antigens to determine the presence of Leishmania infection. Blood samples from cured visceral (N = 18), and cutaneous (N = 20) leishmaniasis cases, and 20 healthy controls were tested. The IFN-? responses to Leishmania major H2B and Leishmania infantum H2B antigens were detected from the majority of treated old viscer...

  4. Identificación de una secuencia de ADN genómico de Leishmania especifica del subgénero Leishmania / Identification of a genomic DNA sequence of Leishmania, specific of Leishmania subgenus

    Scientific Electronic Library Online (English)

    Andrea, Orué; Nancy Y, De Abreu; Clara, Martínez; Alexis, Mendoza-León.

    2008-06-01

    Full Text Available Leishmania es el agente causante de la compleja enfermedad conocida como leishmaniasis. Las distintas especies de este parásito protozoario se encuentran agrupadas en dos subgéneros, Viannia y Leishmania, de acuerdo a su desarrollo en el mosquito vector. Un ensayo de PCR, ?500-PCR, específico del su [...] bgénero Viannia, ha sido desarrollado utilizando la secuencia de ADN genómico denominada ?500. En este trabajo se presenta el aislamiento e identificación de una secuencia genómica de 280 pb, L280, a partir del ADN genómico de Leishmania (Leishmania) mexicana luego de aplicar el ensayo ?500-PCR en condiciones de baja rigurosidad. La secuenciación parcial de L280 permitió diseñar un ensayo de PCR (L280-PCR) que generó un producto de amplificación de 260 pb, en distintas condiciones de rigurosidad, cuando se utilizó el ADN genómico de distintas especies pertenecientes al subgénero Leishmania. El ensayo L280-PCR resultó negativo para el ADN genómico de distintas especies del subgénero Viannia al igual que para el ADN de otros organismos kinetoplastidos o humano. Los resultados sugieren que el ensayo L280-PCR es específico del subgénero Leishmania. Abstract in english Leishmania is the causal agent of the leishmaniasis disease. The different species of this protozoa parasite are grouped in two subgenera, Viannia and Leishmania, according to their development in the sandfly vector. A specific PCR assay, ?500-PCR, has been developed for the Viannia subgenus using t [...] he genomic ?500 DNA sequence. In the present work we present the isolation and identification of a genomic sequence of 280 bp, L280, obtained from genomic DNA of Leishmania (Leishmania) mexicana after application of the ?500-PCR assay at low stringency. After partial sequencing of L280 a PCR assay was generated, L280-PCR, this yielded a product of 260 bp at different conditions of stringency, when genomic DNA of different species of Leishmania subgenus was used. The L280-PCR assay was negative to genomic DNA of species belonging to the Viannia subgenus and also to other kinetoplastid organisms and human. The results suggest specificity of the L280-PCR assay for the Leishmania subgenus.

  5. Characterization of a Monoclonal Antibody Specific for the Parasite Surface Antigen-2 of Leishmania major

    Directory of Open Access Journals (Sweden)

    "AR Khabiri

    2004-08-01

    Full Text Available The Leishmania major Parasite surface Antigen-2 (PSA-2 is a family of glycoinositol phospholipids anchored glycoprotoins expressed in both promastigotes and amastigotes. Promastigote PSA-2 comprises three polypeptides with approximate molecular weight of 96, 80 and 50 kDa. Amastigote express a distinct but closely PSA-2 polypeptide with molecular weight of 50 kDa. In this study fusion of SP2/0 myeloma cells with immunized mice spleenocytes infected with promastigotes of L. major intraperitoneally resulted to a clone of hybridoma producing a specific antibody that only reacts with L. major parasite surface antigen (PSA-2. This mAb showed no crossreactivity with either other Leishmania species including L. tropica, L. donovani and L. infantum or recombinant gp63. Western blot analysis of culture supernatant revealed multiple bands with molecular weight of 50, 58, 80 and 96 kDa only in L. major.

  6. Leishmania (infantum) chagasi in canine urinary sediment / Leishmania (infantum) chagasi em sedimento urinário canino

    Scientific Electronic Library Online (English)

    Ivete Lopes de, Mendonça; Joilson Ferreira, Batista; Leucio Camara, Alves.

    2015-03-01

    Full Text Available A leishmaniose visceral canina (LVC) é uma doença de difícil diagnóstico. Principalmente devido à presença de animais assintomáticos, a diversidade da sintomatologia clínica apresentada e também pela dificuldade em se obter uma prova diagnóstica que reúna alta sensibilidade e especificidade. O objet [...] ivo deste trabalho foi relatar a presença de L. (infantum) chagasi em meio de cultura, utilizando-se sedimento urinário. Foram utilizados neste experimento, 70 cães provenientes do Hospital Veterinário Universitário da Universidade Federal do Piauí e do Centro de Controle de Zoonoses de Teresina, com raça, sexo e idade variada. Foram realizados exames sorológicos: TR DPP® Leishmaniose Visceral Canina (DPP) e Ensaio Imunoenzimático Leishmaniose Visceral Canina (ELISA), exames parasitológicos de amostras de medula e/ou linfonodo e cultura de sedimento urinário. Em 61,0% (43/70) dos animais estudados, observou-se presença de Leishmania em medula e/ou linfonodo, e destes 9,30% (4/43) foram positivos na cultura de sedimento urinário. Nos exames sorológicos, 70,0% (49/70) dos animais apresentavam-se reativos no DPP e 78,2% (55/70) no ELISA. Pode-se concluir, neste estudo, que é possível diagnosticar a LVC por meio da cultura de sedimento urinário. Abstract in english Canine visceral leishmaniasis (CVL) is difficult to diagnosis, mainly due to the presence of asymptomatic animals, the diversity of clinical symptoms and the difficulty in obtaining diagnostic evidence of high sensitivity and specificity. The purpose of this study was to diagnose CVL in urinary sedi [...] ment of 70 dogs of different breeds, sexes and ages from the veterinary hospital of the Federal University of Piauí and Zoonosis Control Center of Teresina, Brazil. The serological tests were TR DPP® for CVL and enzyme-linked immunosorbent assay (ELISA) for CVL, parasitological exams of bone marrow and lymph nodes and urine sediment cultures. Leishmania was detected in the bone marrow and/or lymph node of 61.0% of the animals (43/70), and urine sediment culture was positive in 9.30% (4/43) of these animals. In the serological exams, 70.0% (49/70) were reactive using the DPP and 78.2% (55/70) were reactive using ELISA. The goal of this study was to diagnose the presence of L. (infantum) chagasi in a culture of urinary sediment.

  7. Phenotypic characterization of Leishmania spp. causing cutaneous leishmaniasis in the lower Amazon region, western Pará state, Brazil, reveals a putative hybrid parasite, Leishmania (Viannia) guyanensis × Leishmania (Viannia) shawi shawi

    Science.gov (United States)

    Jennings, Yara Lins; de Souza, Adelson Alcimar Almeida; Ishikawa, Edna Aoba; Shaw, Jeffrey; Lainson, Ralph; Silveira, Fernando

    2014-01-01

    We phenotypically characterized 43 leishmanial parasites from cutaneous leishmaniasis by isoenzyme electrophoresis and the indirect immunofluorescence antibody test (23 McAbs). Identifications revealed 11 (25.6%) strains of Leishmania (V.) braziliensis, 4 (9.3%) of L. (V.) shawi shawi, 7 (16.3%) of L. (V.) shawi santarensis, 6 (13.9%) of L. (V.) guyanensis and L. (V.) lainsoni, 2 (4.7%) of L. (L.) amazonensis, and 7 (16.3%) of a putative hybrid parasite, L. (V.) guyanensis/L. (V.) shawi shawi. McAbs detected three different serodemes of L. (V.) braziliensis: I-7, II-1, and III-3 strains. Among the strains of L. (V.) shawi we identified two populations: one (7 strains) expressing the B19 epitope that was previously considered to be species-specific for L. (V.) guyanensis. We have given this population sub-specific rank, naming it L. (V.) s. santarensis. The other one (4 strains) did not express the B19 epitope like the L. (V.) shawi reference strain, which we now designate as L. (V.) s. shawi. For the first time in the eastern Brazilian Amazon we register a putative hybrid parasite (7 strains), L. (V.) guyanensis/L. (V.) s. shawi, characterized by a new 6PGDH three-band profile at the level of L. (V.) guyanensis. Its PGM profile, however, was very similar to that of L. (V.) s. shawi. These results suggest that the lower Amazon region – western Pará state, Brazil, represents a biome where L. (V.) guyanensis and L. (V.) s. shawi exchange genetic information. PMID:25083790

  8. Chronic interstitial pneumonitis in dogs naturally infected with Leishmania (Leishmania) chagasi: a histopathological and morphometric study / Pneumonia intersticial crônica em cães naturalmente infectados com Leishmania (Leishmania) chagasi: estudo histopatológico e morfométrico

    Scientific Electronic Library Online (English)

    Ricardo, Gonçalves; Washington Luiz, Tafuri; Maria Norma de, Melo; Pedro, Raso; Wagner Luiz, Tafuri.

    2003-06-01

    Full Text Available Neste estudo, foram utilizados dezoito cães naturalmente infectados com Leishmania (Leishmania) chagasi, sem raça e idade definidos, cedidos pelo Centro de Contole de Zoonoses da Cidade de Belo Horizonte. Quatro cães foram utilizados como controle. Durante a necropsia, amostras do pulmão foram colet [...] adas, e imediatamente fixadas em formalina. A histopatologia mostrou principalmente uma pneumonite intersticial crônica, produtiva, difusa e intensa. Os septos inter-alveolares espessados, eram constituídos principalmente por um infiltrado de células inflamatórias, em sua maioria, macrófagos, linfócitos e plasmócitos, além de fibroblastos e fibras colágenas. A análise morfométrica do espessamento septal mostrou diferença significativa entre animais infectados e controle. De fato, a análise morfométrica do colágeno, corado pela prata, confirmou um aumento na deposição de colágeno em todos os animais infectados. O exame parasitológico foi ralizado durante a análise histopatológica, em todos os casos estudados. Contudo neste trabalho, não observamos qualquer correlação entre os achados histopatológicos e morfométricos quando relacionados aos cães em diferentes formas clínicas. Concluímos que as lesões, encontradas nos cães naturalmente infectados, tem correlação com a doença, e que o método de análise morfométrica utilizado apresentou resultados satisfatórios quanto à análise do espessamento septal e ao aumento da deposição de colágeno, confirmando a fibrose. Abstract in english Eighteen mongrel dogs of unknown age and naturally infected with Leishmania (Leishmania) chagasi, were obtained from the City Hall of Belo Horizonte, Brazil. Four dogs were used as control. Lung samples were obtained and immediately fixed in formalin. The histopathological picture of all lung tissue [...] sections was a chronic and diffuse interstitial pneumonitis. The thickened inter-alveolar septa were characterized by the cellular exudate (mostly macrophages, lymphocytes and plasmocytes) associated with collagen deposition. Morphometric analysis showed greater septal thickness in the infected animals than in controls. In fact, the morphometric study of collagen stained with ammoniac silver confirmed a larger deposition of collagen in the infected animals. The parasitologic method was carried out during the study of the lesions on the slides. However, we did not observe any correlation between the histopathologic and morphometric data and the clinical status of the animals. We conclude that the pulmonary lesions observed in all naturally infected dogs were correlated with the disease and that the morphometric method used was satisfactory for the analysis of septal thickness and of increased collagen deposition, confirming the presence of fibrosis.

  9. Leishmania infantum leishmaniasis in corticosteroid – treated patients

    OpenAIRE

    De Marco Michele; Ghirga Piero; Spinazzola Francesco; Nicastri Emanuele; Pittalis Silvia; Paglia Maria; Narciso Pasquale

    2006-01-01

    Abstract Background The number of leishmaniasis cases associated with immunosuppression has increased regularly over the past 20 years. Immunosuppression related to HIV infection, immunosuppressive treatment, organ transplantation, and neoplastic diseases increases the risk for Leishmania-infected people to develop visceral illness. Case presentation Three cases of Leishmania infantum leishmaniasis in corticosteroid (CS)-treated patients are reported: an isolated lingual leishmaniasis in a fa...

  10. Susceptibility of laboratory-reared female Lutzomyia longipalpis (Lutz & Neiva, 1912) to infection by different species and strains of Leishmania Ross, 1903

    Scientific Electronic Library Online (English)

    Ana Lúcia F. F. da, Silva; Paul, Williams; Maria Norma, Melo; Wilson, Mayrink.

    1990-12-01

    Full Text Available A study was undertaken to compare the susceptibility of laboratory-reared female Lutzomyia longipalpis to infection by different species or strains of New World Leishmania. The sand flies proved to be highly susceptible to infection by a strain of Le. guyanensis, with flagellates developing in all ( [...] 18/18) of the specimens examined. A lower infection rate of 37 per cents (10/27) was recorded in flies exposed to infection by a strain of Le. amazonensis. Flagellates developed in 13 per cents (6/46) of the sand flies that glood fed on dogs in the earlly stage of experimental infection with an old laboratory strain of Le. chagasi. In contrast, promastigotes did not develop in sand flies that blood fed on dogs with naturally acquired Le. chagasi. The naturally infected dogas were in an advanced stage of disease. Flagellates developed in 9// (3/32) of the sand flies that blood fed on lesions of hamsters infected with a strain of Le. braziliensis and in 9 per cents (3/34) of those that fed on hamsters with lesions due to a parasite fo the mexicana complex (strain MHOM/BR/73/BH121). Sand flies did not develop flagellate infections after blood feeding on hamsters bearing lesions induced by strain MHOM/BR/71/BR49. Factors influencing the susceptibility of Lu. longipalpis to infection by New World species of Leishmania are discussed.

  11. Susceptibility of laboratory-reared female Lutzomyia longipalpis (Lutz & Neiva, 1912 to infection by different species and strains of Leishmania Ross, 1903

    Directory of Open Access Journals (Sweden)

    Ana Lúcia F. F. da Silva

    1990-12-01

    Full Text Available A study was undertaken to compare the susceptibility of laboratory-reared female Lutzomyia longipalpis to infection by different species or strains of New World Leishmania. The sand flies proved to be highly susceptible to infection by a strain of Le. guyanensis, with flagellates developing in all (18/18 of the specimens examined. A lower infection rate of 37 per cents (10/27 was recorded in flies exposed to infection by a strain of Le. amazonensis. Flagellates developed in 13 per cents (6/46 of the sand flies that glood fed on dogs in the earlly stage of experimental infection with an old laboratory strain of Le. chagasi. In contrast, promastigotes did not develop in sand flies that blood fed on dogs with naturally acquired Le. chagasi. The naturally infected dogas were in an advanced stage of disease. Flagellates developed in 9// (3/32 of the sand flies that blood fed on lesions of hamsters infected with a strain of Le. braziliensis and in 9 per cents (3/34 of those that fed on hamsters with lesions due to a parasite fo the mexicana complex (strain MHOM/BR/73/BH121. Sand flies did not develop flagellate infections after blood feeding on hamsters bearing lesions induced by strain MHOM/BR/71/BR49. Factors influencing the susceptibility of Lu. longipalpis to infection by New World species of Leishmania are discussed.

  12. Infecção por Leishmania (Leishmania chagasi em crianças de uma área endêmica de leishmaniose visceral americana na Ilha de São Luis-MA, Brasil

    Directory of Open Access Journals (Sweden)

    Caldas Arlene J.M.

    2001-01-01

    Full Text Available Realizou-se estudo prospectivo com 648 crianças de zero a cinco anos no município da Raposa-MA, de julho/97 a junho/98, com o objetivo de avaliar as características da infecção por L.(L.chagasi e verificar se existe associação entre desnutrição e infecção assintomática. Utilizou-se questionário com dados socioeconômicos, ambientais e hábitos de vida; realizou-se Intradermorreação de Montenegro(IDRM com antígeno de L. amazonensis e Enzyme Linked Immunosorbant Assay(ELISA para detectar infecção, e exame antropométrico. A prevalência inicial, final e incidência da infecção foram 18,6%, 20,6% e 10,8% pelo IDRM, e 13,5%, 34,4% e 28% pelo ELISA, respectivamente. A prevalência da desnutrição crônica (altura/idade foi 26%. Não houve associação estatisticamente significante entre desnutrição e infecção assintomática por L. (L. chagasi. A forma assintomática da doença está presente nas áreas estudadas, necessitando de medidas de controle mais efetivas.

  13. Transfer of the myrmecophilous soft scale insect Neolecanium amazonensis Foldi to Foldilecanium gen. nov. (Hemiptera: Coccidae), with description of a new species from Colombia

    OpenAIRE

    Kondo,Takumasa

    2011-01-01

    Neolecanium amazonensis Foldi is redescribed and illustrated and is transferred to the new genus Foldilecanium Kondo as Foldilecanium amazonensis (Foldi) comb. nov. A new species, Foldilecanium multisetosus Kondo, is described and illustrated based on specimens collected in Cali, Colombia, on Cananga odorata (Lam.) Hook.f. and Thomson (Annonaceae). An updated taxonomic key to New World Myzolecaniinae and a key to separate the two species of Foldilecanium are provided.

  14. Detección de Leishmania spp. en base al gen que codifica la proteína HSP20 / Detection of Leishmania spp. based on the gene encoding HSP20

    Scientific Electronic Library Online (English)

    Ana M., Montalvo; Jorge, Fraga; Omaira, Rodríguez; Orestes, Blanco; Alejandro, Llanos-Cuentas; Ana L., García; Braulio M., Valencia; Carlos, Muskus; Gert, Van der Auwera; José M., Requena.

    2014-12-01

    Full Text Available Objetivos. Explorar una nueva diana para el diagnóstico molecular de Leishmania. Materiales y métodos. Se evaluó la utilidad del gen que codifica la proteína de choque térmico de 20kDa (hsp20) para la detección de Leishmania por medio de la reacción en cadena de la polimerasa (PCR).Se normalizó la P [...] CR y se determinaron los parámetros analíticos, así como la validez y seguridad diagnóstica y la concordancia con la PCR-18S. Se evaluó la PCR-hsp20 con ADN obtenido de un grupo de muestras clínicas de distinta procedencia. Resultados. Los parámetros analíticos resultaron adecuados. La sensibilidad obtenida fue de 86% y la especificidad del 100%, la concordancia con el método de referencia resultó buena (? = 0,731), lo que apoya su posible uso para el diagnóstico. La posibilidad de identificación posterior de la especie mediante secuenciación del producto amplificado le confiere una ventaja adicional. Conclusiones. Se demuestra la utilidad de este gen como una nueva diana para la detección del género Leishmania. Debido a su potencial, se recomienda mejorar la sensibilidad del procedimiento y realizar su evaluación en diversas regiones endémicas. Abstract in english Objectives. Explore a new target for molecular diagnosis of Leishmania. Materials and methods. We evaluated the utility of the gene that encodes the heat shock protein 20-kDa (Hsp20) for detecting Leishmania by polymerase chain reaction (PCR). PCR was normalized and analytical parameters were determ [...] ined, as well as the validity and diagnostic accuracy, and concordance with the PCR - 18S. PCR-Hsp20 with DNA was obtained from a group of clinical samples from different sources. Results. The analytical parameters were adequate. The sensitivity obtained was 86% and the specificity was 100%. The concordance with the reference method was good (? = 0.731), which supports its potential use for diagnosis. The possibility of subsequent identification of the species by sequencing the amplified product gives an additional advantage. Conclusions. The usefulness of this gene as a new target for the detection of Leishmania was demonstrated. Because of its potential, it is recommended to improve the sensitivity of the method and to evaluate it in different endemic regions.

  15. Detección de Leishmania spp. en base al gen que codifica la proteína HSP20 / Detection of Leishmania spp. based on the gene encoding HSP20

    Scientific Electronic Library Online (English)

    Ana M, Montalvo; Jorge, Fraga; Omaira, Rodríguez; Orestes, Blanco; Alejandro, Llanos-Cuentas; Ana L, García; Braulio M, Valencia; Carlos, Muskus; Gert, Van der Auwera; José M, Requena.

    2014-10-01

    Full Text Available Objetivos. Explorar una nueva diana para el diagnóstico molecular de Leishmania. Materiales y métodos. Se evaluó la utilidad del gen que codifica la proteína de choque térmico de 20kDa (hsp20) para la detección de Leishmania por medio de la reacción en cadena de la polimerasa (PCR).Se normalizó la P [...] CR y se determinaron los parámetros analíticos, así como la validez y seguridad diagnóstica y la concordancia con la PCR-18S. Se evaluó la PCR-hsp20 con ADN obtenido de un grupo de muestras clínicas de distinta procedencia. Resultados. Los parámetros analíticos resultaron adecuados. La sensibilidad obtenida fue de 86% y la especificidad del 100%, la concordancia con el método de referencia resultó buena (? = 0,731), lo que apoya su posible uso para el diagnóstico. La posibilidad de identificación posterior de la especie mediante secuenciación del producto amplificado le confiere una ventaja adicional. Conclusiones. Se demuestra la utilidad de este gen como una nueva diana para la detección del género Leishmania. Debido a su potencial, se recomienda mejorar la sensibilidad del procedimiento y realizar su evaluación en diversas regiones endémicas. Abstract in english Objectives. Explore a new target for molecular diagnosis of Leishmania. Materials and methods. We evaluated the utility of the gene that encodes the heat shock protein 20-kDa (Hsp20) for detecting Leishmania by polymerase chain reaction (PCR). PCR was normalized and analytical parameters were determ [...] ined, as well as the validity and diagnostic accuracy, and concordance with the PCR - 18S. PCR-Hsp20 with DNA was obtained from a group of clinical samples from different sources. Results. The analytical parameters were adequate. The sensitivity obtained was 86% and the specificity was 100%. The concordance with the reference method was good (? = 0.731), which supports its potential use for diagnosis. The possibility of subsequent identification of the species by sequencing the amplified product gives an additional advantage. Conclusions. The usefulness of this gene as a new target for the detection of Leishmania was demonstrated. Because of its potential, it is recommended to improve the sensitivity of the method and to evaluate it in different endemic regions.

  16. Identification of a differentially expressed mRNA in axenic Leishmania panamensis amastigotes

    OpenAIRE

    José Arturo Gutiérrez; Fabiola Puentes; Alberto Moreno; Manuel Elkin Patarroyo; Luis Angel Murillo

    2001-01-01

    Differential display technique was applied in order to identify transcripts which are present in axenic amastigotes but not in promastigotes of the Leishmania panamensis parasites. One of them was cloned and the sequence reveals an open reading frame of 364 amino acids (aprox. 40 kDa). The deduced protein is homologous to the serine/threonine protein kinases and specially to the mitogen activates protein kinases from eukaryotic species. Southern blot analysis suggest that this transcript, nam...

  17. Detection of Leishmania infantum in the smegma of infected dogs / Detecção de Leishmania infantum no esmegma de cães infectados

    Scientific Electronic Library Online (English)

    L.C., Silva; V.P., Assis; V.M., Ribeiro; W.L., Tafuri; J.C., Toledo Júnior; S.O., Silva; M.N., Melo; M.A., Rachid; G.R., Valle.

    2014-06-01

    Full Text Available Tendo em vista a transmissão venérea da leishmaniose visceral do cão para a cadela, o objetivo deste estudo foi verificar se a superfície peniana e o esmegma de cães infectados poderiam ser a fonte de parasitas para a fêmea. Amostras de sêmen e esmegma de 12 cães infectados com Leishmania infantum f [...] oram submetidas à identificação do DNA do parasita por PCR. As incidências de positividade no sêmen (41,7%) e no esmegma (50,0%) foram semelhantes (P>0,05; teste exato de Fisher), sendo 58,3% dos cães positivos para sêmen e/ou esmegma. A positividade para sêmen e esmegma juntos ocorreu em 33,3%, mas em 8,3% dos casos apenas no sêmen, e em 16,7% apenas no esmegma, o que revela uma concordância moderada entre os testes (K=0,5; índice Kappa). Conclui-se que a Leishmania infantum está presente no esmegma de cães contaminados, podendo ser a fonte de parasitas para o sêmen e a cadela. Abstract in english Considering the venereal transmission of visceral leishmaniasis from dogs to bitches, the aim of this study was to verify if the penile surface and smegma from infected dogs can be the source of parasites in bitches. Twelve Leishmania infantum infected dogs had semen and smegma samples collected for [...] submission to PCR identification of the DNA of the parasite. Semen (41.7%) and smegma (50.0%) have similar positive incidence (P>0.05; Fisher's exact test), with 58.3% of the dogs positive for semen and/or smegma samples. The proportion of positivity for both semen and smegma was 33.3%, but 8.3% was positive only for semen, and 16.7% only for smegma, revealing a moderate agreement between tests (K=0.5; Kappa index). It was concluded that Leishmania infantum is present in the smegma of contaminated dogs and it can be a source of parasites for the semen and the bitch.

  18. Relationship between canine visceral leishmaniosis and the Leishmania (Leishmania) chagasi burden in dermal inflammatory foci.

    Science.gov (United States)

    Giunchetti, R C; Mayrink, W; Genaro, O; Carneiro, C M; Corrêa-Oliveira, R; Martins-Filho, O A; Marques, M J; Tafuri, W L; Reis, A B

    2006-01-01

    The skin is the first point of contact with organisms of the genus Leishmania from sand fly vectors, and apparently normal skin of sick dogs harbours amastigote forms of Leishmania chagasi. In relation to canine visceral leishmaniosis (CVL), the ear skin was examined in 10 uninfected dogs (UDs) and in 31 dogs dogs naturally infected with L. chagasi. The infected animals consisted of 10 symptomless dogs (SLDs), 12 mildly affected dogs (MADs) and nine affected dogs (ADs). A higher parasite burden was demonstrated in ADs than in SLDs by anti-Leishmania immunohistochemistry (Pdisease. PMID:16959259

  19. Nematodes of Amazonian birds, with a description of Hoazinstrongylus amazonensis n.gen.n.sp. (Trichostrongylidae, Libyostrongylinae)

    OpenAIRE

    R. Magalhães Pinto; Delir Corrêa Gomes

    1985-01-01

    During recent studies of the parasites of birds from the Amazonian Regio, the following nematodes were recovered: Hoazinstrongylus amazonensis n.gen.n.sp. from Opisthocomus hoazin (Muller, 1776); Ascaridia columbae (Gmel., 1790) Travassos, 1913, from Leptotila r. rufaxilla (Richard & Bernard, 1712) representing a new host record; Inglisakis ibanezi Freitas, Vicente & Santos, 1969, Cyrnea (C.) semilunaris (Molin, 1860) Seurat, 1914 and Thelazia digitata Travassos, 1918. A compelte description ...

  20. Detection of Leishmania infantum in Lutzomyia longipalpis captured in Campo Grande, MS / Detecção de Leishmania infantum em Lutzomyia longipalpis capturados em Campo Grande, MS

    Scientific Electronic Library Online (English)

    Rodrigo Casquero, Cunha; Renato, Andreotti; Marlon Cezar, Cominetti; Elaine Araújo, Silva.

    2014-04-01

    Full Text Available A Leishmaniose é uma zoonose causada por protozoários do gênero Leishmania (Ross 1903), objetos de considerável atenção em medicina humana e veterinária. Na cidade de Campo Grande – MS, o agente etiológico da Leishmaniose Visceral é Leishmania infantum (= L. chagasi), e o principal vetor é a espécie [...] Lutzomyia longipalpis, que representa cerca de 92,9% da população de flebotomíneos. Este trabalho teve como objetivo avaliar a PCR em tempo real como ferramenta para a detecção de kDNA de L. infantum em flebotomíneos, comparando-se com PCR convencional. Flebotomíneos dessa espécie foram capturados, somando 38 amostras de 1 a 4 espécimens cada, distribuídas em 13 bairros, divididos entre as 7 regiões urbanas da cidade de Campo Grande – MS, e armazenados a ?70 °C até a extração de ADN e amplificação por PCR e PCR em tempo real. Das 38 amostras testadas, foram encontradas 3 amostras positivas pela PCR convencional e 11 pela PCR em Tempo Real. Na otimização da PCR em tempo real, a temperatura de dissociação do amplificado foi de 82, 89 °C. Neste estudo, utilizando-se a técnica da PCR em tempo real, foi possível detectar a presença desse agente em 6 dos 13 bairros amostrados, todos na periferia da cidade, indicando o maior potencial enzoótico dessas regiões, que têm maior proximidade com reservas de matas naturais. Conclui-se que a PCR em tempo real pode ser utilizada para estudo epidemiológico de L. infantum. Abstract in english Leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania (Ross, 1903) and is the focus of considerable attention in human and veterinary medicine. In the city of Campo Grande, MS, the causative agent of visceral leishmaniasis is Leishmania infantum (= L. chagasi) primary vector [...] , comprising approximately 92.9% of the local sandfly population, is Lutzomyia longipalpis. The aim of this work was to compare real-time PCR with PCR as a tool for the detection of the kinetoplast DNA (kDNA) of L. infantum in sandflies. Sandflies of this species were caught, and a total of 38 samples with 1-4 individuals in each sample were obtained; these were distributed across 13 districts and divided between seven urban areas of the city of Campo Grande, MS. Three positive samples were found by PCR and, when using real-time PCR, this was able to detect the presence of this agent in 6 of the 13 districts sampled, which were all located on the outskirts of the city, where indicates the greater enzootic potential of these regions, as they are closer to natural forest reserves. We conclude that real-time PCR can be used for epidemiological studies of L. infantum.

  1. Susceptibility of spiny rats (Proechimys semispinosus to Leishmania (Viannia panamensis and Leishmania (Leishmania chagasi

    Directory of Open Access Journals (Sweden)

    BL Travi

    2002-09-01

    Full Text Available The role of Proechimys semispinosus as reservoir of Leishmania (Viannia panamensis on the Colombian Pacific coast was experimentally evaluated. The susceptibility to L. chagasi also was assessed to determine the utility of this rodent as a model for studying reservoir characteristics in the laboratory. Wild-caught animals were screened for natural trypanosomatid infections, and negative individuals were inoculated intradermally (ID in the snout or feet with 10(7 promastigotes of L. panamensis. L. chagasi was inoculated intracardially (10(7 promastigotes or ID in the ear (10(8 promastigotes. PCR-hybridization showed that 15% of 33 spiny rats were naturally infected with L. Viannia sp. Animals experimentally infected with L. panamensis developed non-ulcerated lesions that disappeared by the 7th week post-infection (p.i. and became more resistant upon reinfection. Infectivity to sand flies was low (1/20-1/48 infected/fed flies and transient, and both culture and PCR-hybridization showed that L. panamensis was cleared by the 13th week p.i. Animals inoculated with L. chagasi became subclinically infected and were non-infective to sand flies. Transient infectivity to vectors of spiny rats infected with L. panamensis, combined with population characteristics, e.g., abundance, exploitation of degraded habitats and high reproductive rates, could make them epidemiologically suitable reservoirs.

  2. Description of Leishmania (Leishmania) forattinii sp. n., a new parasite infecting opossums and rodents in Brazil

    OpenAIRE

    Elizaide L. A. Yoshida; Cuba Cuba, Cesar A.; Raquel da Silva Pacheco; Elisa Cupolillo; Celso Cruz Tavares; Gerzia M. C. Machado; Hooman Momen; Gabriel Grimaldi Junior

    1993-01-01

    A new parasite species of Leishmania is described, L. (Leishmania) forattinii sp. n., which was isolated from a pooled triturate of liver and spleen of a opossum (Didelphis marsupialis aurita) and from skin samples from a rodent (Proechmys iheringi denigratus), captured in primary forest on the Atlantic Cost of Brazil. Our results on the basis of biological and molecular criteria indicate that this taxonomically distinct parasite ias a new species of the L. mexicana complex, but closely relat...

  3. Comorbidity of Leishmania major with cutaneous sarcoidosis

    Directory of Open Access Journals (Sweden)

    Hamideh Moravvej

    2014-01-01

    Full Text Available Background: leishmaniasis infection might manifest as sarcoidosis; on the other hand, some evidences propose an association between sarcoidosis and leishmaniasis. Most of the times, it is impossible to discriminate idiopathic sarcoidosis from leishmaniasis by conventional histopathologic exam. Aim: We performed a cross-sectional study to examine the association of sarcoidosis with leishmaniasis in histopathologically diagnosed sarcoidal granuloma biopsy samples by polymerase chain reaction (PCR. Materials and Methods: We examined paraffin-embedded skin biopsy samples obtained from patients with clinical and histopathological diagnosis as naked sarcoidal granuloma, referred to Skin Research Center of Shaheed Beheshti Medical University from January 2001 to March 2010, in order to isolate Leishmania parasite. The samples were reassessed by an independent dermatopathologist. DNA extracted from all specimens was analyzed by the commercially available PCR kits (DNPTM Kit, CinnaGen, Tehran, Iran to detect endemic Leishmania species, namely leishmania major (L. major. Results: L. major was positive in PCR of Eight out of twenty-five examined samples. Conclusion: Cutaneous leishmaniasis may be misinterpreted as sarcoidosis; in endemic areas, when conventional methods fail to detect Leishmania parasite, PCR should be utilized in any granulomatous skin disease compatible with sarcoidosis, regardless of the clinical presentation or histopathological interpretation.

  4. Leishmania donovani and Cutaneous Leishmaniasis, Sri Lanka

    OpenAIRE

    Siriwardana, H. V. Yamuna D.; Noyes, Harry A.; Beeching, Nicholas J; Chance, Michael L.; Karunaweera, Nadira D; Bates, Paul A.

    2007-01-01

    To investigate the relationship of cutaneous leishmaniasis isolates from Sri Lanka to known species, we performed DNA sequencing and microsatellite analyses. We identified Leishmania donovani as the agent of Sri Lanka cutaneous leishmaniasis and showed that these parasites are closely related to those causing visceral leishmaniasis in the Indian subcontinent.

  5. Role of Leishmania (Leishmania chagasi amastigote cysteine protease in intracellular parasite survival: studies by gene disruption and antisense mRNA inhibition

    Directory of Open Access Journals (Sweden)

    Kucknoor Ashwini S

    2005-02-01

    Full Text Available Abstract Background The parasitic protozoa belonging to Leishmania (L. donovani complex possess abundant, developmentally regulated cathepsin L-like cysteine proteases. Previously, we have reported the isolation of cysteine protease gene, Ldccys2 from Leishmania (L. chagasi. Here, we have further characterized this cysteine protease gene and demonstrated its role during infection and survival of Leishmania (L. chagasi within the U937 macrophage cells. Results The amastigote specific Ldccys2 genes of L. (L. chagasi and L. (L. donovani have identical gene organization, as determined by southern blots. In vivo expression analyses by Northern blots showed that Ldccys2 is amastigote specific. Western blot using anti-Ldccys2 antibody confirmed the amastigote specific protein expression. Recombinant expression of Ldccys2, a 30 kDA protein, was functionally active in a gelatin assay. Results from Ldccys2 heterozygous knockout mutants showed its role during macrophage infection and in intra-macrophage survival of the parasites. Since attempts to generate null mutants failed, we used antisense RNA inhibition to regulate Ldcccys2 gene expression. Not surprisingly, the results from antisense studies further confirmed the results from heterozygous knockout mutants, reiterating the importance of amastigote specific cysteine proteases in Leishmania infection and pathogenesis. Conclusions The study shows that Ldccys2 is a developmentally regulated gene and that Ldccys2 is expressed only in infectious amastigote stages of the parasite. The collective results from both the heterozygous knockout mutants and antisense mRNA inhibition studies shows that Ldccys2 helps in infection and survival of L. (L. chagasi amastigotes within the macrophage cells. Finally, antisense RNA technique can be used as an alternate approach to gene knockout, for silencing gene expression in L. (L. chagasi, especially in cases such as this, where a null mutant cannot be achieved by homologous recombination.

  6. Leishmania mexicana: chemistry and biochemistry of sodium stibogluconate (Pentostam)

    International Nuclear Information System (INIS)

    The chemical properties of the primary antileishmanial agent sodium stibogluconate (Pentostam), and the interaction of Pentostam with Leishmania mexicana amastigotes, have been investigated with the aid of [125Sb]Pentostam. The molecular weight by P2 chromatography showed [125Sb]Pentostam to be of multiple species of MW = 100-4000 Da, rather than the one species of 746 Da predicted by the commonly hypothesized structural formula. Nonradioactive Pentostam had a lower osmolarity (789 mOsm for a 100 mg Sb/ml solution) than predicted (1644 mOsm), which indicates that the multiple components of Pentostam (Sb and derivatives of gluconic acid) are more closely complexed with each other than previously thought. When incubated with L. mexicana amastigotes, labeled drug was bound to at least six polypeptides of molecular weights ranging from 14,000 to 68,000 Da as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Interaction with the polypeptides is presumed to contribute to the antileishmanial action of Pentostam

  7. Implications of a Neotropical Origin of the Genus Leishmania

    OpenAIRE

    Harry Noyes

    1998-01-01

    The hypothesis of a Neotropical origin of the Leishmania/Endotrypanum clade is reviewed. The position of the L. (Sauroleishmania) external to the subgenus L. (Leishmania) is not consistent with the Neotropical origin of the latter subgenus. It is suggested that this may be a consequence of a faster evolutionary rate in the L. (Sauroleishmania). The implications for the classsification of the phlebotomine sandflies of the hypothesis for a Neotropical origin of the Leishmania is also considered...

  8. Capacidad infectiva de promastigotes en fase estacionaria de leishmania (viannia) braziliensis y leishmania (viannia) peruviana, en línea celular dh82 / Infectivity of promastigotes in stationary phase of Leishmania (viannia) braziliensis and leishmania (viannia) peruviana, In cell line dh82

    Scientific Electronic Library Online (English)

    Karen Daphne, Calvay-Sánchez; Nyshon Máximo, Rojas-Palomino; Aidé Clorinda, Sandoval-Juarez; Alberto, Cisneros-Tarmeño; Cristian, Obregón-Cahuaya; Gloria Sonia, Minaya-Gómez.

    2015-03-01

    Full Text Available Objetivos. Determinar la capacidad infectiva de los promastigotes de Leishmania (V.) peruviana y Leishmania (V.) braziliensis en la línea celular macrófago-monocítica de Canis familiaris DH82. Materiales y métodos. Se realizó un estudio experimental durante los meses de enero a diciembre de 2013. Se [...] utilizaron cepas referenciales de Leishmania (V.) braziliensis MHOM/PE/84/LC53 y Leishmania (V.) peruviana MHOM/PE/84/LC26. La línea celular fue infectada con promastigotes en fase estacionaria y la capacidad infectiva fue determinada como el producto del porcentaje de macrófagos infectados por el promedio de amastigotes por macrófago infectado, observado al microscopio de epifluorescencia. Resultados. El 13% de formas metacíclicas para Leishmania (V.) braziliensis correspondió al día 17,5 posinoculación y para Leishmania (V.) peruviana un porcentaje de 9,5% en el día 14,5. No se encontró diferencia significativa entre la capacidad infectiva de los promastigotes en fase estacionaria de ambas especies. Conclusiones. Se recomienda evaluar la capacidad infectiva de los promastigotes metacíclicos de cepas de Leishmania (V.) peruviana y Leishmania (V.) braziliensis en líneas celulares, a fin de determinar el modelo de infección in vitro más adecuado, que permita efectuar estudios de susceptibilidad a las drogas leishmanicidas de mayor eficacia para el control de la enfermedad Abstract in english Objectives. To determine the infectivity of promastigotes of Leishmania (V.) peruviana and Leishmania (V.) braziliensis in monocyte-macrophage cell line DH82 of Canis familiaris. Materials and methods. Was conducted a experimental study during the months of january to december 2013. Were used strain [...] s of Leishmania were used (V.) braziliensis MHOM / PE / 84 / LC53 and Leishmania (V.) peruviana MHOM / PE / 84 / LC26. The cell line was infected with stationary phase promastigotes and infectivity was determined as the product of percent infected macrophages average amastigotes per macrophage observed in epifluorescence microscope.Results. 13% of metacyclic forms to Leishmania (V.) braziliensis corresponded to 17.5 days post inoculation and Leishmania (V.) peruviana a percentage of 9.5% on the day 14.5. No significant difference was found between infectivity of stationary phase promastigotes of both species. Conclusions. It is recommended assess the infectivity of metacyclic promastigotes peruviana strains of Leishmania (V.) and Leishmania (V.) braziliensis cell lines in order to determine the most appropriate model in vitro infection, allowing leishmanicidas make the drug more effective susceptibility studies for disease control.

