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Sample records for para leishmania spp

  1. Ensayos metodologicos para la investigacion de reservorios de Leishmania spp en los Andes venezolanos Methodological assay for research of reservoirs of Leishmania spp. in the Venezuelan Andes

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    Ana Lugo Yarbuh

    1982-12-01

    Full Text Available Se describen dos técnicas, presuntiva y confirmativa, para la investigación de mamíferos que pudieran ser reservorios de Leishmania que parasitan al hombre. Se investigan los cambios en los títulos de inmovilización y aglutinación de promastigotos de cultivo por los sueros de animales normales y expuestos una o varias veces a la inoculación intradérmica de pequeñas dosis de promastigotos vivos. Se registra una caída de los títulos de aglutinación en los sueros de hamsteres, de Holochilus venezuelae y de Didelphis marsupialis después de la inoculación con L. mexicana mexicana de Panamá y de L. gamhami de la región de los Andes venezolanos. Se discute la natureza de estos fenómenos. Se han hecho xenodiagnósticos con Lutzomyia townsendi en Holochilus venezuelae y Sigmodon hispidus infectados experimentalmente com L. mexicana mexicana, L. mexicana amazonensis, L. braziliensis y L. garnhami. Las pruebas fueron leidas mediante el examen microscópico de las gotitas de heces excretadas entre las 108 y 132 horas después de la ingesta infectante, tras colorearlas con Giemsa. Se obtuvieron resultados positivos en 23% de los experimentos usando mamíferos con lesiones localizadas, dejando a los flebótomos ingurgitarse libremente sobre animales anestesiados que poseian una hasta varias lesiones localizadas.Presumptive and confirmative techniques for searching mammals which could be reservoirs for Leishmania parasites from man are described. The changes of immobilising and agglutinating titers for promastigotes from culture by sera from normal and exposed mammals after single or repeated intradermal inoculation of promastigotes are described. A fall in titers of agglunation is observed in sera from hamsters, Holochilus venezuelae and Didelphis marsupialis after inoculation with L. mexicana mexicana from Panama and L. garnhami from the Venezuelan Andes region. The nature of this phenomenon is discussed. Xenodiagnoses were made with

  2. Bases experimentales para la identificacion de Leishmania spp. de America por morfometria de amastigotos Experimental basis for the identification of Leishmania spp. of America by morphometry of amastigotes

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    Haideé Urdaneta

    1982-12-01

    Full Text Available Estudios morfométricos sobre los amastigotos de dieciocho poblaciones de cuatro aislados pertenecientes a dos especies venezolanas de Leishmania (L. braziliensis y L. garnhami indican que la posición del einetoplasto nose modifica en modo estadísticamente significativo cuando los parásitos son sometidos a pasajes por hamsteres y ratones,cultivos en medio NNN o por infección en un vector (Lu. townsendi. La posición del cinetoplasto, medido como la distancia entre el extremo posterior del amastigoto al cinetoplasto, dividido entre la distancia del organoide al extremo anterior de la célula, permite diferenciar a L. braziliensis de L. mexicana y L. garnhami. Los otros parámetros morfométricos no son tan confiables.Morphometric studies on amastigotes of eigtheen populations from four isolates of two species of Venezuelan Leishmania (L. braziliensis and L. garnhami indicate that the position of the kinetoplast is not significantly modified when parasites are submitted to pasages into hamsters, and culture in NNN medium or infection in a vector (Lu. townsendi. The location of the kinetoplast measured as the distance from the posterior end of the amastigote to the kinetoplast divided by the distance of the same organoid to the anterior end of the amastigote allows to differenciate L. braziliensis from L. mexicana and L. garnhami. Other morphometric parameters are not so confiable.

  3. Understanding serine proteases implications on Leishmania spp lifecycle.

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    Alves, Carlos Roberto; Souza, Raquel Santos de; Charret, Karen Dos Santos; Côrtes, Luzia Monteiro de Castro; Sá-Silva, Matheus Pereira de; Barral-Veloso, Laura; Oliveira, Luiz Filipe Gonçalves; da Silva, Franklin Souza

    2018-01-01

    Serine proteases have significant functions over a broad range of relevant biological processes to the Leishmania spp lifecycle. Data gathered here present an update on the Leishmania spp serine proteases and the status of these enzymes as part of the parasite degradome. The serine protease genes (n = 26 to 28) in Leishmania spp, which encode proteins with a wide range of molecular masses (35 kDa-115 kDa), are described along with their degrees of chromosomal and allelic synteny. Amid 17 putative Leishmania spp serine proteases, only ∼18% were experimentally demonstrated, as: signal peptidases that remove the signal peptide from secretory pre-proteins, maturases of other proteins and with metacaspase-like activity. These enzymes include those of clans SB, SC and SF. Classical inhibitors of serine proteases are used as tools for the characterization and investigation of Leishmania spp. Endogenous serine protease inhibitors, which are ecotin-like, can act modulating host actions. However, crude or synthetic based-natural serine protease inhibitors, such as potato tuber extract, Stichodactyla helianthus protease inhibitor I, fukugetin and epoxy-α-lapachone act on parasitic serine proteases and are promising leishmanicidal agents. The functional interrelationship between serine proteases and other Leishmania spp proteins demonstrate essential functions of these enzymes in parasite physiology and therefore their value as targets for leishmaniasis treatment. Copyright © 2017 Elsevier Inc. All rights reserved.

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

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    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

  5. TRANSCRIPTIONAL INHIBITION OF INTERLEUKIN-12 PROMOTER ACTIVITY IN LEISHMANIA SPP.-INFECTED MACROPHAGES

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    Jayakumar, Asha; Widenmaier, Robyn; Ma, Xiaojing; McDowell, Mary Ann

    2009-01-01

    To establish and persist within a host, Leishmania spp. parasites delay the onset of cell-mediated immunity by suppressing interleukin-12 (IL-12) production from host macrophages. Although it is established that Leishmania spp.-infected macrophages have impaired IL-12 production, the mechanisms that account for this suppression remain to be completely elucidated. Using a luciferase reporter assay assessing IL-12 transcription, we report here that Leishmania major, Leishmania donovani, and Leishmania chagasi inhibit IL-12 transcription in response to interferon-gamma, lipopolysaccharide, and CD40 ligand and that Leishmania spp. lipophosphoglycan, phosphoglycans, and major surface protein are not necessary for inhibition. In addition, all the Leishmania spp. strains and life-cycle stages tested inhibited IL-12 promoter activity. Our data further reveal that autocrine-acting host factors play no role in the inhibitory response and that phagocytosis signaling is necessary for inhibition of IL-12. PMID:18372625

  6. Peptone-yeast autolysate-fetal bovine serum 10, a simple, inexpensive liquid medium for cultivation of Leishmania spp.

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    Palomino, J C

    1982-01-01

    A simple liquid medium for the cultivation of Leishmania parasites is described. Leishmania brasiliensis and Leishmania peruviana cultured in this medium reached cell densities greater than 10(7) promastigotes per ml within 7 days. This medium compares very favorably with the more complex media used to cultivate Leishmania spp. and other hemoflagellates.

  7. First evidence of Leishmania infection in European brown hare (Lepus europaeus) in Greece: GIS analysis and phylogenetic position within the Leishmania spp.

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    Tsokana, C N; Sokos, C; Giannakopoulos, A; Mamuris, Z; Birtsas, P; Papaspyropoulos, K; Valiakos, G; Spyrou, V; Lefkaditis, M; Chatzopoulos, D C; Kantere, M; Manolakou, K; Touloudi, A; Burriel, A Rodi; Ferroglio, E; Hadjichristodoulou, C; Billinis, C

    2016-01-01

    Although the existence of a sylvatic transmission cycle of Leishmania spp., independent from the domestic cycle, has been proposed, data are scarce on Leishmania infection in wild mammals in Greece. In this study, we aimed to investigate the presence of Leishmania infection in the European brown hare in Greece, to infer the phylogenetic position of the Leishmania parasites detected in hares in Greece, and to identify any possible correlation between Leishmania infection in hares with environmental parameters, using the geographical information system (GIS). Spleen samples from 166 hares were tested by internal transcribed spacer-1 (ITS-1)-nested PCR for the detection of Leishmania DNA. Phylogenetic analysis was performed on Leishmania sequences from hares in Greece in conjunction with Leishmania sequences from dogs in Greece and 46 Leishmania sequences retrieved from GenBank. The Leishmania DNA prevalence in hares was found to be 23.49 % (95 % confidence interval (CI) 17.27-30.69). The phylogenetic analysis confirmed that the Leishmania sequences from hares in Greece belong in the Leishmania donovani complex. The widespread Leishmania infection in hares should be taken into consideration because under specific circumstances, this species can act as a reservoir host. This study suggests that the role of wild animals, including hares, in the epidemiology of Leishmania spp. in Greece deserves further elucidation.

  8. Potential role for dog fleas in the cycle of Leishmania spp.

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    Ferreira, Marilia Gabriele Prado Albuquerque; Fattori, Karina Reinaldo; Souza, Fausto; Lima, Valéria Marçal Felix

    2009-10-28

    Several species of Leishmania spp. cause diseases in humans that range from self-healing cutaneous lesions to fatal visceral leishmaniosis. It has been observed that besides being transmitted by sand flies, Leishmania spp. may also be transmitted by arthropods such as ticks and fleas. To investigate the possible role of dog fleas in the transmission of Leishmania spp., Ctenocefalides felis were removed from 22 dogs which were positive according to ELISA and rK-39 tests. A C. felis sample from each of the 22 dogs was used to infect a hamster. The 22 hamsters were euthanized 4 months after infection with the fleas and the blood was subjected to ELISA to detect antibody anti-Leishmania spp., and the spleen samples were submitted to PCR for detection of Leishmania spp. DNA. PCR and ELISA were both positive in 18.1% (4/22), with PCR alone being positive in 45% (10/22) and ELISA alone in only 9% (2/22). These results suggest the participation of dog fleas in the Leishmania spp. cycle. Confirmation that C. felis indeed transmit leishmaniosis to dogs requires new strategies against leishmaniosis to be enforced by public health authorities and which focus on better ways to keep dogs free of fleas.

  9. Detection of Leishmania spp. in Bats from an Area of Brazil Endemic for Visceral Leishmaniasis.

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    de Rezende, M B; Herrera, H M; Carvalho, C M E; Carvalho Anjos, E A; Ramos, C A N; de Araújo, F R; Torres, J M; de Oliveira, C E

    2017-12-01

    The multihost parasites Leishmania spp. infect a broad range of wild mammalian species including bats. Several species of bats have adapted to a variety of food resources and shelters in urban areas. This study aimed to detect Leishmania spp. DNA in bats present in forest fragments located in metropolitan areas endemic for leishmaniasis in Campo Grande, Mato Grosso do Sul (MS), Brazil. Blood samples were obtained from 80 individuals, including eight species of Phyllostomidae and one species of Vespertilionidae. Thirty of the 80 bats were positive for Leishmania spp. using conventional PCR, all belonging to the family Phyllostomidae. Eighteen samples tested by real-time PCR (qPCR) using specific primers for the kDNA of Leishmania infantum were positive. To the best of our knowledge, this is the first report detecting Leishmania spp. in Platyrrhinus incarum in addition to being the first reported detection of L. infantum in the bat species Phyllostomus discolor, Platyrrhinus lineatus, Artibeus planirostris and Artibeus lituratus. Our results show that bats can host Leishmania spp. in areas endemic for leishmaniasis, which must be taken into account in disease control operations by public health authorities. © 2017 Blackwell Verlag GmbH.

  10. Dynamics of sterol synthesis during development of Leishmania spp. parasites to their virulent form.

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    Yao, Chaoqun; Wilson, Mary E

    2016-04-12

    The Leishmania spp. protozoa, the causative agents of the "neglected" tropical disease leishmaniasis, are transmitted to mammals by sand fly vectors. Within the sand fly, parasites transform from amastigotes to procyclic promastigotes, followed by development of virulent (metacyclic) promastigote forms. The latter are infectious to mammalian hosts. Biochemical components localized in the parasite plasma membrane such as proteins and sterols play a pivotal role in Leishmania pathogenesis. Leishmania spp. lack the enzymes for cholesterol synthesis, and the dynamics of sterol acquisition and biosynthesis in parasite developmental stages are not understood. We hypothesized that dynamic changes in sterol composition during metacyclogenesis contribute to the virulence of metacyclic promastigotes. Sterols were extracted from logarithmic phase or metacyclic promastigotes grown in liquid culture with or without cholesterol, and analyzed qualitatively and quantitatively by gas chromatograph-mass spectrometry (GC-MS). TriTrypDB was searched for identification of genes involved in Leishmania sterol biosynthetic pathways. In total nine sterols were identified. There were dynamic changes in sterols during promastigote metacyclogenesis. Cholesterol in the culture medium affected sterol composition in different parasite stages. There were qualitative and relative quantitative differences between the sterol content of virulent versus avirulent parasite strains. A tentative sterol biosynthetic pathway in Leishmania spp. promastigotes was identified. Significant differences in sterol composition were observed between promastigote stages, and between parasites exposed to different extracellular cholesterol in the environment. These data lay the foundation for further investigating the role of sterols in the pathogenesis of Leishmania spp. infections.