  9. Synthesis, characterization and study of activity inhibitory of new dialkylphosphorylhdrazones on the growth of trypanosomatids; Sintese, caracterizacao e estudo da atividade inibitoria de novas dialquilfosforilarilidrazonas sobre o crescimento de tripanossomatideos

    Energy Technology Data Exchange (ETDEWEB)

    Nogueira, Andrea Janaina M.; Lima, Marco Edilson F. de; DaCosta, Joao Batista N., E-mail: dacosta@ufrrj.br [Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropedica, RJ (Brazil). Inst. de Ciencias Exatas. Dept. de Quimica; Alves, Eliomara Sousa Sobral; Anjos, Danielle Oliveira dos; Vannier-Santos, Marcos Andre; Lanfredi-Rangel, Adriana [Fundacao Oswaldo Cruz, Salvador, BA (Brazil). Centro de Pesquisas Goncalo Moniz

    2011-07-01

    A new series of dialkylphosphorylhydrazones was synthesized through the condensation of aromatic aldehydes with different phosphorylhydrazines. All synthesized compounds were characterized by IR, {sup 1}H-NMR, {sup 13}C-NMR and {sup 31}P-NMR spectroscopies. The in vitro investigation of the activity of these compounds against Leishmania amazonensis promastigotes and epimastigotes of T. cruzi, showed an efficient inhibition of proliferation, at non toxic concentrations to mammalian cells. The results have shown some derivatives as potential antiparasitic agents against trypanosomatids. (author)

  10. LA BIOLOGÍA MOLECULAR DE LEISHMANIA SPP. COMO PUNTO DE PARTIDA PARA NUEVAS ALTERNATIVAS DE TRATAMIENTO / AMOLECULAR BIOLOGY LEISHMANIA SPP NEW DRUGS RESISTANCE / A BIOLOGIA MOLECULAR DE LEISHMANIA SPP. COMO PONTO DE PARTIDA PARA NOVAS ALTERNATIVAS DE TRATAMENTO

    Scientific Electronic Library Online (English)

    Martha Cecilia, Beltrán Cifuentes; Patricia, Durán Ospina; Luisa Fernanda, Corredor Arias.

    2008-04-01

    Full Text Available Introdução: Na Colômbia existem grupos de pesquisas em Leishmaniasis que têm somado esforços para a identificação do genoma da Leishmania spp. Como profs-sionais da saúde, é uma prioridade conecé-los para entender os mecanismos de resistência a fármacos. Métodos: As bases de dados empregadas para es [...] ta busca foram, entre outras: NCBI PubMed, MEDLINE, Science Direct, Nucleic Acids Research e Molecular and Biochemical Parasitology, publicações da revista Biomédica e de varias Universidades. Resultados: existem muitas espécies de vetores distribuídos em todo o país. As multi-resistências criadas por Leishmania sp. baseiam-se especialmente em proteínas de membrana e mutações pontuais no DNA do parasita e seus vetores. Conclusões: As técnicas de PCR deveriam ser implementadas na clínica para estudar resistências a fármacos. Os protocolos atuais contra Leishmaniasis não incluem novas alternativas. Os tratamentos imuno-moduladores são uma nova es-perança para o tratamento desta enfermidade re-emergente. Abstract in spanish Introducción: en Colombia existen grupos de investigación en Leishmaniasis que han aunado esfuerzos para lograr la identificación del genoma de la Leishmania spp. Como profesionales de la salud es una prioridad conocerlos para entender los mecanismos de resistencia a fármacos. Métodos: las bases de [...] datos empleadas para esta búsqueda fueron entre otras: NCBI PubMed, MEDLINE, Science Direct, Nucleic Acids Research y Molecular and Biochemical Parasitology, publicaciones de la revista Biomédica y de varias Universidades. Resultados: existen muchas especies de vectores distribuidos en todo el país. Las multiresistencias creadas por Leishmania sp. se basan especialmente en proteínas de membrana y mutaciones puntuales en el DNA del parásito y sus vectores. Conclusiones: las técnicas de PCR deberían ser implementadas en la clínica para estudiar resistencias a fármacos. Los protocolos actuales contra Leishmaniasis no incluyen nuevas alternativas. Los tratamientos inmunomoduladores son una nueva esperanza para el tratamiento de esta enfermedad re-emergente. Abstract in english Introduction: In Colombia there are research groups in Leishmaniasis that have joined forces to achieve the identification of the genome of Leishmania spp. As health professionals know this is a priority to understand the mechanisms of drug resistance. Methods: The databases used for this search wer [...] e among others: NCBI PubMed MEDLINE, Science Direct, Nucleic Acids Research and Biochemical and Molecular Parasitology, as a publication of the journal Biomedical and several universities. Results: There are many species and vectors distributed throughout the country. The multiresistents created by Leishmania spp. rely especially in membrane proteins and mutations in the DNA of the parasite and their delivery systems. Conclusions: The PCR techniques should be implemented at the clinic to study resistance to drugs. The current protocols against Leishmaniasis not include additional alternatives. The treatments with immunomodulators are the new hope for treating this re-emerging disease.

  11. Plants used in the treatment of leishmanial ulcers due to Leishmania (Viannia) braziliensis in an endemic area of Bahia, Brazil

    Scientific Electronic Library Online (English)

    Flávio, França; Ednaldo L., Lago; Philip D., Marsden.

    1996-06-01

    Full Text Available Este trabalho relata as plantas usadas no tratamento da leishmaniose cutânea, causada por Leishmania (Viannia) braziliensis (L(V)b), na população rural da faixa litorânea produtora de cacau do estado da Bahia, Brasil. Um inquérito realizado entre 100 pacientes, identificou 49 espécies de plantas usa [...] das para tratar úlceras de pele causadas por esta espécie de Leishmânia. As principais plantas usadas foram o cajueiro-branco (Anacardium occidentale - Anacardiaceae) usado por 65% da população, a folha-fogo (Clidemia hirta - Melastomataceae) 39%, a alfavaca-grossa (Plectranthus amboinicus - Lamiaceae) 33%, o mastruz (Chenopodium ambrosioides - henopodiaceae) 31%, a erva-de-santa-maria (Solanum americanum - Solanaceae) 25% e a transagem (Plantago major - Plantaginaceae) 2%. Abstract in english This paper records the plants used in the treatment of cutaneous leishmaniasis due to Leishmania (Viannia) braziliensis (L(V)b) among the rural population of a cocoa- producing coastal area of Bahia state, Brazil. An enquiry conducted among a hundred patients identified 49 plant species used to trea [...] t skin ulceration caused by this Leishmania species. The principal plants used are caju-branco (Anacardium occidentale - Anacardiaceae), used by 65% of the population, folha-fogo (Clidemia hirta - Melastomataceae) 39%, alfavaca-grossa (Plectranthus amboinicus - Lamiaceae) 33%, mastruz (Chenopodium ambrosioides - Chenopodiaceae) 31%, erva-de-santa-maria (Solatium americanum - Solanaceae) (25%) and transagem (Plantago major - Plantaginaceae.) 2%.

  12. Plants used in the treatment of leishmanial ulcers due to Leishmania (Viannia braziliensis in an endemic area of Bahia, Brazil

    Directory of Open Access Journals (Sweden)

    Flávio França

    1996-06-01

    Full Text Available This paper records the plants used in the treatment of cutaneous leishmaniasis due to Leishmania (Viannia braziliensis (L(Vb among the rural population of a cocoa- producing coastal area of Bahia state, Brazil. An enquiry conducted among a hundred patients identified 49 plant species used to treat skin ulceration caused by this Leishmania species. The principal plants used are caju-branco (Anacardium occidentale - Anacardiaceae, used by 65% of the population, folha-fogo (Clidemia hirta - Melastomataceae 39%, alfavaca-grossa (Plectranthus amboinicus - Lamiaceae 33%, mastruz (Chenopodium ambrosioides - Chenopodiaceae 31%, erva-de-santa-maria (Solatium americanum - Solanaceae (25% and transagem (Plantago major - Plantaginaceae. 2%.Este trabalho relata as plantas usadas no tratamento da leishmaniose cutânea, causada por Leishmania (Viannia braziliensis (L(Vb, na população rural da faixa litorânea produtora de cacau do estado da Bahia, Brasil. Um inquérito realizado entre 100 pacientes, identificou 49 espécies de plantas usadas para tratar úlceras de pele causadas por esta espécie de Leishmânia. As principais plantas usadas foram o cajueiro-branco (Anacardium occidentale - Anacardiaceae usado por 65% da população, a folha-fogo (Clidemia hirta - Melastomataceae 39%, a alfavaca-grossa (Plectranthus amboinicus - Lamiaceae 33%, o mastruz (Chenopodium ambrosioides - henopodiaceae 31%, a erva-de-santa-maria (Solanum americanum - Solanaceae 25% e a transagem (Plantago major - Plantaginaceae 2%.

  13. Description of Leishmania (Leishmania forattinii sp. n., a new parasite infecting opossums and rodents in Brazil

    Directory of Open Access Journals (Sweden)

    Elizaide L. A. Yoshida

    1993-09-01

    Full Text Available A new parasite species of Leishmania is described, L. (Leishmania forattinii sp. n., which was isolated from a pooled triturate of liver and spleen of a opossum (Didelphis marsupialis aurita and from skin samples from a rodent (Proechmys iheringi denigratus, captured in primary forest on the Atlantic Cost of Brazil. Our results on the basis of biological and molecular criteria indicate that this taxonomically distinct parasite ias a new species of the L. mexicana complex, but closely related to L. (L. aristidesi Laison & shaw, 1979, as revelated by phenetic and phylogenetic numerical analyses of the enzyme data. L. forattinii was clearly distinguishable from other Leishmania species of the genus usisng enzyme electrophoresis, monoclonal antibodies, molecular karyotypes, analysis of restriction enzyme digestion patterns of kinetoplast DNA (kDNA, as well as the use of kDNA hybridization procedures.

  14. Description of Leishmania (Leishmania) forattinii sp. n., a new parasite infecting opossums and rodents in Brazil

    Scientific Electronic Library Online (English)

    Elizaide L. A., Yoshida; Cesar A Cuba, Cuba; Raquel da Silva, Pacheco; Elisa, Cupolillo; Celso Cruz, Tavares; Gerzia M. C., Machado; Hooman, Momen; Gabriel, Grimaldi Junior.

    1993-09-01

    Full Text Available A new parasite species of Leishmania is described, L. (Leishmania) forattinii sp. n., which was isolated from a pooled triturate of liver and spleen of a opossum (Didelphis marsupialis aurita) and from skin samples from a rodent (Proechmys iheringi denigratus), captured in primary forest on the Atla [...] ntic Cost of Brazil. Our results on the basis of biological and molecular criteria indicate that this taxonomically distinct parasite ias a new species of the L. mexicana complex, but closely related to L. (L.) aristidesi Laison & shaw, 1979, as revelated by phenetic and phylogenetic numerical analyses of the enzyme data. L. forattinii was clearly distinguishable from other Leishmania species of the genus usisng enzyme electrophoresis, monoclonal antibodies, molecular karyotypes, analysis of restriction enzyme digestion patterns of kinetoplast DNA (kDNA), as well as the use of kDNA hybridization procedures.

  15. Genetic and clinical characterization of canine leishmaniasis caused by Leishmania (Leishmania) infantum in northeastern Argentina.

    Science.gov (United States)

    Barroso, Paola A; Nevot, M Cecilia; Hoyos, Carlos L; Locatelli, Fabricio M; Lauthier, Juan J; Ruybal, Paula; Cardozo, Rubén M; Russo, Pablo D; Vassiliades, Carola N; Mora, María C; Estévez, J Octavio; Hashiguchi, Yoshihisa; Korenaga, Masataka; Basombrío, Miguel A; Marco, Jorge D

    2015-10-01

    Leishmaniases comprise zoonotic diseases caused by protozoan flagellates of the Leishmania genus. They are endemic to South America, and the visceral form has been recently reported in Argentina. Dogs can play different roles in the Leishmania transmission cycles, depending mainly on the species of parasite involved. Here we focused on the clinical characterization of canine leishmaniasis (CanL) in Northeast Argentina and on the molecular typing of its etiological agent. The nested polymerase chain reaction and sequence analysis of the Leishmania cytochrome b (cyt b) gene was performed on DNA templates purified from lymph nodes, bone marrow or spleen aspirates obtained from 48 dogs previously diagnosed by the observation of Leishmania amastigotes on smears from these aspirates. Their clinical and epidemiological data were also recorded. Systemic abnormalities were observed in 46 subjects (95.8%), most frequently lymphadenopathy, and emaciation (89.6 and 75%). Furthermore, 87% also presented tegumentary abnormalities, such as alopecia (54.2%) or secondary skin lesions (47.9%), among others. Twenty three dogs were positive for cyt b amplification. The sequence analysis showed the presence of two genotypes, LiA1 and LiA2, assigned to Leishmania (Leishmania) infantum, with 99.9 and 100% homology with the reference strain MHOM/TN/80/IPT1 respectively. LiA1 was identified in 18 cases (78.3%) and LiA2 in five (21.7%). Two cyt b variants of L. (L.) infantum were incriminated as the causative agents of CanL cases from three cities: Posadas, Garupá, and Ituzaingó. All three cities are located in the northeastern area of the country, where these parasites seem to be spreading in urban areas. PMID:26277067

  16. Herbal extract targets in Leishmania tropica.

    Science.gov (United States)

    Mohammad, Bassim I; Al Shammary, Maani N; Abdul Mageed, Roaa H; Yousif, Nasser Ghaly

    2015-12-01

    The present study aims to investigate the effect of some herbal extract such as phenolic compounds on the viability of Leishmania tropica promastigotes in vitro. Four tested chemical agents (caffeic acid (CA), ferulic acid (FA), syringic acid (SA) and 4-hydroxybenzoic acid (4-HBA)) were used in this study. The viability of Leishmania tropica promastigotes was investigated under five different concentrations (10, 15, 20, 25 and 30 mg/ml) of each agent after (72 h). CA was the most active agent on the promastigotes viability after 72 h exposure to 30 mg/ml concentration so that the parasiticidal effect reach (53 × 10(4)) promastigote/ml. FA is the second agent in parasiticidal effect that parasiticidal effect reach to (50 × 10(4) promastigote/ml) at a concentration (30 mg/ml), 4-HBA is the third agent in parasiticidal effect that reach to (48 × 10(4) promastigote/ml) at a concentration (30 mg/ml), SA is the weakest agent in parasiticidal activity that reach to (44 × 10(4) promastigote/ml) at a concentration (30 mg/ml). It can be concluded that (CA, FA, SA and 4-HBA) possess acidal effect on the Leishmania tropica promastigotes in vitro. PMID:26688631

  17. Qualidade espermática de sêmen de cães naturalmente infectados por Leishmania sp: / Semen quality of dogs naturally infected by Leishmania sp

    Scientific Electronic Library Online (English)

    É., Labat; J.T., Carreira; B.H., Matsukuma; M.T.A., Martins; V.M.F., Lima; S.R.M., Bomfim; S.H.V., Perri; M.B., Koivisto.

    2010-06-01

    Full Text Available Avaliaram-se alterações espermáticas associadas à infecção por leishmaniose no sêmen de cães naturalmente infectados, utilizando-se, durante oito semanas consecutivas, ejaculados de seis cães soronegativos e seis cães soropositivos. As amostras foram colhidas uma vez por semana e avaliadas quanto ao [...] volume, concentração, motilidade, vigor, morfologia espermática, integridade da cromatina, avaliação simultânea da integridade da membrana plasmática, acrossoma e potencial mitocondrial. Concomitantemente foram dosadas a proteína total do plasma seminal e sanguíneo. A leishmaniose visceral causou aumento dos defeitos maiores e menores nos espermatozoides dos animais acometidos pelo estágio moderado a severo da doença. Em estágios mais avançados da enfermidade, a integridade das membranas acrossomal e plasmática foi afetada negativamente. Não foi possível estabelecer um critério quanto à avaliação do potencial mitocondrial. A incidência de alterações morfológicas nos animais acometidos não promoveu aumento de injurias à cromatina. Todos os animais com leishmaniose apresentaram hiperproteinemia do sêmen. Abstract in english The spermatic changes associated with the natural infection in dogs by Leishmania sp was evaluated during eight consecutive weeks, using ejaculates of six seronegative and six seropositive dogs. The samples were collected once a week and evaluated for volume, concentration, motility, vigor, sperm mo [...] rphology, chromatin integrity, simultaneous evaluation of the plasmatic membrane integrity, acrosome, and mitochondrial potential. The total proteins of the seminal plasma and blood were measured. The visceral leishmaniasis caused increase of major and minor defects in spermatozoa of animals attacked by moderate to severe stages of the disease. In more advanced stages of the illness, the acrosomal and plasmatic membranes integrity was adversely affected. It was not possible to establish a pattern refering the evaluation of the mitochondrial potential. The incidence of morphological changes in the seropositive animals did not promote an increase of injuries to the chromatin. All animals with leishmaniasis presented hyperproteinemia of the semen.

  18. Localized mucosal leishmaniasis due to Leishmania (Leishmania) infantum: clinical and microbiologic findings in 31 patients.

    Science.gov (United States)

    Aliaga, Luis; Cobo, Fernando; Mediavilla, Juan Diego; Bravo, Juan; Osuna, Antonio; Amador, José Manuel; Martín-Sánchez, Joaquina; Cordero, Elisa; Navarro, José María

    2003-05-01

    The clinical and microbiologic characteristics of 31 patients with mucosal leishmaniasis due to Leishmania (Leishmania) infantum are described. Twenty-eight (90%) patients were male. Mean age at presentation was 48 +/- 14 years. Thirteen (42%) patients had no underlying disease, while 18 (58%) patients had several other medical conditions. Fifteen (48%) patients were immunocompromised, 7 patients were infected with human immunodeficiency virus (HIV), and 3 were graft recipients. The primary location of lesions was the larynx in 11 (35%) patients, oral mucosa in 10 (32%) patients, and the nose in 5 (16%) patients. Mucosal lesions were painless in all patients but 2 and consisted of whitish, red, or violaceous nodular swelling or tumorlike masses. Ulceration was reported in 6 patients. Pathologically, the lesions showed a chronic inflammatory infiltrate. Granuloma may be seen. The localization of the lesions determined the symptomatology of the disease. Symptoms included hoarseness, difficulty swallowing, and nasal obstruction. The disease presentation was usually protracted, with a mean time from the onset of symptoms to diagnosis of 13 months (range, 3 wk-4.5 yr), and the clinical diagnosis was usually mistaken for neoplasia of the upper aerodigestive tract. No laboratory abnormalities were found in these patients due to the localized disease, apart from those attributed to underlying diseases. Parasites were easily identified in smears or sections by Giemsa stain or hematoxylin-eosin stain. Leishmania was grown in culture in 12 (60%) patients; culture was negative in 8 (40%) patients. Leishmania (Leishmania) infantum was identified in only 9 instances. The following zymodemes were reported: MON-1 (2 patients), MON-24 (2 patients), MON-27 (1 patient), and MON-34 (1 patient). Serologic test results were known in 25 patients. Serology was usually positive at low titer; 6 (24%) patients had negative serologic test results. Twenty patients were treated with antimonial compounds for between 3 and 36 days. Three patients were given drugs other than antimonial drugs. Five patients were treated only locally, by surgery (3 patients) or topical medical therapy. One patient received no therapy, and treatment was not reported in 2 cases. Patients were cured in 25 (89%) cases, and sequelae were uncommon (14%). Relapse was detected in 2 individuals and 1 patient developed visceral leishmaniasis after treatment. Two HIV-coinfected patients died of causes unrelated to leishmaniasis. The results of the present report stress the clinical importance of searching for the presence of Leishmania in patients with suspected neoplasia of the upper respiratory tract if they have visited or resided in zones endemic for Leishmania (Leishmania) infantum. The treatment of choice for these patients is not established yet, but most patients respond to antimonial compounds given for 28 days or less. PMID:12792301

  19. DNA Transformation of Leishmania infantum Axenic Amastigotes and Their Use in Drug Screening

    OpenAIRE

    Sereno, Denis; Roy, Gaétan; Lemesre, Jean Loup; Papadopoulou, Barbara; Ouellette, Marc

    2001-01-01

    Protocols for DNA electroporation in Leishmania promastigote cells are well established. More recently, in vitro culture of axenic Leishmania amastigotes became possible. We have established conditions for DNA transformation of axenically grown Leishmania infantum amastigotes. Parameters for DNA electroporation of Leishmania axenic amastigotes were systematically studied using luciferase-mediated transient transfection. Cell lines expressing stable luciferase activity were then selected, and ...

  20. Leishmania (Leishmania) infantum chagasi em canídeos silvestres mantidos em cativeiro, no Estado de Mato Grosso / Leishmania (Leishmania) infantum chagasi in wild canids kept in captivity in the State of Mato Grosso

    Scientific Electronic Library Online (English)

    Nely Pinheiro, Souza; Arleana do Bom Parto Ferreira de, Almeida; Tatiana Pádua Tavares de, Freitas; Regina Celia Rodrigues da, Paz; Valéria, Dutra; Luciano, Nakazato; Valéria Régia Franco, Sousa.

    2010-06-01

    Full Text Available INTRODUÇÃO: Leishmaniose visceral é uma zoonose que acomete diversos mamíferos tendo os canídeos domésticos como principais reservatórios em ambiente urbano. A presente nota descreve a infecção de canídeos silvestres por Leishmania (Leishmania) infantum chagasi mantidos em cativeiro no Estado de Mat [...] o Grosso, Brasil. MÉTODOS: De seis raposas (Cerdocyon thous) e um cachorro vinagre (Spheotos venaticus), foram coletadas amostras de pele, medula óssea e linfonodo para detecção e caracterização de Leishmania sp pela técnica de PCR-RFLP. RESULTADOS: Todos as animais pesquisados apresentaram-se positivos para Leishmania (L.) infantum chagasi. CONCLUSÕES: Destaca-se a importância de monitoramento adequado dos mesmos, além do maior controle desta enfermidade já que estes animais estão em ambientes de recreação pública. Abstract in english INTRODUCTION: Visceral leishmaniasis is a zoonosis that affects many mammals, and domestic canids are the main reservoirs in urban environments. This note describes infection by Leishmania (Leishmania) infantum chagasi among wild canids kept in captivity in the State of Mato Grosso, Brazil. METHODS: [...] Skin, bone marrow and lymph node samples were collected from six crab-eating foxes (Cerdocyon thous) and one bush dog (Spheotos venaticus), in order to detect and characterize Leishmania using the PCR-RFLP technique. RESULTS: All the animals studied were positive for Leishmania (L.) infantum chagasi. CONCLUSIONS: This study highlights the importance of adequate monitoring of these animals, as well as greater control of this disease, given that these animals are in a public recreation environment.

  1. Leishmania (Leishmania infantum chagasi em canídeos silvestres mantidos em cativeiro, no Estado de Mato Grosso Leishmania (Leishmania infantum chagasi in wild canids kept in captivity in the State of Mato Grosso

    Directory of Open Access Journals (Sweden)

    Nely Pinheiro Souza

    2010-06-01

    Full Text Available INTRODUÇÃO: Leishmaniose visceral é uma zoonose que acomete diversos mamíferos tendo os canídeos domésticos como principais reservatórios em ambiente urbano. A presente nota descreve a infecção de canídeos silvestres por Leishmania (Leishmania infantum chagasi mantidos em cativeiro no Estado de Mato Grosso, Brasil. MÉTODOS: De seis raposas (Cerdocyon thous e um cachorro vinagre (Spheotos venaticus, foram coletadas amostras de pele, medula óssea e linfonodo para detecção e caracterização de Leishmania sp pela técnica de PCR-RFLP. RESULTADOS: Todos as animais pesquisados apresentaram-se positivos para Leishmania (L. infantum chagasi. CONCLUSÕES: Destaca-se a importância de monitoramento adequado dos mesmos, além do maior controle desta enfermidade já que estes animais estão em ambientes de recreação pública.INTRODUCTION: Visceral leishmaniasis is a zoonosis that affects many mammals, and domestic canids are the main reservoirs in urban environments. This note describes infection by Leishmania (Leishmania infantum chagasi among wild canids kept in captivity in the State of Mato Grosso, Brazil. METHODS: Skin, bone marrow and lymph node samples were collected from six crab-eating foxes (Cerdocyon thous and one bush dog (Spheotos venaticus, in order to detect and characterize Leishmania using the PCR-RFLP technique. RESULTS: All the animals studied were positive for Leishmania (L. infantum chagasi. CONCLUSIONS: This study highlights the importance of adequate monitoring of these animals, as well as greater control of this disease, given that these animals are in a public recreation environment.

  2. An improved purification procedure for Leishmania RNA virus (LRV)

    Science.gov (United States)

    de Souza, Marcos Michel; Manzine, Livia Regina; da Silva, Marcos Vinicius G.; Bettini, Jefferson; Portugal, Rodrigo Vilares; Cruz, Angela Kaysel; Arruda, Eurico; Thiemann, Otavio Henrique

    2014-01-01

    Leishmania RNA Virus (LRV, Totiviridae) infect Leishmania cells and subvert mice immune response, probably promoting parasite persistence, suggesting significant roles for LRV in host-parasite interaction. Here we describe a new LRV1-4 purification protocol, enabling capsid visualization by negatively stained electron microscopy representing a significant contribution to future LRV investigations. PMID:25242960

  3. Detection of Leishmania donovani and L. tropicain Ethiopian wild rodents.

    Czech Academy of Sciences Publication Activity Database

    Kassahun, A.; Sádlová, J.; Dvo?ák, V.; Koš?álová, T.; Rohoušová, I.; Frynta, D.; Aghová, Tatiana; Yasur-Landau, D.; Lemma, W.; Hailu, A.; Baneth, G.; Warburg, A.; Volf, P.; Votýpka, J.

    2015-01-01

    Ro?. 145, May 2015 (2015), s. 39-44. ISSN 0001-706X EU Projects: European Commission(XE) 261504 Institutional support: RVO:68081766 Keywords : Leishmania donovani * Leishmania tropica * Phlebotomine sand fly * Rodents * kDNA * ITS1 Subject RIV: EG - Zoology Impact factor: 2.270, year: 2014

  4. An improved purification procedure for Leishmania RNA virus (LRV)

    Scientific Electronic Library Online (English)

    Marcos Michel de, Souza; Livia Regina, Manzine; Marcos Vinicius G. da, Silva; Jefferson, Bettini; Rodrigo Vilares, Portugal; Angela Kaysel, Cruz; Eurico, Arruda; Otavio Henrique, Thiemann.

    2014-06-01

    Full Text Available Leishmania RNA Virus (LRV, Totiviridae) infect Leishmania cells and subvert mice immune response, probably promoting parasite persistence, suggesting significant roles for LRV in host-parasite interaction. Here we describe a new LRV1-4 purification protocol, enabling capsid visualization by negative [...] ly stained electron microscopy representing a significant contribution to future LRV investigations.

  5. Recombinant NAD-dependent SIR-2 Protein of Leishmania donovani: Immunobiochemical Characterization as a Potential Vaccine against Visceral Leishmaniasis

    OpenAIRE

    Baharia, Rajendra K.; Tandon, Rati; Sharma, Tanuj; Suthar, Manish K; Das, Sanchita; Siddiqi, Mohammad Imran; Saxena, Jitendra Kumar; Sunder, Shyam; Dube, Anuradha

    2015-01-01

    Visceral Leishmaniasis (VL) is the most fatal form of leishmaniasis disease in Indian subcontinent. Through proteomic approaches, NAD-dependent Silent information regulator-2 was identified as one of the potent immunostimulatory proteins. Herein, it was first reported the cloning, expression, purification and immunobiochemical characterization of a NAD+-dependent protein from Leishmania donovani. The gene encodes a monomeric protein (LdSir2RP) of approximately 45 kDa and showed NAD+-dependent...

  6. Nematodes of Amazonian birds, with a description of Hoazinstrongylus amazonensis n.gen.n.sp. (Trichostrongylidae, Libyostrongylinae

    Directory of Open Access Journals (Sweden)

    R. Magalhães Pinto

    1985-06-01

    Full Text Available During recent studies of the parasites of birds from the Amazonian Regio, the following nematodes were recovered: Hoazinstrongylus amazonensis n.gen.n.sp. from Opisthocomus hoazin (Muller, 1776; Ascaridia columbae (Gmel., 1790 Travassos, 1913, from Leptotila r. rufaxilla (Richard & Bernard, 1712 representing a new host record; Inglisakis ibanezi Freitas, Vicente & Santos, 1969, Cyrnea (C. semilunaris (Molin, 1860 Seurat, 1914 and Thelazia digitata Travassos, 1918. A compelte description is restrained to the new genus and new species here proposed. The other known and well described species are listed and accounted.Durante recentes estudos dos parasitas de aves capturadas na Região Amazônica, foram identificadas as seguintes espécies de nematóides: Hoazinstrongylus amazonensis n.gen.n.sp, de Opisthocomus hoazin (Muller, 1776, Ascaridia columbae (Gmel., 1790 Travassos, 1912, de Leptotila r. rufaxilla (Richard & Bernard, 1712 que se constitui em um novo hospedeiro para a espécie, Inglisakis ibanezi Freitas, Vicente & Santos, 1969, Cyrnea (C. semilunaris (Molin, 1860 Seurat, 1914 e Thelazia digitata Travassos, 1918.

  7. Nematodes of Amazonian birds, with a description of Hoazinstrongylus amazonensis n.gen.n.sp. (Trichostrongylidae, Libyostrongylinae)

    Scientific Electronic Library Online (English)

    R. Magalhães, Pinto; Delir Corrêa, Gomes.

    1985-06-01

    Full Text Available Durante recentes estudos dos parasitas de aves capturadas na Região Amazônica, foram identificadas as seguintes espécies de nematóides: Hoazinstrongylus amazonensis n.gen.n.sp, de Opisthocomus hoazin (Muller, 1776), Ascaridia columbae (Gmel., 1790) Travassos, 1912, de Leptotila r. rufaxilla (Richard [...] & Bernard, 1712) que se constitui em um novo hospedeiro para a espécie, Inglisakis ibanezi Freitas, Vicente & Santos, 1969, Cyrnea (C.) semilunaris (Molin, 1860) Seurat, 1914 e Thelazia digitata Travassos, 1918. Abstract in english During recent studies of the parasites of birds from the Amazonian Regio, the following nematodes were recovered: Hoazinstrongylus amazonensis n.gen.n.sp. from Opisthocomus hoazin (Muller, 1776); Ascaridia columbae (Gmel., 1790) Travassos, 1913, from Leptotila r. rufaxilla (Richard & Bernard, 1712) [...] representing a new host record; Inglisakis ibanezi Freitas, Vicente & Santos, 1969, Cyrnea (C.) semilunaris (Molin, 1860) Seurat, 1914 and Thelazia digitata Travassos, 1918. A compelte description is restrained to the new genus and new species here proposed. The other known and well described species are listed and accounted.

  8. Synthesis and in vitro Screening of 29, 30-Dibromo-28-oxoallobetulin against Parasitic Protozoans, Leishmania donovani and Leishmania Major

    Science.gov (United States)

    Ghosh, P.; Mandal, A.; Dey, S.; Pal, C.

    2015-01-01

    A simple synthesis and in vitro antileishmanial activity of 29,30-dibromo-28-oxoallobetulin against the parasitic protozoans, Leishmania donovani and Leishmania major is described. The structure of the compound is established on the basis of spectral data (IR, NMR, MS). Both the antiproliferative effect and the cell cycle progression were studied. PMID:26009654

  9. Synthesis and in vitro screening of 29, 30-dibromo-28-oxoallobetulin against parasitic protozoans, leishmania donovani and leishmania major

    Directory of Open Access Journals (Sweden)

    P Ghosh

    2015-01-01

    Full Text Available A simple synthesis and in vitro antileishmanial activity of 29,30-dibromo-28-oxoallobetulin against the parasitic protozoans, Leishmania donovani and Leishmania major is described. The structure of the compound is established on the basis of spectral data (IR, NMR, MS. Both the antiproliferative effect and the cell cycle progression were studied.

  10. Synthesis and in vitro screening of 29, 30-dibromo-28-oxoallobetulin against parasitic protozoans, leishmania donovani and leishmania major

    OpenAIRE

    P. Ghosh; Mandal, A.; Dey, S.; Pal, C

    2015-01-01

    A simple synthesis and in vitro antileishmanial activity of 29,30-dibromo-28-oxoallobetulin against the parasitic protozoans, Leishmania donovani and Leishmania major is described. The structure of the compound is established on the basis of spectral data (IR, NMR, MS). Both the antiproliferative effect and the cell cycle progression were studied.

  11. [Importance of amastigote forms morphology to differentiate Leishmania infantum and Leishmania major species].

    Science.gov (United States)

    Aoun, K; Chahed, M K; Mokni, M; Harrat, Z; Bouratbine, A

    2003-01-01

    The microscopic study of the dermal smears of 62 cases of cutaneous leishmaniose, 27 infected by Leishmania (L.) infantum and 35 by L. major, showed that the amastigotes of L. infantum are meaningfully smaller (p < 0.001). This criteria is a simple pary alternative to distinguish these 2 species which have completely different epidemiology, recovery delay and prophylactic dispositions. PMID:15941068

  12. Nitric oxide production by Peromyscus yucatanicus (Rodentia infected with Leishmania (Leishmania mexicana

    Directory of Open Access Journals (Sweden)

    Elsy Nalleli Loría-Cervera

    2013-04-01

    Full Text Available Peromyscus yucatanicus (Rodentia: Cricetidae is a primary reservoir of Leishmania (Leishmania mexicana (Kinetoplastida: Trypanosomatidae. Nitric oxide (NO generally plays a crucial role in the containment and elimination of Leishmania. The aim of this study was to determine the amount of NO produced by P. yucatanicus infected with L. (L. mexicana. Subclinical and clinical infections were established in P. yucatanicus through inoculation with 1 x 10 2 and 2.5 x 10 6 promastigotes, respectively. Peritoneal macrophages were cultured alone or co-cultured with lymphocytes with or without soluble Leishmania antigen. The level of NO production was determined using the Griess reaction. The amount of NO produced was significantly higher (p ? 0.0001 in co-cultured macrophages and lymphocytes than in macrophages cultured alone. No differences in NO production were found between P. yucatanicus with subclinical L. (L. mexicana infections and animals with clinical infections. These results support the hypothesis that the immunological mechanisms of NO production in P. yucatanicus are similar to those described in mouse models of leishmaniasis and, despite NO production, P. yucatanicus is unable to clear the parasite infection.

  13. Atypical epidermal alterations in chronic Leishmania mexicana mexicana lesions fo C3H mice

    Scientific Electronic Library Online (English)

    G., Grimaldi Junior; A. C., Queiroz; P. L., Moriearty.

    1982-12-01

    Full Text Available Camundongos C3H cronicamente infectados com Leishmania mexicana mexicana, e em alguns grupos tratados com BCG ou levamisole, apresentaram alterações epidérmicas atípicas, incluindo hiperplasia pseudo+epiteliomatosa, displasia e hiperceratose. Estas alterações foram mais intensas em frequência e inte [...] nsidade durante o curso da infecção, porém não foram relacionadas com o tamanho da lesão ou com a carga parasitária tissular. Animais controles, respectivamente, normais com a mesma idade, tratados com BCG e levamisole, examinados simultaneamente, não mostraram tais modificações epidérmicas. Nos camundongos infectados, a derme e a hipoderme apresentaram um infiltrado inflamatório contendo histiócitos, linfócitos e plasmócitos, acompanhado por vezes de neutrófilos e eosinófilos, o qual não variou com a duração da infecção. Abstract in english C3H mice chronically infected with Leishmania m. mexicana, and in some groups treated with BCG or levamisole, presented atypical epidermal alterations, including pseudoepitheliomatous hyperplasia, hyperkeratosis and dysplasia. These alterations increased in frequency and intensity during the course [...] of infection, but were not related to lesion size or tissue parasite load. Age matched normal, BCG and levamisole treated control mice, examined simultaneously, did not show epidermal modifications. In infected mice the dermis and hypodermis presented an inflammatory infiltrate of histiocytes, lymphocytes and plasma cells, accompanied at times by neutrophils and eosinophils, which did not vary with duration of infection.

  14. Lutzomyia longipalpis naturally infected by Leishmania (L.) chagasi in Várzea Grande, Mato Grosso State, Brazil, an area of intense transmission of visceral leishmaniasis / Lutzomyia longipalpis naturalmente infectado por Leishmania (L.) chagasi em Várzea Grande, Mato Grosso, Brasil, uma área de transmissão intensa de leishmaniose visceral

    Scientific Electronic Library Online (English)

    Nanci A., Missawa; Érika Monteiro, Michalsky; Consuelo Latorre, Fortes-Dias; Edelberto, Santos Dias.

    2010-12-01

    Full Text Available A leishmaniose visceral americana (LVA) é causada por parasitos pertencentes ao gênero Leishmania (Trypanosomatidae) e transmitida ao homem através da picada de certas espécies de flebotomíneos, previamente infectados. Neste trabalho, investigamos o índice de infecção natural de Lutzomyia longipalpi [...] s, principal vetor da LVA no Brasil, em Várzea Grande, Estado do Mato Grosso. De julho de 2004 a junho de 2006, foram feitas capturas de flebotomíneos em áreas peridomésticas utilizando armadilhas de luz CDC. Foram capturadas 420 espécimens de Lu. longipalpis. Quarenta e dois grupos, formados por 10 espécimens de Lu. longipalpis cada um, foram submetidos à extração de DNA genômico e amplificação por PCR (reação em cadeia da polimerase). DNA de Leishmania spp. foi detectado em 3 dos 42 grupos testados, resultando em um índice mínimo de infecção de 0,71%. A análise de polimorfismos de fragmentos de restrição (RFLP) indicou Leishmania (L.) chagasi como a espécie infectante nos grupos positivos. Abstract in english The American visceral leishmaniasis (AVL) is caused by parasites belonging to the genus Leishmania (Trypanosomatidae) and is transmitted to humans through the bite of certain species of infected phlebotomine sand flies. In this study, we investigated the natural infection ratio of Lutzomyia longipal [...] pis, the main vector species of AVL in Brazil, in Várzea Grande, Mato Grosso State. Between July 2004 and June 2006, phlebotomine sand flies were captured in peridomestic areas using CDC light-traps. Four hundred and twenty (420) specimens of Lu. longipalpis were captured. 42 pools, containing 10 specimens of Lu. longipalpis each, were used for genomic DNA extraction and PCR (polymerase chain reaction) amplification. Leishmania spp. DNA was detected in three out of the 42 pools tested, resulting in a minimal infection ratio of 0.71%. Restriction fragment length polymorphism (RFLP) analysis indicated that Leishmania (L.) chagasi was the infective agent in the positive pools.

  15. The SNARE protein family of Leishmania major

    Directory of Open Access Journals (Sweden)

    Mottram Jeremy C

    2006-10-01

    Full Text Available Abstract Background Leishmania major is a protozoan parasite with a highly polarised cell shape that depends upon endocytosis and exocytosis from a single area of the plasma membrane, the flagellar pocket. SNAREs (soluble N-ethylmaleimide-sensitive factor adaptor proteins receptors are key components of the intracellular vesicle-mediated transports that take place in all eukaryotic cells. They are membrane-bound proteins that facilitate the docking and fusion of vesicles with organelles. The recent availability of the genome sequence of L. major has allowed us to assess the complement of SNAREs in the parasite and to investigate their location in comparison with metazoans. Results Bioinformatic searches of the L. major genome revealed a total of 27 SNARE domain-containing proteins that could be classified in structural groups by phylogenetic analysis. 25 of these possessed the expected features of functional SNAREs, whereas the other two could represent kinetoplastid-specific proteins that might act as regulators of the SNARE complexes. Other differences of Leishmania SNAREs were the absence of double SNARE domain-containing and of the brevin classes of these proteins. Members of the Qa group of Leishmania SNAREs showed differential expressions profiles in the two main parasite forms whereas their GFP-tagging and in vivo expression revealed localisations in the Golgi, late endosome/lysosome and near the flagellar pocket. Conclusion The early-branching eukaryote L. major apparently possess a SNARE repertoire that equals in number the one of metazoans such as Drosophila, showing that the machinery for vesicle fusion is well conserved throughout the eukaryotes. However, the analysis revealed the absence of certain types of SNAREs found in metazoans and yeast, while suggesting the presence of original SNAREs as well as others with unusual localisation. This study also presented the intracellular localisation of the L. major SNAREs from the Qa group and reveals that these proteins could be useful as organelle markers in this parasitic protozoon.

  16. Pathology of dogs in Campo Grande, MS, Brazil naturally co-infected with Leishmania infantum and Ehrlichia canis / Patologia de cães naturalmente coinfectados por Leishmania infantum e Ehrlichia canis em Campo Grande, MS, Brasil

    Scientific Electronic Library Online (English)

    Gisele Braziliano, Andrade; Wanessa Teixeira Gomes, Barreto; Luciana Ladislau dos, Santos; Laura Raquel Rios, Ribeiro; Gabriel Carvalho de, Macedo; Keyla Carstens Marques de, Sousa; Marcos Rogério, André; Rosangela Zacarias, Machado; Heitor Miraglia, Herrera.

    2014-12-01

    Full Text Available A infecção simultânea por parasitas de diferentes espécies pode resultar em alterações imprevisíveis. O presente estudo avaliou a patologia de cães naturalmente coinfectados por Leishmania infantum e Ehrlichia canis. A saúde dos cães foi investigada pelas análises histopatológicas, hematológicas e b [...] ioquímicas de 21 cães infectados somente por L. infantum e 22 cães coinfectados por L. infantum e E. canis. Observou-se uma reação inflamatória crônica, predominantemente linfohistioplasmocítica, na pele dos dois grupos. A plasmocitose, encontrada nos tecidos linfóides, provavelmente estava relacionada com a hipergamaglobulinemia observada em todos os cães amostrados. A desorganização da matriz extracelular da derme da região inguinal e da orelha, demonstrada pela substituição das fibras de colágeno espessas por fibras finas, foi relacionada com o grau de reação inflamatória, independente da presença de parasitas. Ainda, observamos duas vezes mais animais do grupo coinfectado apresentando formas amastigotas na pele de orelha pela histopatologia comparado ao número de cães infectados apenas por Leishmania, tornando-os desta forma mais infectivos aos vetores. Nossos resultados ressaltam que a saúde de cães coinfectados estava severamente comprometida devido aos altos níveis de proteína plasmática total, globulinas, fosfatase alcalina, creatina quinase e anemia acentuada. Abstract in english Different parasites that commonly occur concomitantly can influence one another, sometimes with unpredictable effects. We evaluated pathological aspects of dogs naturally co-infected with Leishmania infantum and Ehrlichia canis. The health status of the dogs was investigated based on histopathologic [...] al, hematological and biochemical analyses of 21 animals infected solely with L. infantum and 22 dogs co- infected with L. infantum and E. canis. The skin of both groups showed chronic, predominantly lymphohistioplasmacytic inflammatory reaction. The plasmacytosis in the lymphoid tissues was likely related with the hypergammaglobulinemia detected in all the dogs. The disorganization of extracellular matrix found in the reticular dermis of the inguinal region and ear, characterized by the substitution of thick collagen fibers for thin fibers, was attributed to the degree of inflammatory reaction, irrespective of the presence of parasites. In addition, the histopathological analysis revealed that twice as many dogs in the co-infected group presented Leishmania amastigotes in the ear skin than those infected solely with Leishmania, increasing the possibility of becoming infected through sand fly vectors. Our findings highlight the fact that the health of dogs infected concomitantly with L. infantum and E. canis is severely compromised due to their high levels of total plasma protein, globulins, alkaline phosphatase and creatine kinase, and severe anemia.

  17. Charaterization of Leishmania major Friedlin Telomeric Terminus

    Directory of Open Access Journals (Sweden)

    Chiurillo Miguel Angel

    2002-01-01

    Full Text Available Here we have characterized Leishmania major (Friedlin telomeric terminus (the very end using recombinants obtained by a vector-adaptor cloning protocol. As in L. donovani, the last nine nucleotides of L. major terminus are 5'-GGTTAGGGT-OH 3', differing from Trypanosoma cruzi and T. brucei terminus 5'GGGTTAGGG-OH 3', thus indicating that these sequences are genus specific. We have also made a comparative analysis between L. major and L. donovani telomere-associated sequences, and described a novel non-repeated telomeric associated sequence common to L. major low molecular weight chromosomal bands.

  18. Charaterization of Leishmania major Friedlin Telomeric Terminus

    Scientific Electronic Library Online (English)

    Miguel Angel, Chiurillo; José Luis, Ramírez.

    2002-04-01

    Full Text Available Here we have characterized Leishmania major (Friedlin) telomeric terminus (the very end) using recombinants obtained by a vector-adaptor cloning protocol. As in L. donovani, the last nine nucleotides of L. major terminus are 5'-GGTTAGGGT-OH 3', differing from Trypanosoma cruzi and T. brucei terminus [...] 5'GGGTTAGGG-OH 3', thus indicating that these sequences are genus specific. We have also made a comparative analysis between L. major and L. donovani telomere-associated sequences, and described a novel non-repeated telomeric associated sequence common to L. major low molecular weight chromosomal bands.

  19. Identification and distribution of New World Leishmania species characterized by serodeme analysis using monoclonal antibodies.

    Science.gov (United States)

    Grimaldi, G; David, J R; McMahon-Pratt, D

    1987-03-01

    Five hundred thirty stocks of Leishmania isolated from human and domestic and wild reservoir hosts, representing a wide geographic distribution of endemic foci of American cutaneous (ACL) and visceral leishmaniases (AVL) were characterized and identified at species and/or subspecies levels based on their reactivity to a cross-panel of specific monoclonal antibodies using a radioimmune binding assay. This study confirms and extends our preliminary results on the high specificity of some of these monoclonals for the L. braziliensis, L. mexicana, and L. donovani complexes. This study also demonstrates the relative stability of these molecular markers and the general usefulness of the method for parasite identification. Two hundred ninety-two of 420 isolates of ACL were classified as members of the L. braziliensis complex. Two hundred twenty-seven were L. b. braziliensis; these showed the widest geographical distribution (Brazil: Amazonas, Bahia, Ceara, Espirito Santo, Goias, Minas Gerais, Para, Paraiba, Rio de Janeiro, and Sao Paulo; Honduras: Santa Barbara and Yoko; Peru: Ancash, Piura, and Ucayali; and Venezuela: Cojedes, Distrito Federal, Lara, Portuguesa, Vale Hondo, Yaracuy, and Zulia). Forty-one stocks were identified as L. b. guyanensis (from North Brazil: Amazonas, Amapa, Para, and Rondonia). Twenty-one stocks were identified as L. b. panamensis (from Costa Rica: Alajuela, Guanacasten, Limon, Puntarenas, and San Jose; and Honduras: El Paraiso, and Olancho). Out of 128 isolates classified as members of the L. mexicana complex, 74 were differentiated as L. m. amazonensis (from Bolivia; Brazil: Amazonas, Bahia, Ceara, Goias, Maranhao, Mato Grosso do Norte, and Para; Peru: Pasco Forest and Van Humboldt; and Venezuela: Carabobo, Guarico, and Merida). Forty-four stocks were identified as L. m. venezuelensis (from Venezuela: Lara). Six stocks were L. m. mexicana (from Belize; and Mexico: Campeche [corrected] and Quintana Roo, Yucatan). One hundred ten isolates from AVL were identified as L. donovani chagasi (from Brazil: Bahia, Ceara, Maranhao, Minas Gerais, Mato Grosso do Sul, Piaui, Rio de Janeiro, and Sergipe; and Honduras: Valle). The implications of these results with respect to both the clinical and epidemiological data (including the detection of seven unusual characterized stocks) are discussed. PMID:3826486

  20. Cross reactive molecules of human lymphatic filaria Brugia malayi inhibit Leishmania donovani infection in hamsters.

    Science.gov (United States)

    Verma, Richa; Joseph, Sujith K; Kushwaha, Vikas; Kumar, Vikash; Siddiqi, M I; Vishwakarma, Preeti; Shivahare, Rahul; Gupta, Suman; Murthy, P K

    2015-12-01

    Coinfections are common in natural populations and the outcome of their interactions depends on the immune responses of the host elicited by the parasites. Earlier we showed that immunization with BmAFII (Sephadex G-200 eluted) fraction of human lymphatic filaria Brugia malayi inhibited progression of Leishmania donovani infection in golden hamsters. In the present study we identified cross reactive molecules of B. malayi, and investigated their effect on L. donovani infection and associated immune responses in the host. The sequence alignment and sharing of linear T- and B-cell epitopes in protein molecules of B. malayi and L. donovani counterparts were studied in silico. Hamsters were immunized with robustly cross reactive SDS-PAGE resolved fractions F6 (54.2-67.8kDa) and F9 (41.3-45.0kDa) of B. malayi and subsequently inoculated with amastigotes of L. donovani intracardially. F6 inhibited (?72%) L. donovani infection and upregulated Th1 cytokine expression, lymphoproliferation, IgG2, IgG2/3 levels and NO production, and downregulated Th2 cytokine expression. Sequences in HSP60 and EF-2 of F6 and L. donovani counterparts were conserved and B- and T-cell epitopes in the proteins shared antigenic regions. In conclusion, leishmania-cross reactive molecules of filarial parasite considerably inhibited leishmanial infection via Th1-mediated immune responses and NO production. Common B- and T-cell epitope regions in HSP60 and EF-2 of the parasites might have contributed to the inhibitory effect on the L. donovani infection. Thus, leishmania-cross reactive filarial parasite molecules may help in designing prophylactic(s) against L. donovani. PMID:26341753

  1. The spiny rat Proechimys guyannensis (Rodentia: Echimydae) fails to respond to intradermal inoculation with Leishmania (Viannia) braziliensis / O rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) falhou para inoculação intradérmica com Leishmania (Viannia) braziliensis.

    Scientific Electronic Library Online (English)

    Stela Rechinelli, PASSOS; Ana Paula, MADUREIRA; Sayanne Luns Hatum de, ALMEIDA; Marcos Santos, ZANINI.

    2015-06-01

    Full Text Available A leishmaniose é uma doença de ocorrência mundial causada por protozoários do gênero Leishmania. No Brasil, a Leishmania (Viannia) braziliensis é o principal parasita responsável pela leishmaniose tegumentar americana. Os principais hospedeiros deste protozoário são pequenos mamíferos selvagens em p [...] articular marsupiais e roedores. O objetivo deste estudo foi avaliar o papel do rato espinhoso Proechimys guyannensis (Rodentia: Echimydae) no ciclo da leishmaniose tegumentar americana. Para isto, formas promastigotas (estágio flagelado) de Leishmania (Viannia) braziliensis foram inoculadas em sete ratos espinhosos (Proechimys guyannensis). Após a inoculação intradérmica no pavilhão auricular, focinho e área plantar, os ratos foram monitorizados durante 180 dias. Amostras de tecido colhidas aos 90 e 180 dias dos ratos revelaram-se negativas para a presença de material genético do parasita. Após eutanásia, tecidos coletados dos ratos também falharam para crescimento em meio de cultura demonstrando que não houve infecção. Estes resultados demonstram que o rato espinhoso não tem papel no ciclo da leishmaniose tegumentar americana causada por L. (V.) braziliensis. Abstract in english Leishmaniasis a disease of worldwide occurrence is caused by protozoa of the Leishmania genus. In Brazil, Leishmania (Viannia) braziliensis is the main parasite responsible for the American cutaneous leishmaniasis. Main hosts of this protozoa are small wild mammals particularly marsupials and rodent [...] s. The aim of this study was to evaluate if spiny rat Proechimys guyannensis (Rodentia: Echimydae) has role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis. Thus, promastigotes (the flagellate stage) of Leishmania (Viannia) braziliensis were used to inoculate seven spiny rats (Proechimys guyannensis). After inoculated intradermal at the ear pinna, nose and plantar pad, the rats were monitored for 180 days. Tissue samples collected at 90 and 180 days from the rats proved to be negative for the presence of genetic material from the parasite. After euthanasia, the protozoa also failed to growth in culture medium containing tissue samples collected from the rats showing that there was no infection. These results fail to prove that spiny rat has a role in the cycle of the American cutaneous leishmaniasis caused by L. (V.) braziliensis.

  2. DISTINCT CELLULAR MIGRATION INDUCED BY Leishmania infantum chagasi AND SALIVA FROM Lutzomyia longipalpis IN A HEMORRHAGIC POOL MODEL / Migração celular distinta induzida por Leishmania infantum chagasi e saliva de Lutzomyia longipalpis em um modelo de pool hemorrágico

    Scientific Electronic Library Online (English)

    Camila Oliveira, Vasconcelos; Zirlane C. Branco, Coelho; Cristina de Souza, Chaves; Clarissa Romero, Teixeira; Margarida M. Lima, Pompeu; Maria Jania, Teixeira.

    2014-01-01

    Full Text Available O recrutamento de uma população de células específicas após infecção por Leishmania pode influenciar o resultado da doença. A migração celular em resposta a Leishmania ou saliva do vetor tem sido reportada utilizando o modelo da bolsa de ar subcutânea, entretanto, a migração celular induzida por Lei [...] shmania associada com o sangue do hospedeiro e saliva do vetor neste modelo ainda não foi descrita. Neste trabalho foi investigada a migração celular no modelo da bolsa de ar subcutânea em hamster após a estimulação com a combinação de L. chagasi, sangue do hospedeiro e saliva de Lutzomyia longipalpis. A migração induzida por saliva foi três vezes maior do que a induzida por L. chagasi sozinha. Adicionalmente, L. chagasi associada com sangue e saliva induziu significativamente ainda mais leucócitos no exsudato inflamatório do que o estímulo com Leishmania sozinha. L. chagasi recrutou uma população de células distintas, no entanto, a maioria dessas células parece não ter migrado para o exsudato inflamatório, permanecendo no tecido da bolsa de ar. Estes resultados indicam que L. chagasi pode reduzir o acúmulo de leucócitos para o local inicial da infecção e que quando associada à saliva do vetor e na presença de componentes do sangue aumenta o influxo de mais neutrófilos do que macrófagos, sugerindo que o parasito desenvolveu uma estratégia para minimizar a resposta inflamatória inicial, permitindo uma progressão ilimitada dentro do hospedeiro. Este trabalho reforça a importância de mais estudos sobre os componentes da saliva dos vetores das leishmanioses no processo de transmissão e no estabelecimento da infecção. Abstract in english Recruitment of a specific cell population after Leishmania infection can influence the outcome of the disease. Cellular migration in response to Leishmania or vector saliva has been reported in air pouch model, however, cellular migration induced by Leishmania associated with host's blood and vector [...] saliva in this model has not been described. Herein we investigated cellular migration into air pouch of hamster after stimulation with combination of L. chagasi and host's blood and Lutzomyia longipalpis saliva. Migration induced by saliva was 3-fold more than those induced by L. chagasi alone. Additionally, L. chagasi associated with blood and saliva induced significantly even more leukocytes into air pouch than Leishmania alone. L. chagasi recruited a diverse cell population; however, most of these cells seem to have not migrated to the inflammatory exudate, remaining in the pouch lining tissue. These results indicate that L. chagasi can reduce leukocyte accumulation to the initial site of infection, and when associated with vector saliva in the presence of blood components, increase the influx of more neutrophils than macrophages, suggesting that the parasite has developed a strategy to minimize the initial inflammatory response, allowing an unlimited progression within the host. This work reinforces the importance of studies on the salivary components of sand fly vectors of leishmaniasis in the transmission process and the establishment of the infection.

  3. The interactions and essential effects of intrinsic insulin-like growth factor-I on Leishmania (Leishmania) major growth within macrophages

    OpenAIRE

    L.C. Reis; Ramos-Sanchez, E M; Goto, H.

    2013-01-01

    Previously, we showed in Leishmania infections that extrinsic insulin-like growth factor (IGF)-I favored Leishmania proliferation and leishmaniasis development. In this study, the interaction of intrinsically expressed IGF-I and Leishmania (Leishmania) major in macrophages was addressed, and a key finding was the observation, using confocal microscopy, of the co-localization of IGF-I and parasites within macrophages. Following stimulation with interferon-γ (IFN-γ), which is known to inhibit I...

  4. Endoplasmic reticulum stress responses in Leishmania.

    Science.gov (United States)

    Dolai, Subhankar; Adak, Subrata

    2014-10-01

    Perturbation of endoplasmic reticulum (ER) homeostasis can lead to an accumulation of misfolded proteins within the ER lumen causing initiation of ER stress. To reestablish homeostasis and mitigate the stress, a series of adaptive intracellular signaling pathways termed the unfolded protein response (UPR) are activated. ER stress is of considerable interest to parasitologists because it takes place in parasites subjected to adverse environmental conditions. During a digenetic lifestyle, Leishmania parasites encounter and adapt to harsh environmental conditions that provide potential triggers of ER stress. These include nutrient deficiency, hypoxia, oxidative stress, changing pH, and shifts in temperature. Protozoan human pathogens, including the causative agents of trypanosomiasis, leishmaniasis, toxoplasmosis and malaria, contain a minimal conventional UPR network relative to higher eukaryotic cells. Three different signaling pathways in the ER stress response have been described in trypanosomatids: these pathways involve (i) the down-regulation of translation by a protein kinase RNA-like ER kinase (PERK), (ii) the ER-associated degradation (ERAD) pathway, and (iii) the spliced leader silencing (SLS) pathway and its target mRNAs. Under short-term ER stress, signaling from PERK activates autophagy, a cell survival response. But both chronic and unresolved ER stresses lead to initiation of apoptotic events and eventual cell death. This review presents the current understanding of the ER stress response in Leishmania with an emphasis on protein folding and ER quality control, unfolded protein response, autophagy as well as apoptosis in reference to the mammalian system. PMID:25224909

  5. Cyclic nucleotide specific phosphodiesterases of Leishmania major

    Directory of Open Access Journals (Sweden)

    Linder Markus

    2006-03-01

    Full Text Available Abstract Background Leishmania represent a complex of important human pathogens that belong to the systematic order of the kinetoplastida. They are transmitted between their human and mammalian hosts by different bloodsucking sandfly vectors. In their hosts, the Leishmania undergo several differentiation steps, and their coordination and optimization crucially depend on numerous interactions between the parasites and the physiological environment presented by the fly and human hosts. Little is still known about the signalling networks involved in these functions. In an attempt to better understand the role of cyclic nucleotide signalling in Leishmania differentiation and host-parasite interaction, we here present an initial study on the cyclic nucleotide-specific phosphodiesterases of Leishmania major. Results This paper presents the identification of three class I cyclic-nucleotide-specific phosphodiesterases (PDEs from L. major, PDEs whose catalytic domains exhibit considerable sequence conservation with, among other, all eleven human PDE families. In contrast to other protozoa such as Dictyostelium, or fungi such as Saccharomyces cerevisiae, Candida ssp or Neurospora, no genes for class II PDEs were found in the Leishmania genomes. LmjPDEA contains a class I catalytic domain at the C-terminus of the polypeptide, with no other discernible functional domains elsewhere. LmjPDEB1 and LmjPDEB2 are coded for by closely related, tandemly linked genes on chromosome 15. Both PDEs contain two GAF domains in their N-terminal region, and their almost identical catalytic domains are located at the C-terminus of the polypeptide. LmjPDEA, LmjPDEB1 and LmjPDEB2 were further characterized by functional complementation in a PDE-deficient S. cerevisiae strain. All three enzymes conferred complementation, demonstrating that all three can hydrolyze cAMP. Recombinant LmjPDEB1 and LmjPDEB2 were shown to be cAMP-specific, with Km values in the low micromolar range. Several PDE inhibitors were found to be active against these PDEs in vitro, and to inhibit cell proliferation. Conclusion The genome of L. major contains only PDE genes that are predicted to code for class I PDEs, and none for class II PDEs. This is more similar to what is found in higher eukaryotes than it is to the situation in Dictyostelium or the fungi that concomitantly express class I and class II PDEs. Functional complementation demonstrated that LmjPDEA, LmjPDEB1 and LmjPDEB2 are capable of hydrolyzing cAMP. In vitro studies with recombinant LmjPDEB1 and LmjPDEB2 confirmed this, and they demonstrated that both are completely cAMP-specific. Both enzymes are inhibited by several commercially available PDE inhibitors. The observation that these inhibitors also interfere with cell growth in culture indicates that inhibition of the PDEs is fatal for the cell, suggesting an important role of cAMP signalling for the maintenance of cellular integrity and proliferation.

  6. Characterization and regulation of Leishmania major 3-hydroxy-3-methylglutaryl-CoA reductase

    DEFF Research Database (Denmark)

    Montalvetti, A; Pena Diaz, Javier

    2000-01-01

    In eukaryotes the enzyme 3-hydroxy-3-methylglutaryl CoA (HMG-CoA) reductase catalyses the synthesis of mevalonic acid, a common precursor to all isoprenoid compounds. Here we report the isolation and overexpression of the gene coding for HMG-CoA reductase from Leishmania major. The protein from Leishmania lacks the membrane domain characteristic of eukaryotic cells but exhibits sequence similarity with eukaryotic reductases. Highly purified protein was achieved by ammonium sulphate precipitation followed by chromatography on hydroxyapatite. Kinetic parameters were determined for the protozoan reductase, obtaining K(m) values for the overall reaction of 40.3+/-5.8 microM for (R,S)-HMG-CoA and 81.4+/-5.3 microM for NADPH; V(max) was 33.55+/-1.8 units x mg(-1). Gel-filtration experiments suggested an apparent molecular mass of 184 kDa with subunits of 46 kDa. Finally, in order to achieve a better understanding of the role of this enzyme in trypanosomatids, the effect of possible regulators of isoprenoid biosynthesis in cultured promastigote cells was studied. Neither mevalonic acid nor serum sterols appear to modulate enzyme activity whereas incubation with lovastatin results in significant increases in the amount of reductase protein. Western- and Northern-blot analyses indicate that this activation is apparently performed via post-transcriptional control.

  7. Evaluacion por western blot, inmunofluorescencia indirecta y elisa de perros infectados con leishmania (leishmania) infantum

    OpenAIRE

    Jimmy J Vargas-Duarte; López-Páez, Myriam C.; Escovar-Castro, Jesús E.; Fernández-Manrique, José

    2012-01-01

    Objetivo Evaluar el desempeño de las pruebas empleadas en Colombia para el diagnóstico de la leishmaniasis visceral canina y adaptar una técnica de Western blot empleando animales experimental y naturalmente infectados.  Metodología Se obtuvieron sueros de 10 perros infectados experimentalmente con L. infantum, 5 perros infectados naturalmente, 16 perros sanos, 26 de reacción cruzada (infectados con Babesia canis, Erhlichia canis, Dirofilaria immitis, Trypanosoma cruzi, Leishmania (Viannia) s...

  8. Reconstrucción quirúrgica tras destrucción nasal por Leishmania Panamensis / Surgical reconstruction after nasal destruction by Leishmania Panamensis

    Scientific Electronic Library Online (English)

    F., Vélez Bernal; J.A., Vélez Trujillo; I.D., Vélez Bernal.

    2013-03-01

    Full Text Available Algunas especies de Leishmania del subgénero Viannia, especialmente Leishmania braziliensis y Leishmania panamensis, pueden invadir la mucosa naso-orofaríngea al diseminarse por vía linfática y sanguínea a partir de una lesión cutánea y ocasionar lesiones en el tabique nasal, paladar blando, úvula, [...] pilares amigdalinos, laringe, faringe, dorso nasal, labios y pómulos, que pueden conducir a la desfiguración. La mucosa más frecuentemente afectada es la del tabique nasal, principalmente en su parte anterior. La invasión de la mucosa puede ocurrir simultáneamente con lesiones cutáneas activas, aunque más frecuentemente aparecen 1 o 2 años después de la lesión en la piel; sin embargo, en el 16 % de los casos no hay antecedentes de lesiones cutáneas, lo que sugiere que con la picadura del insecto vector se produjo una infección primaria asintomática u oligosintomática y luego se produjo la diseminación del parasito a la mucosas. En este artículo presentamos 2 casos clínicos de leishmaniosis mucosa producidos por L. panamensis y los procedimientos quirúrgicos reconstructivos que se realizaron. Se hace además un recuento de los diagnósticos diferenciales en tejidos oronasales. Abstract in english Species of Leishmania of Viannia subgenus, mainly L. braziliensis and L. panamensis, may invade the nasooro-pharyngeal mucosal after spread from the skin lesion via lymph and blood, causing lesions in the nasal septum, soft palate, uvula, tonsillar pillars, larynx, pharynx, nasal dorsum, lips and ch [...] eeks. The mucosal membrane most frequently affected is the nasal septum, mainly in the anterior region. The invasion of mucosa may occur simultaneously with active skin lesions, but most often appear 1 or 2 years after the skin lesion; nevertheless, in 16 % of cases there is no history of skin lesions suggesting that the primary infection coursed with few symptoms and then was spread to mucosal membranes. In this article 2 cases of L panamensis mucosal leishmaniasis and the reconstructive surgery used are presented. A discussion of the differential diagnosis of oro-nasal tissues is also presented.

  9. Antigenic profile of heat-killed versus thimerosal-treated Leishmania major using sodium dodecyl sulfate-polyacrylamide gel electrophoresis

    Directory of Open Access Journals (Sweden)

    Reza Arjmand

    2015-01-01

    Full Text Available Background: Leishmania is a parasitic protozoan of trypanosomatidae family which causes a wide spectrum of diseases ranging from self-healing cutaneous lesions to deadly visceral forms. In endemic areas, field trials of different preparations of Leishmania total antigen were tested as leishmaniasis vaccine. Two preparations of killed Leishmania major were produced In Iran, which were heat-killed vaccine called autoclaved L. major (ALM and thimerosal-treated freeze-thawed vaccine called killed L. major (KLM. In this study, the protein content of both ALM and KLM were compared with that of freshly harvested intact L. major promastigotes using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE. Materials and Methods: L. major (MRHO/IR/75/ER from pre-infected Balb/c mice was isolated with modified Novy-MacNeal-Nicolle (NNN medium and then subcultured in liquid RPMI 1640 medium supplemented with fetal calf serum (FCS 20% for mass production. Two preparations of KLM and ALM were produced by Razi Vaccine and Serum Research Institute, Iran, under WHO/TDR supervision. Electrophoresis was performed by SDS-PAGE method and the gel was stained by Coomassie brilliant blue dye. The resultant unit bands were compared using standard molecular proteins. Results: Electrophoresis of the two preparations produced many bands from 10 kDa to 100 kDa. KLM bands were much like those of freshly harvested intact L. major. Conclusion: It is concluded that although there are similar bands in the three forms of Leishmania antigens, there are some variations which might be considered for identification and purification of protective immunogens in a total crude antigen, and detection of their stability is essential for the production and marketing of a putative vaccine.

  10. Leishmaniose tegumentar, visceral e doença de Chagas caninas em municípios do Triângulo Mineiro e Alto Paranaíba, Minas Gerais, Brasil

    OpenAIRE

    Maywald Paula Guardenho; Machado Maria Inês; Costa-Cruz Julia Maria; Gonçalves-Pires Maria do Rosário de Fátima

    1996-01-01

    Inquérito envolvendo leishmaniose e doença de Chagas, por meio da Reação de Imunofluorescência Indireta, foi realizado com soros de 331 cães de Uberlândia e Coromandel, Municípios do Estado de Minas Gerais, Brasil. Para tal inquérito, utilizaram-se, como antígenos, Leishmania amazonensis e Trypanosoma cruzi. No que tange a Uberlândia, examinaram-se 230 soros, sendo 200 da área urbana com 4,5% de positividade, e 30 da área rural, dos quais, 6,6% positivos para a RIFI com antígeno L. amazonensi...

  11. Leishmaniose tegumentar, visceral e doença de Chagas caninas em municípios do Triângulo Mineiro e Alto Paranaíba, Minas Gerais, Brasil Canine cutaneous and visceral leishmaniasis and Chagas' disease from counties in the Triângulo Mineiro and Alto Paranaíba regions, Minas Gerais State, Brazil

    OpenAIRE

    Paula Guardenho Maywald; Maria Inês Machado; Julia Maria Costa-Cruz; Maria do Rosário de Fátima Gonçalves-Pires

    1996-01-01

    Inquérito envolvendo leishmaniose e doença de Chagas, por meio da Reação de Imunofluorescência Indireta, foi realizado com soros de 331 cães de Uberlândia e Coromandel, Municípios do Estado de Minas Gerais, Brasil. Para tal inquérito, utilizaram-se, como antígenos, Leishmania amazonensis e Trypanosoma cruzi. No que tange a Uberlândia, examinaram-se 230 soros, sendo 200 da área urbana com 4,5% de positividade, e 30 da área rural, dos quais, 6,6% positivos para a RIFI com antígeno L. amazonensi...

  12. Implications of a Neotropical Origin of the Genus Leishmania

    Directory of Open Access Journals (Sweden)

    Noyes Harry

    1998-01-01

    Full Text Available The hypothesis of a Neotropical origin of the Leishmania/Endotrypanum clade is reviewed. The position of the L. (Sauroleishmania external to the subgenus L. (Leishmania is not consistent with the Neotropical origin of the latter subgenus. It is suggested that this may be a consequence of a faster evolutionary rate in the L. (Sauroleishmania. The implications for the classsification of the phlebotomine sandflies of the hypothesis for a Neotropical origin of the Leishmania is also considered. The classification of Galati (1995 is proposed to be most consistent with the hypothesis of a Neotropical origin of the Leishmania, whilst classifications which place the New and Old World species in separate taxa are inconsistent with this hypothesis.

  13. A comparison of molecular markers to detect Lutzomyia longipalpis naturally infected with Leishmania (Leishmania) infantum

    Scientific Electronic Library Online (English)

    Kárita Cláudia, Freitas-Lidani; Iara J de, Messias-Reason; Edna Aoba Y, Ishikawa.

    2014-07-03

    Full Text Available The aim of the present study was to detect natural infection by Leishmania (Leishmania) infantum in Lutzomyia longipalpis captured in Barcarena, state of Pará, Brazil, through the use of three primer sets. With this approach, it is unnecessary to previously dissect the sandfly specimens. DNA of 280 [...] Lu. longipalpis female specimens were extracted from the whole insects. PCR primers for kinetoplast minicircle DNA (kDNA), the mini-exon gene and the small subunit ribosomal RNA (SSU-rRNA) gene of Leishmania were used, generating fragments of 400 bp, 780 bp and 603 bp, respectively. Infection by the parasite was found with the kDNA primer in 8.6% of the cases, with the mini-exon gene primer in 7.1% of the cases and with the SSU-rRNA gene primer in 5.3% of the cases. These data show the importance of polymerase chain reaction as a tool for investigating the molecular epidemiology of visceral leishmaniasis by estimating the risk of disease transmission in endemic areas, with the kDNA primer representing the most reliable marker for the parasite.

  14. In vitro evaluation of new terpenoid derivatives against Leishmania infantum and Leishmania braziliensis

    Scientific Electronic Library Online (English)

    Inmaculada, Ramírez-Macías; Clotilde, Marín; Rachid, Chahboun; Francisco, Olmo; Ibtisam, Messouri; Oscar, Huertas; María Jose, Rosales; Ramón, Gutierrez-Sánchez; Enrique, Alvarez-Manzaneda; Manuel, Sánchez-Moreno.

    2012-05-01

    Full Text Available The activity of five (1-5) abietane phenol derivatives against Leishmania infantum and Leishmania braziliensis was studied using promastigotes and axenic and intracellular amastigotes. Infectivity and cytotoxicity tests were performed with J774.2 macrophage cells using Glucantime as a reference drug [...] . The mechanisms of action were analysed by performing metabolite excretion and transmission electron microscopy ultrastructural studies. Compounds 1-5 were more active and less toxic than Glucantime. The infection rates and mean number of parasites per cell observed in amastigote experiments showed that derivatives 2, 4 and 5 were the most effective against both L. infantum and L. braziliensis. The ultrastructural changes observed in the treated promastigote forms confirmed that the greatest cell damage was caused by the most active compound (4). Only compound 5 caused changes in the nature and amounts of catabolites excreted by the parasites, as measured by ¹H nuclear magnetic resonance spectroscopy. All of the assayed compounds were active against the two Leishmania species in vitro and were less toxic in mammalian cells than the reference drug.

  15. A comparison of molecular markers to detect Lutzomyia longipalpis naturally infected with Leishmania (Leishmania infantum

    Directory of Open Access Journals (Sweden)

    Kárita Cláudia Freitas-Lidani

    2014-07-01

    Full Text Available The aim of the present study was to detect natural infection by Leishmania (Leishmania infantum in Lutzomyia longipalpis captured in Barcarena, state of Pará, Brazil, through the use of three primer sets. With this approach, it is unnecessary to previously dissect the sandfly specimens. DNA of 280 Lu. longipalpis female specimens were extracted from the whole insects. PCR primers for kinetoplast minicircle DNA (kDNA, the mini-exon gene and the small subunit ribosomal RNA (SSU-rRNA gene of Leishmania were used, generating fragments of 400 bp, 780 bp and 603 bp, respectively. Infection by the parasite was found with the kDNA primer in 8.6% of the cases, with the mini-exon gene primer in 7.1% of the cases and with the SSU-rRNA gene primer in 5.3% of the cases. These data show the importance of polymerase chain reaction as a tool for investigating the molecular epidemiology of visceral leishmaniasis by estimating the risk of disease transmission in endemic areas, with the kDNA primer representing the most reliable marker for the parasite.

  16. Reduced Antimony Accumulation in ARM58-Overexpressing Leishmania infantum

    OpenAIRE

    Schäfer, Carola; Tejera Nevado, Paloma; Zander, Dorothea; Clos, Joachim

    2014-01-01

    Antimony-based drugs are still the mainstay of chemotherapy against Leishmania infections in many countries where the parasites are endemic. The efficacy of antimonials has been compromised by increasing numbers of resistant infections, the basis of which is not fully understood and likely involves multiple factors. By using a functional cloning strategy, we recently identified a novel antimony resistance marker, ARM58, from the parasite Leishmania braziliensis that protects the parasites aga...

  17. Pathways of iron acquisition and utilization in Leishmania

    OpenAIRE

    Flannery, Andrew R.; Renberg, Rebecca L.; Andrews, Norma W

    2013-01-01

    Iron is essential for many metabolic pathways, but is toxic in excess. Recent identification of the ferric iron reductase LFR1, the ferrous iron transporter LIT1, and the heme transporter LHR1 greatly advanced our understanding of how Leishmania parasites acquire iron and regulate its uptake. LFR1 and LIT1 have close orthologs in plants, and are required for Leishmania virulence. Consistent with the lack of heme biosynthesis in trypanosomatids, LHR1 and LABCG5, a protein involved in heme salv...

  18. Application of the microscopic method in cutaneous leishmania diagnosis

    OpenAIRE

    Mohammed Wael Daboul

    2011-01-01

    Introduction: Cutaneous leishmania is spreading fast. This study aims at developing the microscopic method to achieve a full detection of all positive cases of leishmania.Methods: 50 human cases have been studied by applying microscopic smears stained with Wright stain. Microscopic photos were taken for the presumed unfamiliar figures.Results: Mononuclear cells with tails are present at a rate of (98%). They are associated with Leishman Donovan (LD) bodies in 50% of the cases. The polygonal f...

  19. Chronic interstitial pneumonitis in dogs naturally infected with Leishmania (Leishmania) chagasi: a histopathological and morphometric study Pneumonia intersticial crônica em cães naturalmente infectados com Leishmania (Leishmania) chagasi: estudo histopatológico e morfométrico

    OpenAIRE

    Ricardo Gonçalves; Washington Luiz Tafuri; Maria Norma de Melo; Pedro Raso; Wagner Luiz Tafuri

    2003-01-01

    Eighteen mongrel dogs of unknown age and naturally infected with Leishmania (Leishmania) chagasi, were obtained from the City Hall of Belo Horizonte, Brazil. Four dogs were used as control. Lung samples were obtained and immediately fixed in formalin. The histopathological picture of all lung tissue sections was a chronic and diffuse interstitial pneumonitis. The thickened inter-alveolar septa were characterized by the cellular exudate (mostly macrophages, lymphocytes and plasmocytes) associa...

  20. Leishmania (Viannia) Infection in the Domestic Dog in Chaparral, Colombia

    OpenAIRE

    Santaella, Julián; Clara B. Ocampo; Nancy G Saravia; Méndez, Fabián; Góngora, Rafael; Gomez, Maria Adelaida; Leonard E Munstermann; Quinnell, Rupert J.

    2011-01-01

    Peridomestic transmission of American cutaneous leishmaniasis is increasingly reported and dogs may be a reservoir of Leishmania (Viannia) in this setting. We investigated the prevalence of infection in dogs in Chaparral County, Colombia, the focus of an epidemic of human cutaneous leishmaniasis caused by Leishmania (Viannia) guyanensis. Two (0.72%) of 279 dogs had lesions typical of cutaneous leishmaniasis that were biopsy positive by kinetoplast DNA polymerase chain reaction–Southern blotti...

  1. Miltefosine Induces Apoptosis-Like Death in Leishmania donovani Promastigotes

    OpenAIRE

    Paris, Caroline; Philippe M. Loiseau; Bories, Christian; Bréard, Jaqueline

    2004-01-01

    Miltefosine (hexadecylphosphocholine [HePC]) has proved to be a potent oral treatment for human visceral leishmaniasis due to Leishmania donovani. The molecular mechanisms that contribute to the antileishmanial activity of HePC are still unknown. We report that in wild-type promastigotes of Leishmania donovani HePC is able to induce a cell death process with numerous cytoplasmic, nuclear, and membrane features of metazoan apoptosis, including cell shrinkage, DNA fragmentation into oligonucleo...

  2. Preliminary phytochemical and antileishmanial studies of the ethanolic extracts of Pterodon pudescens / Estudos preliminares sobre a fitoquimica e a atividade anti-leishmania de extratos etanólicos de Pterodon pudescens

    Scientific Electronic Library Online (English)

    W.W., Arrais-Silva; P.S.G., Nunes; J.D., Carvalho; M.W., Brune; C., Arrais-Lima; C., Batalini.

    2014-09-01

    Full Text Available Antimoniais pentavalentes são a primeira escolha para o tratamento das leishmanioses humanas. No entanto, no interior brasileiro plantas tradicionais são usadas para o tratamento dessas lesões. De fato, recentes trabalhos tem relatado o potencial terapêutico de produtos naturais, especialmente deriv [...] ados de plantas. Neste estudo avaliamos a atividade leishmanicida de Pterodon pubescens, uma árvore nativa, distribuída pela região central brasileira e usada em infusões para tratamento de inflamações. Foi realizada a análise fitoquímica e o ensaio in vitro em macrófagos infectados com Leishmania amazonensis em concentrações de 150, 300, 450, 600 µg/ml do extrato etanólico de folhas de Pterodon pudescens (PPE) para comprovar o uso tradicional desta planta como terapia para as leishmanioses. Os testes fitoquímicos indicaram a presença de taninos catequímicos, flavonas, esteroides, triterpenoides, flavonoides e xantonas. Os ensaios biológicos revelaram que o PPE foi capaz de controlar a carga parasitária em macrófagos de maneira dose dependente. Estes resultados corroboram com o potencial terapêutico de compostos de Pterodon pudescens e, junto com sua ampla distribuição no Brasil, podem representar promissor agente leishmanicida. Abstract in english Pentavalent antimonials are the first choice for the treatment of human leishmaniasis. However in rural areas the traditional plants may be preferred for the treatment of lesions. In recent years a number of papers are published related to the natural products especially plant derivative with infect [...] ious diseases. The present work was undertaken to evaluate the antileishmanial activity of Pterodon pubescens which is a native tree widely distributed over the central region of Brazil and used in folk medicine as wine infusions to treat inflammatory disease. The phytochemical screening and the biological essay of ethanolic extract of Pterodon pudescens (PPE) leaves at the concentrations of 150, 300, 450, 600 µg/ml were tested in vitro in Leishmania amazonensis-infected macrophages to support its traditional medicinal use as a leishmaniasis remedy. Phytochemical screening of PPE has shown the presence of catechemical tannins, steroids, triterpenoids and flavonoids. The biological test suggests that PPE were found to control parasite burden of cell cultures in dose-dependent manner. These findings highlight the fact that the apparent potency of Pterodon pudescens compounds, together with their widely distribution over Latin America and Brazil, may represent a promising antileishmanial agent.

  3. immune response in human leishmania infections Respuesta inmune en infecciones humanas por Leishmania spp

    Directory of Open Access Journals (Sweden)

    Sara María Robledo Restrepo

    2000-03-01

    Full Text Available This review summarizes relevant information about the immune response triggered during leishmaniosis, a disease of great importance from the epidemiological point of view, since it is endemic in Colombia and other countries. We emphasize on human leishmaniosis; nevertheless, some important findings in the murine model are also mentioned. This information allows to conclude that Leishmania infection is a complex and coordinated process, which includes adhesion and entrance of the parasite into the host cells and its survival inside them. Events that mediate the infection process may influence its result in terms of elimination of the parasite or development of the disease, through induction or not of an effective specific immune response which involves host cell activation and parasite destruction. La presente revisión tiene como objetivo resumir la información más relevante acerca de la respuesta inmune que se desencadena durante la leishmaniosis, una enfermedad de gran importancia desde el punto de vista epidemiológico dado que es endémica en Colombia y otros países. Aunque la respuesta inmune en la leishmaniosis es un tema que se ha estudiado ampliamente en las infecciones por especies de Leishmania del Viejo Mundo, particularmente Leishmania major y Leishmania donovani y en el modelo murino, la presente revisión hace énfasis en la leishmaniosis humana. Algunos hallazgos importantes en el modelo murino también se mencionan. La información contenida en la revisión, en su mayoría, proviene de publicaciones derivadas de investigaciones, las cuales se seleccionaron con base en la calidad del trabajo realizado y en los aportes de sus resultados en el avance del conocimiento sobre las infecciones en humanos. La síntesis de la información seleccionada nos permite concluir que la infección por Leishmania es un proceso complejo y coordinado que incluye la adherencia y entrada del parásito a la célula hospedera y su posterior supervivencia en el interior de la célula infectada. Los eventos que median el proceso de infección influyen en su resultado en términos de eliminación del parásito o desarrollo de la enfermedad, a través de la inducción o no de una respuesta inmune específica efectiva que lleve a la activación de la célula hospedera y la muerte y destrucción del parásito.