  11. Serological survey of dogs from Egypt for antibodies to Leishmania spp.

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    Leishmaniasis is an insect-transmitted parasitic disease with worldwide distribution. Leishmania spp. infections cause a broad spectrum of clinical signs ranging from skin lesions to fatal visceral disease. Dogs are a major reservoir host for visceral leishmaniasis in humans. Leishmaniasis is endemi...

  12. Antibodies to Toxoplasma gondii and Leishmania spp. in domestic cats from Luanda, Angola

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    Toxoplasma gondii and Leishmania spp. are zoonotic agents of importance to public health, with domestic cats as potential reservoirs for both protozoal infections. The present study aimed at assessing for the first time the seroprevalence of these zoonotic parasites in a domestic feline population l...

  13. Antibodies to Toxoplasma gondii and Leishmania spp. in domestic cats from Luanda, Angola

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    Lopes, Ana Patrícia; Oliveira, Ana Cristina; Granada, Sara; Rodrigues, Filipa T.; Papadopoulos, Elias; Schallig, Henk; Dubey, Jitender P.; Cardoso, Luís

    2017-01-01

    Toxoplasma gondii and Leishmania spp. are zoonotic protozoa of importance to animal and public health. The present study aimed to assess for the first time the seroprevalence of these zoonotic parasites in a domestic feline population living in Luanda, Angola. One hundred and two cats were sampled

  14. Molecular and serological detection of Leishmania spp. in captive wild animals from Ilha Solteira, SP, Brazil Detecção sorológica e molecular de Leishmania spp. em animais selvagens do zoológico de Ilha Solteira, SP, Brasil

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    Márcia Mariza Gomes Jusi

    2011-09-01

    Full Text Available 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.Leishmaniose é uma doença zoonótica que afeta cerca de 12 milhões de pessoas no mundo todo. Várias espécies mamíferas podem servir de reservatório para a doença. Os cães são considerados os principais reservatórios para a leishmaniose visceral em áreas urbanas, o que tem se tornado um sério problema de saúde pública no Brasil. O objetivo deste trabalho foi avaliar a presença de Leishmania spp. em animais selvagens mantidos no zoológico de Ilha Solteira, São Paulo, Brasil. Foram coletados amostras de sangue e tecidos de cinco espécies diferentes: Speothos venaticus, Chrysocyon brachyurus, Cerdocyon thous, Pseudalopex vetulus, e Procyon

  15. Histochemical and molecular evaluation of the prevalence of Leishmania spp. in hematophagous insects

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    Willian Marinho Dourado Coelho

    2016-06-01

    Full Text Available The prevalence study of Leishmania spp. in hematophagous insects captured from the environment in bat roosts and pigeon nests, or feeding their hosts (cattle, pigs, horses, dogs and humans in urban, peri-urban and rural areas, between 2012 and 2014. For this study, the amastigotes present in these insects were detected by histochemical and PCR techniques. Positive gene amplification for Leishmania was found in two horseflies of the species Tabanus importunus collected in the environment, and amastigote forms of Leishmania spp., as well as erythrocytes and leukocytes, were histochemically detected in one of that insect. The other analyzed insects were not positive by PCR our by direct parasitological examination. Only horseflies captured in urban and peri-urban areas were positive. During the collection, no phlebotomine sand flies were captured in rural areas far from the city limits. It can be concluded that the discovery of horseflies positive for Leishmania spp. in urban and peri-urban areas indicates the likelihood that urban areas and their surroundings provide vector parasites with an environment suitable for the spread and consequent perpetuation of the biological cycle of this protozoan.

  16. Identification of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus actively infesting dogs.

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    Viol, Milena Araúz; Guerrero, Felix D; de Oliveira, Bruno César Miranda; de Aquino, Monally Conceição Costa; Loiola, Saulo Hudson; de Melo, Guilherme Dias; de Souza Gomes, Aparecida Helena; Kanamura, Cristina Takami; Garcia, Marcos Valério; Andreotti, Renato; de Lima, Valéria Marçal Félix; Bresciani, Katia Denise Saraiva

    2016-09-01

    Sand flies are recognized as the major vector of canine visceral leishmaniasis. However, in some areas of Brazil where sand flies do not occur, this disease is found in humans and dogs. There has been speculation that ticks might play a role in transmission of canine visceral leishmaniasis and the DNA of Leishmania spp. has been reported in whole ticks. We investigated the presence of Leishmania spp. promastigotes in the intestines, ovaries, and salivary glands of Rhipicephalus sanguineus ticks collected from tick-infested dogs in two cities of Brazil. We used 66 dogs that tested positive and 33 that tested negative for Leishmania spp. according to direct cytological examination assays. Ten ticks were collected from each dog and dissected to collect the intestines, ovaries, and salivary glands for immunohistochemistry (IHC) and diagnostic real-time polymerase chain reaction (RT-PCR). IHC results showed Leishmania spp. in 98, 14, and 8 % of the intestines, ovaries, and salivary glands, respectively. Real-time PCR showed that 89, 41, and 33 % of the tick intestine, ovary, and salivary glands, respectively, were positive for Leishmania spp. The verification of promastigotes of Leishmania spp. by two independent techniques in ticks collected from these urban region dogs showed that there is need for clarification of the role of ticks in the transmission of canine visceral leishmaniasis in Brazil.

  17. 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)

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

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    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

  19. Identification of Tunisian Leishmania spp. by PCR amplification of cysteine proteinase B (cpb) genes and phylogenetic analysis.

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    Chaouch, Melek; Fathallah-Mili, Akila; Driss, Mehdi; Lahmadi, Ramzi; Ayari, Chiraz; Guizani, Ikram; Ben Said, Moncef; Benabderrazak, Souha

    2013-03-01

    Discrimination of the Old World Leishmania parasites is important for diagnosis and epidemiological studies of leishmaniasis. We have developed PCR assays that allow the discrimination between Leishmania major, Leishmania tropica and Leishmania infantum Tunisian species. The identification was performed by a simple PCR targeting cysteine protease B (cpb) gene copies. These PCR can be a routine molecular biology tools for discrimination of Leishmania spp. from different geographical origins and different clinical forms. Our assays can be an informative source for cpb gene studying concerning drug, diagnostics and vaccine research. The PCR products of the cpb gene and the N-acetylglucosamine-1-phosphate transferase (nagt) Leishmania gene were sequenced and aligned. Phylogenetic trees of Leishmania based cpb and nagt sequences are close in topology and present the classic distribution of Leishmania in the Old World. The phylogenetic analysis has enabled the characterization and identification of different strains, using both multicopy (cpb) and single copy (nagt) genes. Indeed, the cpb phylogenetic analysis allowed us to identify the Tunisian Leishmania killicki species, and a group which gathers the least evolved isolates of the Leishmania donovani complex, that was originated from East Africa. This clustering confirms the African origin for the visceralizing species of the L. donovani complex. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Leishmania spp. Epidemiology of Canine Leishmaniasis in the Yucatan Peninsula

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    A. López-Céspedes

    2012-01-01

    Full Text Available Canine Leishmaniasis is widespread in various Mexican states, where different species of Leishmania have been isolated from dogs. In the present study, we describe the detection of L. braziliensis, L. infantum, and L. mexicana in serum of dogs from the states of Yucatan and Quintana Roo in the Yucatan Peninsula (Mexico. A total of 412 sera were analyzed by ELISA using the total extract of the parasite and the iron superoxide dismutase excreted by different trypanosomatids as antigens. We found the prevalence of L. braziliensis to be 7.52%, L. infantum to be 6.07%, and L. mexicana to be 20.63%, in the dog population studied. The results obtained with ELISA using iron superoxide dismutase as the antigen were confirmed by western blot analysis with its greater sensitivity, and the agreement between the two techniques was very high.

  1. Leishmania spp. epidemiology of canine leishmaniasis in the Yucatan Peninsula.

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    López-Céspedes, A; Longoni, S S; Sauri-Arceo, C H; Sánchez-Moreno, M; Rodríguez-Vivas, R I; Escobedo-Ortegón, F J; Barrera-Pérez, M A; Bolio-González, M E; Marín, C

    2012-01-01

    Canine Leishmaniasis is widespread in various Mexican states, where different species of Leishmania have been isolated from dogs. In the present study, we describe the detection of L. braziliensis, L. infantum, and L. mexicana in serum of dogs from the states of Yucatan and Quintana Roo in the Yucatan Peninsula (Mexico). A total of 412 sera were analyzed by ELISA using the total extract of the parasite and the iron superoxide dismutase excreted by different trypanosomatids as antigens. We found the prevalence of L. braziliensis to be 7.52%, L. infantum to be 6.07%, and L. mexicana to be 20.63%, in the dog population studied. The results obtained with ELISA using iron superoxide dismutase as the antigen were confirmed by western blot analysis with its greater sensitivity, and the agreement between the two techniques was very high.

  2. Seroepidemiology of Leishmania spp. in dogs residing in Telêmaco Borba, Paraná, Brazil

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    Caroline Constantino

    2014-12-01

    Full Text Available Leishmaniasis is an important metazoonosis caused by protozoa of the genus Leishmania and has a heteroxenic life cycle involving invertebrate and vertebrate hosts. Transmission occurs during the blood meal of infected phlebotomine sand flies in wild species, domestic animals, and humans. The dog is a reservoir for the parasite causing visceral leishmaniasis (VL, whereas in American tegumentary leishmaniasis (ATL, dogs are erratic hosts that are accidentally infected, as in humans. Dogs are considered an important indicator of the parasite and its vectors in the environment, thus highlighting the importance of diagnosis in these animals. This study aimed to assess the seroepidemiology of Leishmania spp. in dogs in the municipality of Telêmaco Borba that were part of a castration campaign. Blood samples from 191 dogs were collected, and their owners were surveyed on various epidemiological variables. Serological analysis was performed using indirect immunofluorescence (IIF and rapid immunochromatography (DPP®. Screening by IIF identified 13 (6.81% positive animals, none of which were positive for the DPP® test, which is specific for VL. Statistical analysis of the questionnaire responses indicated a significant association between seropositivity and the presence of stacked or composting leaves in the backyard (p = 0.0498, forest areas (squares, woods, parks near the residence (p = 0.0015, and poorly healing ulcerated or nodular epidermal lesions in the dog (p = 0.0138. This study revealed the presence of anti-Leishmania spp. IgG antibodies in dogs residing in Telêmaco Borba, suggesting the presence of the parasite and vector in the environment. In addition, the existence of stacked or composting leaves in the backyard, forest areas near the residence, and epidermal lesions in dogs are factors associated with Leishmania spp. infection in pet dogs.

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

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    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.

  4. Development and evaluation of zinc phthalocyanine nanoemulsions for use in photodynamic therapy for Leishmania spp.

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    Betzler de Oliveira de Siqueira, Luciana; da Silva Cardoso, Verônica; Almeida Rodrigues, Igor; Lúcia Vazquez-Villa, Ana; Pereira dos Santos, Elisabete; da Costa Leal Ribeiro Guimarães, Bruno; dos Santos Cerqueira Coutinho, Cristal; Vermelho, Alane Beatriz; Ricci Junior, Eduardo

    2017-02-01

    Photodynamic therapy (PDT) combines light with photosensitizers (PS) for production of reactive oxygen species (ROS) that can kill infectious microorganisms such as bacteria, fungi and protozoa. The application of nanotechnology has enabled the advancement of PDT because many PS are insoluble in water, necessitating a nanocarrier as a physiologically acceptable carrier. Nanoemulsions are efficient nanocarriers for solubilizing liposoluble drugs, like the PS, in water. Cutaneous (CL) and mucocutaneous leishmaniasis (ML) are caused by different species of the genus Leishmania, transmitted to humans by sandfly bites. Parasites are hosted in skin macrophages producing ulcerative lesions. Thus, a topical treatment, effective and inexpensive, for CL and ML is preferable to systemic interventions. There are topical treatments like paromomycin and amphotericin B, but they have many local side effects or a very high cost, limiting their use. This work aimed to develop a zinc phthalocyanine (photosensitizer) oil-in-water nanoemulsion, essential clove oil and polymeric surfactant (Pluronic® F127) for the formulation of a topical delivery system for use in PDT against Leishmania amazonensis and Leishmania infantum. The nanoemulsion was produced by a high-energy method and characterized by size, polydispersity, morphology, pH, content and stability studies. The toxicity in the dark and the photobiological activity of the formulations were evaluated in vitro for Leishmania and macrophages. The formulation presented was pH compatible with topical use, approximately 30 nm in size, with a polydispersity index ≤0.1 and remained stable at room and refrigerator temperature during the stability study (60 days). The zinc phthalocyanine nanoemulsion is effective in PDT against Leishmania spp.; use against skin infections can be a future application of this topical formulation, avoiding the use of oral or injectable medications, decreasing systemic adverse effects.

  5. PCR-RFLP y RAPD para la tipificación de Leishmania neotropical

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    Ana Margarita Montalvo

    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.

  6. Antibodies to Toxoplasma gondii and Leishmania spp. in domestic cats from Luanda, Angola.

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    Lopes, Ana Patrícia; Oliveira, Ana Cristina; Granada, Sara; Rodrigues, Filipa T; Papadopoulos, Elias; Schallig, Henk; Dubey, Jitender P; Cardoso, Luís

    2017-05-30

    Toxoplasma gondii and Leishmania spp. are zoonotic protozoa of importance to animal and public health. The present study aimed to assess for the first time the seroprevalence of these zoonotic parasites in a domestic feline population living in Luanda, Angola. One hundred and two cats were sampled at a veterinary medical centre, from May 2014 to February 2016. The age of the cats ranged from 2.5 to 143 months (median: 12 months; interquartile range: 7.5-24). Serum samples were tested for immunoglobulin (Ig) G antibodies to T. gondii at two-fold dilutions of 1:20 to 1:2560 with a modified agglutination test (MAT) commercial kit. The direct agglutination test (DAT) for titration of IgG antibodies specific to Leishmania spp. used a standard freeze-dried antigen at a concentration of 5×10 7 promastigotes per milliliter, following a predefined protocol. Two-fold dilution series ranging from 1:25 to 1:800 were tested, with a cut-off titre of 100 chosen for seropositivity. Four out of 102 cats (3.9%; 95% confidence interval [CI]: 1.1-9.7) had antibodies to T. gondii: one had a titer of 20, one a titer of 160, and two had a titer≥2560. No cat (0.0%; CI: 0.0-3.5) was found seropositive for Leishmania spp. A statistically significant difference was found between T. gondii seroprevalence and Leishmania spp. seroprevalence (p=0.043). The odds of a cat being seropositive to T. gondii increased by an average factor of 1.58 for each 1-year increase in age (p=0.003). The sampled cats were well-cared animals and may not represent the overall feline population of Angola at the national and city levels. The fact that only 12 out of the 102 sampled cats ate or had access to raw or undercooked meat and/or viscera may have reduced the likelihood of finding seropositive results. Under these circumstances, additional studies, including a larger number of cats, are necessary for a more comprehensive assessment of the zoonotic risk posed by these animals in Angola. Copyright © 2017

  7. Coinfection by Toxoplasma gondii and Leishmania spp. in domestic cats (Felis catus in State of Mato Grosso do Sul

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    Audrey Rennó Campos Braga

    2014-12-01

    Full Text Available Introduction Leishmaniasis and toxoplasmosis are important to public health. Methods Antibodies for Toxoplasma gondii and Leishmania spp. were evaluated in cats from Campo Grande, State of Mato Grosso do Sul, Brazil, a region endemic for canine visceral leishmaniasis. Serum samples from 50 asymptomatic cats were titrated for T. gondii by the immunofluorescence antibody test and modified agglutination test and for Leishmania spp. by the immunofluorescence antibody test. Results These two agents coinfected two (4% of the 50 tested animals. Conclusions These findings demonstrate the concomitant presence of two important zoonoses in cats from Brazilian endemic regions for canine visceral leishmaniasis.

  8. In vitro and in vivo efficacy of ether lipid edelfosine against Leishmania spp. and SbV-resistant parasites.

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    Rubén E Varela-M

    Full Text Available BACKGROUND: The leishmaniases are a complex of neglected tropical diseases caused by more than 20 Leishmania parasite species, for which available therapeutic arsenal is scarce and unsatisfactory. Pentavalent antimonials (SbV are currently the first-line pharmacologic therapy for leishmaniasis worldwide, but resistance to these compounds is increasingly reported. Alkyl-lysophospoholipid analogs (ALPs constitute a family of compounds with antileishmanial activity, and one of its members, miltefosine, has been approved as the first oral treatment for visceral and cutaneous leishmaniasis. However, its clinical use can be challenged by less impressive efficiency in patients infected with some Leishmania species, including L. braziliensis and L. mexicana, and by proneness to develop drug resistance in vitro. METHODOLOGY/PRINCIPAL FINDINGS: We found that ALPs ranked edelfosine>perifosine>miltefosine>erucylphosphocholine for their antileishmanial activity and capacity to promote apoptosis-like parasitic cell death in promastigote and amastigote forms of distinct Leishmania spp., as assessed by proliferation and flow cytometry assays. Effective antileishmanial ALP concentrations were dependent on both the parasite species and their development stage. Edelfosine accumulated in and killed intracellular Leishmania parasites within macrophages. In vivo antileishmanial activity was demonstrated following oral treatment with edelfosine of mice and hamsters infected with L. major, L. panamensis or L. braziliensis, without any significant side-effect. Edelfosine also killed SbV-resistant Leishmania parasites in in vitro and in vivo assays, and required longer incubation times than miltefosine to generate drug resistance. CONCLUSIONS/SIGNIFICANCE: Our data reveal that edelfosine is the most potent ALP in killing different Leishmania spp., and it is less prone to lead to drug resistance development than miltefosine. Edelfosine is effective in killing Leishmania

  9. Lulo cell line derived from Lutzomyia longipalpis (Diptera: Psychodidae): a novel model to assay Leishmania spp. and vector interaction.

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    Côrtes, Luzia Mc; Silva, Roger Mm; Pereira, Bernardo As; Guerra, Camila; Zapata, Angela C; Bello, Felio J; Finkelstein, Léa C; Madeira, Maria F; Brazil, Reginaldo P; Côrte-Real, Suzana; Alves, Carlos R

    2011-11-14

    Leishmania (Vianna) braziliensis, Leishmania (Leishmania) amazonensis and Leishmania (Leishmania) chagasi are important parasites in the scenario of leishmaniasis in Brazil. During the life cycle of these parasites, the promastigote forms adhere to the midgut epithelial microvillii of phlebotomine insects to avoid being secreted along with digestive products. Lulo cells are a potential model that will help to understand the features of this adhesion phenomenon. Here, we analyze the interaction between Leishmania spp. promastigotes and Lulo cells in vitro, specifically focusing on adhesion events occurring between three Leishmania species and this cell line. Confluent monolayers of Lulo cells were incubated with promastigotes and adhesion was assessed using both light microscopy and scanning electron microscopy. The results indicate that species from the subgenera Leishmania and Viannia have great potential to adhere to Lulo cells. The highest adherence rate was observed for L. (L.) chagasi after 24 h of incubation with Lulo cells (27.3 ± 1.8% of cells with adhered promastigotes), followed by L. (L.) amazonensis (16.0 ± 0.7%) and L. (V.) braziliensis (3.0 ± 0.7%), both after 48 h. In the ultrastructural analysis, promastigote adherence was also assessed by scanning electron microscopy, showing that, for parasites from both subgenera, adhesion occurs by both the body and the flagellum. The interaction of Lulo cells with Leishmania (L.) chagasi showed the participation of cytoplasmic projections from the former closely associating the parasites with the cells. We present evidence that Lulo cells can be useful in studies of insect-parasite interactions for Leishmania species.

  10. Detection of Leishmania spp in silvatic mammals and isolation of Leishmania (Viannia braziliensis from Rattus rattus in an endemic area for leishmaniasis in Minas Gerais State, Brazil.

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    Agnes Antônia Sampaio Pereira

    Full Text Available Knowledge of potential reservoirs of Leishmania spp. in an anthropic environment is important so that surveillance and control measures can be implemented. The aim of this study was to investigate the infection by Leishmania in small mammals in an area located in Minas Gerais, Brazil, that undergoes changes in its natural environment and presents autochthonous human cases of cutaneous leishmaniasis (CL and visceral leishmaniasis (VL. For the capture of the animals, Sherman and Tomahawk traps were used and distributed in the peridomicile of houses with reports of autochthonous cases of CL or VL. Six catches were carried out on two consecutive nights with intervals of two months during one year and samples of spleen, liver, tail skin, ear skin and bone marrow of the animals were obtained. Parasitological and molecular methods were used to detect the infection. Identification of the Leishmania species was performed by PCR RFLPhsp70. Twenty five animals of four species were captured: ten Rattus rattus, nine Didelphis albiventris, five Cerradomys subflavus and one Marmosops incanus. In the PCR-hsp70, five animals were positive (20%. The Leishmania species identified in PCR-RFLPhsp70 were: Leishmania braziliensis in D. albiventris (2, C. subflavus (1 and R. rattus (1 and Leishmania infantum in R. rattus (1. The highest positivity rate for L. braziliensis was obtained in the liver samples. The spleen was the only tissue positive for L. infantum. It was isolated in culture medium L. braziliensis from two samples (liver and spleen of R. rattus. This is the first record of isolation of L. braziliensis from R. rattus in the southeastern region of Brazil. These results are relevant to the knowledge of the epidemiology of leishmaniasis in the region, mainly in the investigation of the presence of hosts and possible reservoirs of the parasite.

  11. An Innovative Field-Applicable Molecular Test to Diagnose Cutaneous Leishmania Viannia spp. Infections.

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    Omar A Saldarriaga

    2016-04-01

    Full Text Available Cutaneous and mucosal leishmaniasis is widely distributed in Central and South America. Leishmania of the Viannia subgenus are the most frequent species infecting humans. L. (V. braziliensis, L. (V. panamensis are also responsible for metastatic mucosal leishmaniasis. Conventional or real time PCR is a more sensitive diagnostic test than microscopy, but the cost and requirement for infrastructure and trained personnel makes it impractical in most endemic regions. Primary health systems need a sensitive and specific point of care (POC diagnostic tool. We developed a novel POC molecular diagnostic test for cutaneous leishmaniasis caused by Leishmania (Viannia spp. Parasite DNA was amplified using isothermal Recombinase Polymerase Amplification (RPA with primers and probes that targeted the kinetoplast DNA. The amplification product was detected by naked eye with a lateral flow (LF immunochromatographic strip. The RPA-LF had an analytical sensitivity equivalent to 0.1 parasites per reaction. The test amplified the principal L. Viannia species from multiple countries: L. (V. braziliensis (n = 33, L. (V. guyanensis (n = 17, L. (V. panamensis (n = 9. The less common L. (V. lainsoni, L. (V. shawi, and L. (V. naiffi were also amplified. No amplification was observed in parasites of the L. (Leishmania subgenus. In a small number of clinical samples (n = 13 we found 100% agreement between PCR and RPA-LF. The high analytical sensitivity and clinical validation indicate the test could improve the efficiency of diagnosis, especially in chronic lesions with submicroscopic parasite burdens. Field implementation of the RPA-LF test could contribute to management and control of cutaneous and mucosal leishmaniasis.

  12. Leishmania infantum, Dirofilaria spp. and other endoparasite infections in kennel dogs in central Italy

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    Sauda, Federica; Malandrucco, Livia; Macrì, Gladia; Scarpulla, Manuela; De Liberato, Claudio; Terracciano, Giuliana; Fichi, Gianluca; Berrilli, Federica; Perrucci, Stefania

    2018-01-01

    Prevalence and risk factors of Leishmania infantum, Dirofilaria spp. and other potentially zoonotic or canine-specific endoparasite infections were assessed in 639 kennel dogs from central Italy. To this end, individual blood and fecal samples were examined using parasitological, immunological and molecular techniques. The presence of compatible clinical pictures, as well as age and gender were considered as putative risks factors. To evaluate risk factors, multivariable analysis with logistic regression and univariable analysis with a Chi square test and a Fischer’s exact test were performed. Overall, 52.6% of dogs (95% CI 48.6-56.5) were found positive, while 39.6% of dogs (95% CI 35.8-43.5) were infected by potentially zoonotic species. Leishmania infantum and Dirofilaria repens showed prevalences of 2.5% (95% CI 1.5-4.1) and 2.8% (95% CI 1.7-4.5), respectively. The prevalence of cardiorespiratory parasites was 7.8% (95% CI 5.9-10.3) and included the species Angiostrongylus vasorum, Eucoleus aerophilus, Eucoleus boehmi and D. immitis; the latter showed a prevalence of 0.2% (95% CI 0.001-1). Intestinal parasites were significantly prevalent (38.8%, 95% CI 35-42.7) and they consisted mainly of species of major zoonotic concern, including ancylostomatids, Toxocara canis, Giardia duodenalis, Dipylidium caninum, Taeniidae, Strongyloides stercoralis and Cryptosporidium parvum. Endoparasites were significantly prevalent in clinically suspected dogs. Leishmania infantum and cardiorespiratory nematodes were prevalent in older dogs, while intestinal parasites were prevalent in younger dogs. Results show high dog and public health risks in kennels in central Italy, and suggest the need for more effective control measures. PMID:29388550

  13. Leishmania infantum, Dirofilaria spp. and other endoparasite infections in kennel dogs in central Italy

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    Sauda Federica

    2018-01-01

    Full Text Available Prevalence and risk factors of Leishmania infantum, Dirofilaria spp. and other potentially zoonotic or canine-specific endoparasite infections were assessed in 639 kennel dogs from central Italy. To this end, individual blood and fecal samples were examined using parasitological, immunological and molecular techniques. The presence of compatible clinical pictures, as well as age and gender were considered as putative risks factors. To evaluate risk factors, multivariable analysis with logistic regression and univariable analysis with a Chi square test and a Fischer’s exact test were performed. Overall, 52.6% of dogs (95% CI 48.6-56.5 were found positive, while 39.6% of dogs (95% CI 35.8-43.5 were infected by potentially zoonotic species. Leishmania infantum and Dirofilaria repens showed prevalences of 2.5% (95% CI 1.5-4.1 and 2.8% (95% CI 1.7-4.5, respectively. The prevalence of cardiorespiratory parasites was 7.8% (95% CI 5.9-10.3 and included the species Angiostrongylus vasorum, Eucoleus aerophilus, Eucoleus boehmi and D. immitis; the latter showed a prevalence of 0.2% (95% CI 0.001-1. Intestinal parasites were significantly prevalent (38.8%, 95% CI 35-42.7 and they consisted mainly of species of major zoonotic concern, including ancylostomatids, Toxocara canis, Giardia duodenalis, Dipylidium caninum, Taeniidae, Strongyloides stercoralis and Cryptosporidium parvum. Endoparasites were significantly prevalent in clinically suspected dogs. Leishmania infantum and cardiorespiratory nematodes were prevalent in older dogs, while intestinal parasites were prevalent in younger dogs. Results show high dog and public health risks in kennels in central Italy, and suggest the need for more effective control measures.