  4. Diagnosis of Leishmania (Leishmania) chagasi infection in dogs and the relationship with environmental and sanitary aspects in the municipality of Palmas, state of Tocantins, Brazil / Identificação de cães infectados por Leishmania (Leishmania) chagasi e sua relação com aspectos ambientais e sanitários no município de Palmas, estado do Tocantins, Brasil

    Scientific Electronic Library Online (English)

    Julio Gomes, Bigeli; Waldesse Piragé de, Oliveira Júnior; Natália Melquie Monteiro, Teles.

    2012-02-01

    Full Text Available INTRODUÇÃO: O estudo foi realizado com o objetivo de identificar, através da PCR, a presença da infecção por Leishmania (Leishmania) chagasi em cães no município de Palmas, no Estado do Tocantins, Brasil, de modo a elencar os hot spots de cães infectados no município e associar sua ocorrência a alte [...] rações ambientais marcantes nos locais de captura. MÉTODOS: O DNA foi extraído do sangue dos cães e as reações de PCR foram realizadas com os primers RV1/RV2. Após o screening da população estudada, foram detectadas as regiões do município que apresentavam as maiores ocorrências da infecção canina. Esses locais foram visitados, e parâmetros de distúrbio ecológico com origem antrópica foram avaliados. RESULTADOS: Algumas características importantes foram constantes entre as regiões visitadas, entre elas a baixa urbanização, inexistência de coleta pública de esgoto, coleta publica de lixo pouco abrangente, lotes vagos com vegetação alta, e matéria orgânica em decomposição, com destaque para criação de cães soltos, e aves nas residências. CONCLUSÕES: A metodologia adotada para screening da população se mostrou bastante eficiente, sobretudo na avaliação de um grande número de indivíduos em tempo reduzido, com alto grau de automatização. Os resultados apresentados indicam associação entre os parâmetros observados e a ocorrência da infecção em cães. O modelo apresentado no município é ideal para estudos do desenvolvimento da doença, bem como sua expansão, além da avaliação das medidas de controle adotadas para a leishmaniose visceral canina. Abstract in english INTRODUCTION: The aim of the present study was to identify the presence of Leishmania (Leishmania) chagasi infection in dogs in the City of Palmas, Tocantins, Brazil, using the PCR technique to list the hot spots of infected dogs in the city and associate their occurrence to significant environmenta [...] l changes at capture sites. METHODS: DNA was extracted from blood of dogs, and the PCR were performed with primers RV1/RV2. After screening the population studied, the regions of the city that had the highest occurrence of canine infection were detected. These sites were visited, and ecological parameters denoting anthropogenic disturbance were evaluated. RESULTS: Some important features were listed in the regions visited, such as low urbanization, lack of public collection of sewage, limited garbage collection, vacant lots with tall vegetation, decaying organic matter, and, most importantly, the occurrence of stray dogs and poultry in homes. CONCLUSIONS: The methodology for screening the population was very efficient, especially in evaluating a large number of individuals in a short time, with a high degree of automation. The results indicate an association between the observed parameters and the occurrence of infection in dogs. The model presented in the city is ideal for studies of disease progression and expansion and for the evaluation of control measures adopted for canine VL.

  5. Leishmaniosis (Leishmania infantum infection) in dogs.

    Science.gov (United States)

    Gharbi, M; Mhadhbi, M; Rejeb, A; Jaouadi, K; Rouatbi, M; Darghouth, M A

    2015-08-01

    The authors present an overview of canine leishmaniosis due to Leishmania infantum. This protozoan is transmitted by sandflies and the disease is frequently characterised by chronic evolution. Cutaneous and visceral clinical signs appear as the infection progresses. Lymph node enlargement, emaciation and skin lesions are the main signs observed in the classical forms of the disease. Control is difficult since infected dogs remain carriers for years and may relapse at any time. The mass screening of infected animals and their treatment or euthanasia represent the best way to reduce the prevalence of this disease in endemic regions. Further research is needed to improve the efficiency of the vaccines available to protect dogs against infection. This disease is zoonotic; in humans, clinical cases are reported mainly in elderly people, the young and those whose immune systems have been compromised. PMID:26601461

  6. How clonal are Trypanosoma and Leishmania?

    Science.gov (United States)

    Tibayrenc, Michel; Ayala, Francisco J

    2013-06-01

    The clonal theory of parasitic protozoa has been recently challenged by researchers stating that recombination in Kinetoplastida is much more frequent than previously believed, or that selfing and homogamy should be distinguished from 'strict' clonality. These researchers and many others show that the concept of clonality proposed by us is not correctly understood. A recapitulation of the clonal theory will thus be addressed herein. Comparisons with various other pathogens evidence general features among them and enhance our understanding of Trypanosoma and Leishmania population genetics. The relevance is considerable not only for our knowledge of the basic biology of these organisms but also for applied research: molecular epidemiology (strain-typing), clinical research, vaccine and drug design, and experimental evolution. PMID:23602631

  7. ATP generation in Leishmania donovani amastigote form

    Directory of Open Access Journals (Sweden)

    Anup Kumar Roy

    2013-06-01

    Full Text Available Leishmania is the causative agent of various forms of leishmaniasis, a significant cause of morbidity and mortality. The clinical manifestations of the disease range from selfhealing cutaneous and mucocutaneous skin ulcers to a fatal visceral form named visceral leishmaniasis or kala-azar. The differentiation of Leishmania parasites from the insect stage, the promastigote, towards the pathogenic mammalian stage, the amastigote, is triggered primarily by the rise in ambient temperature encountered during the insect to mammal transmission. The survival of amastigote stage is dependent on that of the host. Regarding energy metabolism, which is an essential factor for the survival, parasites adapt to the environment under low oxygen tension in the host using metabolic systems which are very different from that of the host mammals. The amastigote form of L. donovani is independent on oxidative phosphorylation for ATP production. Indeed, its cell growth was not inhibited by 20-fold excess oligomycin and dicyclohexylcarbodiimide, which are the most specific inhibitors of the mitochondrial FoF1-ATP synthase. In contrast, mitochondrial complex I inhibitor rotenone and complex III inhibitor antimycin A inhibited amastigote cell growth, suggesting the role of complex I and complex III in cell survival. Complex II appeared to have no role in cell survival. To further investigate the site of ATP production, we studied the substrate level phosphorylation, which was involved in the synthesis of ATP. Succinate-pyruvate couple showed the highest substrate level phosphorylation, whereas NADHfumarate and NADH-pyruvate couples failed to produce ATP. In contrast, NADPH-fumarate showed the highest rate of ATP formation in promastigotes. We conclude that substrate level phosphorylation is essential for the growth of L. donovani amastigotes.

  8. Evidência de transmissão de leishmaniose visceral por Lutzomyia cruzi no município de Jaciara, Estado de Mato Grosso, Brasil / Evidence of transmission of visceral leishmaniasis by Lutzomyia cruzi in the municipality of Jaciara, State of Mato Grosso, Brazil

    Scientific Electronic Library Online (English)

    Nanci Akemi, Missawa; Márcia Aurélia Esser, Veloso; Giovana Belem Moreira Lima, Maciel; Érika Monteiro, Michalsky; Edelberto Santos, Dias.

    2011-02-01

    Full Text Available INTRODUÇÃO: O município de Jaciara foi classificado em 2003, como área de transmissão de leishmaniose visceral em situação de surto. O trabalho objetivou determinar evidência de transmissão de Leishmania (Leishmania) infantum chagasi por Lutzomyia cruzi no município de Jaciara, Estado de Mato Grosso [...] , Brasil. MÉTODOS: O município situa-se a 127km da capital Cuiabá e é um importante ponto de atração para os praticantes de eco-turismo. Fêmeas de Lutzomyia cruzi, capturadas com armadilha de CDC, foram dissecadas para confirmação da espécie e armazenadas a -20ºC em pools de 10 indivíduos para extração de DNA, PCR genérico, RFLP específico e eletroforese em gel de poliacrilamida. RESULTADOS: O levantamento entomológico demonstrou a ocorrência abundante de Lutzomyia cruzi e ausência de Lutzomyia longipalpis, principal vetora da Leishmania (Leishmania) infantum chagasi. Uma das três amostras analisadas apresentou banda característica de DNA de Leishmania (120pb) em PCR genérico. Para confirmação da espécie de Leishmania, na RFLP utilizaram-se controles positivos de Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis e Leishmania (Leishmania) infantum chagasi digeridas com enzima de restrição HaeIII. Constatou-se um padrão de bandas semelhante à Leishmania (Leishmania) infantum chagasi em uma amostra, confirmando a detecção de infecção natural de Leishmania (Leishmania) infantum chagasi em Lutzomyia cruzi. CONCLUSÕES: A ocorrência de casos humanos e cães positivos, a presença da Lutzomyia cruzi e a ausência de Lutzomyia longipalpis, bem como a detecção de infecção natural por Leishmania (Leishmania) infantum chagasi, evidenciam a participação de Lutzomyia cruzi na transmissão da leishmaniose visceral em Jaciara, Estado de Mato Grosso, Brasil. Abstract in english INTRODUCTION: The municipality of Jaciara was classified in 2003 as a transmission area for visceral leishmaniasis in outbreak situations. This study aimed to establish evidence of transmission of Leishmania (Leishmania) infantum chagasi by Lutzomyia cruzi in the municipality of Jaciara, State of Ma [...] to Grosso, Brazil. METHODS: This municipality is located 127 km from the state capital (Cuiabá) and is an important center for ecotourism practitioners. Females of Lutzomyia cruzi, captured using CDC traps, were dissected to confirm the species and stored at -20ºC in pools of 10 individuals for DNA extraction, generic PCR, specific RFLP and electrophoresis on polyacrylamide gel. RESULTS: The entomological survey showed abundant occurrence of Lutzomyia cruzi and absence of Lutzomyia longipalpis (the main vector for Leishmania (Leishmania) infantum chagasi). One of the three samples showed a characteristic DNA band from Leishmania (120 bp) in generic PCR. To confirm the Leishmania species via RFLP, positive controls for Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis and Leishmania (Leishmania) infantum chagasi digested with restriction enzyme HaeIII were used. A pattern of bands similar to Leishmania (Leishmania) infantum chagasi was found in one sample, thus confirming the detection of natural infection with Leishmania (Leishmania) infantum chagasi in Lutzomyia cruzi. CONCLUSIONS: The occurrences of positive cases in humans and dogs, the presence of Lutzomyia cruzi and the absence of Lutzomyia longipalpis together with the detection of natural infection with Leishmania (Leishmania) infantum chagasi, indicate that Lutzomyia cruzi participates in visceral leishmaniasis transmission in Jaciara, State of Mato Grosso, Brazil.

  9. Parasitological analysis of synovial fluid from dogs naturally infected with Leishmania sp. / Análise parasitológica do líquido sinovial de cães naturalmente infectados por Leishmania sp

    Scientific Electronic Library Online (English)

    Mayara Nobrega Gomes da, Silva; Deyverson Thiago Prates, Pereira; Maria Ligia de Arruda, Mistieri; Irina, Lubeck.

    2014-07-01

    Full Text Available O objetivo deste estudo foi verificar a viabilidade da análise do líquido sinovial (LS) para o diagnóstico parasitológico de leishmaniose em cães naturalmente infectados. Artrocenteses assépticas das articulações do tarso e do carpo de 40 cães infectados (totalizando 158 amostras articulares) foram [...] realizadas após a eutanásia. Os esfregaços LS foram avaliados sob microscopia óptica e formas amastigotas de Leishmania sp. foram detectadas no LS de 35 cães (87,5%). 93 amostras articulares foram positivas, sendo 53% obtidas a partir das articulações tarsais e 47% obtidas de articulações carpais. Formas amastigotas foram confirmadas em uma (17%), duas (26%), três (31%) ou todas as quatro articulações investigadas (26%). Este trabalho destaca a viabilidade e a alta sensibilidade da avaliação LS para o diagnóstico parasitológico de leishmaniose em cães sintomáticos. Abstract in english The aim of this study was to verify the feasibility of synovial fluid (SF) analysis for the parasitological diagnosis of leishmaniasis in naturally infected dogs. Aseptic arthrocentesis of the tarsal and carpal joints of 40 infected dogs (158 Leishmania sp samples) was conducted after euthanasia. T [...] he SF smears were evaluated under optic microscopy and amastigotes were detected in the SF of 35 dogs (87.5%). Of the 93 positive samples, 53% were obtained from the tarsal joints, and 47% were obtained from the carpal joints. Amastigote forms were confirmed in one (17%), two (26%), three (31%) or all four investigated joints (26%). This research highlights the feasibility and high sensitivity of SF evaluation for the parasitological diagnosis of leishmaniasis in symptomatic dogs.

  10. Presença de formas amastigotas de Leishmania chagasi e perfil leucocitário no aparelho reprodutivo de cães / Presence of amastigotes forms the Leishmania chagasi and profile the leucocytes cells in the reproductive tract of dogs

    Scientific Electronic Library Online (English)

    Ariane P., Benites; Carlos E., Fernandes; Karine B., Brum; Maria Auxiliadora G.S., Abdo.

    2011-01-01

    Full Text Available A leishmaniose visceral (LV) é uma zoonose causada pelo protozoário Leishmania (Leishmania) chagasi. A leishmaniose visceral canina (LVC) é a doença de maior relevância zoonótica. Usualmente, a infecção ocorre entre um hospedeiro invertebrado para um hospedeiro vertebrado, entretanto, a transmissão [...] na ausência do vetor já é conhecida. O objetivo principal deste estudo foi identificar a presença de formas amastigotas, quantificar as células leucocitárias, estimar o risco relativo da presença de formas amastigotas no aparelho reprodutivo de cães sorologicamente positivos com e sem sinais clínicos. Para isso, foram utilizados cães sem raça definida, sexualmente maduros e testados sorologicamente para LVC (com sinais clínicos, n=25; sem sinais clínicos, n=25), que após eutanásia, tiveram fragmentos de testículo, epidídimo (cabeça, corpo e cauda) e glândula prostática (selecionados ao acaso) impressos em lâminas. Um grupo de 20 cãs sorologicamente negativos e sem sinais clínicos foi usado como controle. Amostras do baço foram incluídas como controle parasitológico positivo. O percentual de linfócitos foi superior (P Abstract in english Visceral leishmaniasis (VL) is a zoonosis caused by Leishmania (Leishmania) chagasi. Canine visceral leishmaniasis (VLC) is most important. The infection occurs usually between the invertebrate host and vertebrate host; however, transmission in the absence of the vector has been reported. The aim of [...] this study was to identify the presence of amastigote forms, quantify the leucocyte cells and to estimate the presence (odds ratio) of the amastigotes in the reproductive tract of dogs serologically positive with and without clinical signs. Sexually mature Mongrel dogs, serologically tested to VLC (symptomatic, n=25; asymptomatic, n=25), were used. After euthanasia, testes, epidydimal (caput, corpus and cauda) and prostate gland fragments (randomized) were recovered and impressed on slides. Twenty animals serologically negative and asymptomatic were used as control group. Samples of spleen were included as parasitological positive controls. Lymphocyte percentages were higher (P

  11. Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil

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    Patrícia Flávia Quaresma

    2012-06-01

    Full Text Available 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.

  12. Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil

    Scientific Electronic Library Online (English)

    Patrícia Flávia, Quaresma; Gustavo Mayr de Lima, Carvalho; Mariana Campos das Neves Farah, Ramos; José Dilermando, Andrade Filho.

    2012-06-01

    Full Text Available 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.

  13. Identification of geographically distributed sub-populations of Leishmania (Leishmania major by microsatellite analysis

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    Schwenkenbecher Jan

    2008-06-01

    Full Text Available Abstract Background Leishmania (Leishmania major, one of the agents causing cutaneous leishmaniasis (CL in humans, is widely distributed in the Old World where different species of wild rodent and phlebotomine sand fly serve as animal reservoir hosts and vectors, respectively. Despite this, strains of L. (L. major isolated from many different sources over many years have proved to be relatively uniform. To investigate the population structure of the species highly polymorphic microsatellite markers were employed for greater discrimination among it's otherwise closely related strains, an approach applied successfully to other species of Leishmania. Results Multilocus Microsatellite Typing (MLMT based on 10 different microsatellite markers was applied to 106 strains of L. (L. major from different regions where it is endemic. On applying a Bayesian model-based approach, three main populations were identified, corresponding to three separate geographical regions: Central Asia (CA; the Middle East (ME; and Africa (AF. This was congruent with phylogenetic reconstructions based on genetic distances. Re-analysis separated each of the populations into two sub-populations. The two African sub-populations did not correlate well with strains' geographical origin. Strains falling into the sub-populations CA and ME did mostly group according to their place of isolation although some anomalies were seen, probably, owing to human migration. Conclusion The model- and distance-based analyses of the microsatellite data exposed three main populations of L. (L. major, Central Asia, the Middle East and Africa, each of which separated into two sub-populations. This probably correlates with the different species of rodent host.

  14. Identification and functional evaluation of Leishmania braziliensis Nicotinamide Mononucleotide Adenylyltransferase.

    Science.gov (United States)

    Contreras, Luis E; Neme, Rafik; Ramírez, María H

    2015-11-01

    The progressive increase in Leishmania resistance to current control approaches prompts the need to develop therapeutic strategies based on comprehensive knowledge of the parasite's biology. The enzyme Nicotinamide Mononucleotide Adenylyltransferase (NMNAT, EC 2.7.7.1) catalyzes the central step in nicotinamide adenine dinucleotide (NAD(+)) biosynthesis, making it essential for the survival of all organisms. NAD(+) metabolism is related to the maintenance of several biochemical, cellular, and physiological processes; consequently, the characterization and analysis of the enzymes involved in its biosynthesis represent key steps in the development of control strategies. In this study, the NMNAT enzymes of different Leishmania species were identified using bioinformatics procedures. The sequences were used to construct structural homology models that revealed characteristic elements common to NMNATs. The open reading frame of Leishmania braziliensis NMNAT was cloned from complementary DNA and the enzymatic activity of the corresponding recombinant protein was confirmed through enzymatic assays. Primary structure analysis revealed a Leishmania-specific amino-terminal insertion in NMNAT. The deletion of this insertion is negatively correlated with in vitro enzymatic activity. From our observations, we suggest the amino-terminal insertion of Leishmania NMNATs as a promising pharmacological target for the development of specific control strategies. PMID:26318236

  15. Molecular characterization of the MRPA transporter and antimony uptake in four New World Leishmania spp. susceptible and resistant to antimony?

    OpenAIRE

    Moreira, Douglas S.; Monte Neto, Rubens L.; Andrade, Juvana M; Santi, Ana Maria M.; Reis, Priscila G.; Frézard, Frédéric; Murta, Silvane M.F.

    2013-01-01

    •Level of expression of Pgp is increased in the SbIII-resistant L. guyanensis and L. amazonensis lines.•Incorporation of antimony was reduced in the SbIII-resistant L. guyanensis, L. amazonensis and L. braziliensis lines.•Down-regulation of AQP1 protein was observed in the SbIII-resistant L. guyanensis and L. amazonensis lines.•Rates of SbIII efflux are higher in the SbIII-resistant lines of L. guyanensis and L. braziliensis.•Mechanisms of antimony-resistance of the MRPA gene are different am...

  16. Cutaneous leishmaniasis in northeastern Brazil: a critical appraisal of studies conducted in State of Pernambuco / Leishmaniose cutânea no nordeste do Brasil: uma avaliação crítica dos estudos realizados no Estado de Pernambuco

    Scientific Electronic Library Online (English)

    Maria Edileuza Felinto de, Brito; Maria Sandra, Andrade; Filipe, Dantas-Torres; Eduardo Henrique Gomes, Rodrigues; Milena de Paiva, Cavalcanti; Alzira Maria Paiva de, Almeida; Sinval Pinto, Brandão-Filho.

    2012-08-01

    Full Text Available A leishmaniose cutânea americana (LCA) é uma doença complexa com características clínicas e epidemiológicas que podem variar de região para região. De fato, pelo menos, sete diferentes espécies de Leishmania, incluindo Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania (V [...] iannia) lainsoni, Leishmania (Viannia) naiffi, Leishmania (Viannia) shawi, Leishmania (Viannia) lindenbergi e Leishmania (Leishmania) amazonensis, têm sido incriminadas na etiologia da LCA no Brasil, e numerosas espécies de flebotomíneos do gênero Lutzomyia, foram considerados vetores suspeitos ou comprovados. Devido ao seu caráter focal, a compreensão da dinâmica da LCA a nível local é imprescindível para a implementação de medidas de controle eficazes. Este trabalho consiste de uma revisão narrativa sobre a epidemiologia da LCA em Pernambuco, nordeste do Brasil. Além disso, se enfatiza a necessidade de maior efetividade no diagnóstico, tratamento, controle e estratégias de prevenção para as populações afetadas. Este artigo fornecerá aos pesquisadores uma avaliação crítica da LCA em Pernambuco. Espera-se também contribuir com as autoridades de saúde pública no aprimoramento das estratégias atuais de controle da LCA nos níveis estaduais e nacional. Abstract in english American cutaneous leishmaniasis (ACL) is a complex disease with clinical and epidemiological features that may vary from region to region. In fact, at least seven different Leishmania species, including Leishmania (Viannia) braziliensis, Leishmania (Viannia) guyanensis, Leishmania (Viannia) lainson [...] i, Leishmania (Viannia) naiffi, Leishmania (Viannia) shawi, Leishmania (Viannia) lindenbergi, and Leishmania (Leishmania) amazonensis, have been implicated in the etiology of ACL in Brazil, and numerous phlebotomine sandfly species of the genus Lutzomyia have been regarded as putative or proven vectors. Because ACL is a focal disease, understanding the disease dynamics at the local level is essential for the implementation of more effective control measures. The present paper is a narrative review about the ACL epidemiology in Pernambuco, northeastern Brazil. Furthermore, the need for more effective diagnosis, treatment, control and prevention strategies for the affected populations is highlighted. This paper will provide researchers with a critical appraisal of ACL in Pernambuco. Hopefully, it will also be helpful for public health authorities to improve current control strategies against ACL at the state and country levels.

  17. Partial purification, immunogenicity and putative new localization of a native Leishmania heat shock protein 70

    Scientific Electronic Library Online (English)

    ELIETE CAVALCANTI, DA SILVA; CYNTHIA D, RAYOL; PATRICIA M. M. F, MOURA; PAULO P, ANDRADE.

    2008-12-01

    Full Text Available Neste trabalho estudamos urna proteína recombinante (S7) contendo aproximadamente 230 aminoácidos da extremidade carboxi-terminal da proteína de choque térmico de 70 kDa (HSP70) de Leishmania chagasi. As proteínas de choque térmico estâo entre os antígenos parasitarios mais abundantes e mais conserv [...] ados na natureza. Esta familia pertence a urna das classes de proteínas mais imunogênicas, presentes em organismos patogênicos. Aproteína S7 foi parcialmente purificada por eletroeluição, e em seguida precipitada em acetona. A eletroeluição não modificou suas propriedades imunológicas e antigênicas, pois a proteína continuou a ser reconhecida (tanto no ELISA como no Western blot) por soros positivos para leishmaniose visceral e pelo sistema imunológico de coelhos durante a imunização. Aproducção de soros policlonais com urna concentração antigênica muito inferior a dose máxima, reforca a idéia de que as proteínas desta familia sâo altamente antigênicas e imunogénicas. Nossos resultados com os soros policlonais no ensaio de aglutinação direta (DAT) permitem concluir que a HSP70 nativa de L. chagasi está presente na superficie do parásita. Abstract in english In this work we focused on a recombinant protein, containing approximately 230 aminoacids from the carboxy-terminal extremity of the Leishmania chagasi heat shock protein 70. The heat shock proteins are among the most abundant parasite antigens and conserved proteins in nature, and this family is on [...] e of the most immunogenic proteins present within pathogenic organisms. The recombinant protein has been partially purified by electroelution and further precipitation in acetone. The electroelution process did not modify its immunological and antigenic properties, as it continued to be recognized by visceral leishmaniasis positive sera and by the immunological system of rabbits during the immunization, both in ELISA and Western blots. The production of polyclonal sera with an antigen concentration that is far from the maximum dose, strengthens the idea that the proteins of this family are highly antigenic and immunogenic. Our results with these polyclonal sera in the Direct Agglutination Assay allow the conclusion that the Leishmania chagasi native heat shock protein 70 is distributed on the surface of the parasite.

  18. Seasonal transmission of Leishmania (Leishmania mexicana in the state of Campeche, Yucatan Peninsula, Mexico

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    Andrade-Narvaez Fernando J

    2003-01-01

    Full Text Available In the Yucatan Peninsula, Mexico, localized cutaneous leishmaniasis (LCL caused by Leishmania (Leishmania mexicana is a typical wild zoonosis restricted to the forest, and humans are only accidentally involved. The transmission of L. (L. mexicana has been related to the patient's occupation: "chicleros"(gum collectors and agricultural workers. The objective of this study was to document L. (L. mexicana seasonally of transmission in endemic areas of LCL in the state of Campeche, Yucatan Peninsula, Mexico. The timing of incidence of LCL in humans during 1993-1994, as well as the rate and time of infection in rodents and sand flies between February 1993 and March 1995 were analyzed. Rodents and sand flies were found infected between November and March, when men carried out their field activities and are exposed. Based on results analyzed, it is concluded that L. (L. mexicana in the endemic area of LCL in the state of Campeche, Yucatan Peninsula, Mexico, presents a seasonal transmission restricted to the months of November to March. The knowledge of the timing of the transmission cycle in an endemic area of leishmaniasis is very important because intervention measures on the high-risk focus and population might be restricted.

  19. FIRST REPORT OF CUTANEOUS LEISHMANIASIS CAUSED BY Leishmania (Leishmania infantum chagasi IN AN URBAN AREA OF RIO DE JANEIRO, BRAZIL

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    Marcelo Rosandiski LYRA

    2015-10-01

    Full Text Available SUMMARY American tegumentary leishmaniasis (ATL is an infectious disease caused by protozoa of the genus Leishmania, and transmitted by sandflies. In the state of Rio de Janeiro, almost all of the cases of American tegumentary leishmaniasis (ATL are caused by Leishmania (Viannia braziliensis, while cases of visceral leishmaniasis (VL are caused by Leishmania (Leishmania infantum chagasi. The resurgence of autochthonous VL cases in Rio de Janeiro is related to the geographic expansion of the vector Lutzomyia longipalpis and its ability to adapt to urban areas. We report the first case of leishmaniasis with exclusively cutaneous manifestations caused by L. (L. infantum chagasi in an urban area of Rio de Janeiro. An eighty-one-year-old woman presented three pleomorphic skin lesions that were not associated with systemic symptoms or visceromegalies. Multilocus enzyme electrophoresis identified L. (L. infantum chagasi, but direct smear and PCR of bone narrow were negative for Leishmania sp. (suggesting exclusively cutaneous involvement. We discuss the different dermatological presentations of viscerotropic leishmaniasis of the New and Old World, and the clinical and epidemiological importance of the case. Etiologic diagnosis of ATL based upon exclusive clinical criteria may lead to incorrect conclusions. We should be aware of the constant changes in epidemiological patterns related to leishmaniases.

  20. FIRST REPORT OF CUTANEOUS LEISHMANIASIS CAUSED BY Leishmania (Leishmania) infantum chagasi IN AN URBAN AREA OF RIO DE JANEIRO, BRAZIL.

    Science.gov (United States)

    Lyra, Marcelo Rosandiski; Pimentel, Maria Inês Fernandes; Madeira, Maria de Fátima; Antonio, Liliane de Fátima; Lyra, Janine Pontes de Miranda; Fagundes, Aline; Schubach, Armando de Oliveira

    2015-10-01

    American tegumentary leishmaniasis (ATL) is an infectious disease caused by protozoa of the genus Leishmania, and transmitted by sandflies. In the state of Rio de Janeiro, almost all of the cases of American tegumentary leishmaniasis (ATL) are caused by Leishmania (Viannia) braziliensis, while cases of visceral leishmaniasis (VL) are caused by Leishmania (Leishmania) infantum chagasi. The resurgence of autochthonous VL cases in Rio de Janeiro is related to the geographic expansion of the vector Lutzomyia longipalpis and its ability to adapt to urban areas. We report the first case of leishmaniasis with exclusively cutaneous manifestations caused by L. (L.) infantum chagasi in an urban area of Rio de Janeiro. An eighty-one-year-old woman presented three pleomorphic skin lesions that were not associated with systemic symptoms or visceromegalies. Multilocus enzyme electrophoresis identified L. (L.) infantum chagasi, but direct smear and PCR of bone narrow were negative for Leishmania sp. (suggesting exclusively cutaneous involvement). We discuss the different dermatological presentations of viscerotropic leishmaniasis of the New and Old World, and the clinical and epidemiological importance of the case. Etiologic diagnosis of ATL based upon exclusive clinical criteria may lead to incorrect conclusions. We should be aware of the constant changes in epidemiological patterns related to leishmaniases. PMID:26603237

  1. Molecular and immunological characterisation of the glucose regulated protein 78 of Leishmania donovani

    DEFF Research Database (Denmark)

    Jensen, A T; Curtis, J; Montgomery, J; Handman, E; Theander, T G

    2001-01-01

    assessed in mice vaccine experiments. A GRP78 DNA vaccine primed for an immune response that protected C57Bl/6 and C3H/He mice against infection with L. major. Similarly vaccination with a recombinant form of GRP78 purified from Escherichia coli and administered with Freund's as adjuvant induced protective......To identify novel potential Leishmania vaccine antigens, antibodies from patients with visceral leishmaniasis (VL) were used to isolate clones from a cDNA expression library of L. donovani amastigotes. Glucose Regulated Protein (GRP78), a member of the 70 kDa heat-shock protein family was...

  2. Immunosuppressive Activity of Leishmania major Culture Supernatant

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    M Abolhassani

    2006-07-01

    Full Text Available BALB/c mice are highly susceptible to infection with the protozoan parasite Leishmania major. This susceptibility has been attributed in part to the expansion of Th2 cells and production of their cytokines, IL-4 and IL-10 and down-regulation of Th1 cytokine, INF-?. The inability of susceptible hosts to mount the immune response necessary to activate macrophage and destroy the parasites can be due to the parasite-specific proteins that are able to modulate the immune system. In this report, we monitored the culture supernatant of the parasite in order to show the existence of immunosuppressive factor. The L. major parasites were isolated from lesion of infected BALB/c mice. Amastigotes were grown at 35 °C and promastigotes at 22-25°C in the presence of 10% FCS. Culture supernatants were harvested after 72 h of incubation and used for lymphocyte proliferation assay using lymph node and spleen cells stimulated with mitogens. We have shown that the excretory materials of promastigotes and axenic amastigotes have a potent immunosuppressive activity. Culture supernatants can suppress the proliferation of mitogen-stimulated lymph node lymphocytes (74% or spleen cells (90% in a dose-dependent manner. This result may show that parasite by excretory materials can influence INF-? production by T cells and macrophage activation in order to survive within the macrophage and propagating inside the phagolysosome.

  3. The polymerase chain reaction can reveal the occurrence of naturally mixed infections with Leishmania parasites

    DEFF Research Database (Denmark)

    Ibrahim, M E; Smyth, A J; Ali, M H; Barker, D C; Kharazmi, A

    1994-01-01

    On isolation and characterization of Leishmania parasites from Sudanese patients with visceral leishmaniasis (VL), four cases of mixed infections were found. Three of those cases were from the Eastern Sudan focus of VL. In one case the patient was found to be concomitantly infected with Leishmania donovani and Leishmania aethiopica, while the remaining three patients possessed mixed infections of Leishmania donovani and Leishmania major. Mixed infections were identified by PCR amplification of L...

  4. Fishing for anti-leishmania drugs: principles and problems.

    Science.gov (United States)

    Handman, Emanuela; Kedzierski, Lukasz; Uboldi, Alessandro D; Goding, James W

    2008-01-01

    To date, there are no vaccines against any of the major parasitic diseases including leishmaniasis, and chemotherapy is the main weapon in our arsenal. Current drugs are toxic and expensive, and are losing their effectiveness due to parasite resistance. The availability of the genome sequence of two species of Leishmania, Leishmania major and Leishmania infantum, as well as that of Trypanosoma brucei and Trypanosoma cruzi should provide a cornucopia of potential new drug targets. Their exploitation will require a multi-disciplinary approach that includes protein structure and function and high throughput screening of random and directed chemical libraries, followed by in vivo testing in animals and humans. We outline the opportunities that are made possible by recent technologies, and potential problems that need to be overcome. PMID:18365658

  5. Presence of amastigotes in the central nervous system of hamsters infected with Leishmania sp. Presença de amastigotas em sistema nervoso central de hamster infectado com Leishmania sp.

    Directory of Open Access Journals (Sweden)

    Elisangela de Oliveira

    2011-06-01

    Full Text Available Visceral leishmaniasis (VL is a severe chronic disease caused by Leishmania (Leishmania infantum chagasi. Better knowledge on the effects caused by this disease can help develop adequate clinical management and treatment. Parasitological and immunohistochemical studies were performed golden hamsters Mesocricetus auratus infected with bone marrow from individuals with VL in the State of Mato Grosso do Sul, central-west Brazil. The effects of parasitism in the spleen, liver, kidneys, lungs, heart and brain of the animals were examined. Eighteen hamsters were inoculated intraperitoneally, and six healthy animals were used as negative controls. The animals were kept in the animal house and checked for clinical signs. Specimens of each organ were examined for the presence of amastigotes. Immunohistochemical technique was performed in all brain specimens and organs negative on the direct examination of parasites. Direct examination of amastigotes was positive in the spleen and liver of all infected animals; 33.3% showed the parasite in the kidneys and lungs, and 16.7% in the heart. Parasitic forms were seen in 83.3% (15/18 of the brain examined. Immunohistochemistry confirmed the results of the direct examination, except in two specimens of lung tissue and in the brain specimens. Other studies are needed to further clarify the effect of the parasite in the central nervous system.A leishmaniose visceral (LV é uma doença crônica grave, causada pelo parasito Leishmania (Leishmania infantum chagasi. Esclarecer as alterações provocadas pela doença é fundamental para que se adotem condutas clínicas e de tratamento adequadas. Com o objetivo de analisar a infecção experimental em hamsters da linhagem golden, Mesocricetus auratus, infectados com tecido de medula óssea de pacientes com LV no Estado de Mato Grosso do Sul, foram realizados estudos parasitológicos e de imunomarcação. Foi verificada a distribuição do parasitismo no baço, fígado, rim, pulmão, coração e encéfalo desses animais. Foram utilizados 18 hamsters experimentalmente inoculados via intra-peritoneal, e seis animais sadios como controles negativos. Os animais foram mantidos em biotério de experimentação e observados, em busca de alterações clínicas. Com fragmentos de cada órgão, procedeu-se a confecção de lâminas por aposição para pesquisa de amastigotas. Nos órgãos com resultado negativo na pesquisa direta do parasito, e em todas as amostras de encéfalo, foi realizada a técnica de imunohistoquímica. A pesquisa direta de amastigotas foi positiva no baço e fígado de todos os animais infectados; 33,3% apresentaram o parasito em rim e pulmão, e 16,7% no coração. Quando realizada a pesquisa em encéfalo, formas parasitárias foram observadas em 83,3% (15/18 dos animais. A imunomarcação confirmou os resultados da pesquisa direta, exceto em duas amostras de tecido pulmonar e nas amostras de encéfalo. Mais estudos são necessários, para esclarecer o real papel do parasito no sistema nervoso central.

  6. Presence of amastigotes in the central nervous system of hamsters infected with Leishmania sp. / Presença de amastigotas em sistema nervoso central de hamster infectado com Leishmania sp.

    Scientific Electronic Library Online (English)

    Elisangela de, Oliveira; Elisa Teruya, Oshiro; Rebeca Vieira, Pinto; Bruna Corrêa de, Castro; Karla Borges, Daniel; Janaina Michelle de, Oliveira; Manoel Sebastião da Costa, Lima Júnior; Euripedes Batista, Guimarães; Jesiel Mamedes, Silva; Maria Elizabeth Cavalheiros, Dorval.

    2011-06-01

    Full Text Available A leishmaniose visceral (LV) é uma doença crônica grave, causada pelo parasito Leishmania (Leishmania) infantum chagasi. Esclarecer as alterações provocadas pela doença é fundamental para que se adotem condutas clínicas e de tratamento adequadas. Com o objetivo de analisar a infecção experimental em [...] hamsters da linhagem golden, Mesocricetus auratus, infectados com tecido de medula óssea de pacientes com LV no Estado de Mato Grosso do Sul, foram realizados estudos parasitológicos e de imunomarcação. Foi verificada a distribuição do parasitismo no baço, fígado, rim, pulmão, coração e encéfalo desses animais. Foram utilizados 18 hamsters experimentalmente inoculados via intra-peritoneal, e seis animais sadios como controles negativos. Os animais foram mantidos em biotério de experimentação e observados, em busca de alterações clínicas. Com fragmentos de cada órgão, procedeu-se a confecção de lâminas por aposição para pesquisa de amastigotas. Nos órgãos com resultado negativo na pesquisa direta do parasito, e em todas as amostras de encéfalo, foi realizada a técnica de imunohistoquímica. A pesquisa direta de amastigotas foi positiva no baço e fígado de todos os animais infectados; 33,3% apresentaram o parasito em rim e pulmão, e 16,7% no coração. Quando realizada a pesquisa em encéfalo, formas parasitárias foram observadas em 83,3% (15/18) dos animais. A imunomarcação confirmou os resultados da pesquisa direta, exceto em duas amostras de tecido pulmonar e nas amostras de encéfalo. Mais estudos são necessários, para esclarecer o real papel do parasito no sistema nervoso central. Abstract in english Visceral leishmaniasis (VL) is a severe chronic disease caused by Leishmania (Leishmania) infantum chagasi. Better knowledge on the effects caused by this disease can help develop adequate clinical management and treatment. Parasitological and immunohistochemical studies were performed golden hamste [...] rs Mesocricetus auratus infected with bone marrow from individuals with VL in the State of Mato Grosso do Sul, central-west Brazil. The effects of parasitism in the spleen, liver, kidneys, lungs, heart and brain of the animals were examined. Eighteen hamsters were inoculated intraperitoneally, and six healthy animals were used as negative controls. The animals were kept in the animal house and checked for clinical signs. Specimens of each organ were examined for the presence of amastigotes. Immunohistochemical technique was performed in all brain specimens and organs negative on the direct examination of parasites. Direct examination of amastigotes was positive in the spleen and liver of all infected animals; 33.3% showed the parasite in the kidneys and lungs, and 16.7% in the heart. Parasitic forms were seen in 83.3% (15/18) of the brain examined. Immunohistochemistry confirmed the results of the direct examination, except in two specimens of lung tissue and in the brain specimens. Other studies are needed to further clarify the effect of the parasite in the central nervous system.

  7. Time-course analysis of the Shewanella amazonensis SB2B proteome in response to sodium chloride shock

    Energy Technology Data Exchange (ETDEWEB)

    Parnell, John J.; Callister, Stephen J.; Rompato, Giovanni; Nicora, Carrie D.; Pasa-Tolic, Ljiljana; Williamson, Ashley; Pfrender, Michael E.

    2011-06-29

    Organisms in the genus Shewanella have become models for response to environmental stress. One of the most important environmental stresses is change in osmolarity. In this study, we experimentally determine the response mechanisms of Shewanella amazonensis SB2B during osmotic stress. Osmotic stress in SB2B was induced through exposure to NaCl, and the time-course proteomics response was measured using liquid chromatography mass spectrometry. Protein trends were qualitatively compared to gene expression trends and to phenotypic characterization. Osmotic stress affects motility, and has also been associated with a change in the membrane fatty acid composition (due to induction of branched chain amino acid degradation pathways); however, we show this is not the case for SB2B. Although proteins and genes involved with branched chain amino acid degradation are induced, fatty acid degradation pathways are not induced and no change in the fatty acid profile occurs in SB2B as a result of osmotic shock. The most extensive response of SB2B over the time course of acclimation to high salt involves an orchestrated sequence of events comprising increased expression of signal transduction associated with motility and restricted cell division and DNA replication. After SB2B has switched to increased branched chain amino acid degradation, motility, and cellular replication proteins return to pre-perturbed levels.