  14. Molecular screening of Leishmania spp. infection and bloodmeals in sandflies from a leishmaniasis focus in southwestern Turkey.

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    Karaku Ş, M; Pekağ Irba Ş, M; Demir, S; Eren, H; Töz, S; Özbel, Y

    2017-06-01

    Leishmaniasis is an arthropod-borne disease that affects approximately 2 million people worldwide annually. The aims of this study were to detect the presence of Leishmania (Kinetoplastida: Trypanosomatidae) DNA and the feeding preferences of probable vector species in an endemic focus of Leishmania infantum in Turkey. Entomological sampling was performed in August and October 2015 in Aydın province, where cases of human and canine leishmaniasis have been reported previously. A total of 1059 sandfly specimens comprising nine species belonging to two genera, Phlebotomus and Sergentomyia (both: Diptera: Psychodidae), and five subgenera of the Phlebotomus genus (Phlebotomus, Paraphlebotomus, Larroussius, Adlerius and Transphlebotomus) were collected in five villages. Among all Phlebotomus specimens, Phlebotomus neglectus (39%) was noted as the most abundant species, followed by Phlebotomus tobbi (18%). Leishmania DNA was detected in pools from P. neglectus, P. tobbi and Sergentomyia dentata by kDNA polymerase chain reaction (PCR). Leishmania DNA from Phlebotomus specimens was identified as L. infantum, but Leishmania DNA from Sergentomyia spp. could not be identified to species level by ITS-1 real-time PCR. The detection of Leishmania DNA in wild-caught P. neglectus and the high percentage (24.2%) of human DNA in engorged specimens suggests that P. neglectus is probably an important vector species for L. infantum in Aydın province. © 2016 The Royal Entomological Society.

  15. Detection and Differentiation of Leishmania spp. in Clinical Specimens by Use of a SYBR Green-Based Real-Time PCR Assay.

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    de Almeida, Marcos E; Koru, Ozgur; Steurer, Francis; Herwaldt, Barbara L; da Silva, Alexandre J

    2017-01-01

    Leishmaniasis in humans is caused by Leishmania spp. in the subgenera Leishmania and Viannia Species identification often has clinical relevance. Until recently, our laboratory relied on conventional PCR amplification of the internal transcribed spacer 2 (ITS2) region (ITS2-PCR) followed by sequencing analysis of the PCR product to differentiate Leishmania spp. Here we describe a novel real-time quantitative PCR (qPCR) approach based on the SYBR green technology (LSG-qPCR), which uses genus-specific primers that target the ITS1 region and amplify DNA from at least 10 Leishmania spp., followed by analysis of the melting temperature (T m ) of the amplicons on qPCR platforms (the Mx3000P qPCR system [Stratagene-Agilent] and the 7500 real-time PCR system [ABI Life Technologies]). We initially evaluated the assay by testing reference Leishmania isolates and comparing the results with those from the conventional ITS2-PCR approach. Then we compared the results from the real-time and conventional molecular approaches for clinical specimens from 1,051 patients submitted to the reference laboratory of the Centers for Disease Control and Prevention for Leishmania diagnostic testing. Specimens from 477 patients tested positive for Leishmania spp. with the LSG-qPCR assay, specimens from 465 of these 477 patients also tested positive with the conventional ITS2-PCR approach, and specimens from 10 of these 465 patients had positive results because of retesting prompted by LSG-qPCR positivity. On the basis of the T m values of the LSG-qPCR amplicons from reference and clinical specimens, we were able to differentiate four groups of Leishmania parasites: the Viannia subgenus in aggregate; the Leishmania (Leishmania) donovani complex in aggregate; the species L (L) tropica; and the species L (L) mexicana, L (L) amazonensis, L (L) major, and L (L) aethiopica in aggregate. Copyright © 2016 American Society for Microbiology.

  16. Molecular Identification of Leishmania spp. in Sand Flies (Diptera: Psychodidae, Phlebotominae) From Ecuador

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    Cevallos, Varsovia; Morales, Diego; Baldeón, Manuel E; Cárdenas, Paúl; Rojas-Silva, Patricio; Ponce, Patricio

    2017-01-01

    Abstract The detection and identification of natural infections in sand flies by Leishmania protozoan species in endemic areas is a key factor in assessing the risk of leishmaniasis and in designing prevention and control measures for this infectious disease. In this study, we analyzed the Leishmania DNA using nuclear ribosomal internal transcript spacer (ITS) sequences. Parasite DNA was extracted from naturally infected, blood-fed sand flies collected in nine localities considered leishmaniasis-endemic foci in Ecuador. The species of parasites identified in sand flies were Leishmania major-like, Leishmania naiffi, Leishmania mexicana, Leishmania lainsoni, and “Leishmania sp. siamensis”. Sand fly specimens of Brumptomyia leopoldoi, Mycropigomyia cayennensis, Nyssomyia yuilli yuilli, Nyssomyia trapidoi, Pressatia triacantha, Pressatia dysponeta, Psychodopygus carrerai carrerai, Psychodopygus panamensis, and Trichophoromyia ubiquitalis were found positive for Leishmania parasite. These findings contribute to a better understanding of the epidemiology and transmission dynamics of the disease in high-risk areas of Ecuador. PMID:28981860

  17. Serosurvey for Leishmania spp., Toxoplasma gondii, Trypanosoma cruzi and Neospora caninum in neighborhood dogs in Curitiba-Paraná, Brazil

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    Caroline Constantino

    Full Text Available Abstract Neighborhood dogs may act as reservoirs for several zoonotic protozoan infections, particularly in urban areas, thus constituting a potential public health threat. Accordingly, the aim of the present study was to evaluate the exposure of neighborhood dogs to four protozoan pathogens in public areas with high levels of human movement in Curitiba, southern Brazil. Blood samples from 26 neighborhood dogs were screened by means of the indirect immunofluorescent antibody test (IFAT for Leishmania spp., Toxoplasma gondii, Trypanosoma cruzi and Neospora caninum, and a questionnaire was answered by the respective keeper. A total of 8/26 dogs (30.7% seroreactive to T. gondii, 3/26 (11.5% to N. caninum and 2/26 (7.7% to both were identified. All the samples were seronegative for T. cruzi and Leishmania spp. Pathogen seroreactivity was not associated with the daily human movements or other epidemiological variables investigated (p > 0.05. In conclusion, the low seroprevalence for T. gondii and N. caninum indicated low environmental and food risk for animal infection and the seronegativity for Leishmania spp. and T. cruzi may reflect the absence of these pathogens in urban areas of Curitiba. Moreover, neighborhood dogs may be used as environmental sentinels for the presence of protozoan pathogens and their vectors.

  18. Identification by real-time PCR with SYBR Green of Leishmania spp. and Serratia marcescens in canine 'sterile' cutaneous nodular lesions.

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    Cornegliani, Luisa; Corona, Antonio; Vercelli, Antonella; Roccabianca, Paola

    2015-06-01

    Noninfectious, non-neoplastic, nodular to diffuse, so-called 'sterile' granulomatous/pyogranulomatous skin lesions (SGPSLs) are infrequently identified in dogs and may represent a diagnostic challenge. Their correct identification is based on history, histopathology and absence of intralesional foreign bodies and micro-organisms. The aim of this study was to investigate the presence of Leishmania spp., Mycobacterium spp., Serratia marcescens and Nocardia spp. by real-time PCR in canine nodular skin lesions histologically diagnosed as putatively sterile. Formalin-fixed skin biopsies were collected from 40 dogs. All samples were associated with an SGPSL diagnosis characterized by multifocal, nodular to diffuse, periadnexal and perifollicular pyogranulomas/granulomas. Neither micro-organisms nor foreign bodies were detected with haematoxylin and eosin staining, under polarized light. Further analyses included periodic acid Schiff, Ziehl-Neelsen, Fite Faraco, Giemsa and Gram histochemical stains; anti-Bacillus Calmette-Guérin (BCG) and Leishmania spp. immunohistochemistry; and real-time PCR analysis for Leishmania spp., Mycobacterium spp., S. marcescens and Nocardia spp. Special stains and BCG/immunohistochemistry were negative in all samples. Real-time PCR was positive for Leishmania spp. in four of 40 biopsies and for S. marcescens in two of 40 samples. Real-time PCR for Mycobacterium spp. and Nocardia spp. was negative. No correlation between real-time PCR positivity and a specific histological pattern was identified. Leishmania spp. have been previously identified as possible agents of certain SGPSLs, while the involvement of S. marcescens has not been investigated previously. According to our findings, Serratia spp. should be included in the list of agents possibly associated with a subgroup of granulomatous/pyogranulomatous skin lesions in dogs. © 2015 ESVD and ACVD.

  19. Detection and identification of Leishmania spp.: application of two hsp70-based PCR-RFLP protocols to clinical samples from the New World.

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    Montalvo, Ana M; Fraga, Jorge; Tirado, Dídier; Blandón, Gustavo; Alba, Annia; Van der Auwera, Gert; Vélez, Iván Darío; Muskus, Carlos

    2017-07-01

    Leishmaniasis is highly prevalent in New World countries, where several methods are available for detection and identification of Leishmania spp. Two hsp70-based PCR protocols (PCR-N and PCR-F) and their corresponding restriction fragment length polymorphisms (RFLP) were applied for detection and identification of Leishmania spp. in clinical samples recruited in Colombia, Guatemala, and Honduras. A total of 93 cases were studied. The samples were classified into positive or suspected of leishmaniasis according to parasitological criteria. Molecular amplification of two different hsp70 gene fragments and further RFLP analysis for identification of Leishmania species was done. The detection in parasitologically positive samples was higher using PCR-N than PCR-F. In the total of samples studied, the main species identified were Leishmania panamensis, Leishmania braziliensis, and Leishmania infantum (chagasi). Although RFLP-N was more efficient for the identification, RFLP-F is necessary for discrimination between L. panamensis and Leishmania guyanesis, of great importance in Colombia. Unexpectedly, one sample from this country revealed an RFLP pattern corresponding to Leishmania naiffi. Both molecular variants are applicable for the study of clinical samples originated in Colombia, Honduras, and Guatemala. Choosing the better tool for each setting depends on the species circulating. More studies are needed to confirm the presence of L. naiffi in Colombian territory.

  20. Detection of different Leishmania spp. and Trypanosoma cruzi antibodies in cats from the Yucatan Peninsula (Mexico) using an iron superoxide dismutase excreted as antigen.

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    Longoni, Silvia S; López-Cespedes, Angeles; Sánchez-Moreno, Manuel; Bolio-Gonzalez, Manuel E; Sauri-Arceo, Carlos H; Rodríguez-Vivas, Roger I; Marín, Clotilde

    2012-09-01

    Although human leishmaniasis has been reported in 20 states in Mexico, no case of leishmaniasis has been reported in cats to date. In the Yucatan Peninsula, it has been found that dogs may act as reservoirs for at least three Leishmania species (Leishmania mexicana, Leishmania braziliensis, and Leishmania panamensis). In this study we identified specific antibodies against these three Leishmania spp. and Trypanosoma cruzi in the sera from 95 cats from two States on the Yucatan Peninsula, namely Quintana Roo and Yucatan, by ELISA and Western blot techniques using whole extract and an iron superoxide dismutase excreted by the parasites as antigens. As well as demonstrating the presence of trypanosomatid antibodies in the feline population on the Yucatan Peninsula, we were also able to confirm the high sensitivity and specificity of the iron superoxide dismutase antigen secreted by them, which may prove to be very useful in epidemiological studies. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Surveillance for antibodies to Leishmania spp. in dogs from Sri Lanka and India

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    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...