  8. Movement of Heterorhabditis amazonensis and Steinernema arenarium in search of corn fall armyworm larvae in artificial conditions

    Directory of Open Access Journals (Sweden)

    Vanessa Andaló

    2012-06-01

    Full Text Available Spodoptera frugiperda (Smith, 1797 (Lepidoptera: Noctuidae is considered to be the main pest of maize crops in Brazil. Entomopathogenic nematodes (EPN may be used to control this pest and exhibit different, unique abilities to search for their hosts. The movement of EPN in relation to S. frugiperda was evaluated. To test for horizontal movement, a styrofoam enclosure filled with sand was divided into segments, nematodes were placed at the entrance to the enclosure and a larva was placed at the end of each division. The same approach was used to evaluate vertical movement; however, PVC pipes were used in this case. In general, the mortality was inversely proportional to the initial distance between host and nematodes. In the vertical displacement test, both nematodes were able to kill the larvae up to a distance of 25 cm. Therefore, the infective juveniles of H. amazonensis and S. arenarium can search out, infect and kill larvae of S. frugiperda at distances of up to 60 cm and 25 cm of horizontal and vertical displacement, respectively.

  9. Leishmania (L.) mexicana infected bats in Mexico: novel potential reservoirs.

    Science.gov (United States)

    Berzunza-Cruz, Miriam; Rodríguez-Moreno, Ángel; Gutiérrez-Granados, Gabriel; González-Salazar, Constantino; Stephens, Christopher R; Hidalgo-Mihart, Mircea; Marina, Carlos F; Rebollar-Téllez, Eduardo A; Bailón-Martínez, Dulce; Balcells, Cristina Domingo; Ibarra-Cerdeña, Carlos N; Sánchez-Cordero, Víctor; Becker, Ingeborg

    2015-01-01

    Leishmania (Leishmania) mexicana causes cutaneous leishmaniasis, an endemic zoonosis affecting a growing number of patients in the southeastern states of Mexico. Some foci are found in shade-grown cocoa and coffee plantations, or near perennial forests that provide rich breeding grounds for the sand fly vectors, but also harbor a variety of bat species that live off the abundant fruits provided by these shade-giving trees. The close proximity between sand flies and bats makes their interaction feasible, yet bats infected with Leishmania (L.) mexicana have not been reported. Here we analyzed 420 bats from six states of Mexico that had reported patients with leishmaniasis. Tissues of bats, including skin, heart, liver and/or spleen were screened by PCR for Leishmania (L.) mexicana DNA. We found that 41 bats (9.77%), belonging to 13 species, showed positive PCR results in various tissues. The infected tissues showed no evidence of macroscopic lesions. Of the infected bats, 12 species were frugivorous, insectivorous or nectarivorous, and only one species was sanguivorous (Desmodus rotundus), and most of them belonged to the family Phyllostomidae. The eco-region where most of the infected bats were caught is the Gulf Coastal Plain of Chiapas and Tabasco. Through experimental infections of two Tadarida brasiliensis bats in captivity, we show that this species can harbor viable, infective Leishmania (L.) mexicana parasites that are capable of infecting BALB/c mice. We conclude that various species of bats belonging to the family Phyllostomidae are possible reservoir hosts for Leishmania (L.) mexicana, if it can be shown that such bats are infective for the sand fly vector. Further studies are needed to determine how these bats become infected, how long the parasite remains viable inside these potential hosts and whether they are infective to sand flies to fully evaluate their impact on disease epidemiology. PMID:25629729

  10. Histological changes and immunolabeling of Leishmania infantum in kidneys and urinary bladder of dogs / Alterações histológicas e imunomarcação de Leishmania infantum em rins e bexiga de cães

    Scientific Electronic Library Online (English)

    Jamile Prado dos, Santos; Leucio Camara, Alves; Rafael Antonio Nascimento, Ramos; Danillo de Souza, Pimentel; Gilcia Aparecida de, Carvalho; Maria Fernanda Melo, Monteiro; Maria Aparecida da Gloria, Faustino.

    2013-09-01

    Full Text Available Objetivou-se neste estudo avaliar as alterações histológicas e realizar a imunomarcação de Leishmania infantum em rins e bexiga de cães naturalmente infectados. Vinte e cinco amostras de bexiga e rins de animais sorologicamente (ELISA S7® Biogene and IFAT ? 1:40 - Biomanguinhos/Fiocruz) positivas fo [...] ram analisadas histologicamente e por meio da técnica de imuno-histoquímica. Os resultados revelaram cistite em 44% (11/25) das amostras de bexiga e glomerulonefrite membranoproliferativa em 92% (23/25) das amostras de rins. A imunomarcação do parasito revelou 32% (8/25) e 8% (2/25) de positividade em bexiga e rins, respectivamente. Conclui-se que a técnica de imunohistoquímica é uma útil ferramenta para detecção de formas amastigotas de L. infantum em órgãos de cães infectados. Além disso, o presente trabalho reporta a primeira descrição de formas amastigotas de L. infantum em bexiga de cães. Abstract in english The aim of this study was to assess histological alterations and perform immunolabeling of Leishmania infantum in the kidneys and urinary bladder of naturally infected dogs. Twenty-five urinary bladder and kidney samples of serologically positive animals (ELISA S7® Biogene and IFAT ? 1:40 - Biomangu [...] inhos/Fiocruz) were analyzed by means of immunohistochemical and histological techniques. Cystitis was found in 44% (11/25) of the bladder samples and membranoproliferative glomerulonephritis in 92% (23/25) of the kidney samples. Immunolabeling of the parasite revealed that 32% (8/25) of the bladders and 8% (2/25) of the kidneys were positive. In conclusion, the immunohistochemical technique is a useful tool for detecting amastigote forms of L. infantum in organs of infected dogs. In addition, this was the first report of detection of amastigote forms of L. infantum in the bladders of dogs.

  11. Further support for a palaearctic origin of Leishmania

    Directory of Open Access Journals (Sweden)

    Sara F Kerr

    2000-08-01

    Full Text Available The fossil record and systematics of murid rodents, reservoirs of zoonotic cutaneous leishmaniasis in the Palaearctic, Oriental, African, Nearctic and Neotropical, strongly support a Palaearctic origin of Leishmania. The fossil record and systematics of phlebotomine sand flies reinforce this idea. Interpretations of molecular data that place the origin of Leishmania in the Neotropical are inconsistent with the natural histories of reservoirs and vectors. The evolutionary pattern of New World rats (Sigmodontinae indicates that they may be the most important reservoirs of zoonotic cutaneous leishmaniasis throughout their range.

  12. Riesgo de transmisión de Leishmania (Kinetoplastida: Trypanosomatidae en Mérida Venezuela

    Directory of Open Access Journals (Sweden)

    Elsa Nieves

    2014-09-01

    Full Text Available La leishmaniasis es una enfermedad causada por la infección de un parásito protozoario del género Leishmania, transmitido por la picada de insectos hematófagos conocidos como flebotominos. El estudio tiene como objetivo determinar la presencia de flebotominos en los Distritos Sanitarios del estado Mérida y diseñar un mapa de riesgo de transmisión entomológico. Se utilizaron cuatro métodos de captura de flebotominos, los ejemplares se identificaron y se les determinó la infección natural por Leishmania. Se estimó la riqueza de especies, y se realizó un proceso analítico Jerárquico. Los resultados muestran la presencia de diversas especies de flebotominos en los Distritos Sanitarios del estado Mérida, siendo las especies de mayor frecuencia L. youngi, L. gomezi, L. ovallesi y L. walkeri. Se detectó 2,1% de infección natural con Leishmania, la cual se encontró en las 4 especies más frecuentes. Se presenta un mapa de riesgo de transmisión entomológico para el estado Mérida. El conocimiento de la situación actual de los vectores de Leishmania en el estado Mérida y el riesgo de transmisión son relevantes a la hora de considerar la prevención y posible surgimiento de nuevos brotes de leishmaniasis. Abstract (english The leishmaniasis is a disease caused by infection with a protozoan parasite of the genus Leishmania, transmitted by the bite of blood-sucking insects known as sandflies. The study aims to determine the presence of sandflies in Merida state health districts and design a map of entomological risk of transmission. Four methods capture sandflies were used, the specimens were identified and natural Leishmania infection was determined. The richness species was estimated and analityc Hierarchie procesess was performed. The results show the presence of various species of sandflies in Merida state health districts, L. youngi, L. gomezi, L. ovallesi and L. walkeri were most abundant species. The 2.1% of natural infection with Leishmania was detected, which were found in the 4 most abundant species. A map of risk of transmission to Mérida is presented. Knowledge of the current status of Leishmania vectors in Mérida and the risk of transmission are relevant when considering the prevention and possible emergence of new outbreaks of leishmaniasis.

  13. First isolation of a new species of Leishmania responsible for human cutaneous leishmaniasis in Ghana and classification in the Leishmania enriettii complex.

    Science.gov (United States)

    Kwakye-Nuako, Godwin; Mosore, Mba-Tihssommah; Duplessis, Christopher; Bates, Michelle D; Puplampu, Naiki; Mensah-Attipoe, Israel; Desewu, Kwame; Afegbe, Godwin; Asmah, Richard H; Jamjoom, Manal B; Ayeh-Kumi, Patrick F; Boakye, Daniel A; Bates, Paul A

    2015-09-01

    An active case detection approach with PCR diagnosis was used in the Ho District of the Volta Region, Ghana that identified individuals with active cutaneous leishmaniasis. Three isolates were successfully cultured and DNA sequences from these were analysed (ribosomal RNA internal transcribed spacer 1; ribosomal protein L23a intergenic spacer; RNA polymerase II large subunit), showing them to be Leishmania, identical to each other but different from all other known Leishmania spp. Phylogenetic analysis showed the parasites to be new members of the Leishmania enriettii complex, which is emerging as a possible new subgenus of Leishmania parasites containing human pathogens. PMID:26099650

  14. Identification of a differentially expressed mRNA in axenic Leishmania panamensis amastigotes

    Scientific Electronic Library Online (English)

    José Arturo, Gutiérrez; Fabiola, Puentes; Alberto, Moreno; Manuel Elkin, Patarroyo; Luis Angel, Murillo.

    2001-08-01

    Full Text Available Differential display technique was applied in order to identify transcripts which are present in axenic amastigotes but not in promastigotes of the Leishmania panamensis parasites. One of them was cloned and the sequence reveals an open reading frame of 364 amino acids (aprox. 40 kDa). The deduced p [...] rotein is homologous to the serine/threonine protein kinases and specially to the mitogen activates protein kinases from eukaryotic species. Southern blot analysis suggest that this transcript, named lpmkh, is present in the genome of the parasite as a single copy gene. These results could imply that lpmkh could be involved in the differentiation process or the preservation of amastigotes in axenic conditions.

  15. Identification of a differentially expressed mRNA in axenic Leishmania panamensis amastigotes

    Directory of Open Access Journals (Sweden)

    José Arturo Gutiérrez

    2001-08-01

    Full Text Available Differential display technique was applied in order to identify transcripts which are present in axenic amastigotes but not in promastigotes of the Leishmania panamensis parasites. One of them was cloned and the sequence reveals an open reading frame of 364 amino acids (aprox. 40 kDa. The deduced protein is homologous to the serine/threonine protein kinases and specially to the mitogen activates protein kinases from eukaryotic species. Southern blot analysis suggest that this transcript, named lpmkh, is present in the genome of the parasite as a single copy gene. These results could imply that lpmkh could be involved in the differentiation process or the preservation of amastigotes in axenic conditions.

  16. Leishmaniasis in Bolivia: II. The involvement of Psychodopygus yucumensis and Psychodopygus llanosmartinsi in the selvatic transmission cycle of Leishmania braziliensis braziliensis in a lowland subandean region

    Scientific Electronic Library Online (English)

    F., Le Pont; Philippe, Desjeux.

    1986-09-01

    Full Text Available Uma pesquisa epidemiológica sobre os vetores da leishmaniose tegumentar (do tipo espúndia) foi empreendida na região do Alto Beni, na Bolívia, uma zona de contrafortes andinos no limite Este das baixas planícies amazônicas. O clima é tipicamente tropical úmido (15- de latitude Sul). As espécies de f [...] lebótomos antropofílicos foram amostradas em vinte estações, todas na floresta. A importância das espécies do grupo Psychodopygus, já suspeitas como vetores na transmissão da Leishmania do complexo braziliensis, foi comprovada por: 1) a agressividade e a variedade das espécies encontradas (83% das capturas e nove espécies); 2) a descoberta de uma nova espécie antropofílica, P. yucumensis e 3) o isolamento de uma amostra de Leishmania braziliensis braziliensis indistinguível das amostras humanas da mesma região, de duas espécies, P. llanosmartinsi e P. yucumensis. Abstract in english An epidemiological survey of the vectors of cutaneous leishmaniasis ("espúndia" type) was caried out in the Alto Beni region of Bolivia, an area of Andean foothills at the Eastern limit of the Amazonian lowlands. The climate is typical wet tropical (15ºS latitude). Anthropophilic phlebotomine sandfl [...] y species were sampled at 20 sites, all forested. The importance of species from the Psychodopygus group, already suspected as a vector in the transmission of Leishmania from the braziliensis complex, was confirmed by: 1) the aggressiveness and diversity of the species encountered (83% of catches, nine species), 2) the discovery of a new anthropophilic species, P. yucumensis and 3) the isolation of a strain of Leishmania braziliensis braziliensis indistinguishable from human strains from the same area, from two species, P. llanosmartinsi and P. yucumensis.

  17. Vaccination using live attenuated Leishmania donovani centrin deleted parasites induces protection in dogs against Leishmania infantum.

    Science.gov (United States)

    Fiuza, Jacqueline Araújo; Gannavaram, Sreenivas; Santiago, Helton da Costa; Selvapandiyan, Angamuthu; Souza, Daniel Menezes; Passos, Lívia Silva Araújo; de Mendonça, Ludmila Zanandreis; Lemos-Giunchetti, Denise da Silveira; Ricci, Natasha Delaqua; Bartholomeu, Daniella Castanheira; Giunchetti, Rodolfo Cordeiro; Bueno, Lilian Lacerda; Correa-Oliveira, Rodrigo; Nakhasi, Hira L; Fujiwara, Ricardo Toshio

    2015-01-01

    Live attenuated Leishmania donovani parasites such as LdCen(-/-) have been shown elicit protective immunity against leishmanial infection in mice and hamster models. Previously, we have reported on the induction of strong immunogenicity in dogs upon vaccination with LdCen(-/-) including an increase in immunoglobulin isotypes, higher lymphoproliferative response, higher frequencies of activated CD4(+) and CD8(+) T cells, IFN-? production by CD8(+) T cells, increased secretion of TNF-? and IL-12/IL-23p40 and, finally, decreased secretion of IL-4. To further explore the potential of LdCen(-/-) parasites as vaccine candidates, we performed a 24-month follow up of LdCen(-/-) immunized dogs after challenge with virulent Leishmania infantum, aiming determination of parasite burden by qPCR, antibody production (ELISA) and cellular responses (T cell activation and cytokine production) by flow cytometry and sandwich ELISA. Our data demonstrated that vaccination with a single dose of LdCen(-/-) (without any adjuvant) resulted in the reduction of up to 87.3% of parasite burden after 18 months of virulent challenge. These results are comparable to those obtained with commercially available vaccine in Brazil (Leishmune(®)). The protection was associated with antibody production and CD4(+) and CD8(+) proliferative responses, as well as T cell activation and significantly higher production of IFN-?, IL-12/IL-23p40 and TNF-?, which was comparable to responses induced by immunization with Leishmune(®), with significant differences when compared to control animals (Placebo). Moreover, only animals immunized with LdCen(-/-) expressed lower levels of IL-4 when compared to animals vaccinated either with Leishmune(®) or PBS. Our results support further studies aiming to demonstrate the potential of genetically modified live attenuated L. donovani vaccine to control L. infantum transmission in endemic areas for CVL. PMID:25475955

  18. Leishmania development in sand flies: parasite-vector interactions overview

    Directory of Open Access Journals (Sweden)

    Dostálová Anna

    2012-12-01

    Full Text Available Abstract Leishmaniases are vector-borne parasitic diseases with 0.9 – 1.4 million new human cases each year worldwide. In the vectorial part of the life-cycle, Leishmania development is confined to the digestive tract. During the first few days after blood feeding, natural barriers to Leishmania development include secreted proteolytic enzymes, the peritrophic matrix surrounding the ingested blood meal and sand fly immune reactions. As the blood digestion proceeds, parasites need to bind to the midgut epithelium to avoid being excreted with the blood remnant. This binding is strictly stage-dependent as it is a property of nectomonad and leptomonad forms only. While the attachment in specific vectors (P. papatasi, P. duboscqi and P. sergenti involves lipophosphoglycan (LPG, this Leishmania molecule is not required for parasite attachment in other sand fly species experimentally permissive for various Leishmania. During late-stage infections, large numbers of parasites accumulate in the anterior midgut and produce filamentous proteophosphoglycan creating a gel-like plug physically obstructing the gut. The parasites attached to the stomodeal valve cause damage to the chitin lining and epithelial cells of the valve, interfering with its function and facilitating reflux of parasites from the midgut. Transformation to metacyclic stages highly infective for the vertebrate host is the other prerequisite for effective transmission. Here, we review the current state of knowledge of molecular interactions occurring in all these distinct phases of parasite colonization of the sand fly gut, highlighting recent discoveries in the field.

  19. Sand fly evolution and its relationship to Leishmania transmission

    Directory of Open Access Journals (Sweden)

    PD Ready

    2000-08-01

    Full Text Available The evolutionary relationships of sand flies and Leishmania are discussed in this report, which draws distinctions between co-association, co-evolution and co-speciation (or co-cladogenesis. Examples focus on Phlebotomus vectors of Le. infantum and Le. major in the Mediterranean subregion.

  20. Leishmania tropica infection in golden jackals and red foxes, Israel.

    Science.gov (United States)

    Talmi-Frank, Dalit; Kedem-Vaanunu, Noa; King, Roni; Bar-Gal, Gila Kahila; Edery, Nir; Jaffe, Charles L; Baneth, Gad

    2010-12-01

    During a survey of wild canids, internal transcribed spacer 1 real-time PCR and high-resolution melt analysis identified Leishmania tropica in samples from jackals and foxes. Infection was most prevalent in ear and spleen samples. Jackals and foxes may play a role in the spread of zoonotic L. tropica. PMID:21122235

  1. Molecular Modeling of Cathepsin B protein in different Leishmania strains

    Directory of Open Access Journals (Sweden)

    Pawan Kumar Jayaswal

    2011-02-01

    Full Text Available Cathepsin B like cysteine proteases representing a major component of the lysosomal proteolytic repertoire plays an important role in intracel-lular protein degradation. Comparative models of cathepsin B (CatB protein of six different Leishmania strains were developed using MOD-ELLER. The modeled three-dimensional (3-D structure has the correct stereochemistry as gauged from the Ramachandran plot and good 3-D structure compatibility as assessed by PROCHECK and the DOPE score (DS2.1, Accelrys. The modeled proteins were energy minimized and validated using standard dynamic cascade protocol (DS 2.1. Seven different disulfide bonding sites are predicted in CatB protein of Leishma-nia. Two domains were identified and different motifs are present in catB protein of Leishmania like aspargine glycosylation sites, protein ki-nase phosphorylation sites, protein kinase C activation sites and N-myristoylation sites. Considering that cathepsin B is essential for survival of Leishmania, including for virulence to the mammalian host, it may be viewed as an attractive drug target.

  2. Leishmania parasite detection and quantification using PCR-ELISA.

    Czech Academy of Sciences Publication Activity Database

    Kobets, Tetyana; Badalová, Jana; Grekov, Igor; Havelková, Helena; Lipoldová, Marie

    2010-01-01

    Ro?. 5, ?. 6 (2010), s. 1074-1080. ISSN 1754-2189 R&D Projects: GA ?R GA310/08/1697; GA MŠk(CZ) LC06009 Institutional research plan: CEZ:AV0Z50520514 Keywords : polymerase chain reaction * Leishmania major infection * parasite quantification Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 8.362, year: 2010

  3. Genetic Diversity in Natural Populations of New World Leishmania

    Scientific Electronic Library Online (English)

    Elisa, Cupolillo; Hooman, Momen; Gabriel, Grimaldi Jr.

    1998-09-01

    Full Text Available Our results have shown the wide diversity of parasites within New World Leishmania. Biochemical and molecular characterization of species within the genus has revealed that much of the population heterogeneity has a genetic basis. The source of genetic diversity among Leishmania appears to arise fro [...] m predominantly asexual, clonal reproduction, although occasional bouts of sexual reproduction can not be ruled out. Genetic variation is extensive with some clones widely distributed and others seemingly unique and localized to a particular endemic focus. Epidemiological studies of leishmaniasis has been directed to the ecology and dynamics of transmission of Leishmania species/variants, particularly in localized areas. Future research using molecular techniques should aim to identify and follow Leishmania types in nature and correlate genetic typing with important clinical characteristics such as virulence, pathogenicity, drug resistance and antigenic variation. The epidemiological significance of such variation not only has important implications for the control of the leishmaniases, but would also help to elucidate the evolutionary biology of the causative agents.

  4. Genetic Diversity in Natural Populations of New World Leishmania

    Directory of Open Access Journals (Sweden)

    Elisa Cupolillo

    1998-09-01

    Full Text Available Our results have shown the wide diversity of parasites within New World Leishmania. Biochemical and molecular characterization of species within the genus has revealed that much of the population heterogeneity has a genetic basis. The source of genetic diversity among Leishmania appears to arise from predominantly asexual, clonal reproduction, although occasional bouts of sexual reproduction can not be ruled out. Genetic variation is extensive with some clones widely distributed and others seemingly unique and localized to a particular endemic focus. Epidemiological studies of leishmaniasis has been directed to the ecology and dynamics of transmission of Leishmania species/variants, particularly in localized areas. Future research using molecular techniques should aim to identify and follow Leishmania types in nature and correlate genetic typing with important clinical characteristics such as virulence, pathogenicity, drug resistance and antigenic variation. The epidemiological significance of such variation not only has important implications for the control of the leishmaniases, but would also help to elucidate the evolutionary biology of the causative agents.

  5. Zosteriform cutaneous leishmaniasis due to Leishmania ( Viannia ) panamensis and Leishmania ( Viannia ) braziliensis: Report of three cases / Leishmaniasis cutánea zosteriforme causada por Leishmania ( Viannia) panamensis y Leishmania ( Viannia ) braziliensis: reporte de tres casos

    Scientific Electronic Library Online (English)

    Camilo Andrés, Morales; Juliana, Palacio; Gerzaín, Rodríguez; Yenny Carolina, Camargo.

    2014-09-01

    Full Text Available La leishmaniasis zosteriforme es una variante clínica de la leishmaniasis cutánea, infrecuente y poco conocida en Colombia. Clínicamente se caracteriza por una o varias placas conformadas por pápulas y pseudovesículas que siguen un patrón lineal, y por lesiones satelitales que comprometen uno o vari [...] os dermatomas sin sobrepasar la línea media. Se presentan tres casos de leishmaniasis cutánea zosteriforme en los que se identificaron Leishmania panamensis y Leishmania braziliensis como especies infectantes. La sospecha epidemiológica derivada de la procedencia de los pacientes, así como la sospecha clínica a partir del reconocimiento de una presentación infrecuente de la enfermedad, permitieron hacer el diagnóstico. Abstract in english In Colombia, zosteriform leishmaniasis is a little-known and infrequent clinical variant of cutaneous leishmaniasis. Its clinical features include one or more plaques made up of papules and pseudo-vesicles, which conform to a lineal pattern, as well as satellite lesions that affect one or more derma [...] tomes, without crossing the median line. We present three zosteriform cutaneous leishmaniasis cases in which Leishmania panamensis and Leishmania braziliensis were identified as the infective species. In light of the fact that the disease occurs infrequently, diagnosis was reached by taking into account epidemiological and clinical suspicion.

  6. Leishmania serology in the diagnosis of cutaneous leishmaniasis

    International Nuclear Information System (INIS)

    Background: The gold standard to diagnose cutaneous leishmaniasis is histopathology, but there has always been a need of a rapid, reliable, cheap and convenient laboratory investigation. Serological tests fulfill the above criteria. Objective: The objective of the study was to determine the sensitivity and specificity of enzyme linked immunosorbent assay (ELISA) in detection of leishmania antibodies, in comparison with the histopathology. Place and duration of study: The study was conducted in Military Hospital Rawalpindi from 1st November 2010 to 30th June 2011. Patients and methods: The study population included the patients who were clinically diagnosed with cutaneous leishmaniasis. All of them were biopsied and serum was sent for leishmania serology. Results: A total of 47 patients were included. They were all adult males. The histopathology was positive in 31/47 patients (65.95%), while the leishmania serology was positive in 36/47 cases (76.59%). The sensitiuites was 74.19%, specificity was 18.75%, positive predictive value has 63.88%, negative predicative value was 27% and accuracy was 55%. Conclusion: In the light of sensitivity analysis, it may be concluded that leishmania serology has moderate sensitivity and low specificity; hence it is not a reliable test for cutaneous leishmaniasis. (author)

  7. Tetracycline-inducible gene expression system in Leishmania mexicana.

    Czech Academy of Sciences Publication Activity Database

    Kraeva, N.; Ishemgulova, A.; Lukeš, Julius; Yurchenko, Vyacheslav

    2014-01-01

    Ro?. 198, ?. 1 (2014), s. 11-13. ISSN 0166-6851 R&D Projects: GA MŠk(CZ) EE2.3.30.0032 Institutional support: RVO:60077344 Keywords : Leishmania mexicana * Gene expression * Tet-inducible system Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.787, year: 2014

  8. Vectorborne Transmission of Leishmania infantum from Hounds, United States.

    Science.gov (United States)

    Schaut, Robert G; Robles-Murguia, Maricela; Juelsgaard, Rachel; Esch, Kevin J; Bartholomay, Lyric C; Ramalho-Ortigao, Marcelo; Petersen, Christine A

    2015-12-01

    Leishmaniasis is a zoonotic disease caused by predominantly vectorborne Leishmania spp. In the United States, canine visceral leishmaniasis is common among hounds, and L. infantum vertical transmission among hounds has been confirmed. We found that L. infantum from hounds remains infective in sandflies, underscoring the risk for human exposure by vectorborne transmission. PMID:26583260

  9. Molecular detection of Leishmania infantum in naturally infected

    Directory of Open Access Journals (Sweden)

    C. Maia, M.O. Afonso, L. Neto, L. Dionísio & L. Campino

    2009-12-01

    Full Text Available Background & objectives: In Portugal, Phlebotomus perniciosus and P. ariasi, (SubgenusLarroussius; Diptera: Psychodidae are the proven vectors of leishmaniasis caused by Leishmaniainfantum. The Algarve Region in southern Portugal has been considered an endemic focus ofleishmaniasis since 1980s. The main objective of the present study was to validate a molecularapproach to detect Leishmania infection in phlebotomines based on DNA extraction from thefemale sandfly whole body, minus genitalia, followed by PCR for application on epidemiologicalsurveys.Methods: In Algarve Region, from early May until early November 2006, sandflies were capturedby CDC miniature light-traps. kDNA-PCR and ITS1-PCR were used to screen the presence ofLeishmania DNA in female sandflies after species identification by entomological keys.Results: A total of 474 sandflies were collected in 108 biotopes. One female of P. perniciosus, thepredominant species, was found infected with L. infantum reflecting an overall infection rate of0.47%.Interpretation & conclusion: PCR associated with morphological characterization of the sandflieswill be a powerful epidemiological tool for the determination of the number of phlebotominesinfected with Leishmania spp in nature. In addition, the simultaneous occurrence of dogs and P.perniciosus infected with L. infantum shows that Algarve continues to be an endemic focus ofcanine leishmaniasis. Furthermore, as P. sergenti and P. papatasi which transmit L. tropica and L.major, respectively were present, the future introduction of these two Leishmania species in southernregion of Portugal should not be neglected.

  10. Characterization of anti-silencing factor 1 in Leishmania major

    Scientific Electronic Library Online (English)

    Ricardo, Scher; Juliana Bório Ferreira, Garcia; Bruno, Pascoalino; Sergio, Schenkman; Angela Kaysel, Cruz.

    2012-05-01

    Full Text Available Anti-silencing factor 1 (ASF1) is a histone chaperone that contributes to the histone deposition during nucleosome assembly in newly replicated DNA. It is involved in chromatin disassembly, transcription activation and in the cellular response to DNA damage. In Leishmania major the ASF1 gene (LmASF1 [...] ) is located in chromosome 20 and codes for a protein showing 67% of identity with the Trypanosoma brucei TbASF1a. Compared to orthologous proteins, LmASF1 conserves the main residues relevant for its various biological functions. To study ASF1 in Leishmania we generated a mutant overexpressing LmASF1 in L. major. We observed that the excess of LmASF1 impaired promastigotes growth rates and had no impact on cell cycle progress. Differently from yeast, ASF1 overproduction in Leishmania did not affect expression levels of genes located on telomeres, but led to an upregulation of proteins involved in chromatin remodelling and physiological stress, such as heat shock proteins, oxidoreductase activity and proteolysis. In addition, we observed that LmASF1 mutant is more susceptible to the DNA damaging agent, methyl methane sulphonate, than the control line. Therefore, our study suggests that ASF1 from Leishmania pertains to the chromatin remodelling machinery of the parasite and acts on its response to DNA damage.

  11. Characterization of anti-silencing factor 1 in Leishmania major

    Directory of Open Access Journals (Sweden)

    Ricardo Scher

    2012-05-01

    Full Text Available Anti-silencing factor 1 (ASF1 is a histone chaperone that contributes to the histone deposition during nucleosome assembly in newly replicated DNA. It is involved in chromatin disassembly, transcription activation and in the cellular response to DNA damage. In Leishmania major the ASF1 gene (LmASF1 is located in chromosome 20 and codes for a protein showing 67% of identity with the Trypanosoma brucei TbASF1a. Compared to orthologous proteins, LmASF1 conserves the main residues relevant for its various biological functions. To study ASF1 in Leishmania we generated a mutant overexpressing LmASF1 in L. major. We observed that the excess of LmASF1 impaired promastigotes growth rates and had no impact on cell cycle progress. Differently from yeast, ASF1 overproduction in Leishmania did not affect expression levels of genes located on telomeres, but led to an upregulation of proteins involved in chromatin remodelling and physiological stress, such as heat shock proteins, oxidoreductase activity and proteolysis. In addition, we observed that LmASF1 mutant is more susceptible to the DNA damaging agent, methyl methane sulphonate, than the control line. Therefore, our study suggests that ASF1 from Leishmania pertains to the chromatin remodelling machinery of the parasite and acts on its response to DNA damage.

  12. The FML (Fucose Mannose Ligand) of Leishmania donovani: a new tool in diagnosis, prognosis, transfusional control and vaccination against human kala-azar

    Scientific Electronic Library Online (English)

    Claris B. Palatnik de, Sousa; Elza M., Gomes; Edilma Paraguai de, Souza; Wania R. dos, Santos; Sirley R. de, Macedo; Linnete V. de, Medeiros; Kleber, Luz.

    1996-04-01

    Full Text Available O FML (Ligame de Fucose-Manose) de Leishmania donovani é uma fração glicoproteica complexa. O seu potencial no diagnóstico da leishmaniose visceral humana foi testado com soros provenientes de Natal, Rio Grande do Norte, Brasil. O teste de FML-ELISA mostrou 100% de sensibilidade e 96% de especificid [...] ade, identificando pacientes com calazar declarado (p Abstract in english The Fucose-Mannose Ligand (FML) of Leishmania donovani is a complex glycoproteic fraction. Its potential use as a tool for diagnosis of human visceral leishmaniasis was tested with human sera from Natal, Rio Grande do Norte, Brazil. The FML-ELISA test, showed 100% sensitivity and 96% specificity, id [...] entifying patients with overt kala-azar (p

  13. Dissecting Leishmania infantum Energy Metabolism - A Systems Perspective.

    Science.gov (United States)

    Subramanian, Abhishek; Jhawar, Jitesh; Sarkar, Ram Rup

    2015-01-01

    Leishmania infantum, causative agent of visceral leishmaniasis in humans, illustrates a complex lifecycle pertaining to two extreme environments, namely, the gut of the sandfly vector and human macrophages. Leishmania is capable of dynamically adapting and tactically switching between these critically hostile situations. The possible metabolic routes ventured by the parasite to achieve this exceptional adaptation to its varying environments are still poorly understood. In this study, we present an extensively reconstructed energy metabolism network of Leishmania infantum as an attempt to identify certain strategic metabolic routes preferred by the parasite to optimize its survival in such dynamic environments. The reconstructed network consists of 142 genes encoding for enzymes performing 237 reactions distributed across five distinct model compartments. We annotated the subcellular locations of different enzymes and their reactions on the basis of strong literature evidence and sequence-based detection of cellular localization signal within a protein sequence. To explore the diverse features of parasite metabolism the metabolic network was implemented and analyzed as a constraint-based model. Using a systems-based approach, we also put forth an extensive set of lethal reaction knockouts; some of which were validated using published data on Leishmania species. Performing a robustness analysis, the model was rigorously validated and tested for the secretion of overflow metabolites specific to Leishmania under varying extracellular oxygen uptake rate. Further, the fate of important non-essential amino acids in L. infantum metabolism was investigated. Stage-specific scenarios of L. infantum energy metabolism were incorporated in the model and key metabolic differences were outlined. Analysis of the model revealed the essentiality of glucose uptake, succinate fermentation, glutamate biosynthesis and an active TCA cycle as driving forces for parasite energy metabolism and its optimal growth. Finally, through our in silico knockout analysis, we could identify possible therapeutic targets that provide experimentally testable hypotheses. PMID:26367006

  14. Dissecting Leishmania infantum Energy Metabolism - A Systems Perspective

    Science.gov (United States)

    Subramanian, Abhishek; Jhawar, Jitesh; Sarkar, Ram Rup

    2015-01-01

    Leishmania infantum, causative agent of visceral leishmaniasis in humans, illustrates a complex lifecycle pertaining to two extreme environments, namely, the gut of the sandfly vector and human macrophages. Leishmania is capable of dynamically adapting and tactically switching between these critically hostile situations. The possible metabolic routes ventured by the parasite to achieve this exceptional adaptation to its varying environments are still poorly understood. In this study, we present an extensively reconstructed energy metabolism network of Leishmania infantum as an attempt to identify certain strategic metabolic routes preferred by the parasite to optimize its survival in such dynamic environments. The reconstructed network consists of 142 genes encoding for enzymes performing 237 reactions distributed across five distinct model compartments. We annotated the subcellular locations of different enzymes and their reactions on the basis of strong literature evidence and sequence-based detection of cellular localization signal within a protein sequence. To explore the diverse features of parasite metabolism the metabolic network was implemented and analyzed as a constraint-based model. Using a systems-based approach, we also put forth an extensive set of lethal reaction knockouts; some of which were validated using published data on Leishmania species. Performing a robustness analysis, the model was rigorously validated and tested for the secretion of overflow metabolites specific to Leishmania under varying extracellular oxygen uptake rate. Further, the fate of important non-essential amino acids in L. infantum metabolism was investigated. Stage-specific scenarios of L. infantum energy metabolism were incorporated in the model and key metabolic differences were outlined. Analysis of the model revealed the essentiality of glucose uptake, succinate fermentation, glutamate biosynthesis and an active TCA cycle as driving forces for parasite energy metabolism and its optimal growth. Finally, through our in silico knockout analysis, we could identify possible therapeutic targets that provide experimentally testable hypotheses. PMID:26367006

  15. Susceptibility of Cebus apella monkey (Primates: Cebidae) to experimental Leishmania (L.) infantum chagasi-infection / Susceptibilidade do macaco Cebus apella (Primata: Cebidae) à infecção experimental por Leishmania (L.) infantum chagasi

    Scientific Electronic Library Online (English)

    Liliane Almeida, Carneiro; Fernando Tobias, Silveira; Marliane Batista, Campos; Maria do Carmo de Oliveira, Brígido; Claudia Maria C., Gomes; Carlos E.P., Corbett; Márcia D., Laurenti.

    2011-02-01

    Full Text Available Na Amazônia Brasileira, o macaco Cebus apella (Primata: Cebidae) tem sido associado com o ciclo enzoótico da Leishmania (V.) shawi, um parasito dermotrópico causador da Leishmaniose Tegumentar Americana (LTA). Ele tem sido também empregado com sucesso como modelo experimental para estudo da leishman [...] iose tegumentar. Neste trabalho, foi investigada sua susceptibilidade à infecção experimental por Leishmania (L.) infantum chagasi, o agente etiológico da Leishmaniose Visceral Americana (LVA). Foram usados dez espécimes de C. apella oito adultos e dois jovens, quatro machos e seis fêmeas, todos nascidos e criados em cativeiro. Dois protocolos de infecção experimental foram feitos: i) seis macacos foram inoculados por via intradérmica (ID), na base da cauda com 2x10(6) formas promastigotas em fase estacionária de crescimento; ii) outros quatro macacos foram inoculados com 3x10(7) formas amastigotas de infecção visceral de hamsteres por duas vias diferentes: a) dois por via intravenosa (IV) e, b) outros dois pela via intraperitoneal (IP). A avaliação da infecção incluiu parâmetros: clínico: exame físico do abdômen, peso e temperatura corporal; b) parasitológico: aspirado de medula óssea por agulha para procura de amastigotas (esfregaço corado por Giemsa) e formas promastigotas (meio de cultura); c) imunológico: Reação de Imunofluorescência Indireta (RIFI) e, resposta de hipersensibilidade tardia (DTH). Nos seis macacos inoculados ID (formas promastigotas) todos os parâmetros de avaliação da infecção foram negativos durante o período de 12 meses. Entre os quatro macacos inoculados com formas amastigotas, dois inoculados IV mostraram parasitos na medula óssea do primeiro ao sexto mês p.i. e em seguida houve a resolução da infecção, no entanto os outros dois inoculados IP foram totalmente negativos. Esses quatro macacos apresentaram resposta específica de anticorpo IgG desde o terceiro mês p.i. (IP: 1/80 e IV: 1/320) até o décimo segundo mês (IP: 1/160 e IV: 1/5120). A conversão DTH ocorreu em apenas um macaco inoculado IV com uma forte reação na pele (30 mm). Considerando esses resultados, nós não recomendamos o uso do macaco C. apella como modelo animal para estudo da LVA. Abstract in english In Amazonian Brazil, the Cebus apella monkey (Primates: Cebidae) has been associated with the enzootic cycle of Leishmania (V.) shawi, a dermotropic parasite causing American cutaneous leishmaniasis (ACL). It has also been successfully used as animal model for studying cutaneous leishmaniasis. In th [...] is work, there has been investigated its susceptibility to experimental Leishmania (L.) infantum chagasi-infection, the etiologic agent of American visceral leishmaniasis (AVL). There were used ten C. apella specimens, eight adult and two young, four males and six females, all born and raised in captivity. Two experimental infection protocols were performed: i) six monkeys were inoculated, intra-dermal via (ID), into the base of the tail with 2 x 10(6) promastigotes forms from the stationary phase culture medium; ii) other four monkeys were inoculated with 3 x 10(7) amastigotes forms from the visceral infection of infected hamsters by two different via: a) two by intravenous via (IV) and, b) other two by intra-peritoneal via (IP). The parameters of infection evaluation included: a) clinical: physical exam of abdomen, weigh and body temperature; b) parasitological: needle aspiration of the bone-marrow for searching of amastigotes (Giemsa-stained smears) and promastigotes forms (culture medium); c) immunological: Indirect fluorescence antibody test (IFAT) and, Delayed-type hypersensitivity (DTH). In the six monkeys ID inoculated (promastigotes forms) all parameters of infection evaluation were negative during the 12 months period of follow-up. Among the four monkeys inoculated with amastigotes forms, two IV inoculated showed the parasite in the bone-marrow from the first toward to the sixth month p.i. and following that they cleared the infection, whereas the oth

  16. Respuesta inmune a la infección por Leishmania infantum en caninos - Immune response to infection by Leishmania infantum in dogs

    Directory of Open Access Journals (Sweden)

    Rodríguez Villamizar, Irlene Evelyne

    2009-11-01

    Full Text Available ResumenLa leishmaniasis visceral canina (LVC es una enfermedad infecciosaclásicamente asociada al protozoo Leishmania spp, que se manifiestacon un amplio espectro patológico, desde infecciones asintomáticashasta procesos viscerales fatales. El control de la infección implica el desarrollo de una respuesta inmune protectora, que en el caso de los perros es dicotómica, caracterizada tanto por mecanismos humoralescomo celulares. Los mecanismos humorales involucran anticuerpos ycomplemento mientras que el mecanismo celular implica la activación demacrófagos y linfocitos T con la producción de citocinas. El objetivo de esta revisión es ofrecer un panorama general de los mecanismosinmunológicos, células y moléculas que intervienen durante elestablecimiento de la infección por Leishmania en perros.SummaryCanine Visceral Leishmaniosis (CVL is an infectious disease classicallyassociated with the protozoan Leishmania spp, which cause a broadpathological spectrum, ranging from asymptomatic infection to fatalvisceral processes. Infection control involves the development of aprotective immune response, which in the case of dogs is dichotomous,characterized by both humoral and cellular mechanisms. Humoralmechanisms include antibodies and complement but cellular mechanisminvolved alveolar macrophages and T lymphocytes activation with theproduction of cytokines. The aim of this review is to provide an overview about the immunological mechanisms, cells and moleculesinvolved during the establishment of the Leishmania infection in dogs.

  17. Alta prevalencia de infección por Leishmania (Kinetoplastidae: Trypanosomatidae) en perros del norte de Colombia / High prevalence of infection with Leishmania (Kinetoplastea: Trypanosomatidae) in dogs in northern Colombia

    Scientific Electronic Library Online (English)

    Margaret Paternina, Gómez; Yirys, Díaz-Olmos; Luis Enrique, Paternina; Eduar Elías, Bejarano.

    2013-09-01

    Full Text Available Introducción. Aunque el perro se considera el principal reservorio doméstico de Leishmania infantum en varios países de Latinoamérica, se desconocen las tasas de infección por parásitos del género Leishmania en perros de algunos focos de leishmaniasis visceral del norte de Colombia. Objetivo. Establ [...] ecer las tasas de infección por Leishmania spp. en poblaciones caninas del departamento de Sucre en el norte de Colombia. Materiales y métodos. Se estudiaron perros de zonas rurales de los municipios de Sincelejo, Sampués y Ovejas. Los parásitos del género Leishmania se detectaron mediante la amplificación de un segmento de ADN de la subunidad ribosómica pequeña (rSSU). Se analizó la relación entre variables como sexo, grupos etarios y presencia de signos clínicos, respecto a la infección por Leishmania . Resultados. Se amplificó un fragmento de 561 pb de la rSSU de Leishmania spp. en 34,9 % (29/83), 35,7 % (15/42) y 11,1 % (1/9) de los perros analizados de Sincelejo, Sampués y Ovejas, respectivamente. La prevalencia total de infección por Leishmania spp. fue de 33,6 %. En dos perros de Sincelejo, dos de Sampués y uno de Ovejas, se obtuvo una banda de 650 pb, característica de parásitos del género Trypanosoma . El 64,2 % de los perros que mostraron al menos un signo clínico indicativo de leishmaniasis canina, fue positivo por PCR. Se halló una relación estadísticamente significativa entre los perros mayores de 33 meses de edad y el resultado positivo para Leishmania spp. (p=0,043). No se encontró relación entre el sexo del perro y el resultado positivo para Leishmania spp. (p=0,85) . Conclusión. Un tercio de la población canina analizada se encontró infectada con parásitos del género Leishmania , lo cual podría sustentar su posible papel como fuente de infección para el insecto vector. Abstract in english Introduction: Although the dog is considered to be the main domestic reservoir of Leishmania infantum in several Latin American countries, the rate of infection with Leishmania parasites remains unknown in dogs from foci of visceral leishmaniasis of northern of Colombia. Objetive: To determine the p [...] revalence of Leishmania infection in canine population from department of Sucre in northern Colombia. Materials and methods: The canine population of rural areas of the municipalities of Sincelejo, Sampués and Ovejas was studied. Parasites of the genus Leishmania were detected by amplifying a DNA segment of the small ribosomal subunit (rSSU). The relationship among sex, age group and clinical signs with infection by Leishmania was analyzed. Results: A 561 bp fragment of the Leishmania rSSU was amplified in 34.9% (29/83), 35.7% (15/42) and 11.1% (1/9) of the dogs analyzed from Sincelejo, Sampués and Ovejas, respectively. Overall prevalence of Leishmania infection was 33.6%. A 650 bp band, characteristic of parasites of the genus Trypanosoma, was amplified in two dogs from Sincelejo and Sampués and in one dog from Ovejas. The 64.2% of the dogs that showed clinical signs compatible with canine leishmaniasis, was PCRpositive. A statistically significant relationship was found between dogs over 33 months old and positivity to Leishmania (p=0.043). No relationship was observed between sex and positivity to Leishmania (p=0.85) . Conclusion: A third of the canine population studied is infected with Leishmania , which could support its potential role as a source of infection for the insect vector.

  18. An oligonucleotide probe derived from kDNA minirepeats is specific for Leishmania (Viannia)

    Scientific Electronic Library Online (English)

    Octavio, Fernandes; Marcelo, Bozza; Juan M, Pascale; Antonio B de, Miranda; Ulisses G, Lopes; Wim M, Degrave.

    1996-06-01

    Full Text Available Sequence analysis of Leishmania (Viannia) kDNA minicircles and analysis of multiple sequence alignments of the conserved region (minirepeats) of five distinct minicircles from L. (V.) braziliensis species with corresponding sequences derived from other dermotropic leishmanias indicated the presence [...] of a sub-genus specific sequence. An oligonucleotide bearing this sequence was designed and used as a molecular probe, being able to recognize solely the sub-genus Viannia species in hybridization experiments. A dendrogram reflecting the homologies among the minirepeat sequences was constructed. Sequence clustering was obtained corresponding to the traditional classification based on similarity of biochemical, biological and parasitological characteristics of these Leishmania species, distinguishing the Old World dermotropic leishmanias, the New World dermotropic leishmanias of the sub-genus Leishmania and of the sub-genus Viannia.

  19. An oligonucleotide probe derived from kDNA minirepeats is specific for Leishmania (Viannia

    Directory of Open Access Journals (Sweden)

    Octavio Fernandes

    1996-06-01

    Full Text Available Sequence analysis of Leishmania (Viannia kDNA minicircles and analysis of multiple sequence alignments of the conserved region (minirepeats of five distinct minicircles from L. (V. braziliensis species with corresponding sequences derived from other dermotropic leishmanias indicated the presence of a sub-genus specific sequence. An oligonucleotide bearing this sequence was designed and used as a molecular probe, being able to recognize solely the sub-genus Viannia species in hybridization experiments. A dendrogram reflecting the homologies among the minirepeat sequences was constructed. Sequence clustering was obtained corresponding to the traditional classification based on similarity of biochemical, biological and parasitological characteristics of these Leishmania species, distinguishing the Old World dermotropic leishmanias, the New World dermotropic leishmanias of the sub-genus Leishmania and of the sub-genus Viannia.

  20. Computational Prediction of Protein-Protein Interactions in Leishmania Predicted Proteomes

    OpenAIRE

    Rezende, Antonio M; Folador, Edson L; Resende, Daniela de M.; Ruiz, Jeronimo C.

    2012-01-01

    The Trypanosomatids parasites Leishmania braziliensis, Leishmania major and Leishmania infantum are important human pathogens. Despite of years of study and genome availability, effective vaccine has not been developed yet, and the chemotherapy is highly toxic. Therefore, it is clear just interdisciplinary integrated studies will have success in trying to search new targets for developing of vaccines and drugs. An essential part of this rationale is related to protein-protein interaction netw...

  1. Effects of Sheep and Mouse Urine on the Growth Pattern of Leishmania major Promastigotes

    OpenAIRE

    Abdolhossein Dalimi; Habibollah Paykari; Vahid Nasiri; Gholamreza Karimi; Fatemeh Ghaffarifar

    2013-01-01

    The protozoan parasites of the genus Leishmania are the causative agents of different clinical diseases. Fetal calf serum (FCS) is the main part and the most expensive ingredient of the Leishmania culture media. Here, the efficacies of different concentrations (1%, 2.5%, 5%, and 10%) of the filtered and autoclaved sheep and mouse urine were evaluated as a growth stimulator in Leishmania culture procedure. The results indicated that culture media enriched with the filtered sheep and mouse urin...

  2. Molecular Cloning and Expression of Iranian Leishmania major Pteridine Reductase 1

    OpenAIRE

    Mosaffa, N; M . Mohebali; F. Mahboudi; Haghighi, A.; M. Bandehpour; Kheirandish, F; B Kazemi

    2008-01-01

    Background: Leishmaniasis is an endemic disease in 88 countries. Reports on Leishmania drug resistance are growing in number. The mechanism of unresponsiveness against glucantime in Iranian cutaneous leishmaniasis has not yet been characterized. To begin the first step in finding an anti-Leishmania chemotherapy, we prepared recombinant L. major PTR1 enzyme and characterized its activity by enzymatic assay. Methods: Leishmania promastigote DNA was extracted and the ptr1 gene amplified using sp...

  3. Selective Fusion of Azurophilic Granules with Leishmania-containing Phagosomes in Human Neutrophils*

    OpenAIRE

    Mollinedo, Faustino; Janssen, Hans; de la Iglesia-Vicente, Janis; Villa-Pulgarin, Janny A.; Calafat, Jero

    2010-01-01

    Leishmania parasites use polymorphonuclear neutrophils as intermediate hosts before their ultimate delivery to macrophages following engulfment of parasite-infected neutrophils. This leads to a silent and unrecognized entry of Leishmania into the macrophage host cell. Neutrophil function depends on its cytoplasmic granules, but their mobilization and role in how Leishmania parasites evade intracellular killing in neutrophils remain undetermined. Here, we have found by ultrastructural approach...

  4. Leishmania infection in humans, dogs and sandflies in a visceral leishmaniasis endemic area in Maranhão, Brazil

    OpenAIRE

    Ilana Mirian Almeida Felipe; Dorlene Maria Cardoso de Aquino; Oliver Kuppinger; Max Diego Cruz Santos; Maurício Eduardo Salgado Rangel; David Soeiro Barbosa; Aldina Barral; Guilherme Loureiro Werneck; Arlene de Jesus Mendes Caldas

    2011-01-01

    Leishmania infection in humans, dogs and sandflies was examined in the endemic visceral leishmaniasis (VL) municipality of Raposa, state of Maranhão, Brazil. In this study, we examined Leishmania chagasi infection in the blood serum of both humans and Canis familiaris and the natural Leishmania sp. infection rate in the sandfly vector, Lutzomyia longipalpis. Enzyme-linked immunosorbent assay, indirect immunofluorescence reaction and polymerase chain reaction were performed to detect Leishmani...

  5. Las proteínas salivales de los flebotominos en la transmisión de Leishmania y su impacto epidemiológico

    OpenAIRE

    Elsa Nieves; Mireya Sánchez; Maritza Rondón

    2013-01-01

    Los flebotominos son insectos hematófagos transmisores de Leishmania, agente causal de la leishmaniasis, enfermedad que presenta un amplio espectro clínico en los humanos. En el presente trabajo se realiza una revisión sobre la saliva de los flebotominos en la transmisión de Leishmania y su impacto epidemiológico. Se aborda primeramente el potencial de la saliva de los flebotominos como un coctel farmacológico que interviene en la biología del insecto, en el proceso de infección de Leishmania...

  6. Leishmania ?-1,2-mannan is assembled on a mannose-cyclic phosphate primer

    OpenAIRE

    Sernee, M. Fleur; Ralton, Julie E.; Dinev, Zoran; Khairallah, George N.; O’Hair, Richard A.; Williams, Spencer J; McConville, Malcolm J.

    2006-01-01

    Infective stages of the protozoan parasite Leishmania spp. accumulate a class of ?-1,2-mannan oligosaccharides as their major carbohydrate reserve material. Here, we describe the biosynthesis of Leishmania mannan. Mannan precursors were identified by metabolic labeling of Leishmania mexicana promastigotes with [3H]mannose. Label was initially incorporated into a phosphomannose primer and short phosphorylated ?-1,2-mannan oligomers that were two to five residues long. Analysis of the mannan pr...

  7. Serological detection of Toxoplasma gondii, Leishmania infantum and Neospora caninum in cats from an area endemic for leishmaniasis in Brazil / Detecção sorológica de Toxoplasma gondii, Leishmania infantum e Neospora caninum em gatos de uma área endêmica para leishmaniose no Brasil

    Scientific Electronic Library Online (English)

    Keyla Carstens Marques de, Sousa; Heitor Miraglia, Herrera; Iara Helena, Domingos; João Bosco Vilela, Campos; Isabela Maria Campanelli dos, Santos; Haroldo Henrique, Neves; Rosangela Zacarias, Machado; Marcos Rogério, André.

    2014-12-01

    Full Text Available O presente estudo teve como objetivo investigar a ocorrência de anticorpos anti-Toxoplasma gondii, Leishmania infantum e Neospora caninum, em 151 gatos, por meio da Reação de Imunofluorescência Indireta (RIFI). Entre os meses de janeiro e abril de 2013, amostras de soro foram coletadas de 151 gatos [...] domésticos (65 gatos errantes e 86 gatos domiciliados; 55 machos e 96 fêmeas), de Campo Grande, Mato Grosso do Sul, Brasil. Anticorpos IgG anti-T. gondii, anti-L. infantum e anti-N. caninum foram encontrados em 49 (32,5%), 34 (22,5%) e 10 (6,6%) gatos amostrados, respectivamente. Verificou-se uma associação estatisticamente significativa entre as infecções por T. gondii e N. caninum, T. gondii e L. infantum e N. caninum e Leishmania infantum (p Abstract in english An investigation was made into the occurrence of antibodies to Toxoplasma gondii, Leishmania infantum and Neospora caninum in 151 domestic cats, based on the indirect fluorescent antibody test (IFAT). Serum samples were collected from 151 domestic cats (65 free-roaming and 86 domiciled cats; 55 male [...] s and 96 females) in Campo Grande, Mato Grosso do Sul, Brazil between January and April 2013. IgG antibodies to T. gondii, L. infantum and N. caninum were found, respectively, in 49 (32.5%), 34 (22.5%) and 10 (6.6%) sampled cats. A positive correlation was found between T. gondii and N. caninum, T. gondii and L. infantum, and N. caninum and L. infantum (p

  8. Leishmaniose causada por Leishmania viannia braziliensis (Lvb um caso de evolução atípica

    Directory of Open Access Journals (Sweden)

    Jackson Maurício Lopes Costa

    1988-06-01

    Full Text Available Os autores relatam o caso de uma criança portadora de Leishmaniose Tegumentar Americana causada por Leishmania braziliensis braziliensis que foi infectada durante a amamentação, desenvolvendo lesão infiltrativa e nodular nos lábios, com posterior disseminação para os seios da face, fossas nasais e pavilhão auricular e cuja evolução clinica pós-terapêutica caracterizou-se por períodos sucessivos de regressão e de reativação da lesão. Enfatizam a gravidade do caso, e as dificuldades terapêuticas com a utilização dos antimoniais pentavalentes, antimoniato-N-metil glucamina (Glucantime e o stibogluconato de sódio(Pentostam.

  9. Efecto de la diminacina en hámsteres inoculados con Leishmania chagasi

    Scientific Electronic Library Online (English)

    A.N, Burna; M.S, Catuogno; M, Sánchez Negrette; A, Mirad; O.A, Maccio.

    2013-12-01

    Full Text Available La leishmaniosis es una enfermedad causada por protozoarios del género Leishmania, que afecta al hombre y animales a través de la picadura de insectos infectados. La diminacina es una droga con actividad in vitro inhibitoria del crecimiento de Leishmania donovani; también ha sido utilizada para el t [...] ratamiento contra tripanosomosis y babesiosis. El objetivo del presente trabajo fue comprobar el efecto de diminacina in vivo como tratamiento antileishmaniásico en hámsteres, y en caso afirmativo transpolarla como alternativa terapéutica para el control y posible erradicación de la leishmaniosis en caninos. Se utilizaron 26 hámsteres agrupados en cuatro lotes de 6-7 animales cada uno: (a) inyectados con diminacina, (b) inoculados con Leishmania chagasi, (c) inoculados con L. chagasi e inyectados con diminacina, y (d) controles. La inoculación se efectuó con un macerado de bazo extraído de un canino naturalmente infectado con L. chagasi. La diminacina fue administrada a razón de 3,5 mg/kg vía IM. Los frotis y cortes histopatológicos de los hámsteres infectados revelaron amastigotes de L. chagasi. Los ejemplares infectados y tratados con diminacina mantuvieron un mejor estado de salud que aquéllos privados de tratamiento, lo cual sugiere que la droga tendría un efecto benéfico en el organismo del animal infectado. Abstract in english Leishmaniosis is a disease caused by protozoa of the genus Leishmania, which is transmitted to human beings by the bite of infected insects. Diminazine is a drug with proven in vitro growth inhibition activity of Leishmania donovani and has also been used for the treatment against trypanosomosis and [...] other parasitic diseases. The aim of this study was to determine the effect of diminazine in vivo as an antileishmanial drug in hamsters for the possible extrapolation and thus potential use as a therapeutic alternative in dogs. Twenty six hamsters were divided into four groups, two groups of 6 animals each, and two groups of 7 each. Some animals were inoculated IM with macerated spleen from a dog naturally infected with Leishmania chagasi. The diminazine treated group and the control animals inoculated with the protozoa presented amastigotes of L. chagasi in smears and in histopathological sections. Diminazine treated animals remained in better health state unlike the control group inoculated with L. chagasi. Results suggest that diminazine would have a benefcial effect in infected dogs.

  10. Selective Fusion of Azurophilic Granules with Leishmania-containing Phagosomes in Human Neutrophils*

    Science.gov (United States)

    Mollinedo, Faustino; Janssen, Hans; de la Iglesia-Vicente, Janis; Villa-Pulgarin, Janny A.; Calafat, Jero

    2010-01-01

    Leishmania parasites use polymorphonuclear neutrophils as intermediate hosts before their ultimate delivery to macrophages following engulfment of parasite-infected neutrophils. This leads to a silent and unrecognized entry of Leishmania into the macrophage host cell. Neutrophil function depends on its cytoplasmic granules, but their mobilization and role in how Leishmania parasites evade intracellular killing in neutrophils remain undetermined. Here, we have found by ultrastructural approaches that neutrophils ingested Leishmania major promastigotes, and azurophilic granules fused in a preferential way with parasite-containing phagosomes, without promoting parasite killing. Azurophilic granules, identified by the granule marker myeloperoxidase, also fused with Leishmania donovani-engulfed vacuoles in human neutrophils. In addition, the azurophilic membrane marker CD63 was also detected in the vacuole surrounding the parasite, and in the fusion of azurophilic granules with the parasite-engulfed phagosome. Tertiary and specific granules, involved in vacuole acidification and superoxide anion generation, hardly fused with Leishmania-containing phagosomes. L. major interaction with neutrophils did not elicit production of reactive oxygen species or mobilization of tertiary and specific granules. By using immunogold electron microscopy approaches in the engulfment of L. major and L. donovani by human neutrophils, we did not find a significant contribution of endoplasmic reticulum to the formation of Leishmania-containing vacuoles. Live Leishmania parasites were required to be optimally internalized by neutrophils. Our data suggest that Leishmania promastigotes modulate their uptake by neutrophils, and regulate granule fusion processes in a rather selective way to favor parasite survival in human neutrophils. PMID:20801889

  11. The first detection of Leishmania major in naturally infected Sergentomyia minuta in Portugal

    Scientific Electronic Library Online (English)

    Lenea, Campino; Sofia, Cortes; Lidia, Dionisio; Luis, Neto; Maria Odete, Afonso; Carla, Maia.

    2013-06-01

    Full Text Available Phlebotomine sandflies of the genus Sergentomyia are widely distributed throughout the Old World. It has been suggested that Sergentomyia spp are involved in the transmission of Leishmania in India and Africa, whereas Phlebotomus spp are thought to be the sole vectors of Leishmania in the Old World. [...] In this study, Leishmania major DNA was detected in one Sergentomyia minuta specimen that was collected in the southern region of Portugal. This study challenges the dogma that Leishmania is exclusively transmitted by species of the genus Phlebotomus in the Old World.

  12. The genome of the kinetoplastid parasite, Leishmania major.

    Science.gov (United States)

    Ivens, Alasdair C; Peacock, Christopher S; Worthey, Elizabeth A; Murphy, Lee; Aggarwal, Gautam; Berriman, Matthew; Sisk, Ellen; Rajandream, Marie-Adele; Adlem, Ellen; Aert, Rita; Anupama, Atashi; Apostolou, Zina; Attipoe, Philip; Bason, Nathalie; Bauser, Christopher; Beck, Alfred; Beverley, Stephen M; Bianchettin, Gabriella; Borzym, Katja; Bothe, Gordana; Bruschi, Carlo V; Collins, Matt; Cadag, Eithon; Ciarloni, Laura; Clayton, Christine; Coulson, Richard M R; Cronin, Ann; Cruz, Angela K; Davies, Robert M; De Gaudenzi, Javier; Dobson, Deborah E; Duesterhoeft, Andreas; Fazelina, Gholam; Fosker, Nigel; Frasch, Alberto Carlos; Fraser, Audrey; Fuchs, Monika; Gabel, Claudia; Goble, Arlette; Goffeau, André; Harris, David; Hertz-Fowler, Christiane; Hilbert, Helmut; Horn, David; Huang, Yiting; Klages, Sven; Knights, Andrew; Kube, Michael; Larke, Natasha; Litvin, Lyudmila; Lord, Angela; Louie, Tin; Marra, Marco; Masuy, David; Matthews, Keith; Michaeli, Shulamit; Mottram, Jeremy C; Müller-Auer, Silke; Munden, Heather; Nelson, Siri; Norbertczak, Halina; Oliver, Karen; O'neil, Susan; Pentony, Martin; Pohl, Thomas M; Price, Claire; Purnelle, Bénédicte; Quail, Michael A; Rabbinowitsch, Ester; Reinhardt, Richard; Rieger, Michael; Rinta, Joel; Robben, Johan; Robertson, Laura; Ruiz, Jeronimo C; Rutter, Simon; Saunders, David; Schäfer, Melanie; Schein, Jacquie; Schwartz, David C; Seeger, Kathy; Seyler, Amber; Sharp, Sarah; Shin, Heesun; Sivam, Dhileep; Squares, Rob; Squares, Steve; Tosato, Valentina; Vogt, Christy; Volckaert, Guido; Wambutt, Rolf; Warren, Tim; Wedler, Holger; Woodward, John; Zhou, Shiguo; Zimmermann, Wolfgang; Smith, Deborah F; Blackwell, Jenefer M; Stuart, Kenneth D; Barrell, Bart; Myler, Peter J

    2005-07-15

    Leishmania species cause a spectrum of human diseases in tropical and subtropical regions of the world. We have sequenced the 36 chromosomes of the 32.8-megabase haploid genome of Leishmania major (Friedlin strain) and predict 911 RNA genes, 39 pseudogenes, and 8272 protein-coding genes, of which 36% can be ascribed a putative function. These include genes involved in host-pathogen interactions, such as proteolytic enzymes, and extensive machinery for synthesis of complex surface glycoconjugates. The organization of protein-coding genes into long, strand-specific, polycistronic clusters and lack of general transcription factors in the L. major, Trypanosoma brucei, and Trypanosoma cruzi (Tritryp) genomes suggest that the mechanisms regulating RNA polymerase II-directed transcription are distinct from those operating in other eukaryotes, although the trypanosomatids appear capable of chromatin remodeling. Abundant RNA-binding proteins are encoded in the Tritryp genomes, consistent with active posttranscriptional regulation of gene expression. PMID:16020728

  13. Leishmania vaccine development: exploiting the host-vector-parasite interface.

    Science.gov (United States)

    Reed, S G; Coler, R N; Mondal, D; Kamhawi, S; Valenzuela, J G

    2016-01-01

    Visceral leishmaniasis (VL) is a disease transmitted by phlebotomine sand flies, fatal if untreated, and with no available human vaccine. In rodents, cellular immunity to Leishmania parasite proteins as well as salivary proteins of the sand fly is associated with protection, making them worthy targets for further exploration as vaccines. This review discusses the notion that a combination vaccine including Leishmania and vector salivary antigens may improve vaccine efficacy by targeting the parasite at its most vulnerable stage just after transmission. Furthermore, we put forward the notion that better modeling of natural transmission is needed to test efficacy of vaccines. For example, the fact that individuals living in endemic areas are exposed to sand fly bites and will mount an immune response to salivary proteins should be considered in pre-clinical and clinical evaluation of leishmaniasis vaccines. Nevertheless, despite remaining obstacles there is good reason to be optimistic that safe and effective vaccines against leishmaniasis can be developed. PMID:26595093

  14. Human cutaneous leishmaniasis caused by Leishmania (Viannia braziliensis in Santiago del Estero, Argentina: identification of parasites by monoclonal antibodies and isoenzymes Leishmaniose cutânea humana causada por Leishmania (Viannia braziliensis na Província de Santiago del Estero, Argentina: identificação dos parasitas por anticorpos monoclonais e isoenzimas

    Directory of Open Access Journals (Sweden)

    C.A. Cuba Cuba

    1996-12-01

    Full Text Available Diagnostic and parasite characterization and identification studies were carried out in human patients with cutaneous leishmaniasis lesions in Santiago del Estero, Northern Province of Argentina. Diagnostic procedures were biopsies of lesions for smears and inoculations in hamster, needle aspirations of material from ulcers for "in vitro" cultures. Immunodiagnostic techniques applied were IFAT-IgG and Montenegro skin test. Primary isolation of eight stocks of leishmanial parasites was achieved from patients with active lesions. All stocks were biologically characterized by their behaviour in hamster, measurements of amastigote and promastigotes and growth "in vitro". Eight stocks were characterized and identified at species level by their reactivity to a cross-panel of sub-genus and specie-specific Monoclonal Antibodies through an Indirect Immunofluorescence technique and a Dot-ELISA. We conclude from the serodeme analysis of Argentina stocks that: stocks MHOM/AR/92/SE-1; SE-2; SE-4; SE-8; SE-8-I; SE-30; SE-34 and SE-36 are Leishmania (Viannia braziliensis. Three Leishmania stocks (SE-1; SE-2 and SE-30 did not react with one highly specie-specific Monoclonal Antibody (Clone: B-18, Leishmania (Viannia braziliensis marker disclosing two serodeme group patterns. Five out of eight soluble extracts of leishmanial promastigotes were electrophoresed on thin-layer starch gels and examined for the enzyme MPI, Mannose Phosphate Isomerase; MDH, Malate Dehydrogenase; 6PGD, 6 Phosphogluconate Dehydrogenase; NH, Nucleoside Hydrolase, 2-deoxyinosinc as substrate; SOD, Superoxide Dismutase; GPI, Glucose Phosphate Isomerase and ES, Esterase. From the isoenzyme studies we concluded that stocks: MHOM/AR/92/SE-1; SE-2; SE-4; SE-8 and SE-8-I are isoenzymatically Leishmania (Viannia braziliensis. We need to analyze more enzymes before assigning them to a braziliensis zymodeme.Estudos de diagnóstico, caracterização parasitária e identificação foram conduzidos em pacientes humanos com lesões cutâneas de leishmaniose na Província de Santiago del Estero, no Norte da Argentina. Os procedimentos de diagnóstico foram: biópsias de lesões para utilização em esfregaços e inoculação em hamster; aspiração (com agulha de úlceras, para cultura "in vitro". As técnicas imunodiagnósticas empregadas foram a IFAT-IgG e o teste intradérmico de Montenegro. Oito cepas de parasitas foram isoladas, sendo estas obtidas de pacientes com lesões ativas. Todas as cepas foram inicialmente caracterizadas biologicamente por seu comportamento na infecção experimental do hamster, mensuração dos amastigotas e promastigotas e crescimento "in vitro". As mesmas oito cepas foram logo identificadas e caracterizadas a nível de espécie, devido a sua reatividade frente a um painel de anticorpos monoclonais subgênero e espécie-específicos. Isso foi realizado utilizando o teste de Imunofluorescência Indireta (IFAT/MAbs e de um procedimento de Dot-ELISA. Nós concluímos a partir da análise de serodema dos isolados argentinos que: MHOM/AR/92/SE-1; SE-2; SE-4; SE-8; SE-8-I; SE-30; SE-34 e SE-36 são Leishmania (Viannia braziliensis. Entretanto, três dos isolados de Leishmania (SE-1; SE-2 e SE-30 não foram reconhecidos quando testados com um anticorpo monoclonal de reconhecida alta espécie-especificidade (clone B-18, marcador consagrado de Leishmania (Viannia braziliensis, revelando a existência de dois tipos de serodemas entre as cepas estudadas. Cinco dos oito extratos solúveis de Leishmania foram submetidos à eletroforese em gel de amido de camada fina e subseqüentemente examinadas a fim de constatar a atividade das enzimas MPI, MDH, 6PGD, NH, NH-D, SOD, GPI e ES. Fundamentados nos estudos dos corridos eletroforéticos obtidos nos ensaios isoenzimáticos chegamos à conclusão que as cepas MHOM/AR/92 SE-1; SE-2; SE-4; SE-8 e SE-8-I são Leishmania (Viannia braziliensis. É necessário analisar mais enzimas antes de enquadrá-los nos zymodema braziliensis

  15. Biological action of inosine analogs in Leishmania and Trypanosoma spp.

    OpenAIRE

    Marr, J J; Berens, R L; Cohn, N K; Nelson, D J; Klein, R.S.

    1984-01-01

    Previous investigations have suggested that inosine analogs would be good models for the development of chemotherapeutic agents active against pathogenic hemoflagellates. We have systematically modified the five-membered heterocyclic ring of six inosine analogs and tested them for their antiprotozoal activities and toxicity to a mammalian cell line. All six analogs were very active against the three protozoan pathogens Leishmania donovani, Trypanosoma cruzi, and Trypanosoma gambiense. Two of ...

  16. Miltefosine Affects Lipid Metabolism in Leishmania donovani Promastigotes?

    OpenAIRE

    Rakotomanga, M.; Blanc, S.; Gaudin, K; Chaminade, P.; Loiseau, P.M.

    2007-01-01

    Miltefosine (hexadecylphosphocholine [HePC]) is the first orally active antileishmanial drug. Transient HePC treatment of Leishmania donovani promastigotes at 10 ?M significantly reduced the phosphatidylcholine content and enhanced the phosphatidylethanolamine (PE) content in parasite membranes, suggesting a partial inactivation of PE-N-methyltransferase. Phospholipase D activity did not seem to be affected by HePC. In addition, the enhancement of the lysophosphatidylcholine content could be ...

  17. Riesgo de transmisión de Leishmania (Kinetoplastida: Trypanosomatidae) en Mérida Venezuela

    OpenAIRE

    Elsa Nieves; Luzmary Oraá; Yorfer Rondón; Mireya Sánchez; Yetsenia Sánchez; Maria Rujano; Maritza Rondón; Masyelly Rojas; Nestor Gonzalez; Dalmiro Cazorla

    2014-01-01

    La leishmaniasis es una enfermedad causada por la infección de un parásito protozoario del género Leishmania, transmitido por la picada de insectos hematófagos conocidos como flebotominos. El estudio tiene como objetivo determinar la presencia de flebotominos en los Distritos Sanitarios del estado Mérida y diseñar un mapa de riesgo de transmisión entomológico. Se utilizaron cuatro métodos de captura de flebotominos, los ejemplares se identificaron y se les determinó la infección natural por L...

  18. Immunostimulatory DNA as an Adjuvant in Vaccination against Leishmania major

    OpenAIRE

    Stacey, Katryn J.; Blackwell, Jenefer M.

    1999-01-01

    Oligodeoxynucleotides (ODN) which contain immunostimulatory CG motifs (CpG ODN) can promote T helper 1 (Th1) responses, an adjuvant activity that is desirable for vaccination against leishmaniasis. To test this, susceptible BALB/c mice were vaccinated with soluble leishmanial antigen (SLA) with or without CpG ODN as adjuvant and then challenged with Leishmania major metacyclic promastigotes. CpG ODN alone gave partial protection when injected up to 5 weeks prior to infection, and longer if th...

  19. Genetic Data Showing Evolutionary Links between Leishmania and Endotrypanum

    OpenAIRE

    Elisa Cupolillo; Luiza OR Pereira; Octávio Fernandes; Marcos P Catanho; Júlio C Pereira; Enrique Medina-Acosta; Gabriel Grimaldi Jr.

    1998-01-01

    Striking similarities at the morphological, molecular and biological levels exist between many trypanosomatids isolated from sylvatic insects and/or vertebrate reservoir hosts that make the identification of medically important parasites demanding. Some molecular data have pointed to the relationship between some Leishmania species and Endotrypanum, which has an important epidemiological significance and can be helpful to understand the evolution of those parasites. In this study, we have dem...

  20. Glycosome turnover in Leishmania major is mediated by autophagy.

    Science.gov (United States)

    Cull, Benjamin; Prado Godinho, Joseane Lima; Fernandes Rodrigues, Juliany Cola; Frank, Benjamin; Schurigt, Uta; Williams, Roderick Am; Coombs, Graham H; Mottram, Jeremy C

    2014-01-01

    Autophagy is a central process behind the cellular remodeling that occurs during differentiation of Leishmania, yet the cargo of the protozoan parasite's autophagosome is unknown. We have identified glycosomes, peroxisome-like organelles that uniquely compartmentalize glycolytic and other metabolic enzymes in Leishmania and other kinetoplastid parasitic protozoa, as autophagosome cargo. It has been proposed that the number of glycosomes and their content change during the Leishmania life cycle as a key adaptation to the different environments encountered. Quantification of RFP-SQL-labeled glycosomes showed that promastigotes of L. major possess ~20 glycosomes per cell, whereas amastigotes contain ~10. Glycosome numbers were significantly greater in promastigotes and amastigotes of autophagy-defective L. major Δatg5 mutants, implicating autophagy in glycosome homeostasis and providing a partial explanation for the previously observed growth and virulence defects of these mutants. Use of GFP-ATG8 to label autophagosomes showed glycosomes to be cargo in ~15% of them; glycosome-containing autophagosomes were trafficked to the lysosome for degradation. The number of autophagosomes increased 10-fold during differentiation, yet the percentage of glycosome-containing autophagosomes remained constant. This indicates that increased turnover of glycosomes was due to an overall increase in autophagy, rather than an upregulation of autophagosomes containing this cargo. Mitophagy of the single mitochondrion was not observed in L. major during normal growth or differentiation; however, mitochondrial remnants resulting from stress-induced fragmentation colocalized with autophagosomes and lysosomes, indicating that autophagy is used to recycle these damaged organelles. These data show that autophagy in Leishmania has a central role not only in maintaining cellular homeostasis and recycling damaged organelles but crucially in the adaptation to environmental change through the turnover of glycosomes. PMID:25484087

  1. Infecção natural de Lutzomyia (Lutzomyia) longipalpis (Diptera: Psychodidae) por Leishmania infantum chagasi em flebotomíneos capturados no município de Janaúba, Estado de Minas Gerais, Brasil / Natural infection with Leishmania infantum chagasi in Lutzomyia (Lutzomyia) longipalpis (Diptera: Psychodidae) sandflies captured in the municipality of Janaúba, State of Minas Gerais, Brazil

    Scientific Electronic Library Online (English)

    Érika Monteiro, Michalsky; Karla de Sena, Guedes; Fabiana de Oliveira, Lara e Silva; João Carlos, França-Silva; Consuelo Latorre Fortes, Dias; Ricardo Andrade, Barata; Edelberto Santos, Dias.

    2011-02-01

    Full Text Available INTRODUÇÃO: A leishmaniose visceral tem sido notificada em quase todos os estados do Brasil, e principalmente no norte de Minas Gerais, onde a doença é endêmica. Este estudo visou detectar a infecção natural de Lutzomyia longipalpis e identificar através da técnica de PCR/RFLP a espécie de Leishmani [...] a encontrada nos flebotomíneos do município de Janaúba. MÉTODOS: Utilizando-se armadilhas luminosas, foram capturadas 1.550 fêmeas de L. longipalpis, que agrupadas em pool de 10 exemplares foram submetidas à extração e amplificação de DNA, através das técnicas de PCR genérico e cacofonia. RESULTADOS: Dos 155 pools, seis apresentaram-se positivos para Leishmania sp., sendo a taxa de infecção do município de 3,9%. Através da PCR/RFLP determinou-se que o padrão de digestão das amostras positivas foi semelhante ao da cepa referência Leishmania chagasi (MHOM/BR/74/PP75). CONCLUSÕES: A detecção de infecção natural associada a estudos sobre a epidemiologia da LV sugere que L. longipalpis esteja envolvida na transmissão de L. infantum chagasi em Janaúba, principalmente nas áreas de intensa transmissão de LV. Abstract in english INTRODUCTION: Visceral leishmaniasis has been notified in nearly all states of Brazil, and particularly in the north of Minas Gerais, where the disease is endemic. The aim of this study was to detect natural infection of Lutzomyia longipalpis and, through the PCR/RFLP technique, identify Leishmania [...] species found in sandflies in the municipality of Janaúba. METHODS: Using light traps, 1,550 females of L. longipalpis were caught and grouped into pools of 10 specimens to be subjected to DNA extraction and amplification, by means of generic PCR and cacophony. RESULTS: Out of the 155 pools, six were positive for Leishmania sp., and thus the infection rate in the municipality was 3.9%. Through PCR/RFLP, the digestion pattern among the positive samples was found to be similar to that of the reference strain of Leishmania chagasi (MHOM/BR/74/PP75). CONCLUSIONS: The detection of natural infection associated with studies on the epidemiology of visceral leishmaniasis suggests that L. longipalpis is involved in transmission of L. infantum chagasi in Janaúba, particularly in areas of intense transmission of visceral leishmaniasis.

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

    Directory of Open Access Journals (Sweden)

    B Bastrenta

    2003-03-01

    Full Text Available 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.

  3. A Historical Overview of the Classification, Evolution, and Dispersion of Leishmania Parasites and Sandflies

    Science.gov (United States)

    Akhoundi, Mohammad; Kuhls, Katrin; Cannet, Arnaud; Votýpka, Jan; Marty, Pierre; Delaunay, Pascal; Sereno, Denis

    2016-01-01

    Background The aim of this study is to describe the major evolutionary historical events among Leishmania, sandflies, and the associated animal reservoirs in detail, in accordance with the geographical evolution of the Earth, which has not been previously discussed on a large scale. Methodology and Principal Findings Leishmania and sandfly classification has always been a controversial matter, and the increasing number of species currently described further complicates this issue. Despite several hypotheses on the origin, evolution, and distribution of Leishmania and sandflies in the Old and New World, no consistent agreement exists regarding dissemination of the actors that play roles in leishmaniasis. For this purpose, we present here three centuries of research on sandflies and Leishmania descriptions, as well as a complete description of Leishmania and sandfly fossils and the emergence date of each Leishmania and sandfly group during different geographical periods, from 550 million years ago until now. We discuss critically the different approaches that were used for Leishmana and sandfly classification and their synonymies, proposing an updated classification for each species of Leishmania and sandfly. We update information on the current distribution and dispersion of different species of Leishmania (53), sandflies (more than 800 at genus or subgenus level), and animal reservoirs in each of the following geographical ecozones: Palearctic, Nearctic, Neotropic, Afrotropical, Oriental, Malagasy, and Australian. We propose an updated list of the potential and proven sandfly vectors for each Leishmania species in the Old and New World. Finally, we address a classical question about digenetic Leishmania evolution: which was the first host, a vertebrate or an invertebrate? Conclusions and Significance We propose an updated view of events that have played important roles in the geographical dispersion of sandflies, in relation to both the Leishmania species they transmit and the animal reservoirs of the parasites. PMID:26937644

  4. Transmission of Leishmania in coffee plantations of Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    Bruce Alexander

    2002-07-01

    Full Text Available Transmission of Leishmania was studied in 27 coffee plantations in the Brazilian State of Minas Gerais. Eighteen females and six males (11.6% of the people tested, aged between 7-65 gave a positive response to the Montenegro skin test. Awareness of sand flies based on the ability of respondents to identify the insects using up to seven predetermined characteristics was significantly greater among inhabitants of houses occupied by at least one Mn+ve individual. Five species of phlebotomine sand fly, including three suspected Leishmania vectors, were collected within plantations under three different cultivation systems. Four of these species i.e., Lu. fischeri (Pinto 1926, Lu. migonei (França 1920, Lu. misionensis (Castro 1959 and Lutzomyia whitmani (Antunes & Coutinho 1939 were collected in an organic plantation and the last of these was also present in the other two plantation types. The remaining species, Lu. intermedia (Lutz & Neiva 1912, was collected in plantations under both the "adensado" and "convencional" systems. The results of this study indicate that transmission of Leishmania to man in coffee-growing areas of Minas Gerais may involve phlebotomine sand flies that inhabit plantations.