  2. Molecular Identification of Leishmania spp. in Sand Flies (Diptera: Psychodidae, Phlebotominae) From Ecuador.

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    Quiroga, Cristina; Cevallos, Varsovia; Morales, Diego; Baldeón, Manuel E; Cárdenas, Paúl; Rojas-Silva, Patricio; Ponce, Patricio

    2017-11-07

    The detection and identification of natural infections in sand flies by Leishmania protozoan species in endemic areas is a key factor in assessing the risk of leishmaniasis and in designing prevention and control measures for this infectious disease. In this study, we analyzed the Leishmania DNA using nuclear ribosomal internal transcript spacer (ITS) sequences. Parasite DNA was extracted from naturally infected, blood-fed sand flies collected in nine localities considered leishmaniasis-endemic foci in Ecuador.The species of parasites identified in sand flies were Leishmania major-like, Leishmania naiffi, Leishmania mexicana, Leishmania lainsoni, and "Leishmania sp. siamensis". Sand fly specimens of Brumptomyia leopoldoi, Mycropigomyia cayennensis, Nyssomyia yuilli yuilli, Nyssomyia trapidoi, Pressatia triacantha, Pressatia dysponeta, Psychodopygus carrerai carrerai, Psychodopygus panamensis, and Trichophoromyia ubiquitalis were found positive for Leishmania parasite. These findings contribute to a better understanding of the epidemiology and transmission dynamics of the disease in high-risk areas of Ecuador. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America.

  3. Factores dermicos que condicionan la infeccion de Lutzomyia townsendi (Ortiz, 1959 por Leishmania spp. de Venezuela Dermal factors which condition infection of Lutzomyia townsendi (Ortiz, 1959 by Leishmania spp. of Venezuela

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    Maritza Carnevali

    1982-12-01

    Full Text Available Se estudia la susceptibilidad de Lutzomyia townsendi a la infección con Leishmania spp. sobre lesiones experimentales de hamsteres. Se estudia la frecuencia y distribución de los amastigotos en la dermis, relacionándola con la profundidad que alcanza el estilete bucal del insecto. Una correlación positiva, con significante coeficiente de correlación, se establece entre la frecuencia de los parásitos a una profundidad de 100-150 nm en la dermis y el éxito de la infección de los flebótomos.The susceptibility of Lutzomyia townsendi to infection with amastigotes of Leishmania spp. from experimental lesions in hamsters is studied. A good correlation is found when the number of infected sandflies are compared with the distribution and density of parasites in the dermis, taking into account the size of the fascicle, particularly the length of labrun and dental depth in maxillae. Considerations are made on the susceptibilities of some anthropophilic sandflies from America that are considered as good vectors.

  4. The role of gallery forests in maintaining Phlebotominae populations: potential Leishmania spp. vectors in the Brazilian savanna

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    Tâmara Dias Oliveira Machado

    Full Text Available BACKGROUND Knowledge on synanthropic phlebotomines and their natural infection by Leishmania is necessary for the identification of potential areas for leishmaniasis occurrence. OBJECTIVE To analyse the occurrence of Phlebotominae in gallery forests and household units (HUs in the city of Palmas and to determine the rate of natural infection by trypanosomatids. METHODS Gallery forests and adjacent household areas were sampled on July (dry season and November (rainy season in 2014. The total sampling effort was 960 HP light traps and eight Shannon traps. Trypanosomatids were detected in Phlebotominae females through the amplification of the SSU rDNA region, and the positive samples were used in ITS1-PCR. Trypanosomatid species were identified using sequencing. FINDINGS A total of 1,527 sand flies representing 30 species were captured in which 949 (28 spp. and 578 (22 spp. were registered in July and November, respectively. In July, more specimens were captured in the gallery forests than in the HUs, and Nyssomyia whitmani was particularly frequent. In November, most of the specimens were found in the HUs, and again, Ny. whitmani was the predominant species. Lutzomyia longipalpis was commonly found in domestic areas, while Bichromomyia flaviscutellata was most frequent in gallery forests. Molecular analysis of 154 pools of females (752 specimens identified Leishmania amazonensis, L. infantum, and Crithidia fasciculata in Ny. whitmani, as well as L. amazonensis in Lu. longipalpis, Trypanosoma sp. and L. amazonensis in Pintomyia christenseni, and L. amazonensis in both Psathyromyia hermanlenti and Evandromyia walkeri. MAIN CONCLUSIONS These results show the importance of gallery forests in maintaining Phlebotominae populations in the dry month, as well as their frequent occurrence in household units in the rainy month. This is the first study to identify Leishmania, Trypanosoma, and Crithidia species in Phlebotominae collected in Palmas, Tocantins

  5. The role of gallery forests in maintaining Phlebotominae populations: potential Leishmania spp. vectors in the Brazilian savanna.

    Science.gov (United States)

    Machado, Tâmara Dias Oliveira; Minuzzi-Souza, Thaís Tâmara Castro; Ferreira, Tauana de Sousa; Freire, Luciana Pereira; Timbó, Renata Velôzo; Vital, Tamires Emanuele; Nitz, Nadjar; Silva, Mariana Neiva; Santos, Alcinei de Souza; Sales, Nathyla Morgana Cunha; Obara, Marcos Takashi; Andrade, Andrey José de; Gurgel-Gonçalves, Rodrigo

    2017-10-01

    Knowledge on synanthropic phlebotomines and their natural infection by Leishmania is necessary for the identification of potential areas for leishmaniasis occurrence. To analyse the occurrence of Phlebotominae in gallery forests and household units (HUs) in the city of Palmas and to determine the rate of natural infection by trypanosomatids. Gallery forests and adjacent household areas were sampled on July (dry season) and November (rainy season) in 2014. The total sampling effort was 960 HP light traps and eight Shannon traps. Trypanosomatids were detected in Phlebotominae females through the amplification of the SSU rDNA region, and the positive samples were used in ITS1-PCR. Trypanosomatid species were identified using sequencing. A total of 1,527 sand flies representing 30 species were captured in which 949 (28 spp.) and 578 (22 spp.) were registered in July and November, respectively. In July, more specimens were captured in the gallery forests than in the HUs, and Nyssomyia whitmani was particularly frequent. In November, most of the specimens were found in the HUs, and again, Ny. whitmani was the predominant species. Lutzomyia longipalpis was commonly found in domestic areas, while Bichromomyia flaviscutellata was most frequent in gallery forests. Molecular analysis of 154 pools of females (752 specimens) identified Leishmania amazonensis, L. infantum, and Crithidia fasciculata in Ny. whitmani, as well as L. amazonensis in Lu. longipalpis, Trypanosoma sp. and L. amazonensis in Pintomyia christenseni, and L. amazonensis in both Psathyromyia hermanlenti and Evandromyia walkeri. These results show the importance of gallery forests in maintaining Phlebotominae populations in the dry month, as well as their frequent occurrence in household units in the rainy month. This is the first study to identify Leishmania, Trypanosoma, and Crithidia species in Phlebotominae collected in Palmas, Tocantins, Brazil.

  6. Molecular detection of Leishmania spp. in road-killed wild mammals in the Central Western area of the State of São Paulo, Brazil.

    Science.gov (United States)

    Richini-Pereira, Virginia Bodelão; Marson, Pamela Merlo; Hayasaka, Enio Yoshinori; Victoria, Cassiano; da Silva, Rodrigo Costa; Langoni, Hélio

    2014-01-01

    Road-killed wild animals have been classified as sentinels for detecting such zoonotic pathogens as Leishmania spp., offering new opportunities for epidemiological studies of this infection. This study aimed to evaluate the presence of Leishmania spp. and Leishmania chagasi DNA by PCR in tissue samples (lung, liver, spleen, kidney, heart, mesenteric lymph node and adrenal gland) from 70 road-killed wild animals. DNA was detected in tissues of one Cavia aperea (Brazilian guinea pig), five Cerdocyon thous (crab-eating fox), one Dasypus septemcinctus (seven-banded armadillo), two Didelphis albiventris (white-eared opossum), one Hydrochoerus hydrochoeris (capybara), two Myrmecophaga tridactyla (giant anteater), one Procyon cancrivorus (crab-eating raccoon), two Sphiggurus spinosus (porcupine) and one Tamandua tetradactyla (lesser anteater) from different locations in the Central Western part of São Paulo state. The Leishmania chagasi DNA were confirmed in mesenteric lymph node of one Cerdocyon thous. Results indicated common infection in wild animals. The approach employed herein proved useful for detecting the environmental occurrence of Leishmania spp. and L. chagasi, as well as determining natural wild reservoirs and contributing to understand the host-parasite interaction.

  7. Detección de Leishmania spp. en base al gen que codifica la proteína HSP20

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    Ana M Montalvo

    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 PCR 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 (κ = 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.

  8. Comparison of Tetrazolium Salt Assays for Evaluation of Drug Activity against Leishmania spp.

    Science.gov (United States)

    Ginouves, Marine; Carme, Bernard; Couppie, Pierre

    2014-01-01

    In French Guiana, leishmaniasis is an essentially cutaneous infection. It constitutes a major public health problem, with a real incidence of 0.2 to 0.3%. Leishmania guyanensis is the causal species most frequently encountered in French Guiana. The treatment of leishmaniasis is essentially drug based, but the therapeutic compounds available have major side effects (e.g., liver damage and diabetes) and must be administered parenterally or are costly. The efficacy of some of these agents has declined due to the emergence of resistance in certain strains of Leishmania. There is currently no vaccine against leishmaniasis, and it is therefore both necessary and urgent to identify new compounds effective against Leishmania. The search for new drugs requires effective tests for evaluations of the leishmanicidal activity of a particular molecule or extract. Microculture tetrazolium assays (MTAs) are colorimetric tests based on the use of tetrazolium salts. We compared the efficacies of three tetrazolium salts—3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide (XTT), and 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium (WST-8)—for quantification of the promastigotes of various species of Leishmania. We found that the capacity of Leishmania to metabolize a tetrazolium salt depended on the salt used and the species of Leishmania. WST-8 was the tetrazolium salt best metabolized by L. guyanensis and gave the best sensitivity. PMID:24719447

  9. Presence of anti-Toxoplasma gondii, -Neospora caninum, -Leishmania spp. and -Ehrlichia canis antibodies in free-ranging maned wolves (Chrysocyon brachyurus in the northeastern region of the state of São Paulo, Brazil

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    Solange Oliveira

    2016-09-01

    Full Text Available O lobo-guará (Chrysocyon brachyurus habita o ecossistema de Cerrado e é considerado o maior canídeo da América do Sul e uma espécie ameaçada de extinção pela "International Union for Conservation of Nature" (IUNC. O objetivo desse estudo foi investigar a presença de anticorpos anti-Toxoplasma gondii, -Neospora caninum, -Leishmania spp. e -Ehrlichia canis em lobos-guará da região nordeste do estado de São Paulo, Brasil. Das 17 amostras de soro testadas por meio da reação de imunofluorescência indireta (RIFI, 88,2% (15/17, 17,6% (3/17 e 52,9% (9/17 apresentaram anticorpos anti-T. gondii, -Leishmania spp. e -E. canis, respectivamente. Todos os animais testados foram soronegativos para N. caninum. Esses resultados indicam a exposição dos lobos-guará dessa região aos agentes pesquisados. A presença de um complexo industrial, agricultura extensiva e fragmentação de habitat na região nordeste do estado de São Paulo, favorece a proximidade desses animais silvestres a ambientes urbanos o que pode contribuir para a transmissão de doenças entre os animais silvestres, domésticos e o homem.

  10. Effect of aliphatic, monocarboxylic, dicarboxylic, heterocyclic and sulphur-containing amino acids on Leishmania spp. chemotaxis.

    Science.gov (United States)

    Diaz, E; Zacarias, A K; Pérez, S; Vanegas, O; Köhidai, L; Padrón-Nieves, M; Ponte-Sucre, A

    2015-11-01

    In the sand-fly mid gut, Leishmania promastigotes are exposed to acute changes in nutrients, e.g. amino acids (AAs). These metabolites are the main energy sources for the parasite, crucial for its differentiation and motility. We analysed the migratory behaviour and morphological changes produced by aliphatic, monocarboxylic, dicarboxylic, heterocyclic and sulphur-containing AAs in Leishmania amazonensis and Leishmania braziliensis and demonstrated that L-methionine (10-12 m), L-tryptophan (10-11 m), L-glutamine and L-glutamic acid (10-6 m), induced positive chemotactic responses, while L-alanine (10-7 m), L-methionine (10-11 and 10-7 m), L-tryptophan (10-11 m), L-glutamine (10-12 m) and L-glutamic acid (10-9 m) induced negative chemotactic responses. L-proline and L-cysteine did not change the migratory potential of Leishmania. The flagellum length of L. braziliensis, but not of L. amazonensis, decreased when incubated in hyperosmotic conditions. However, chemo-repellent concentrations of L-alanine (Hypo-/hyper-osmotic conditions) and L-glutamic acid (hypo-osmotic conditions) decreased L. braziliensis flagellum length and L-methionine (10-11 m, hypo-/hyper-osmotic conditions) decreased L. amazonensis flagellum length. This chemotactic responsiveness suggests that Leishmania discriminate between slight concentration differences of small and structurally closely related molecules and indicates that besides their metabolic effects, AAs play key roles linked to sensory mechanisms that might determine the parasite's behaviour.