  5. Transmission of Leishmania in coffee plantations of Minas Gerais, Brazil

    Scientific Electronic Library Online (English)

    Bruce, Alexander; Emerson Barbosa de, Oliveria; Emily, Haigh; Lourenço Leal de, Almeida.

    2002-07-01

    Full Text Available Transmission of Leishmania was studied in 27 coffee plantations in the Brazilian State of Minas Gerais. Eighteen females and six males (11.6% of the people tested), aged between 7-65 gave a positive response to the Montenegro skin test. Awareness of sand flies based on the ability of respondents to [...] identify the insects using up to seven predetermined characteristics was significantly greater among inhabitants of houses occupied by at least one Mn+ve individual. Five species of phlebotomine sand fly, including three suspected Leishmania vectors, were collected within plantations under three different cultivation systems. Four of these species i.e., Lu. fischeri (Pinto 1926), Lu. migonei (França 1920), Lu. misionensis (Castro 1959) and Lutzomyia whitmani (Antunes & Coutinho 1939) were collected in an organic plantation and the last of these was also present in the other two plantation types. The remaining species, Lu. intermedia (Lutz & Neiva 1912), was collected in plantations under both the "adensado" and "convencional" systems. The results of this study indicate that transmission of Leishmania to man in coffee-growing areas of Minas Gerais may involve phlebotomine sand flies that inhabit plantations.

  6. Canine experimental infection: intradermal inoculation of Leishmania infantum promastigotes

    Scientific Electronic Library Online (English)

    Gabriela M, Santos-Gomes; Lenea, Campino; Pedro, Abranches.

    2000-04-01

    Full Text Available Five mixed breed dogs were inoculated intradermally (ID) with cultured virulent stationary phase promastigotes of Leishmania infantum Nicole, 1908 stocks recently isolated. Parasite transformations in the skin of ID infected dogs were monitored from the moment of inoculation and for 48 h, by skin bi [...] opsies. Anti-Leishmania antibody levels were measured by indirect immunofluorescence assay, counterimmunoelectrophoresis and direct agglutination test, and clinical conditions were examined. Thirty minutes after ID inoculation the first amastigotes were visualised and 3 to 4 h after inoculation the promastigotes were phagocyted by neutrophils and by a few macrophages. These cells parasitised by amastigotes progressively disappeared from the skin and 24 h after inoculation parasites were no longer observed. Local granulomes were not observed, however, serological conversion for antibodies anti-Leishmania was achieved in all dogs. Direct agglutination test was the only technique positive in all inoculated dogs. Amastigotes were found in the popliteal lymph node in one dog three months after inoculation. This work demonstrates that, with this inoculum, the promastigotes were transformed into amastigotes and were up taken by neutrophils and macrophages. The surviving parasites may have been disseminated in the canine organism, eliciting a humoral response in all cases.

  7. Canine experimental infection: intradermal inoculation of Leishmania infantum promastigotes

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    Gabriela M Santos-Gomes

    2000-04-01

    Full Text Available Five mixed breed dogs were inoculated intradermally (ID with cultured virulent stationary phase promastigotes of Leishmania infantum Nicole, 1908 stocks recently isolated. Parasite transformations in the skin of ID infected dogs were monitored from the moment of inoculation and for 48 h, by skin biopsies. Anti-Leishmania antibody levels were measured by indirect immunofluorescence assay, counterimmunoelectrophoresis and direct agglutination test, and clinical conditions were examined. Thirty minutes after ID inoculation the first amastigotes were visualised and 3 to 4 h after inoculation the promastigotes were phagocyted by neutrophils and by a few macrophages. These cells parasitised by amastigotes progressively disappeared from the skin and 24 h after inoculation parasites were no longer observed. Local granulomes were not observed, however, serological conversion for antibodies anti-Leishmania was achieved in all dogs. Direct agglutination test was the only technique positive in all inoculated dogs. Amastigotes were found in the popliteal lymph node in one dog three months after inoculation. This work demonstrates that, with this inoculum, the promastigotes were transformed into amastigotes and were up taken by neutrophils and macrophages. The surviving parasites may have been disseminated in the canine organism, eliciting a humoral response in all cases.

  8. Application of the microscopic method in cutaneous leishmania diagnosis

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    Mohammed Wael Daboul

    2011-10-01

    Full Text Available Introduction: Cutaneous leishmania is spreading fast. This study aims at developing the microscopic method to achieve a full detection of all positive cases of leishmania.Methods: 50 human cases have been studied by applying microscopic smears stained with Wright stain. Microscopic photos were taken for the presumed unfamiliar figures.Results: Mononuclear cells with tails are present at a rate of (98%. They are associated with Leishman Donovan (LD bodies in 50% of the cases. The polygonal figures and the spherical forms are present at the same rate (60% and are associated with LD bodies in 24% of the cases. The small promastigote like forms are seen at a rate of (76% and are associated with LD bodies in 26% of the cases. The giant promastigotes like forms are present in (80% of the cases and are associated with LD bodies in 28% of the cases. Candle flame forms are present in (40% of the cases and are associated with the LD bodies in 21% of the cases.Discussion: It is applicable to use those discovered figures in diagnosing cutaneous leishmania.

  9. Leishmania (Viannia) infection in the domestic dog in Chaparral, Colombia.

    Science.gov (United States)

    Santaella, Julián; Ocampo, Clara B; Saravia, Nancy G; Méndez, Fabián; Góngora, Rafael; Gomez, Maria Adelaida; Munstermann, Leonard E; Quinnell, Rupert J

    2011-05-01

    Peridomestic transmission of American cutaneous leishmaniasis is increasingly reported and dogs may be a reservoir of Leishmania (Viannia) in this setting. We investigated the prevalence of infection in dogs in Chaparral County, Colombia, the focus of an epidemic of human cutaneous leishmaniasis caused by Leishmania (Viannia) guyanensis. Two (0.72%) of 279 dogs had lesions typical of cutaneous leishmaniasis that were biopsy positive by kinetoplast DNA polymerase chain reaction-Southern blotting. Seroprevalence was 2.2% (6 of 279) by enzyme-linked immunosorbent assay. Buffy coat and ear skin biopsy specimens were positive by polymerase chain reaction-Southern blotting in 7.3% (10 of 137) and 11.4% (12 of 105) of dogs, respectively. Overall 20% of dogs (21 of 105) showed positive results for one or more tests. Amplification and sequencing of the Leishmania 7SL RNA gene identified L. guyanensis in one dog and L. braziliensis in two dogs. No association was identified between the risk factors evaluated and canine infection. Dogs may contribute to transmission but their role in this focus appears to be limited. PMID:21540374

  10. Crystal structure of Leishmania tarentolae hypoxanthine-guanine phosphoribosyltransferase

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    Oliva Glaucius

    2007-09-01

    Full Text Available Abstract Background Hypoxanthine-guanine phosphoribosyltransferase (HGPRT (EC 2.4.2.8 is a central enzyme in the purine recycling pathway. Parasitic protozoa of the order Kinetoplastida cannot synthesize purines de novo and use the salvage pathway to synthesize purine bases, making this an attractive target for antiparasitic drug design. Results The glycosomal HGPRT from Leishmania tarentolae in a catalytically active form purified and co-crystallized with a guanosine monophosphate (GMP in the active site. The dimeric structure of HGPRT has been solved by molecular replacement and refined against data extending to 2.1 Ã… resolution. The structure reveals the contacts of the active site residues with GMP. Conclusion Comparative analysis of the active sites of Leishmania and human HGPRT revealed subtle differences in the position of the ligand and its interaction with the active site residues, which could be responsible for the different reactivities of the enzymes to allopurinol reported in the literature. The solution and analysis of the structure of Leishmania HGPRT may contribute to further investigations leading to a full understanding of this important enzyme family in protozoan parasites.

  11. Deception and manipulation: the arms of leishmania, a successful parasite.

    Science.gov (United States)

    Cecílio, Pedro; Pérez-Cabezas, Begoña; Santarém, Nuno; Maciel, Joana; Rodrigues, Vasco; Cordeiro da Silva, Anabela

    2014-01-01

    Leishmania spp. are intracellular parasitic protozoa responsible for a group of neglected tropical diseases, endemic in 98 countries around the world, called leishmaniasis. These parasites have a complex digenetic life cycle requiring a susceptible vertebrate host and a permissive insect vector, which allow their transmission. The clinical manifestations associated with leishmaniasis depend on complex interactions between the parasite and the host immune system. Consequently, leishmaniasis can be manifested as a self-healing cutaneous affliction or a visceral pathology, being the last one fatal in 85-90% of untreated cases. As a result of a long host-parasite co-evolutionary process, Leishmania spp. developed different immunomodulatory strategies that are essential for the establishment of infection. Only through deception and manipulation of the immune system, Leishmania spp. can complete its life cycle and survive. The understanding of the mechanisms associated with immune evasion and disease progression is essential for the development of novel therapies and vaccine approaches. Here, we revise how the parasite manipulates cell death and immune responses to survive and thrive in the shadow of the immune system. PMID:25368612

  12. Evaluación de la Intradermorreacción con Antígenos de Leishmania (Viannia) peruviana y Leishmania (Viannia) braziliensis en Áreas endémicas de Leishmaniasis en el Perú

    Scientific Electronic Library Online (English)

    G, Minaya; Y, Torres; M, Farfán; L, Mendizabal; M, Colchado; E, Arroyo.

    Full Text Available Se elaboró extractos de antígenos solubles de Leishmania (leishmanina), a partir de formas promastigotes de Leishmania (Viannia) peruviana y Leishmania (Viannia) braziliensis. Los antígenos fueron evaluados respecto a su sensibilidad, especificidad y efectos colaterales. Los ensayos se llevaron a ca [...] bo en una área de transmisión de la "uta" (leishmaniasis andina), en un área de transmisión de la "espundia"(leishmaniasis selvática), y en pacientes atendidos en Lima. El antígeno de Leishmania (Viannia) peruviana, 25 µg/ml, resultó adecuado para la detección de la reacción de hipersensibilidad cutánea tardía en pacientes con lesiones cutáneas y/o cutáneo-mucosas activas, encontrando una sensibilidad promedio de 95,9%. La misma sensibilidad, pero con reacciones más intensas (potentes) fue observada con el antígeno homólogo de 30 µg/ml. Los antígenos de Leishmania (Viannia) braziliensis de 25 y 30 µg/ml mostraron similar sensibilidad y potencia a las dosis equivalentes de Leishmania (Viannia) peruviana.La sensibilidad y la potencia de la reacción se incrementaron mínimamente con el incremento en la concentración de proteínas. No se observaron efectos colaterales como vesiculación y ulceración en el sitio de inoculación. Los antígenos demostraron ser altamente específicos. La especificidad en el grupo control alcanzó el 100,0%, mientras que en el grupo control endémico fue 95,7%. Abstract in english Extracts of Leishmania soluble antigens (leishmanina) were made from Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis promastigotes.Antigens were evaluated for sensitivity, specificity and side effects. Assays were perfomed in transmission areas for "uta" (andean leishmaniasis), [...] "espundia" (sylvatic leishmaniasis), and also in patients seen in Lima. Leishmania (Viannia) peruviana antigen, 25 µg/ml, was adequate to detect late skin hypersensitivity reaction among patients with active cutaneous and/or mucocutaneous lesions, with an average sensitivity of 95,9%. The same sensitivity, but with more intense (potency) reactions were observed with the homologous 30 µg/ml antigen. Antigens of Leishmania (Viannia) braziliensis, 25 and 30 µg/ml, showed similar sensitivity and potency to the equivalent doses of Leishmania (Viannia) peruviana. The reaction’s sensitivity and potency increase minimally with increased protein concentration. No side effects, such as vesiculation and ulceration on the site of the innoculation, were observed. Antigens showed high specificity. Specificity in the control group reached 100,0% whereas in the endemic control group, it was 95,7%.

  13. Diagnosis of American cutaneous leishmaniasis by enzyme immunoassay using membrane antigens of Leishmania (Viannia) braziliensis.

    Science.gov (United States)

    Skraba, Cissiara Manetti; Pedroso, Raíssa Bocchi; Fiorini, Adriana; Rosado, Fábio Rogério; Aristides, Sandra Mara Alessi; Lonardoni, Maria Valdrinez Campana; Teixeira, Jorge Juarez Vieira; Silveira, Thaís Gomes Verzignassi

    2014-04-01

    This study evaluated the reactivity of membrane antigens of Leishmania (Viannia) braziliensis for the diagnosis of ACL by enzyme immunoassay (EIA). Promastigotes of L. (V.) braziliensis were grown in medium 199 and lysed in a sonicator. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting showed that specific proteins of L. (V.) braziliensis (apparent molecular weights 36 kDa and 48-56 kDa) were recognized by sera from ACL patients. These proteins were eluted from the SDS-PAGE and tested in EIA-IgG with sera from ACL patients, healthy individuals, patients with toxoplasmosis, paracoccidioidomycosis, syphilis, tuberculosis, leprosy, and Chagas disease. The EIA-IgG with membrane antigens allowed us to distinguish patients with ACL from healthy individuals and patients with other diseases (P results showed good sensitivity and specificity of EIA-IgG with membrane antigens, indicating their potential use for diagnosis of ACL, as well as seroepidemiological surveys and follow-up of clinically cured patients. PMID:24485589

  14. Leishmania (Leishmania) major-infected rhesus macaques (Macaca mulatta) develop varying levels of resistance against homologous re-infections

    Scientific Electronic Library Online (English)

    VF, Amaral; A, Teva; R, Porrozzi; AJ, Silva; MS, Pereira; MP, Oliveira-Neto; G, Grimaldi Jr.

    2001-08-01

    Full Text Available Seven rhesus macaques were infected intradermally with 10(7) promastigotes of Leishmania (Leishmania) major. All monkeys developed a localized, ulcerative, self-healing nodular skin lesion at the site of inoculation of the parasite. Non-specific chronic inflammation and/or tuberculoid-type granuloma [...] tous reaction were the main histopathological manifestations of the disease. Serum Leishmania-specific antibodies (IgG and IgG1) were detected by ELISA in all infected animals; immunoblot analyses indicated that numerous antigens were recognized. A very high degree of variability was observed in the parasite-specific cell-mediated immune responses [as detected by measuring delayed-type hypersensitivity (DTH) reaction, in vitro lymphocyte proliferation, and gamma interferon (IFN-gamma) production] for individuals over time post challenge. From all the recovered monkeys (which showed resolution of the lesions after 11 weeks of infection), 57.2% (4/7) and 28.6% (2/7) animals remained susceptible to secondary and tertiary infections, respectively, but the disease severity was altered (i.e. lesion size was smaller and healed faster than in the primary infection). The remaining monkeys exhibited complete resistance (i.e. no lesion) to each rechallenge. Despite the inability to consistently detect correlates of cell-mediated immunity to Leishmania or correlation between resistance to challenge and DTH, lymphocyte transformation or IFN-gamma production, partial or complete acquired resistance was conferred by experimental infection. This primate model should be useful for measuring vaccine effectiveness against the human disease.

  15. Infección de fibroblastos de piel de animales con distinto grado de susceptibilidad a Leishmania infantum y Leishmania mexicana (Kinetoplastida: Trypanosomatidae)

    Scientific Electronic Library Online (English)

    Miguel Angel, Minero; Misael, Chinchilla; Olga, Marta Guerrero; Alfredo, Castro.

    2004-03-01

    Full Text Available En este estudio se presenta un modelo in vitro de cultivo de fibroblastos de piel de hámster, ratón y rata hecho con el propósito de determinar diferencias en cuanto a la susceptibilidad a la infección por dos especies del género Leishmania (Kinetoplastida: Trypanosomatidae). Se realizó además un es [...] tudio ultraestructural por microscopía electrónica de transmisión con el fin de establecer si las formas intracelulares observadas correspondían a multiplicación interna o a fagocitosis múltiple. Se estudió la multiplicación de los parásitos en los fibroblastos de las tres especies de roedores infectados tanto por Leishmania infantum como por L. mexicana (cepa OCR) y las diferencias entre las tres fueron estadísticamente significativas (p Abstract in english Infection and multiplication of Leishmania infantum and L. mexicana inside of skin fibroblasts from hamsters, mice and rats was achieved. This process was demonstrated either by counting parasites inside the stained cells or by electronic microscopy studies. In addition multiplication rate differenc [...] es in the cells from these rodent species were determined, for L. infantum as well as for L. mexicana. Parasite development in hamsters and mice fibroblasts was evident but there was not multiplication in rat cells showing that apparently they are refractory to Leishmania infection. These results suggest that the parasite affinity for each animal, as well as any intracellular environment resistance, could involve genetic factors in the parasite multiplication. On the other hand, presence of amastigote multiplication inside of parasitophorus vacuole, showed by electronic microscopy images, probes a true parasite transformation. Therefore it is suggested that fibroblasts could work as host cells for parasite survival and permanency in the infected animals

  16. Infección de fibroblastos de piel de animales con distinto grado de susceptibilidad a Leishmania infantum y Leishmania mexicana (Kinetoplastida: Trypanosomatidae

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    Miguel Angel Minero

    2004-03-01

    Full Text Available En este estudio se presenta un modelo in vitro de cultivo de fibroblastos de piel de hámster, ratón y rata hecho con el propósito de determinar diferencias en cuanto a la susceptibilidad a la infección por dos especies del género Leishmania (Kinetoplastida: Trypanosomatidae. Se realizó además un estudio ultraestructural por microscopía electrónica de transmisión con el fin de establecer si las formas intracelulares observadas correspondían a multiplicación interna o a fagocitosis múltiple. Se estudió la multiplicación de los parásitos en los fibroblastos de las tres especies de roedores infectados tanto por Leishmania infantum como por L. mexicana (cepa OCR y las diferencias entre las tres fueron estadísticamente significativas (pInfection and multiplication of Leishmania infantum and L. mexicana inside of skin fibroblasts from hamsters, mice and rats was achieved. This process was demonstrated either by counting parasites inside the stained cells or by electronic microscopy studies. In addition multiplication rate differences in the cells from these rodent species were determined, for L. infantum as well as for L. mexicana. Parasite development in hamsters and mice fibroblasts was evident but there was not multiplication in rat cells showing that apparently they are refractory to Leishmania infection. These results suggest that the parasite affinity for each animal, as well as any intracellular environment resistance, could involve genetic factors in the parasite multiplication. On the other hand, presence of amastigote multiplication inside of parasitophorus vacuole, showed by electronic microscopy images, probes a true parasite transformation. Therefore it is suggested that fibroblasts could work as host cells for parasite survival and permanency in the infected animals

  17. Recombinant Leishmania (Leishmania) infantum Ecto-Nucleoside Triphosphate Diphosphohydrolase NTPDase-2 as a new antigen in canine visceral leishmaniasis diagnosis.

    Science.gov (United States)

    de Souza, Ronny Francisco; Dos Santos, Yaro Luciolo; de Souza Vasconcellos, Raphael; Borges-Pereira, Lucas; Caldas, Ivo Santana; de Almeida, Márcia Rogéria; Bahia, Maria Terezinha; Fietto, Juliana Lopes Rangel

    2013-01-01

    Canine visceral leishmaniasis is an important public health concern. In the epidemiological context of human visceral leishmaniasis, dogs are considered the main reservoir of Leishmania parasites; therefore, dogs must be epidemiologically monitored constantly in endemic areas. Furthermore, dog to human transmission has been correlated with emerging urbanization and increasing rates of leishmaniasis infection worldwide. Leishmania (Leishmania) infantum (L. chagasi) is the etiologic agent of visceral leishmaniasis in the New World. In this work, a new L. (L.) infantum (L. chagasi) recombinant antigen, named ATP diphosphohydrolase (rLic-NTPDase-2), intended for use in the immunodiagnosis of CVL was produced and validated. The extracellular domain of ATP diphosphohydrolase was cloned and expressed in the pET21b-Escherichia coli expression system. Indirect ELISA assays were used to detect the purified rLic-NTPDase-2 antigen using a standard canine sera library. This library contained CVL-positive samples, leishmaniasis-negative samples and samples from Trypanosoma cruzi-infected dogs. The results show a high sensitivity of 100% (95% CI=92.60-100.0%) and a high specificity of 100% (95% CI=86.77-100.0%), with a high degree of confidence (k=1). These findings demonstrate the potential use of this recombinant protein in immune diagnosis of canine leishmaniasis and open the possibility of its application to other diagnostic approaches, such as immunochromatography fast lateral flow assays and human leishmaniasis diagnosis. PMID:23022017

  18. Biodistribution of meglumine antimoniate in healthy and Leishmania (Leishmania) infantum chagasi-infected BALB/c mice

    Scientific Electronic Library Online (English)

    Samanta Etel Treiger, Borborema; João Alberto, Osso Junior; Heitor Franco de, Andrade Junior; Nanci do, Nascimento.

    2013-08-01

    Full Text Available Pentavalent antimonials such as meglumine antimoniate (MA) are the primary treatments for leishmaniasis, a complex disease caused by protozoan parasites of the genus Leishmania . Despite over 70 years of clinical use, their mechanisms of action, toxicity and pharmacokinetics have not been fully eluc [...] idated. Radiotracer studies performed on animals have the potential to play a major role in pharmaceutical development. The aims of this study were to prepare an antimony radiotracer by neutron irradiation of MA and to determine the biodistribution of MA in healthy and Leishmania (Leishmania) infantum chagasi-infected mice. MA (Glucantime(r)) was neutron irradiated inside the IEA-R1 nuclear reactor, producing two radioisotopes, 122Sb and 124Sb, with high radionuclidic purity and good specific activity. This irradiated compound presented anti-leishmanial activity similar to that of non-irradiated MA in both in vitro and in vivo evaluations. In the biodistribution studies, healthy mice showed higher uptake of antimony in the liver than infected mice and elimination occurred primarily through biliary excretion, with a small proportion of the drug excreted by the kidneys. The serum kinetic curve was bi-exponential, with two compartments: the central compartment and another compartment associated with drug excretion. Radiotracers, which can be easily produced by neutron irradiation, were demonstrated to be an interesting tool for answering several questions regarding antimonial pharmacokinetics and chemotherapy.

  19. Biodistribution of meglumine antimoniate in healthy and Leishmania (Leishmania infantum chagasi-infected BALB/c mice

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    Samanta Etel Treiger Borborema

    2013-08-01

    Full Text Available Pentavalent antimonials such as meglumine antimoniate (MA are the primary treatments for leishmaniasis, a complex disease caused by protozoan parasites of the genus Leishmania . Despite over 70 years of clinical use, their mechanisms of action, toxicity and pharmacokinetics have not been fully elucidated. Radiotracer studies performed on animals have the potential to play a major role in pharmaceutical development. The aims of this study were to prepare an antimony radiotracer by neutron irradiation of MA and to determine the biodistribution of MA in healthy and Leishmania (Leishmania infantum chagasi-infected mice. MA (Glucantime(r was neutron irradiated inside the IEA-R1 nuclear reactor, producing two radioisotopes, 122Sb and 124Sb, with high radionuclidic purity and good specific activity. This irradiated compound presented anti-leishmanial activity similar to that of non-irradiated MA in both in vitro and in vivo evaluations. In the biodistribution studies, healthy mice showed higher uptake of antimony in the liver than infected mice and elimination occurred primarily through biliary excretion, with a small proportion of the drug excreted by the kidneys. The serum kinetic curve was bi-exponential, with two compartments: the central compartment and another compartment associated with drug excretion. Radiotracers, which can be easily produced by neutron irradiation, were demonstrated to be an interesting tool for answering several questions regarding antimonial pharmacokinetics and chemotherapy.

  20. Molecular and serological detection of Leishmania spp. in captive wild animals from Ilha Solteira, SP, Brazil.

    Science.gov (United States)

    Jusi, Márcia Mariza Gomes; Starke-Buzetti, Wilma Aparecida; Oliveira, Trícia Maria Ferreira de Sousa; Tenório, Michely da Silva; Sousa, Lúcio de Oliveira de; Machado, Rosangela Zacarias

    2011-01-01

    Leishmaniasis is a zoonotic disease that affects 12 million people worldwide. Several mammalian species can serve as a reservoir for this disease. Dogs are the main reservoir for visceral leishmaniasis in urban areas, which has become a serious public health concern in Brazil. The aim of this study was to evaluate the presence of Leishmania spp. in captive wild animals from Ilha Solteira, São Paulo, Brazil. Blood and various tissues samples were collected from animals of five different species: Speothos venaticus, Chrysocyon brachyurus, Cerdocyon thous, Pseudalopex vetulus, and Procyon cancrivorus. Antibodies against Leishmania spp. were detected in three wild canids by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). PCR analyses of blood and bone marrow from all animals were negative, but Leishmania DNA was found in the tissues and skin of seropositive animals. Positive PCR samples were also positive for Leishmania donovani complex. Analysis of sequenced PCR products showed similarities with different regions of Leishmania (Leishmania) infantum and Leishmania (Leishmania) chagasi kinetoplastids. Measures to control visceral leishmaniasis in wild animals kept in Brazilian zoos should be established, as no disease control programs are currently available. PMID:21961752

  1. Cross-protective efficacy from a immunogen firstly identified in Leishmania infantum against tegumentary leishmaniasis.

    Science.gov (United States)

    Martins, V T; Lage, D P; Duarte, M C; Costa, L E; Chávez-Fumagalli, M A; Roatt, B M; Menezes-Souza, D; Tavares, C A P; Coelho, E A F

    2016-02-01

    Experimental vaccine candidates have been evaluated to prevent leishmaniasis, but no commercial vaccine has been proved to be effective against more than one parasite species. LiHyT is a Leishmania-specific protein that was firstly identified as protective against Leishmania infantum. In this study, LiHyT was evaluated as a vaccine to against two Leishmania species causing tegumentary leishmaniasis (TL): Leishmania major and Leishmania braziliensis. BALB/c mice were immunized with rLiHyT plus saponin and lately challenged with promastigotes of the two parasite species. The immune response generated was evaluated before and 10 weeks after infection, as well as the parasite burden at this time after infection. The vaccination induced a Th1 response, which was characterized by the production of IFN-?, IL-12 and GM-CSF, as well as by high levels of IgG2a antibodies, after in vitro stimulation using both the protein and parasite extracts. After challenge, vaccinated mice showed significant reductions in their infected footpads, as well as in the parasite burden in the tissue and organs evaluated, when compared to the control groups. The anti-Leishmania Th1 response was maintained after infection, being the IFN-? production based mainly on CD4(+) T cells. We described one conserved Leishmania-specific protein that could compose a pan-Leishmania vaccine. PMID:26756314

  2. Surveillance for antibodies to Leishmania spp. in dogs from Sri Lanka and India

    Science.gov (United States)

    The global distribution of leishmaniasis is rapidly expanding into new geographic regions. Dogs are the primary reservoir hosts for human visceral leishmaniasis (VL) caused by infection with Leishmania infantum. Natural infections with other Leishmania species can occur in dogs, but their role as re...

  3. Qualidade espermática de sêmen de cães naturalmente infectados por Leishmania sp: Semen quality of dogs naturally infected by Leishmania sp

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    É. Labat

    2010-06-01

    Full Text Available Avaliaram-se alterações espermáticas associadas à infecção por leishmaniose no sêmen de cães naturalmente infectados, utilizando-se, durante oito semanas consecutivas, ejaculados de seis cães soronegativos e seis cães soropositivos. As amostras foram colhidas uma vez por semana e avaliadas quanto ao volume, concentração, motilidade, vigor, morfologia espermática, integridade da cromatina, avaliação simultânea da integridade da membrana plasmática, acrossoma e potencial mitocondrial. Concomitantemente foram dosadas a proteína total do plasma seminal e sanguíneo. A leishmaniose visceral causou aumento dos defeitos maiores e menores nos espermatozoides dos animais acometidos pelo estágio moderado a severo da doença. Em estágios mais avançados da enfermidade, a integridade das membranas acrossomal e plasmática foi afetada negativamente. Não foi possível estabelecer um critério quanto à avaliação do potencial mitocondrial. A incidência de alterações morfológicas nos animais acometidos não promoveu aumento de injurias à cromatina. Todos os animais com leishmaniose apresentaram hiperproteinemia do sêmen.The spermatic changes associated with the natural infection in dogs by Leishmania sp was evaluated during eight consecutive weeks, using ejaculates of six seronegative and six seropositive dogs. The samples were collected once a week and evaluated for volume, concentration, motility, vigor, sperm morphology, chromatin integrity, simultaneous evaluation of the plasmatic membrane integrity, acrosome, and mitochondrial potential. The total proteins of the seminal plasma and blood were measured. The visceral leishmaniasis caused increase of major and minor defects in spermatozoa of animals attacked by moderate to severe stages of the disease. In more advanced stages of the illness, the acrosomal and plasmatic membranes integrity was adversely affected. It was not possible to establish a pattern refering the evaluation of the mitochondrial potential. The incidence of morphological changes in the seropositive animals did not promote an increase of injuries to the chromatin. All animals with leishmaniasis presented hyperproteinemia of the semen.

  4. Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil

    Scientific Electronic Library Online (English)

    Luiza de Oliveira Ramos, Pereira; Ana Cláudia, Maretti-Mira; Káris Maria, Rodrigues; Rosimar Baptista, Lima; Manoel Paes de, Oliveira-Neto; Elisa, Cupolillo; Claude, Pirmez; Márcia Pereira de, Oliveira.

    2013-08-01

    Full Text Available Leishmania RNA virus (LRV) has been shown to be a symbiotic component of Leishmania parasites in South America. Nested retro-transcription polymerase chain reaction was employed to investigate LRV1 presence in leishmaniasis lesions from Brazil. In endemic areas of Rio de Janeiro (RJ), no LRV1 infec [...] tion was observed even with mucosal involvement. LRV1 was only detected in Leishmania (V.) guyanensis cutaneous lesions from the northern region, which were obtained from patients presenting with disease reactivation after clinical cure of their primary lesions. Our results indicated that the severity of leishmaniasis in some areas of RJ, where Leishmania (V.) brazi-liensis is the primary etiological agent, was not associated with Leishmania LRV1 infection.

  5. Effects of some insecticides on the neutral lipid percentage, survival and infectivity of Steinernema carpocapsae ALL and Heterorhabditis amazonensis JPM 4

    Directory of Open Access Journals (Sweden)

    Paulo Henrique de Siqueira Sabino

    2014-08-01

    Full Text Available Lipids are an important energy source for entomopathogenic nematodes (EPNs and directly influence their infectivity in the host. Some insecticides reduce the infectivity of infective juveniles (IJs while keeping them viable after exposure. Thus, the objective of this study was to correlate the amounts of lipid reserves in the EPN Heterorhabditis amazonensis JPM 4 and Steinernema carpocapsae ALL with their survival and infectivity when exposed to insecticides that keep the nematodes viable but reduced their infective capacity against Galleria mellonella. Among the tested insecticides, Vertimec? and Klorpan? were incompatible (class 2 with the two EPN species because they reduced infectivity. The insecticides Vertimec? and Klorpan? maintained the viability of the IJs but reduced their infectivity and their lipid amounts after insecticide exposure.

  6. Naturally azole-resistant Leishmania braziliensis promastigotes are rendered susceptible in the presence of terbinafine: comparative study with azole-susceptible Leishmania mexicana promastigotes.

    OpenAIRE

    Rangel, H; Dagger, F; Hernandez, A.; A. Liendo; Urbina, J A

    1996-01-01

    Leishmania braziliensis (isolate 2903) was naturally resistant to ketoconazole or the bis-triazole D0870, inhibitors of sterol C-14 demethylase, which produced only moderate effects on the proliferation of promastigotes at 10 microM. In contrast, Leishmania mexicana (isolate NR) was extremely susceptible to the azoles, as complete growth arrest and cell lysis were induced by incubation of the parasites with 0.05 microM concentrations of the drugs for 72 h. The opposite response was observed w...

  7. Characterization of Leishmania (Leishmania) waltoni n.sp. (Kinetoplastida: Trypanosomatidae), the Parasite Responsible for Diffuse Cutaneous Leishmaniasis in the Dominican Republic.

    Science.gov (United States)

    Shaw, Jeffrey; Pratlong, Francine; Floeter-Winter, Lucile; Ishikawa, Edna; El Baidouri, Fouad; Ravel, Christophe; Dedet, Jean-Pierre

    2015-09-01

    Leishmania parasites isolated, between 1979 and 1988 by the late Bryce Walton, from Dominican Republic (DR) patients with diffuse cutaneous leishmaniasis, were characterized using a panel of 12 isoenzymes, 23 monoclonal antibodies, small subunit ribosomal DNA (SSu rDNA), and multilocus sequence analysis (MLSA). The isoenzyme and monoclonal antibody profiles and the MLSA results showed that the Dominican Republic parasites were distinct from other described Leishmania species. This new species belongs to the mexicana complex, which is distributed in central and parts of northern South America. It is suggested that the parasites uniqueness from other members of the mexicana complex is related to it being isolated on an island for millions of years. If Leishmania (Leishmania) waltoni fails to adapt to some imported mammal, such as the house rat, it will be the only Leishmania to be classified as an endangered species. The excessive destruction of habitats on Hispaniola threatens the survival of its vectors and presumed natural reservoirs, such as the rodent hutias and the small insectivorous mammal solenodon. The concept of Leishmania species is discussed in the light of recent evaluations on criteria for defining bacterial species. PMID:26149864

  8. Human mucocutaneous leishmaniasis in Três Braços, Bahia - Brazil: an area of Leishmania braziliensis braziliensis transmission. II. Cutaneous disease. Presentation and evolution

    Scientific Electronic Library Online (English)

    Elmer A., Llanos-Cuentas; Philip D., Marsden; Edinaldo L., Lago; Air C., Barreto; César C., Cuba; Warren D., Johnson.

    1984-12-01

    Full Text Available Foram analisados os dados clínicos de 182 pacientes com leishmaniose cutânea, provavelmente causada por Leishmania braziliensis braziliensis. Sessenta e oito por cento apresentavam uma única lesão, usualmente uma úlcera, na terça parte inferior anterior da tíbia. Todos os grupos etários estavam repr [...] esentados e muitos apresentaram histórico de um a dois meses. Treze por cento apresentavam lesões fechadas de natureza verrucosa ou em placa. Após tratamento, a evolução destas lesões foi relacionada à regularidade da terapia por antimônio. Embora a cura usualmente ocorresse em três meses, o tempo de cicatrização, após o início de tratamento, foi variável e relativo ao tamanho da lesão (p Abstract in english The clinical records of 182 patients with cutaneous leishmaniasis probably due to Leishmania braziliensis braziliensis are analysed. 68% had a single lesion which was usually an ulceron the lower anterior tibial third. Many had short histories of one to two months and all age groups were represented [...] 13% had closed lesions of a verrucose or plaque like nature. Evolution of these skin lesions after treatment was related to the regularity of antimony therapy. Although healing usually occurred in three months, the time to scarring after commencing treatment was variable and related to the size ofthe lesion (p

  9. Densidade de oviposição, e quantificação de Larvas e Pupas de Mansonia Blanchard, 1901 (Diptera: Culicidae), em Eichhornia crassipes Solms. e Pistia stratiotes Linn. Na ilha da Marchantaria, Amazônia Central / Oviposition density and quantification of Larvae and Puppae of Mansonia Blanchard 1901 (Diptera: Culicidae), in Eichhornia crassipes Solms. and Pistia stratiotes Linn, on Marchantaria island in the Central Amazon

    Scientific Electronic Library Online (English)

    Ruth Leila Menezes, FERREIRA.

    1999-03-01

    Full Text Available A população de imaturos de Mansonia em águas brancas da várzea da ilha da Marchantaria, na Amazônia Central se constituiu de 74% de Mansonia (M.) humeralis Dyar & Knab, 1916, 14% de Mansonia (M.) titillans (Walker, 1848) e 12% de Mansonia (M.) amazonensis Theobald, 1901, como foi indicado pela frequ [...] ência, medida por oviposição (posturas). O número de posturas de M. humeralis e M. titillans foi maior em Pistia stratiotes Linn. enquanto que M. amazonensis foi mais abundante em Eichhornia crassipes Solms. O número total de larvas e pupas de Mansonia spp. foi de 6997 e 570 respectivamente, sendo maior em E. crassipes (4372 e 328) que em P. stratiotes (2625 e 242). Ao longo do ano, o maior número de larvas e pupas foi encontrado nos meses da vazante (agosto-outubro), tendo um pico no mês de agosto, que não foi correlacionado significativamente com os fatores abióticos. Abstract in english The population of immature Mansonia in the white-water floodplain of Marchantaria Island, in Central Amazonia was composed by 74% of Mansonia (M.) humeralis Dyar & Knab, 1916, 14% of Mansonia (M.) titillans (Walker, 1848) and 12% of Mansonia (M.) amazonensis Theobald, 1901, as indicated by frequency [...] of oviposition (egg masses). Number of egg masses of M. humeralis and M. titillans was higher on Pistia stratiotes whereas that of M. amazonensis was higher on Eichhornia crassipes. The total number of larvae and pupae of Mansonia spp. was 6997 and 570, respectively, and was higher in E. crassipes (4372 and 328) than in P. stratiotes (2625 and 242). Most larvae and pupae were found in the season of lowest water-level (August-October), with a peak in August, not being significantly correlated with abiotic factors.

  10. Risk factors for Leishmania chagasi infection in an endemic area in Raposa, State of Maranhão, Brazil / Fatores associados à infecção por Leishmania chagasi em uma área endêmica em Raposa, Estado do Maranhão, Brasil

    Scientific Electronic Library Online (English)

    Charlene Barreto, Ponte; Natália Coelho, Souza; Maria Neuza, Cavalcante; Aldina Maria Prado, Barral; Dorlene Maria Cardoso de, Aquino; Arlene de Jesus Mendes, Caldas.

    2011-12-01

    Full Text Available INTRODUÇÃO: A infecção por Leishmania chagasi é a apresentação clínica mais comum de laishmaniose visceral em áreas endêmicas. O município de Raposa é área endêmica no Estado do Maranhão, tendo registrado casos da doença. Por isso, realizou-se um estudo transversal, com o objetivo de estudar os fato [...] res de risco para infecção por L. chagasi detectada pelo teste intradérmico de Montenegro. MÉTODOS: O estudo envolveu 96% dos moradores das localidades de Maresia, Pantoja e Marisol do município da Raposa, totalizando 1.359 indivíduos. O levantamento dos dados foi realizado utilizando um questionário. Para verificar a associação entre as variáveis estudadas e a infecção por L. chagasi, foram utilizados os modelos de regressão logística uni e multivariada. RESULTADOS: Na análise não ajustada, as variáveis associadas à infecção foram: cobertura da casa de palha, paredes de taipa, piso de chão batido, a presença de flebotomíneos dentro ou fora do domicílio e o local do banho fora de casa. Na análise ajustada, a presença de flebotomíneos dentro ou fora do domicílio foi considerada fator de risco e a idade menor que 10 anos revelou-se como fator de proteção para a infecção assintomática. CONCLUSÕES: Evidenciou-se também, o quanto a precariedade das condições de vida da população contribui para o fortalecimento da cadeia epidemiológica da doença. Abstract in english INTRODUCTION: Infection with Leishmania chagasi is the most common clinical presentation for visceral leishmaniaisis in endemic areas. The municipality of Raposa is an endemic area in State of Maranhão, Brazil, and have had registration cases of visceral leishmaniasis disease. For this reason, a cro [...] ss- sectional study was conducted to evaluate the risk factors for infection with L. chagasi detected by Montenegro skin test. METHODS: The sample comprised 96% of the inhabitants of the villages of Maresia, Pantoja, and Marisol located in the municipality of Raposa, corresponding to 1,359 subjects. Data were collected using a questionnaire. Univariate and multivariate logistic regression models were applied to evaluate the association between the variables studied and infection of L. chagasi. RESULTS: The variables associated with infection upon nonadjusted analysis were a straw roof, mud walls, floors of beaten earth, presence of sand flies inside or outside of the dwelling, and bathing outdoors. Adjusted analysis showed that the presence of sand flies inside/outside the dwelling was a risk factor, and age younger than 10 years was a protective factor against asymptomatic infection. CONCLUSIONS: The results highlight the extent to which precarious living conditions of the population strengthen the epidemiological chain of visceral leishmaniasis.