  11. Risk of Infection with Leishmania spp. in the Canine Population in the Netherlands

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    Slappendel RJ

    2002-12-01

    Full Text Available The dog is the main reservoir of Leishmania infantum, the causative agent of visceral leishmaniasis (VL in humans in Southern Europe. In order to identify the risk of dogs from a Leishmania non-endemic area traveling to a Leishmania-endemic area becoming infected and the risk of transmitting infection to humans in non-endemic areas an investigation was performed, in which the results of a questionnaire were combined with the results of a serologic survey. The questionnaire was sent to 1478 at random chosen families in the Netherlands. Of the 59.0% responders 28.0% had one or more dogs and 4.8% of these dogs had visited Southern Europe during the summer period of that year. On a total population of 1,200,000 dogs in the Netherlands, this means that each year some 58,000 dogs are at risk of being exposed to a Leishmania infection in Southern Europe. During the period 1990–1992 blood was collected for serology in 1911 dogs presented to the Utrecht University Clinic because of clinical problems not related to leishmaniasis, of which 434 had been in Southern Europe in the foregoing years. None was serologically positive. From these data it can be deduced that the highest chance to obtain leishmaniasis during a vacation in Southern Europe is mathematically less than 1/434 or less than 0.23%. Serology was also performed during the period 1989–1993 in 597 dogs that had been in Southern Europe and were suspected of leishmaniasis. Titers were positive in 145 of these samples. Sixty-four of these dogs were born in the Mediterranean and had been imported into the Netherlands. Excluding these imported dogs, it was calculated that at least 0.027% of the 58,000 dogs yearly taken to Southern Europe during holidays become infected with Leishmania. In order to establish the risk of disease transmission for people in close contact with an infected dog, serum samples of owners and house mates of 37 dogs with leishmaniasis were tested. All 112 sera tested

  12. Spatial and simultaneous seroepidemiology of anti-Leishmania spp. antibodies in dog owners and their dogs from randomly selected households in a major city of southern Brazil.

    Science.gov (United States)

    Benitez, Aline do Nascimento; Martins, Felippe Danyel Cardoso; Mareze, Marcelle; Nino, Beatriz de Souza Lima; Caldart, Eloiza Teles; Ferreira, Fernanda Pinto; Mitsuka-Breganó, Regina; Freire, Roberta Lemos; Galhardo, Juliana Arena; Martins, Camila Marinelli; Biondo, Alexander Welker; Navarro, Italmar Teodorico

    2018-06-01

    Although leishmaniasis has been described as a classic example of a zoonosis requiring a comprehensive approach for control, to date, no study has been conducted on the spatial distribution of simultaneous Leishmania spp. seroprevalence in dog owners and dogs from randomly selected households in urban settings. Accordingly, the present study aimed to simultaneously identify the seroprevalence, spatial distribution and associated factors of infection with Leishmania spp. in dog owners and their dogs in the city of Londrina, a county seat in southern Brazil with a population of half a million people and ranked 18th in population and 145th in the human development index (HDI) out of 5570 Brazilian cities. Overall, 564 households were surveyed and included 597 homeowners and their 729 dogs. Anti-Leishmania spp. antibodies were detected by ELISA in 9/597 (1.50%) dog owners and in 32/729 (4.38%) dogs, with significantly higher prevalence (p = 0.0042) in dogs. Spatial analysis revealed associations between seropositive dogs and households located up to 500 m from the local railway. No clusters were found for either owner or dog case distributions. In summary, the seroepidemiological and spatial results collectively show a lack of association of the factors for infection, and the results demonstrated higher exposure for dogs than their owners. However, railway areas may provide favorable conditions for the maintenance of infected phlebotomines, thereby causing infection in nearby domiciled dogs. In such an urban scenario, local sanitary barriers should be focused on the terrestrial routes of people and surrounding areas, particularly railways, via continuous vector surveillance and identification of phlebotomines infected by Leishmania spp. Copyright © 2018. Published by Elsevier B.V.

  13. Validación de la PCR en la detección de parásitos de Leishmania (Viannia spp. en Lutzomyia (Diptera: Psychodidae como herramienta en la definición de especies vectores.

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    Erika Santamaría

    2005-06-01

    Full Text Available lntroducción. En leishmaniasis se acepta que la reacción en cadena de la polimerasa (PCR ha simplificado el proceso de incriminación vectorial. Sin embargo, pocas veces se ha determinado la sensibilidad y la especificidad de cada PCR en la detección y la identificación del parasito en los flebótomos. Objetivo. Evaluar la aplicabilidad de la reacción en cadena de la polimerasa (PCR, basada en los iniciadores 81 y 82, en la detecci6n e identificación de parasitos de Leishmania (Viannia en insectos vectores enteros sin disecar. Metodología. Se determinó la sensibilidad y la especificidad de la PCR empleando diluciones de cultivo de parasitos de Leishmania spp. Se estableció el número máximo de hembras de Lutzomyia que pueden ser procesadas a la vez sin disminuir la sensibilidad de la PCR, procesando el ADN de grupos de una a cinco hembras de Lutzomyia en presencia del ADN de las diluciones de cultivo de parasitos. Además, se comparó en grupos de flebótomos infectados experimentalmente, la sensibilidad de esta PCR en la detección de infección por Leishmania (Viannia frente al método de busqueda de flagelados por disección del insecto y examen microscópico. Resultados. La PCR detectó desde un parasito de Leishmania (Viannia y permiti6 el procesamiento de hasta tres insectos enteros sin alterar la sensibilidad. Los porcentajes de infección experimental detectados con las dos tecnicas fueron similares, 33,3% con la PCR y 30% con el examen microscópico. Además, se confirmó que los iniciadores 81 y 82 son especificos para especies del subgenero Leishmania (Viannia. Conclusión. Los resultados obtenidos demuestran la sensibilidad y la especificidad de esta PCR y permiten recomendar su uso en la determinaci6n de infección natural con parasitos de Leishmania (Viannia en poblaciones silvestres de flebótomos.

  14. Hepatozoon canis and Leishmania spp. coinfection in dogs diagnosed with visceral leishmaniasis

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    Fernanda Nazaré Morgado

    Full Text Available Abstract This study describes the occurrence of dogs naturally co-infected with Hepatozoon canis and two Leishmania species: L. infantum or L. braziliensis. Four dogs serologically diagnosed with Visceral Leishmaniasis were euthanized. Liver and spleen samples were collected for histopathological analysis and DNA isolation. H. canis meronts were observed in tissues from all four dogs. H. canis infection was confirmed by PCR followed by sequencing of a fragment of 18S rRNA gene. Leishmania detection and typing was confirmed by ITS1' PCR-RFLP and parasite burden was calculated using ssrRNA quantitative qPCR. A DPP - Dual Path platform test was performed. One out (Dog #2 of four animals was asymptomatic. Dogs #1 and #4 were infected by L. infantum and were DPP test positive. Dogs #2 and #3 were infected by L. braziliensis and were DPP test negative. Furthermore, visceral dissemination was observed in Dogs #2 and #3, since L. braziliensis was detected in liver and spleen samples. The visceral dissemination of L. braziliensis associated with systemic signs suggested that this co-infection could influence the parasite burden and disease progression.

  15. Hepatozoon canis and Leishmania spp. coinfection in dogs diagnosed with visceral leishmaniasis.

    Science.gov (United States)

    Morgado, Fernanda Nazaré; Cavalcanti, Amanda Dos Santos; Miranda, Luisa Helena de; O'Dwyer, Lúcia Helena; Silva, Maria Regina Lucas da; Menezes, Rodrigo Caldas; Andrade da Silva, Aurea Virgínia; Boité, Mariana Côrtes; Cupolillo, Elisa; Porrozzi, Renato

    2016-01-01

    This study describes the occurrence of dogs naturally co-infected with Hepatozoon canis and two Leishmania species: L. infantum or L. braziliensis. Four dogs serologically diagnosed with Visceral Leishmaniasis were euthanized. Liver and spleen samples were collected for histopathological analysis and DNA isolation. H. canis meronts were observed in tissues from all four dogs. H. canis infection was confirmed by PCR followed by sequencing of a fragment of 18S rRNA gene. Leishmania detection and typing was confirmed by ITS1' PCR-RFLP and parasite burden was calculated using ssrRNA quantitative qPCR. A DPP - Dual Path platform test was performed. One out (Dog #2) of four animals was asymptomatic. Dogs #1 and #4 were infected by L. infantum and were DPP test positive. Dogs #2 and #3 were infected by L. braziliensis and were DPP test negative. Furthermore, visceral dissemination was observed in Dogs #2 and #3, since L. braziliensis was detected in liver and spleen samples. The visceral dissemination of L. braziliensis associated with systemic signs suggested that this co-infection could influence the parasite burden and disease progression.

  16. Arrabidaea chica Hexanic Extract Induces Mitochondrion Damage and Peptidase Inhibition on Leishmania spp.

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    Igor A. Rodrigues

    2014-01-01

    Full Text Available Currently available leishmaniasis treatments are limited due to severe side effects. Arrabidaea chica is a medicinal plant used in Brazil against several diseases. In this study, we investigated the effects of 5 fractions obtained from the crude hexanic extract of A. chica against Leishmania amazonensis and L. infantum, as well as on the interaction of these parasites with host cells. Promastigotes were treated with several concentrations of the fractions obtained from A. chica for determination of their minimum inhibitory concentration (MIC. In addition, the effect of the most active fraction (B2 on parasite’s ultrastructure was analyzed by transmission electron microscopy. To evaluate the inhibitory activity of B2 fraction on Leishmania peptidases, parasites lysates were treated with the inhibitory and subinhibitory concentrations of the B2 fraction. The minimum inhibitory concentration of B2 fraction was 37.2 and 18.6 μg/mL for L. amazonensis and L. infantum, respectively. Important ultrastructural alterations as mitochondrial swelling with loss of matrix content and the presence of vesicles inside this organelle were observed in treated parasites. Moreover, B2 fraction was able to completely inhibit the peptidase activity of promastigotes at pH 5.5. The results presented here further support the use of A. chica as an interesting source of antileishmanial agents.

  17. Infection by Leishmania spp. in Free-Ranging Opossums (Didelphis albiventris) in an Environmentally Protected Area Inhabited by Humans in Southeastern Brazil.

    Science.gov (United States)

    Paiz, Laís Moraes; Donalisio, Maria Rita; Richini-Pereira, Virgínia Bodelão; Motoie, Gabriela; Castagna, Claudio Luiz; Tolezano, José Eduardo

    2016-11-01

    There is a growing concern about the participation of wild hosts and reservoirs in the epidemiology of leishmaniasis, particularly within the context of increasingly frequent environmental changes and the expansion of the One Health concept. This work is a molecular research of infection by Leishmania spp. among the wildlife of an environmentally protected area located in the municipality of Campinas, São Paulo, Brazil. The studied area has a history of intense environmental changes, with notifications of human cases of cutaneous leishmaniasis in the 1990s, and a focus of canine visceral leishmaniasis since 2009. Eighty-two wild mammals were sampled by monthly captures in this region over a 1-year period. Blood samples were collected from each animal and subjected to DNA extraction and PCR using primers for the region of the internal transcribed spacer-1. The results of gene sequencing for the first time revealed the infection of opossums (Didelphis albiventris) by Leishmania spp., subgenera Leishmania and Viannia, in Campinas. These findings, in addition to environmental and historical characteristics of the studied area, indicate a possible role of wildlife in the introduction and/or maintenance of natural foci of leishmaniasis transmission.

  18. 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.

  19. 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)

    Cardoso, Andre Gustavo Tempone

    1999-01-01

    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 μg/ml (EC 50 ), 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 EC 50 of 11|μg/ml. Irradiation of LAO with 6 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 50 of 0.67mM. These data could be of help in the development of alternative therapeutic approaches to the treatment of leishmaniasis. (author)

  20. Detection of Leishmania spp. using parasitological, serological and molecular assays in asymptomatic and sick cats from an endemic area of visceral leishmaniosis in Brazil

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    Maria Fernanda Alves-Martin

    2017-11-01

    Full Text Available Objective: To analyze the prevalence of feline leishmaniosis in 55 asymptomatic cats or assess clinical disease in a visceral leishmaniasis-endemic area using parasitological, serological and molecular techniques. Methods: Fifty-five adult domestic and non-purebred cats held at an animal shelter in a rural area of the municipality of Ilha Solteira, São Paulo, Brazil, a region endemic for canine visceral leishmaniosis, were evaluated. A total of 5 mL of blood was collected from each animal, for blood culture, serological (indirect fluorescent antibody test, soluble extract ELISA, ELISA recombinant K39 antigen and molecular tests (PCR. At the time of blood collection, the cats underwent clinical evaluation for the presence of clinical signs. Results: Flagellate protozoa were found in nine blood cultures (16.4%. Seropositivity by indirect fluorescent antibody test was observed in 32 serum samples (62.7%. ELISA-SE showed seropositivity in 37 cats (72.5%, while ELISA-rK39 revealed positive results in 11 cats (21.6%. Leishmania spp. was detected by PCR in whole blood samples from five cats (9.1%. Based on clinical evaluation, it was possible to detect the presence of clinical signs (alopecia, emaciation, pinna lesions, nose lesions, skin lesions in 30 animals (54.5%. Conclusions: To the knowledge of the authors, this is the first report of Leishmania spp. in domestic cats in the epidemiological cycle of leishmaniosis in the studied area, requiring more attention in the establishment of preventive steps to control this disease.

  1. Apagando genes para iluminar la interacción entre el macrófago y Leishmania

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    Carlos Arturo Clavijo-Ramirez

    2016-02-01

    Full Text Available La enfermedad causada por los parásitos del género Leishmania representa un problema de salud pública de importancia a nivel nacional e internacional. El conocimiento actual de la interacción entre el macrófago humano y el parásito aún luce insuficiente para reflejarse en mejoras del tratamiento empleado. El presente manuscrito aspira a ser un texto de divulgación para el púbico general y trata de explicar la importancia de este tipo de investigación científica y la estrategia empleada por nuestro grupo de investigación, en la búsqueda de soluciones al flagelo representado por los diferentes tipos de Leishmaniasis.

  2. 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. © The American Society of Tropical Medicine and Hygiene.

  3. 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)

  4. Caracteres moleculares para la determinación taxonómica de tres especies de Lutzomyia (Diptera: Psychodidae, vectores potenciales de Leishmania presentes en el valle de Aburrá, Colombia Molecular characters for the taxonomic determination of three species of Lutzomyia (Diptera: Psychodidae, potential Leishmania vectors found in the Aburrá valley, Colombia

    Directory of Open Access Journals (Sweden)

    Alveiro Pérez-Doria

    2008-12-01

    Full Text Available En Colombia están registradas 143 especies de Lutzomyia França, pero menos del 7% de éstas se encuentran incriminadas como vectores de Leishmania spp. Debido a la alta semejanza morfológica de algunas especies vectoras con otras no vectoras, se necesitan caracteres taxonómicos alternativos para identificar correctamente los flebotomíneos de cada zona geográfica del país. Con este objetivo, en el presente trabajo se secuenció el extremo 3' del gen mitocondrial que codifica para la proteína citocromo b en tres vectores potenciales de Leishmania presentes en el valle de Aburrá, Colombia, Lutzomyia hartmanni (Fairchild y Hertig, L. columbiana (Ristorcelli y Van Ty y L. tihuiliensis Le Pont, Torrez-Espejo y Dujardin. A partir del alineamiento múltiple de nucleótidos se determinaron los sitios polimórficos, las distancias genéticas pareadas netas (p y la entropía. Las secuencias de nucleótidos fueron trasladadas a aminoácidos para estimar el número de sustituciones sinónimas y no sinónimas. En el alineamiento múltiple de 321 nucleótidos del gen citocromo b de L. columbiana, L. hartmanni y L. tihuiliensis se detectaron 83 sustituciones. En la secuencia parcial de la proteína se encontraron 18 reemplazos de aminoácidos. Las distancias genéticas interespecíficas fluctuaron en un rango mínimo de 0,137 entre L. tihuiliensis y L. columbiana, y un máximo de 0,215 entre L. columbiana y L. hartmanni. Los polimorfismos detectados en la secuencia de nucleótidos del gen y de aminoácidos de la proteína constituyen caracteres moleculares potencialmente útiles para la determinación taxonómica de estas especies de flebotomíneos.To date, 143 species of Lutzomyia França are recorded in Colombia, but less than 7% is incriminated in the transmission of Leishmania spp. Alternative taxonomic characters are necessary to correctly identify the particular sand fly fauna in each Colombian region, and the separation of morphologically

  5. Molecular Identification of Leishmania spp. in Sand Flies (Diptera: Psychodidae: Phlebotominae) in the Lençóis Maranhenses National Park, Brazil.

    Science.gov (United States)

    Pereira-Filho, Adalberto Alves; Fonteles, Raquel Silva; Bandeira, Maria da Conceição Abreu; Moraes, Jorge Luiz Pinto; Rebêlo, José Manuel Macário; Melo, Maria Norma

    2018-02-20

    Sand flies are very common in the region of Lençóis Maranhenses National Park, an important tourist attraction in Brazil. However, the role of some species and their relative importance locally in Leishmania Ross 1903 transmission is unclear. The objective of this study was to identify Leishmania infection in phlebotomine sand flies collected around the Lençóis Maranhenses National Park, an important conservation area and popular international/national tourist destination with a high incidence of leishmaniasis. Sand flies were collected in peridomiciliary areas on the tourist route from September 2012 to August 2013. The captured females were subjected to molecular analyses for the detection of Leishmania DNA. Sand flies were infected with four Leishmania species: Leishmania (Viannia) braziliensis (Vianna, 1911) was found in Lutzomyia whitmani (Antunes and Coutinho, 1939) (2.1%) and Lutzomyia longipalpis (Lutz and Neiva, 1912) (1.7%); Leishmania (Leishmania) infantum (Nicole, 1908) infected Lutzomyia wellcomei (Fraiha, Shaw, and Lainson, 1971) (20%), Lutzomyia sordellii (Shannon and Del Ponte, 1927) (4.3%), Lu. longipalpis (3.7%), and Lu. whitmani (0.8%); Leishmania (Leishmania) amazonensis (Lainson & Shaw, 1972) was found in Lu. whitmani (0.58%), while Leishmania (Viannia) lainsoni infected Lutzomyia evandroi (Costa Lima and Antunes, 1936) (3.4%), Lu. longipalpis (1.06%), and Lu. whitmani (0.29%). The occurrence of these parasites requires control measures to reduce the incidence of cutaneous leishmaniasis and to contain a possible epidemic of visceral leishmaniasis, the most severe form of the disease.

  6. 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

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    Jennings Yara Lins

    2014-01-01

    Full Text Available 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.

  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. Diseño y evaluación de tres oligonucleótidos para la detección de Leishmania por PCR

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    Omar Cáceres Rey

    2002-07-01

    Full Text Available La leishmaniasis afecta la salud pública en 88 países del mundo y representa un serio obstáculo para su desarrollo socioeconómico. Los métodos para diagnosticar la enfermedad toman tiempo y muchas veces son traumáticos para el paciente. Objetivo: Se aplicó PCR como método alternativo para el diagnóstico rápido de esta enfermedad. Materiales y métodos: A diferentes especies de Leishmania se les extrajo ADN genómico. Se diseñaron tres oligonucleótidos (LEISH 1, LEISH 2 y LEISH 3 dirigidos al extremo carboxilo terminal de la histona H2B de Leishmania (V. peruviana cuya secuencia parcial sirvió para diseñar dichos oligos, los cuales fueron probados en un sistema de PCR. Resultados: Los oligonucleótidos amplificaron exitosamente regiones de 123 pb (LEISH 1 / LEISH 2 y 139 pb (LEISH 1 / LEISH 3 de esta secuencia parcial utilizada. La sensibilidad del PCR fue de hasta 1 fg de ADN purificado de L. (V. peruviana y de 2 parásitos cuando se realizó la técnica de manera directa. La especificidad fue 100% (solo reconoció a Leishmania y no a Trypanosoma ni a humano. Los oligonucleótidos diseñados también amplificaron todas las especies de Leishmania evaluadas. Conclusión: El sistema de PCR diseñado puede ser aplicado en la detección del parásito a partir de cualquier tipo de muestra convirtiéndose en un método alternativo de diagnóstico de la enfermedad por su rapidez, especificidad y sensibilidad.

  9. Ciclo de vida y colonización de Lutzomyia ovallesi (Diptera: Psychodidae, vector de Leishmania spp. en América Latina

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    Olga L. Cabrera

    1999-09-01

    Full Text Available Lutzomyia ovallesi (Diptera: Psychodidae, presente en un foco de Leishmania (V braziliensis en Reventones, Cundinamarca, reg- ión ubicada en el centro de Colombia. se crió y mantuvo por catorce generaciones bajo condiciones de laboratorio con una temperatura diaria entre 22-28 ""C y una humedad relativa entre 90 y 98%. Después de la emergencia, se llevaron machos y hembras a vasos de poliestireno (transparente y se mantuvieron en una nevera de icopor. La duración de cada uno de los estadios del ciclo de vida observado en las crías individuales fue: huevo 9-12 días (X = 10, 42, DE = 0,86; larva 17-11 días (X=9,9, DE=3,06; larva 11 7-13 días (X=9,5, DE= 3,06; larva III 4-12 (X=7,4, DE=1,50; larva IV 9-19 días (X=15,16, DE=2,66 y pupa 8-16 días (X=11, DE=1,89. El tiempo promedio para el desarrollo total del ciclo biológico fue de 63,36 días. La longevidad para las hembras presentó un rango entre 5 y 10 días (X=9,36 y para los machos entre 4 y 13 días (X=7,44. Con respecto a la productividad registrada a partir de un promedio de 30 hembras de Lutzomyia por vaso de cría, se obtuvo una producción mínima de 260 flebótomos en la quinta generación y una máxima de 4.51 1 en la décimotercera generación.

  10. Resistencia antimicrobiana de Salmonella spp aislada de alimentos de origen animal para consumo humano

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    Adriana Quesada

    2016-01-01

    Full Text Available Objetivos. Analizar la información disponible sobre la resistencia antimicrobiana de aislamientos de Salmonella spp de alimentos de origen animal para consumo humano en América Latina. Materiales y métodos. Se realizó una revisión sistemática de estudios epidemiológicos observacionales realizados en América Latina entre los años 2003 y 2014 en las bases de datos PubMed y LILACS. Se excluyeron estudios realizados como parte de estudio de brotes o casos de infecciones en humanos. Tres revisores de forma independiente participaron en la selección de estudios. Además, se realizó la evaluación de calidad a los estudios incluidos. Resultados. Un total de 25 estudios cumplieron con los criterios de inclusión. Los estudios incluidos fueron realizados en Brasil, México, Colombia, Argentina y Venezuela. Los aislamientos de Salmonella spp se obtuvieron principalmente de alimentos de origen avícola, porcino y vacuno, siendo Salmonella typhimurium y Salmonella enteritidis los serotipos que se aislaron con mayor frecuencia (17 y 11 estudios, respectivamente. En 23 de los estudios, Salmonella spp fue resistente a más de un antibiótico, incluyendo ácido nalidíxico, estreptomicina, tetraciclina, cloranfenicol, ampicilina, trimetoprim/sulfametoxazol, gentamicina, ciprofloxacina y cefalosporinas. Conclusiones. Los aislamientos de Salmonella spp obtenidos de alimentos de origen animal para consumo humano en los países analizados presentan con frecuencia resistencia a múltiples antibióticos. Es importante que más países en América Latina realicen y publiquen estudios sobre la resistencia de Salmonella spp para establecer y monitorear estrategias de control adecuadas.

  11. Caracteres moleculares para la determinación taxonómica de tres especies de Lutzomyia (Diptera: Psychodidae, vectores potenciales de Leishmania presentes en el valle de Aburrá, Colombia

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    Alveiro PÉREZ-DORIA

    2008-01-01

    Full Text Available En Colombia están registradas 143 especies de Lutzomyia França, pero menos del 7% de éstas se encuentran incriminadas como vectores de Leishmania spp. Debido a la alta semejanza morfológica de algunas especies vectoras con otras no vectoras, se necesitan caracteres taxonómicos alternativos para identificar correctamente los flebotomíneos de cada zona geográfica del país. Con este objetivo, en el presente trabajo se secuenció el extremo 3’ del gen mitocondrial que codifica para la proteína citocromo b en tres vectores potenciales de Leishmania presentes en el valle de Aburrá, Colombia, Lutzomyia hartmanni (Fairchild y Hertig, L. columbiana (Ristorcelli y Van Ty y L. tihuiliensis Le Pont, Torrez-Espejo y Dujardin. A partir del alineamiento múltiple de nucleótidos se determinaron los sitios polimórficos, las distancias genéticas pareadas netas (p y la entropía. Las secuencias de nucleótidos fueron trasladadas a aminoácidos para estimar el número de sustituciones sinónimas y no sinónimas. En el alineamiento múltiple de 321 nucleótidos del gen citocromo b de L. columbiana, L. hartmanni y L. tihuiliensis se detectaron 83 sustituciones. En la secuencia parcial de la proteína se encontraron 18 reemplazos de aminoácidos. Las distancias genéticas interespecíficas fluctuaron en un rango mínimo de 0,137 entre L. tihuiliensis y L. columbiana, y un máximo de 0,215 entre L. columbiana y L. hartmanni. Los polimorfismos detectados en la secuencia de nucleótidos del gen y de aminoácidos de la proteína constituyen caracteres moleculares potencialmente útiles para la determinación taxonómica de estas especies de flebotomíneos.

  12. Eco-epidemiological aspects, natural detection and molecular identification of Leishmania spp. in Lutzomyia reburra, Lutzomyia barrettoi majuscula and Lutzomyia trapidoi

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    Jazzmín Arrivillaga-Henríquez

    2017-03-01

    Conclusion: Natural infection with Le. amazonensis was recorded for the first time in Lu. reburra and Lu. b. majuscula, demonstrating the importance of zoophilic phlebotomines in the maintenance of the Leishmania transmission cycle in endemic foci.

  13. Leishmania hijacking of the macrophage intracellular compartments.

    Science.gov (United States)

    Liévin-Le Moal, Vanessa; Loiseau, Philippe M

    2016-02-01

    Leishmania spp., transmitted to humans by the bite of the sandfly vector, are responsible for the three major forms of leishmaniasis, cutaneous, diffuse mucocutaneous and visceral. Leishmania spp. interact with membrane receptors of neutrophils and macrophages. In macrophages, the parasite is internalized within a parasitophorous vacuole and engages in a particular intracellular lifestyle in which the flagellated, motile Leishmania promastigote metacyclic form differentiates into non-motile, metacyclic amastigote form. This phenomenon is induced by Leishmania-triggered events leading to the fusion of the parasitophorous vacuole with vesicular members of the host cell endocytic pathway including recycling endosomes, late endosomes and the endoplasmic reticulum. Maturation of the parasitophorous vacuole leads to the intracellular proliferation of the Leishmania amastigote forms by acquisition of host cell nutrients while escaping host defense responses. © 2015 FEBS.