  11. Risk factors for Leishmania chagasi infection in an endemic area in Raposa, State of Maranhão, Brazil Fatores associados à infecção por Leishmania chagasi em uma área endêmica em Raposa, Estado do Maranhão, Brasil

    Directory of Open Access Journals (Sweden)

    Charlene Barreto Ponte

    2011-12-01

    Full Text Available INTRODUCTION: Infection with Leishmania chagasi is the most common clinical presentation for visceral leishmaniaisis in endemic areas. The municipality of Raposa is an endemic area in State of Maranhão, Brazil, and have had registration cases of visceral leishmaniasis disease. For this reason, a cross- sectional study was conducted to evaluate the risk factors for infection with L. chagasi detected by Montenegro skin test. METHODS: The sample comprised 96% of the inhabitants of the villages of Maresia, Pantoja, and Marisol located in the municipality of Raposa, corresponding to 1,359 subjects. Data were collected using a questionnaire. Univariate and multivariate logistic regression models were applied to evaluate the association between the variables studied and infection of L. chagasi. RESULTS: The variables associated with infection upon nonadjusted analysis were a straw roof, mud walls, floors of beaten earth, presence of sand flies inside or outside of the dwelling, and bathing outdoors. Adjusted analysis showed that the presence of sand flies inside/outside the dwelling was a risk factor, and age younger than 10 years was a protective factor against asymptomatic infection. CONCLUSIONS: The results highlight the extent to which precarious living conditions of the population strengthen the epidemiological chain of visceral leishmaniasis.INTRODUÇÃO: A infecção por Leishmania chagasi é a apresentação clínica mais comum de laishmaniose visceral em áreas endêmicas. O município de Raposa é área endêmica no Estado do Maranhão, tendo registrado casos da doença. Por isso, realizou-se um estudo transversal, com o objetivo de estudar os fatores de risco para infecção por L. chagasi detectada pelo teste intradérmico de Montenegro. MÉTODOS: O estudo envolveu 96% dos moradores das localidades de Maresia, Pantoja e Marisol do município da Raposa, totalizando 1.359 indivíduos. O levantamento dos dados foi realizado utilizando um questionário. Para verificar a associação entre as variáveis estudadas e a infecção por L. chagasi, foram utilizados os modelos de regressão logística uni e multivariada. RESULTADOS: Na análise não ajustada, as variáveis associadas à infecção foram: cobertura da casa de palha, paredes de taipa, piso de chão batido, a presença de flebotomíneos dentro ou fora do domicílio e o local do banho fora de casa. Na análise ajustada, a presença de flebotomíneos dentro ou fora do domicílio foi considerada fator de risco e a idade menor que 10 anos revelou-se como fator de proteção para a infecção assintomática. CONCLUSÕES: Evidenciou-se também, o quanto a precariedade das condições de vida da população contribui para o fortalecimento da cadeia epidemiológica da doença.

  12. Compatibilidade de Heterorhabditis amazonensis (Rhabditida: Heterorhabditidae) isolado RSC-5 com diferentes carrapaticidas utilizados no controle de Rhipicephalus microplus (Acari: Ixodidae) / Compatibility of Heterorhabditis amazonensis (Rhabditida: Heterorhabditidae) strain RSC-5 with different acaricides used for the control of Rhipicephalus microplus (Acari: Ixodidae)

    Scientific Electronic Library Online (English)

    Caio Márcio de Oliveira, Monteiro; Renata da Silva, Matos; Márcia Cristina de Azevedo, Prata; Elder Simões, Batista; Wendell Marcelo de Souza, Perinotto; Vânia Rita Elias Pinheiro, Bittencourt; John, Furlong; Vanessa Andaló Mendes de, Carvalho.

    2014-03-01

    Full Text Available O estudo teve como objetivo avaliar a viabilidade de Heterorhabditis amazonensis isolado RSC-5 após exposição a diferentes carrapaticidas utilizados no controle de Rhipicephalus microplus. Foram constituídos seis tratamentos, cada um composto por um produto, sendo cada grupo com 75.000 nematoides (N [...] EPs) em suspensão de 20 mL de solução de diferentes carrapaticidas, em concentração comercial. O controle foi formado por 75.000 NEPs e 20 mL de água destilada, e todos os grupos foram mantidos em câmara climatizada a 25°C. A avaliação do percentual de sobrevivência e infectividade em lagartas Galleria mellonella foi realizada 24 e 72 horas após o início do experimento. A mortalidade de lagartas no teste de infectividade foi analisada após 72 e 120 horas. Com 24 horas de exposição, o percentual de sobrevivência de H. amazonensis RSC-5 não foi significativamente reduzido (p > 0,05) somente na exposição ao princípio ativo deltametrina. O mesmo foi observado no período de 72 horas em relação à associação clorpirifós + cipermetrina + butóxido de piperonila + citronelal. Não foi constatada sobrevivência de nenhum juvenil infectivo nos grupos expostos à associação clorfenvinfós + diclorvós. A exposição ao clorfenvinfós resultou em percentual de sobrevivência inferior a 50% após 72 horas. O potencial de infectar lagartas de G. mellonella foi reduzido apenas no grupo tratado com o princípio ativo clorfenvinfós. Dessa forma, é possível concluir que o princípio ativo clorfenvinfós e a associação clorfenvinfós + diclorvós não foram compatíveis com H. Amazonenses RSC-5, causando redução no percentual de sobrevivência e infectividade dos juvenis desse nematoide. Os outros produtos foram compatíveis, não causando redução na infectividade do isolado testado. Abstract in english The aim of this study was to assess the viability of Heterorhabditis amazonensis strain RSC-5 after exposure to different acaricides used for Rhipicephalus microplus control. Six treatment groups were formed, one for each product. Each group was composed of 75,000 nematodes in a 20 mL solution of di [...] fferent acaricides, at commercial concentration. The control group was formed by the same number of nematodes in 20 mL of distilled water. All the groups were kept in a climate-controlled chamber at 25°C. The percentage of survival and infectivity in Galleria mellonella caterpillars were determined 24 and 72 hours after the beginning of the experiment. The mortality of the caterpillars in the infectivity test was assessed 72 and 120 hours. After 24 hours of exposure, only the active ingredient deltamethrin did not significantly reduce the survival percentage of H. amazonensis RSC-5 (p > 0.05). The same was observed after 72 hours of exposure to the combination of chlorpyriphos + cypermethrin + piperonyl butoxide + citronellal. There was no survival of infective juveniles in the groups exposed to the combination of chlorphenvinphos + dichlorvos. The exposure to chlorphenvinphos for 72 hours resulted in 50% of mortality. The potential to infect G. mellonella caterpillars was only reduced in the group treated with the active ingredient chlorphenvinphos. Chlorphenvinphos and the combination of chlorphenvinphos + dichlorvos were not compatible with H. amazonensis RSC-5, causing a reduction in the survival and infectivity of juveniles of this nematode, while the other products were compatible, causing no reduction in the infectivity of this isolate.

  13. Protein kinase A signaling during bidirectional axenic differentiation in Leishmania.

    Science.gov (United States)

    Bachmaier, Sabine; Witztum, Ronit; Tsigankov, Polina; Koren, Roni; Boshart, Michael; Zilberstein, Dan

    2016-02-01

    Parasitic protozoa of the genus Leishmania are obligatory intracellular parasites that cycle between the phagolysosome of mammalian macrophages, where they proliferate as intracellular amastigotes, and the midgut of female sand flies, where they proliferate as extracellular promastigotes. Shifting between the two environments induces signaling pathway-mediated developmental processes that enable adaptation to both host and vector. Developmentally regulated expression and phosphorylation of protein kinase A subunits in Leishmania and in Trypanosoma brucei point to an involvement of protein kinase A in parasite development. To assess this hypothesis in Leishmania donovani, we determined proteome-wide changes in phosphorylation of the conserved protein kinase A phosphorylation motifs RXXS and RXXT, using a phospho-specific antibody. Rapid dephosphorylation of these motifs was observed upon initiation of promastigote to amastigote differentiation in culture. No phosphorylated sites were detected in axenic amastigotes. To analyse the kinetics of (re)phosphorylation during axenic reverse differentiation from L. donovani amastigotes to promastigotes, we first established a map of this process with morphological and molecular markers. Upon initiation, the parasites rested for 6-12h before proliferation of an asynchronous population resumed. After early changes in cell shape, the major changes in molecular marker expression and flagella biogenesis occurred between 24 and 33h after initiation. RXXS/T re-phosphorylation and expression of the regulatory subunit PKAR1 correlated with promastigote maturation, indicating a promastigote-specific function of protein kinase A signaling. This is supported by the localization of PKAR1 to the flagellum, an organelle reduced to a remnant in amastigote forms. We conclude that a significant increase in protein kinase A-mediated phosphorylation is part of the ordered changes that characterise the amastigote to promastigote differentiation. PMID:26460237

  14. pH homeostasis in Leishmania donovani amastigotes and promastigotes.

    OpenAIRE

    Glaser, T A; Baatz, J E; Kreishman, G P; Mukkada, A J

    1988-01-01

    Intracellular pH and pH gradients of Leishmania donovani amastigotes and promastigotes were determined over a broad range of extracellular pH values. Intracellular pH was determined by 31P NMR and by equilibrium distribution studies with 5,5-dimethyloxazolidine-2,4-dione or methylamine. Promastigotes maintain intracellular pH values close to neutral between extracellular pH values of 5.0 and 7.4. Amastigote intracellular pH is maintained close to neutral at external pH values as low as 4.0. B...

  15. Genetic causes involved in Leishmania Chagasi infection innortheastern: Brazil

    Directory of Open Access Journals (Sweden)

    Feitosa Mary Furlan

    1999-01-01

    Full Text Available A sample of 502 individuals from 94 families from Jacobina, State of Bahia, Brazil, was investigated to determine the causal mechanisms involved in Leishmania chagasi (the causal agent of visceral leishmaniasis in the American hemisphere infection, as measured by the intradermic reaction to antigens derived from this parasite, using complex segregation analyses. The results showed evidence of a major genetic mechanism acting on infection, with a frequency of a recessive (or additive susceptibility gene (q of approximately 0.45. A small multifactorial component (H = 0.29 acting in conjunction with a major recessive gene (q = 0.37 is not ruled out as a concomitant causative factor.

  16. Development of Cutaneous Leishmaniasis after Leishmania Skin Test

    OpenAIRE

    Dantas, Marina L.; Sérgio Arruda; Machado, Gustavo U.; de Carvalho, Augusto M.; Paulo R Machado

    2011-01-01

    Thirty-year-old female with a previous history of a cutaneous ulcer suspicious of leishmaniasis 20 years ago presented with a new complaint of a depressed papular lesion 8 × 7?mm in the right lower extremity. The lesion was of 10-day duration. Because early cutaneous leishmaniasis (CL) lesions may have a non-ulcerated appearance, a Leishmania skin test (LST) was performed on the forearm with a strong positive result (38 × 32?mm). After 8 days, the lesion in the leg, which was diag...

  17. Detección por inmunohistoquímica de Leishmania infantum en hámster infectado experimentalmente Detection of Leishmania infantum in an experimentally-infected hamster using immunohistochemistry

    Directory of Open Access Journals (Sweden)

    Lisset Fonseca Géigel

    2011-12-01

    Full Text Available Introducción: la leishmaniasis visceral se considera la forma más severa de las leishmaniasis y puede llegar a ser fatal en ausencia de tratamiento oportuno. En América Latina la infección es causada por Leishmania infantum (syn. Leishmania chagasi. El diagnóstico inequívoco y la selección de un adecuado modelo experimental son una necesidad para emprender estudios con este agente biológico. Objetivo: determinar la utilidad de la técnica de inmunohistoquímica en la identificación de Leishmania. Métodos: los animales se inocularon con promastigotes de Leishmania infantum. Se monitoreó el peso corporal de cada animal y se determinó el peso relativo de bazos e hígados. Para la identificación de amastigotes se prepararon improntas coloreadas con Giemsa y se desarrolló un protocolo de inmunohistoquímica en tejido incluido en parafina. Resultados: se reprodujo la infección en el modelo experimental. La técnica de inmunohistoquímica fue positiva en los cortes de los animales infectados y no reactiva para el grupo control. Al comparar con la tinción mediante Giemsa, esta metodología facilitó la identificación, particularmente en órganos con escasos parásitos. Conclusiones: la inmunohistoquímica es una herramienta específica para la detección de Leishmania, facilita la observación y evita confusiones en la identificación del parásito, lo que mejora la calidad del diagnóstico.Introduction: visceral leishmaniasis is considered the most severe form of this disease and can be fatal if not properly treated. In Latin America, the infection is caused by Leishmania infantum (syn. Leishmania chagasi. The unequivocal diagnosis and the selection of a suitable experimental model are required to undertake studies on this biologic agent. Objective: to determine the advantages of immunohistochemistry in identifying Leishmania. Methods: hamsters were inoculated with Leishmania infantum promastigotes. The body weights of every animal were monitored, and the relative weights of their spleens and livers were estimated. For identification of amastigotes, Giemsa-stained imprints and an immunohistochemistry protocol in paraffin-embedded tissues were developed. Results: the infection was reproduced in the experimental model. The immunohistochemistry was positive in infected animal sections and non-reactive for the control group. When compared with the Giemsa staining, this methodology facilitated the identification, particularly in organs infected with few parasites. Conclusions: immunohistochemistry is a specific tool for detection of Leishmania since it facilitates observation and eliminates any confusion in the identification of the parasite, thus improving the quality of diagnosis.

  18. Detection of Leishmania major In Naturally Infected Sand Flies Using Semi Nested-PCR

    Directory of Open Access Journals (Sweden)

    MA Oshaghi

    2008-12-01

    Full Text Available "nBackground: The aim of this study was to assess Leishmania infection in sand fly species from areas where leishmaniasis is endemic. This is important for prediction of the risk and expansion of the disease."nMethods: In this cross-sectional study we used a PCR-based method for detection of Leishmania minicircle DNA within individual sand flies from Orzoieh, a new endemic leishmaniasis focus in southern Iran."nResults: We detected minicircle DNA in 6 of 92 (6.5% Phlebotomus (Phlebotomus papatasi collected indoor, while all of previous microscopic examination of sand flies specimens was negative for Leishmania promastigotes in the region. The species were identified as Leishmania (Leishmania major by comparison of PCR products with a L. major positive control. All the Leishmania-positive sand flies were confirmed as P. (P. papatasi by using a morphological key of Iranian sand flies."nConclusion: Since PCR method is relatively easy and can process a large number of samples, it will be a powerful tool for the rapid identification of Leishmania species as well as monitoring the infection rate in sand fly populations in areas of low endemicity of leishmaniasis.

  19. First molecular detection of Leishmania tarentolae-like DNA in Sergentomyia minuta in Spain.

    Science.gov (United States)

    Bravo-Barriga, Daniel; Parreira, Ricardo; Maia, Carla; Blanco-Ciudad, Juan; Afonso, Maria Odete; Frontera, Eva; Campino, Lenea; Pérez-Martín, Juan Enrique; Serrano Aguilera, Francisco Javier; Reina, David

    2016-03-01

    Phlebotomine sand flies (Diptera, Psychodidae) are vectors of multiple Leishmania species, among which Leishmania infantum stands out as a being frequently pathogenic to humans and dogs in Mediterranean countries. In this study, Sergentomyia minuta sand flies were collected using CDC miniature light traps in different 431 biotopes from Southwest Spain. A total of 114 females were tested for the presence of Leishmania DNA by targeting ITS-1 and cyt-B sequences by PCR. Leishmania DNA was detected in one S. minuta. Characterization of the obtained DNA sequences by phylogenetic analyses revealed close relatedness with Leishmania tarentolae Wenyon, 1921 as well as with both human and canine pathogenic strains of Asian origin (China), previously described as Leishmania sp. To our knowledge, this is the first report of phlebotomine sand flies naturally infected with L. tarentolae-like in Spain. The possible infection of sand flies with novel Leishmania species should be taken into consideration in epidemiological studies of vector species in areas where leishmaniosis is endemic. PMID:26691858

  20. Presence of Leishmania amastigotes in peritoneal fluid of a dog with leishmaniasis from Alagoas, Northeast Brazil Presença de formas amastigotas de Leishmania em fluido peritoneal de cão com leishmaniose proveniente de Alagoas, nordeste do Brasil

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    Filipe Dantas-Torres

    2006-08-01

    Full Text Available The goal of this short communication is to report the uncommon presence of intracellular amastigotes of Leishmania in peritoneal fluid of a dog with leishmaniasis from Alagoas State, Brazil. Physical examination of an adult male rottweiler suspected to be suffering of leishmaniasis revealed severe loss of weight, ascitis, splenomegaly, moderately enlarged lymph nodes, onychogryphosis, generalized alopecia, skin ulcers on the posterior limbs, and conjunctivitis. Samples of bone marrow, popliteal lymph node, skin ulcer, and peritoneal fluid were collected and smears of each sample were prepared and stained with hematoxylin and eosin. Numerous amastigotes were detected in bone marrow, popliteal lymph node, and skin ulcer smears. Smears of peritoneal fluid revealed the unusual presence of several free and intracellular amastigotes of Leishmania. Future studies are needed to determine whether the cytology of ascitic fluid represents a useful tool for diagnosis Leishmania infection in ascitic dogs, particularly in those living in areas where canine leishmaniasis is enzootic.O objetivo desta comunicação é descrever a presença incomum de formas amastigotas de Leishmania em fluido peritoneal de um cão com leishmaniose proveniente do Estado de Alagoas, nordeste do Brasil. O exame físico de um cão macho adulto da raça rottweiler, apresentando suspeita de leishmaniose, revelou perda de peso severa, esplenomegalia, linfonodos moderadamente aumentados, ascite, onicogrifose, alopecia generalizada, conjuntivite e presença de lesões cutâneas ulceradas localizadas nos membros posteriores. Foram coletadas amostras de medula óssea, linfonodo poplíteo, fluido peritoneal e úlcera cutânea. A partir das amostras, foram elaborados esfregaços, os quais foram corados pela hematoxilina e eosina. Inúmeras formas amastigotas foram detectadas na medula óssea, linfonodo poplíteo e úlcera cutânea. Esfregaços de fluido peritoneal revelaram a presença, não usual, de várias formas amastigotas livres e intracelulares. Futuros estudos serão necessários a fim de determinar se a citologia de líquido ascítico representa uma ferramenta útil para o diagnóstico da infecção por Leishmania em cães com ascite, particularmente naqueles que vivem em áreas onde a leishmaniose canina é enzoótica.

  1. Proteomic analyses of membrane enriched proteins of Leishmania donovani Indian clinical isolate by mass spectrometry.

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    Kumar, Awanish; Misra, Pragya; Sisodia, Brijesh; Shasany, Ajit Kumar; Sundar, Shyam; Dube, Anuradha

    2015-08-01

    Visceral leishmaniasis (VL) is a major fatal disease prevalent in North-East India, caused by a protozoan parasite Leishmania donovani. The parasite multiplies and thrives inside mammalian macrophages and is transmitted by the bite of the sandfly. Due to the unsatisfactory treatment measures, increasing drug resistance and the advent of HIV-Leishmania co-infection there has been an urgent need to develop novel drug/vaccine targets against VL. Target identification is the key step in drug discovery and proteomics seems to be a suitable strategy for it due to the availability of Leishmania major, Leishmania infantum, Leishmania braziliensis, Leishmania donovani, Leishmania mexicana and Leishmania tarentolae genome sequence. Since, majority of proteome analyses of Leishmania have, so far, been performed on whole-cell extracts; this study is dealing with the sub-proteome analysis of the membrane-enriched protein (MEP) fractions of L. donovani. The analysis of 95 protein spots of the MEPs from two dimensional (2-D) gel image through matrix asserted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) endorsed the identification of various relevant functional proteins. Out of 95 the MEP spots 72 have been identified and were classified on the basis of their biological function. Several proteins of unknown function that belong to different classes like cell signaling, transmembrane receptors, and transporters have been identified which could be the new potential therapeutic targets against VL in future. The proteome array of the MEPs contributes to further elucidation of the biological system of L. donovani as well as host-parasite relationships which may be further investigated for their crucial biological role in L. donovani for disease management. PMID:25597695

  2. Complete conservation of an immunogenic gene (lcr1 in Leishmania infantum and Leishmania chagasi isolated from Iran, Spain and Brazi

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    H. Mahmoudzadeh-Niknam , F. Abrishami , M. Doroudian , M. Moradi , M.H. Alimohammadian , P. Parvizi

    2010-12-01

    Full Text Available Background & objectives: Kala-azar is the visceral and most severe form of leishmaniasis thatleads to death if untreated. The causative agents of visceral leishmaniasis (VL are members ofLeishmania (L. donovani complex which includes L. chagasi and L. infantum. Genome sequenceshave raised the question whether L. chagasi and L. infantum are synonymous or different. Thisquestion has important implications for clinical and epidemiological studies, evaluation of vaccinesand drugs, and disease control. LCR1 is an immunogenic molecule discovered from L. chagasiwith potential as a component of a Leishmania subunit vaccine. If this protein has potentials forbeing used in a vaccine or diagnostic testing, there should be little variability in this moleculebetween L. infantum isolates from diverse geographic regions. The aim of this study was to determinewhether lcr1 of an Iranian strain of L. infantum was identical to lcr1 of both L. infantum strainfrom a different geographic region (Spain and that of an L. chagasi isolate from Brazil.Methods: L. infantum isolated from an Iranian kala-azar patient was studied. Lcr1 from this isolatewas PCR amplified, cloned, and studied by restriction digest analysis and sequencing.Results: The sequences of lcr1 of the Iranian L. infantum were completely identical at nucleotidelevel to lcr1 sequences of both the Spanish L. infantum and the Brazilian L. chagasi strains.Conclusion: Complete conservation of the DNA sequence encoding for LCR1 molecule betweengeographically distinct Leishmania species adds credibility to the potential for LCR1 as a componentof a subunit vaccine and diagnostic test for kala-azar.

  3. The sera from adult patients with suggestive signs of autoimmune diseases present antinuclear autoantibodies that cross-react with Leishmania infantum conserved proteins: crude Leishmania histone and Soluble Leishmania antigens [corrected].

    Science.gov (United States)

    Lakhal, Sami; Benabid, Meriem; Sghaier, Ines Ben; Bettaieb, Jihen; Bouratbine, Aïda; Galai, Yousr

    2015-02-01

    Visceral leishmaniasis has been associated with hyper-gammaglobulinemia and antinuclear antibodies and may simulate systemic lupus erythematosus. Sera from patients with visceral leishmaniasis have been shown to strongly react against conserved proteins from the parasite, such as ribosomal and histones. Some of these proteins have also been described as immunogenic in several auto-immune syndromes, and the detection of antibodies against them is considered to be indicative of disorder in the immune system. This study aimed to assess by enzyme-linked immunosorbent assay, test routinely employed in visceral leishmaniasis diagnosis, the recognition of Crude Leishmania histone and Soluble Leishmania antigens proteins from Leishmania infantum by adult patients with suggestive signs of autoimmune diseases. Our results show that the humoral response generated during autoimmune diseases cross-reacts with the parasitic Crude Leishmania histone and Soluble Leishmania antigens. In these cases, higher precautions must be taken to confirm the presence of visceral leishmaniasis in front of positive serology in antinuclear antibodies positive sera, in order to avoid wrong diagnosis. PMID:25395341

  4. Diagnóstico molecular e frequência de anticorpos anti-Leishmania infantum chagasi em cães do município de Belém, Pará / Molecular diagnosis and frequency of anti-Leishmania infantum chagasi antibodies in dogs in Belém/Pará, Brazil

    Scientific Electronic Library Online (English)

    Katiane, Schwanke; Aryane M.M. da, Silva; Adlilton, Pacheco; Michele, Bahia; Fernando T., Silveira; Alessandra, Scofield; Gustavo, Góes-Cavalcante.

    2014-03-01

    Full Text Available A leishmaniose visceral é uma enfermidade cujo agente etiológico no Brasil é o protozoário Leishmania infantum chagasi. Os cães são considerados reservatórios urbanos da doença, sendo indicadores da ocorrência de casos humanos. O presente trabalho teve como objetivo diagnosticar a infecção por L. in [...] fantum chagasi em cães domiciliados e errantes do município de Belém, estado do Pará, através da reação em cadeia da polimerase (PCR) e da reação de imunofluorescência indireta (RIFI), empregando dois antígenos distintos. Amostras de sangue venoso de cães adultos, sem distinção de sexo ou raça, de diferentes bairros e épocas do ano da cidade de Belém-PA, foram colhidas em tubos sem e com anticoagulante para obtenção do soro e do DNA, respectivamente. Esses animais foram divididos em dois grupos: cães errantes capturados pelo Centro de Controle de Zoonoses (Grupo A) e cães domiciliados (Grupo B). Os soros foram analisados através do teste de RIFI para pesquisa de IgG utilizando-se dois antígenos distintos: 1) antígeno do kit Bio-Manguinhos/FIOCRUZ (Ag-PRO) contendo formas promastigotas de Leishmania sp. (complexo major-like); 2) Antígeno do Instituto Evandro Chagas (Ag-AMA) constituído por formas amastigotas de L. infantum chagasi. A avaliação dos dois antígenos foi realizada com as amostras reagentes a partir da titulação 1:80. Já a PCR foi realizada a partir do DNA extraído do sangue total dos animais e amplificado utilizando-se os iniciadores RV1e RV2. Das 335 amostras analisadas, 10,4% (35/335) foram reagentes na RIFI (Ag-PRO) e 0,9% (3/335) reagiram com o Ag-AMA. A distribuição das amostras positivas se deu da seguinte forma: Grupo A 14,8% (25/169) com Ag-PRO e 1,2% (2/169) com Ag-AMA; Grupo B 6% (10/166) com Ag-PRO e 0,6% (1/166) com Ag-AMA; sendo que todas as amostras positivas pelo teste de RIFI com o Ag-AMA também reagiram com o Ag-PRO e em nenhuma das amostras foi detectado o DNA de L. infantum chagasi. Os achados do presente estudo indicam que Belém ainda pode ser considerada área não endêmica para leishmaniose visceral canina e que a natureza do antígeno influencia no resultado da RIFI para a pesquisa de anticorpos anti-L. infantum chagasi em cães, sendo que a RIFI que utiliza formas promastigotas de Leishmania major-like como antígeno deve ser utilizada com cautela como método diagnóstico confirmatório em estudos epidemiológicos em áreas não endêmicas para LVC. Abstract in english Visceral leishmaniasis is a disease whose etiological agent in Brazil is Leishmania infantum chagasi. Dogs are considered urban reservoirs of the disease, being an indicator of the human cases occurrence. The present study aimed to diagnose L. infantum chagasi infection in stray and owned dogs in Be [...] lém, Pará State, by polymerase chain reaction (PCR) and indirect immunofluorescence assay (IFA) using two different antigens. Venous blood samples from adult dogs, regardless of gender or breed, of different neighborhoods in Belém-PA, were collected in tubes with and without anticoagulant to obtain DNA and serum, respectively. These animals were divided into two groups: stray dogs captured by the Center for Zoonosis Control (Group A) and owned dogs (Group B). Sera were analyzed by IFA testing for IgG using two different antigens: 1) Bio-Manguinhos/Fiocruz antigen kit (Ag-PRO) containing promastigotes of Leishmania sp. (Complex Major-Like), 2) Instituto Evandro Chagas Antigen (Ag-AMA) consisting of amastigotes of L. infantum chagasi. The evaluation of the two antigens was performed considering positive the reactions above the 1:80 dilution. Already PCR was performed with DNA isolated from whole blood of animals and amplified with the primers RV1 and RV2. Of the 335 samples analyzed, 10.4% (35/335) were positive by IFA (Ag-PRO) and 0.9% (3/335) with the Ag-AMA. The distribution of positive samples is given as follows: Group A 14.8% (25/169) with Ag-PRO and 1.2% (2/169) with Ag-AMA; Group B 6% (10/166) with Ag-PRO and 0.6% (1/166) with Ag-AMA, being t

  5. Characterization of in Vitro Cultivated Amastigote like of Leishmania major: A Substitution for in Vivo Studies

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    S Kazemian

    2008-04-01

    Full Text Available Background: Promastigotes of Leishmania spp. have been readily cultured, but the axenic culture of amastigotes has been successful in L. donovani, L .infantum L. mexicana and L. pifanoi. However, some species such as L. major, is much less amenable to axenic cultivation. In present study, we describe an in vitro culture system for the generation and propagation of axenic amastigotes form of L. major.Methods: Promastigotes of L. major were cultivated in a biphasic NNN medium. The liquid phase was Schneider's medium, pH 3.5, supplemented by 25% FCS (fetal calf serum. The cultures were maintained at 33-34°C for 120 hours. Results: Fine structure analysis of these in vitro-grown amastigotes by electron microscopy, demonstrated that they have a pear-shaped body with abortive short terminal flagellum. The in vitro-grown cells are agglutinated by peanut lectin. SDS-PAGE pattern of these axenic amastigotes showed a 66-kDa band, which was not present in promastigotes. The axenic grown amastigotes were able to infect peritoneum macrophages of BALB/c mice. In supernatant of culture, biochemical, analysis showed decreased protein and acid phosphate activity. Conclusion: These amastigotes like cells might serve as a suitable strain for the study of amastigote biochemistry, in vitro drug testing, and immunology of L. major.

  6. Cloning and expression of codon-optimized recombinant darbepoetin alfa in Leishmania tarentolae T7-TR.

    Science.gov (United States)

    Kianmehr, Anvarsadat; Golavar, Raziyeh; Rouintan, Mandana; Mahrooz, Abdolkarim; Fard-Esfahani, Pezhman; Oladnabi, Morteza; Khajeniazi, Safoura; Mostafavi, Seyede Samaneh; Omidinia, Eskandar

    2016-02-01

    Darbepoetin alfa is an engineered and hyperglycosylated analog of recombinant human erythropoietin (EPO) which is used as a drug in treating anemia in patients with chronic kidney failure and cancer. This study desribes the secretory expression of a codon-optimized recombinant form of darbepoetin alfa in Leishmania tarentolae T7-TR. Synthetic codon-optimized gene was amplified by PCR and cloned into the pLEXSY-I-blecherry3 vector. The resultant expression vector, pLEXSYDarbo, was purified, digested, and electroporated into the L. tarentolae. Expression of recombinant darbepoetin alfa was evaluated by ELISA, reverse-transcription PCR (RT-PCR), Western blotting, and biological activity. After codon optimization, codon adaptation index (CAI) of the gene raised from 0.50 to 0.99 and its GC% content changed from 56% to 58%. Expression analysis confirmed the presence of a protein band at 40 kDa. Furthermore, reticulocyte experiment results revealed that the activity of expressed darbepoetin alfa was similar to that of its equivalent expressed in Chinese hamster ovary (CHO) cells. These data suggested that the codon optimization and expression in L. tarentolae host provided an efficient approach for high level expression of darbepoetin alfa. PMID:26546410

  7. Elevated ergosterol protects Leishmania parasites against antimony-generated stress.

    Science.gov (United States)

    Mathur, Radhika; Das, Rajeev Patrick; Ranjan, Archana; Shaha, Chandrima

    2015-10-01

    Parasite lipids can serve as signaling molecules, important membrane components, energy suppliers, and pathogenesis factors critical for survival. Functional roles of lipid changes in response to drug-generated stress in parasite survival remains unclear. To investigate this, Leishmania donovani parasites, the causative agents of kala-azar, were exposed to the antileishmanial agent potassium antimony tartrate (PAT) (half-maximal inhibitory concentration ? 284 µg/ml). Analysis of cell extracts using gas chromatography-mass spectrometry showed significant increases in very long-chain fatty acids (VLCFAs) prior to an increase in ergosterol in PAT-treated parasites as compared with vehicle-treated controls. Ergosterol biosynthesis inhibition during PAT treatment decreased cell viability. VLCFA inhibition with specific inhibitors completely abrogated ergosterol upsurge followed by a reduction in cell viability. Following PAT-induced VLCFA increase, an upsurge in reactive oxygen species (ROS) occurred and inhibition of this ROS with antioxidants abrogated ergosterol increase. Genetically engineered parasites expressing low constitutive ergosterol levels showed more susceptibility to PAT as compared with wild-type control cells but ergosterol supplementation during PAT treatment increased cell viability. In conclusion, we propose that during antimony treatment, the susceptibility of parasites is determined by the levels of cellular ergosterol that are regulated by oxidative stress generated by VLCFAs.-Mathur, R., Das, R. P., Ranjan, A., Shaha, C. Elevated ergosterol protects Leishmania parasites against antimony-generated stress. PMID:26116701

  8. Genetic diversity of Leishmania infantum field populations from Brazil

    Scientific Electronic Library Online (English)

    Marcela, Segatto; Lucas Secchim, Ribeiro; Dorcas Lamounier, Costa; Carlos Henrique Nery, Costa; Márcia Rosa de, Oliveira; Sílvio Fernando Guimarães, Carvalho; Andréa Mara, Macedo; Helder Magno Silva, Valadares; Reynaldo, Dietze; Cristiana Ferreira Alves de, Brito; Elenice Moreira, Lemos.

    2012-02-01

    Full Text Available Leishmania infantum (syn. Leishmania chagasi) is the etiological agent of visceral leishmaniasis (VL) in Brazil. The epidemiology of VL is poorly understood. Therefore, a more detailed molecular characterization at an intraspecific level is certainly needed. Herein, three independent molecular metho [...] ds, multilocus microsatellite typing (MLMT), random amplification of polymorphic DNA (RAPD) and simple sequence repeats-polymerase chain reaction (SSR-PCR), were used to evaluate the genetic diversity of 53 L. infantum isolates from five different endemic areas in Brazil. Population structures were inferred by distance-based and Bayesian-based approaches. Eighteen very similar genotypes were detected by MLMT, most of them differed in only one locus and no correlation was found between MLMT profiles, geographical origin or the estimated population structure. However, complex profiles composed of 182 bands obtained by both RAPD and SSR-PCR assays gave different results. Unweighted pair group method with arithmetic mean trees built from these data revealed a high degree of homogeneity within isolates of L. infantum. Interestingly, despite this genetic homogeneity, most of the isolates clustered according to their geographical origin.

  9. Apoptotic mimicry: an altruistic behavior in host/Leishmania interplay

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    Wanderley J.L.M.

    2005-01-01

    Full Text Available Apoptosis is the most common phenotype observed when cells die through programmed cell death. The morphologic and biochemical changes that characterize apoptotic cells depend on the activation of a diverse set of genes. Apoptosis is essential for multicellular organisms since their development and homeostasis are dependent on extensive cell renewal. In fact, there is strong evidence for the correlation between the emergence of multicellular organisms and apoptosis during evolution. On the other hand, no obvious advantages can be envisaged for unicellular organisms to carry the complex machinery required for programmed cell death. However, accumulating evidence shows that free-living and parasitic protozoa as well as yeasts display apoptotic markers. This phenomenon has been related to altruistic behavior, when a subpopulation of protozoa or yeasts dies by apoptosis, with clear benefits for the entire population. Recently, phosphatidylserine (PS exposure and its recognition by a specific receptor (PSR were implicated in the infectivity of amastigote forms of Leishmania, an obligatory vertebrate intramacrophagic parasite, showing for the first time that unicellular organisms use apoptotic features for the establishment and/or maintenance of infection. Here we focus on PS exposure in the outer leaflet of the plasma membrane - an early hallmark of apoptosis - and how it modulates the inflammatory activity of phagocytic cells. We also discuss the possible mechanisms by which PS exposure can define Leishmania survival inside host cells and the evolutionary implications of apoptosis at the unicellular level.

  10. Gene Cloning of Iranian Leishmania major Mannose-1-Phosphate Guanyltransferase

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    R Salehi

    2009-07-01

    Full Text Available "nBackground: Leishmania is an obligatory intracellular protozoan parasite, which infects human be­ings when infected sand fly vector takes a blood meal.  Most efforts are towards designing an effective vaccine to prevent leishmaniasis. In this way, development of candidate antigen for vaccine has spe­cial im­portant. In this study, we cloned mannose-1-phosphate guanyltransferase gene of Iranian L .major in pET32a expression vector. "nMethods: Primers based on L. major mannose-1-phosphate guanyltransferase sequence gene was de­signed and synthesized. DNA of Leishmania promastigotes was extracted and PCR reaction was done. PCR product was cloned into pTZ57R and sub cloned into pET32a expression vector. "nResults: Recombinant plasmid containing 1140 bp as L. major mannose-1-phosphate guanyltrans­ferase gene was extracted and confirmed by restriction analysis. PCR product was sequenced and de­posited to GenBank. There were some differences in amino acid sequences between Iranian L. major mannose-1-phosphate guanyltransferase and others previously accepted in GenBank "nConclusion: We amplified and cloned Iranian L. major mannose-1-phosphate guanyltransferase successfully.

  11. Hemophagocytosis in Experimental Visceral Leishmaniasis by Leishmania donovani.

    Science.gov (United States)

    Morimoto, Ayako; Omachi, Satoko; Osada, Yasutaka; Chambers, James K; Uchida, Kazuyuki; Sanjoba, Chizu; Matsumoto, Yoshitsugu; Goto, Yasuyuki

    2016-03-01

    Hemophagocytosis is a phenomenon in which macrophages phagocytose blood cells. There are reports on up-regulated hemophagocytosis in patients with infectious diseases including typhoid fever, tuberculosis, influenza and visceral leishmaniasis (VL). However, mechanisms of infection-associated hemophagocytosis remained elusive due to a lack of appropriate animal models. Here, we have established a mouse model of VL with hemophagocytosis. At 24 weeks after infection with 1 x 107 Leishmania donovani promastigotes, BALB/cA mice exhibited splenomegaly with an average tissue weight per body weight of 2.96%. In the tissues, 28.6% of macrophages contained phagocytosed erythrocytes. All of the hemophagocytosing macrophages were parasitized by L. donovani, and higher levels of hemophagocytosis was observed in heavily infected cells. Furthermore, more than half of these hemophagocytes had two or more macrophage-derived nuclei, whereas only 15.0% of splenic macrophages were bi- or multi-nuclear. These results suggest that direct infection by L. donovani causes hyper-activation of host macrophages to engulf blood cells. To our knowledge, this is the first report on hemophagocytosis in experimental Leishmania infections and may be useful for further understanding of the pathogenesis. PMID:26942577

  12. Comparison of Leishmania killicki (syn. L. tropica) and Leishmania tropica Population Structure in Maghreb by Microsatellite Typing.

    Science.gov (United States)

    Chaara, Dhekra; Bañuls, Anne-Laure; Haouas, Najoua; Talignani, Loïc; Lami, Patrick; Mezhoud, Habib; Harrat, Zoubir; Dedet, Jean-Pierre; Babba, Hamouda; Pratlong, Francine

    2015-12-01

    Leishmania (L.) killicki (syn. L. tropica), which causes cutaneous leishmaniasis in Maghreb, was recently described in this region and identified as a subpopulation of L. tropica. The present genetic analysis was conducted to explore the spatio-temporal distribution of L. killicki (syn. L. tropica) and its transmission dynamics. To better understand the evolution of this parasite, its population structure was then compared with that of L. tropica populations from Morocco. In total 198 samples including 85 L. killicki (syn. L. tropica) (from Tunisia, Algeria and Libya) and 113 L. tropica specimens (all from Morocco) were tested. Theses samples were composed of 168 Leishmania strains isolated from human skin lesions, 27 DNA samples from human skin lesion biopsies, two DNA samples from Ctenodactylus gundi bone marrow and one DNA sample from a Phlebotomus sergenti female. The sample was analyzed by using MultiLocus Enzyme Electrophoresis (MLEE) and MultiLocus Microsatellite Typing (MLMT) approaches. Analysis of the MLMT data support the hypothesis that L. killicki (syn. L. tropica) belongs to the L. tropica complex, despite its strong genetic differentiation, and that it emerged from this taxon by a founder effect. Moreover, it revealed a strong structuring in L. killicki (syn. L. tropica) between Tunisia and Algeria and within the different Tunisian regions, suggesting low dispersion of L. killicki (syn. L. tropica) in space and time. Comparison of the L. tropica (exclusively from Morocco) and L. killicki (syn. L. tropica) population structures revealed distinct genetic organizations, reflecting different epidemiological cycles. PMID:26645812

  13. Comparison of Leishmania killicki (syn. L. tropica) and Leishmania tropica Population Structure in Maghreb by Microsatellite Typing

    Science.gov (United States)

    Chaara, Dhekra; Bañuls, Anne- Laure; Haouas, Najoua; Talignani, Loïc; Lami, Patrick; Mezhoud, Habib; Harrat, Zoubir; Dedet, Jean-Pierre; Babba, Hamouda; Pratlong, Francine

    2015-01-01

    Leishmania (L.) killicki (syn. L. tropica), which causes cutaneous leishmaniasis in Maghreb, was recently described in this region and identified as a subpopulation of L. tropica. The present genetic analysis was conducted to explore the spatio-temporal distribution of L. killicki (syn. L. tropica) and its transmission dynamics. To better understand the evolution of this parasite, its population structure was then compared with that of L. tropica populations from Morocco. In total 198 samples including 85 L. killicki (syn. L. tropica) (from Tunisia, Algeria and Libya) a