  14. Estudo da competência vetorial de Lutzomyia intermedia (Lutz & Neiva, 1912 para Leishmania (Viannia braziliensis, Vianna, 1911 Study of the vectorial competence of Lutzomyia intermedia (Lutz & Neiva, 1912 to Leishmania (Viannia braziliensis, Vianna, 1911

    Directory of Open Access Journals (Sweden)

    Antonio Carlos da Silva

    2001-04-01

    Full Text Available Estudou-se a competência vetorial de Lutzomyia intermedia (Diptera: Psychodidae do Vale do Ribeira (SP para estirpes de Leishmania (Viannia braziliensis (Kinetoplastida: Trypanosomatidae, mediante pesquisa de infectividade natural; exposições de fêmeas silvestres e colonizadas (F1 às lesões de hamsters experimentalmente infectados e testes de transmissão via picada. A infectividade natural e os testes de transmissão revelaram-se negativos e, nas exposições, foram obtidas positividades de 74% (123+/166 dissecados e 70% (115+/164 dissecados para fêmeas silvestres e colonizadas respectivamente, e o desenvolvimento das formas evolutivas compatíveis com o modelo Peripilaria. A suscetibilidade às estirpes testadas associada aos indicadores epidemiológicos concorrem para a suspeita do papel vetorial de Lutzomyia intermedia na região estudada.This paper investigated the vectorial competence of Lutzomyia intermedia (Diptera: Psychodidae in Vale do Ribeira (SP to strains of Leishmania (Viannia braziliensis (Kinetoplastida: Trypanosomatidae, by means of a search for natural infection; exposure of wild and colonized females (F1 to the lesions of experimentally infected hamsters and transmission tests by bite. The natural infection and the transmission tests were negative. In the exposures of Lu. intermedia to infected lesions we found rates of 74% (123+/166 dissected and 70% (115+/164 dissected for the wild and colonized females respectively. The development of the parasites was compatible with the development model of Peripilaria. The susceptibility of the tested strains associated with the epidemiological indicators contribute to the vectorial role suspicion of Lutzomyia intermedia in the studied region.

  15. [Eco-epidemiological aspects, natural detection and molecular identification of Leishmania spp. in Lutzomyia reburra, Lutzomyia barrettoi majuscula and Lutzomyia trapidoi].

    Science.gov (United States)

    Arrivillaga-Henríquez, Jazzmín; Enríquez, Sandra; Romero, Vanessa; Echeverría, Gustavo; Pérez-Barrera, Jorge; Poveda, Ana; Navarro, Juan-Carlos; Warburg, Alon; Benítez, Washington

    2017-03-29

    The province of Pichincha in Ecuador is an endemic area of cutaneous leishmaniasis, where anthropophilic sand flies with natural infection by Leishmania, have been reported as vectors. However, the role in transmission of zoophilic species has not been evaluated. To evaluate natural infection by Leishmania in two zoophilic phlebotomine sand fly species, Lutzomyia reburra and Lu. barrettoi majuscula, and one anthropophilic species, Lu. trapidoi, as well as the endophagy and synanthropism of these species in the northwest of Pichincha. Phlebotomines were collected using CDC light traps in different habitats and altitudes with presence of cutaneous leishmaniasis. Leishmania infection was detected using genomic DNA from females of the collected sand flies. We amplified the internal transcribed spacer gene of ribosomal RNA I (ITS1), the mitochondrial topoisomerase II gene (mtTOPOII), and the nuclear topoisomerase II gene (TopoII). Percentages of positivity for Leishmania, at spatio-temporal scale, proportion of endophagy and synanthropism index were calculated. Natural infection was determined for Le. amazonensis in Lu. reburra (9.5%) and Lu. b. majuscula (23.8%), while in Lu. trapidoi we detected Le. amazonensis, Le. brazilienis and Le. naiffi-lainsoni. Phlebotomines were asynanthropic and with low endophagy. Natural infection with Le. amazonensis was recorded for the first time in Lu. reburra and Lu. b. majuscula, demonstrating the importance of zoophilic phlebotomines in the maintenance of the Leishmania transmission cycle in endemic foci.

  16. Evaluation of Trichoderma spp. and Clonostachys spp. Strains to Control Fusarium circinatum in Pinus radiata Seedlings Evaluación de Cepas de Trichoderma spp. y Clonostachys spp. para Controlar Fusarium circinatum en Plántulas de Pinus radiata

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    Priscila Moraga-Suazo

    2011-09-01

    os. Debido a la ausencia de métodos eficientes para controlar este patógeno, el propósito de este estudio fue evaluar el potencial uso de cepas de Trichoderma y Clonostachys como control de F. circinatum en plántulas de Pinus radiata D. Don. Ochenta y un cepas de Trichoderma spp. y Clonostachys spp. fueron evaluadas a través de ensayos in vitro para determinar su habilidad para actuar como antagonistas de F. circinatum y 21 cepas fueron probadas por su habilidad para reducir la mortalidad post-emergencia e incrementar la supervivencia de P. radiata bajo condiciones de invernadero. Durante los experimentos in vitro, 15 cepas de Trichoderma inhibieron el crecimiento del patógeno más de un 60% y una cepa de Clonostachys mostró parasitismo de hifas de F. circinatum. En experimentos en invernadero, los antagonistas no controlaron la enfermedad cuando fueron agregados después de la inoculación del patógeno. Sin embargo, cuando los antagonistas fueron sembrados antes del patógeno, cuatro cepas (Clonostachys UDC-32 y UDC-222, Trichoderma UDC-23 y UDC-408 redujeron la mortalidad post-emergencia entre 80 y 100%. Entre estas cepas, sólo Clonostachys UDC-222 incrementó significativamente la supervivencia de plántulas de P. radiata. Estos resultados demuestran que Clonostachys UDC-222 posee gran potencial para ser empleado como agente de biocontrol contra F. circinatum en la producción de plantas de P. radiata.

  17. Molecular detection and identification of Leishmania spp. in naturally infected Phlebotomus tobbi and Sergentomyia dentata in a focus of human and canine leishmaniasis in western Turkey.

    Science.gov (United States)

    Özbel, Yusuf; Karakuş, Mehmet; Arserim, Suha K; Kalkan, Şaban Orçun; Töz, Seray

    2016-03-01

    Human visceral leishmaniasis (VL) is reported from 38 provinces of Turkey and dogs are accepted as main reservoir hosts. Kuşadası town, belonging to Aydın province and located in western part of Turkey, is endemic for human and canine visceral leishmaniasis caused by Leishmania infantum MON1 and MON98. In this study, phlebotomine survey was conducted to determine the vector sand fly species and to identify sand fly blood meal sources. In August and September 2012, 1027 sand fly specimens were caught using CDC light traps. Eight Phlebotomus and two Sergentomyia species with the dominancy of Phlebotomus tobbi (61.34%) were detected. A total of 622 female sand flies (571 Phlebotomus; 51 Sergentomyia) were checked for Leishmania infection by direct dissection of the midgut. The half of the midgut content was inoculated into NNN culture for isolation of the parasite. Leishmania species-specific ITS1 real time PCR, conventional PCR assays of ITS1 and hsp70 genes and subsequent sequencing were performed from extracted DNAs. A region of cytochrome b (cyt-b) gene of vertebrates based PCR was used to determine the source of blood meal of sand flies. In microscopical examinations, two female specimens (0.32%) were found naturally infected with high number and different stages of promastigotes. No growth was observed in NNN culture but Leishmania DNA was obtained from both specimens. First positive specimen was identified as P. tobbi and L. infantum DNA was detected. Second specimen was Sergentomyia dentata, but Leishmania DNA could not be identified on species level. A total of 16 blood-fed female P. tobbi specimens were used for blood meal analysis and eight, three and one specimens were positive for human, dog and mouse, respectively. This is the first detection of Leishmania promastigotes using microscopical examination in P. tobbi and S. dentata in human and canine visceral leishmaniasis endemic area in western part of Turkey. Our results indicate that, (i) P. tobbi is

  18. Uso de cal virgem para o controle de Salmonella spp. e Clostridium spp. em camas de aviário Quicklime for controlling Salmonella spp. and Clostridium spp in litter from floor pens of broilers

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    Marcos Antonio Dai Pra

    2009-07-01

    Full Text Available O objetivo deste trabalho foi avaliar a eficácia do uso de cal virgem (CaO para a redução de Salmonella spp. e Clostridium spp. em cama de aviário. Foram aplicados quatro tratamentos: T1- sem adição de cal virgem (controle, T2- aplicação de cal virgem na dose de 300g m-2, T3- aplicação de cal virgem na dose de 600g m-2 e T4- aplicação de cal virgem na dose de 900g m-2. Os valores médios observados para o pH da cama após o 12° dia de aplicação de cal virgem foram 8,95 e 9,91, 10,75 e 11,11 para os tratamentos 1, 2, 3 e 4, respectivamente. O número mais provável Log10 (UFC de Salmonella spp. e Clostridium spp foi reduzido em 82 e 97% com a aplicação de cal na dosagem de 300g m-2 e 100% na dosagem de 600 e 900g m-2, ambos diferindo significativamente em relação ao controle (antes da aplicação da cal. A atividade de água da cama reduziu progressivamente (de 0,2 a 3,82% com a utilização de níveis crescentes de cal. Conclui-se que o uso da cal na cama de aviário, mesmo nas doses mais baixas, reduz o número mais provável de Salmonella e Clostridium ssp.This study aimed to evaluate the efficacy of quicklime (CaO for reducing Salmonella and Clostridium spp. population in used litter from floor pens of broilers. Four treatments were tested: (T1 control (without quicklime; (T2 300g quicklime m-2; (T3 600g quicklime m-2; and (T4 900g quicklime m-2. The following average pH values were observed 12 days after adding quicklime: 8.95, 9.91, 10.75 and 11.11 for treatments 1, 2, 3 and 4, respectively. An 82 and 97% reduction in the most probable number Log10 (CFU of Salmonella and Clostridium spp, respectively, was observed when 300g m-2 of quicklime was added to the used litter from floor pens of broilers. Additionally, a 100% reduction was obtained when both 600 and 900g m-2 of quicklime were added, differing significantly from control (before adding quicklime. A progressive reduction in water activity (from 0.2 to 3.82% was

  19. IFN-γ, IL-2, IP-10, and MIG as Biomarkers of Exposure to Leishmania spp., and of Cure in Human Visceral Leishmaniasis

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    Ana V. Ibarra-Meneses

    2017-05-01

    Full Text Available New biomarkers are needed for monitoring the effectiveness of treatment for visceral leishmaniasis (VL. They might also improve the detection of the asymptomatic population in Leishmania-endemic areas. This paper examines the IL-2, IFN-γ, IFN-γ-induced protein 10 (IP-10, and monokine-induced-by-IFN-γ (MIG levels in whole blood—stimulated in vitro with soluble Leishmania antigen (SLA—taken from asymptomatic individuals and patients treated for VL living in a post-outbreak (Leishmania infantum area in Spain, and in an endemic (Leishmania donovani area of Bangladesh. IP-10 was found to be an accurate global marker of asymptomatic subjects with positive cellular/humoral tests, while MIG was found to be a better marker of contact with L. donovani than IL-2 but no for those with L. infantum. Determining IP-10, MIG, and IFN-γ levels proved useful in monitoring the cellular immune response following treatment for active disease caused by L. infantum.

  20. Ecology, feeding and natural infection by Leishmania spp. of phlebotomine sand flies in an area of high incidence of American tegumentary leishmaniasis in the municipality of Rio Branco, Acre, Brazil.

    Science.gov (United States)

    de Ávila, Márcia Moreira; Brilhante, Andreia Fernandes; de Souza, Cristian Ferreira; Bevilacqua, Paula Dias; Galati, Eunice Aparecida Bianchi; Brazil, Reginaldo Peçanha

    2018-01-26

    Phlebotomine sand flies (Diptera: Psychodidae) are insects of medical importance due to their involvement in the zoonotic transmission of Leishmania spp. to vertebrates. The aim of this work was to study the ecology of the sand fly fauna of two types of environments, a rural environment (the Transacreana Road) and an urban park (Horto Florestal Park), both located in the municipality of Rio Branco in the state of Acre, Brazil. Additionally, this study intended to investigate Leishmania infection and blood meal sources of these sand flies using molecular techniques. The sand fly fauna was studied in different environments (i.e. forest and peridomestic environments in a rural area, and an urban forest) using Shannon traps and HP light traps to collect sand fly specimens over 13 consecutive months (December 2014 to January 2016). For investigating natural infection by Leishmania and the source of sand fly blood meals, DNA samples were extracted from female sand flies and subjected to polymerase chain reaction targeting ITS1 and cytb genes. DNA sequencing was subsequently used to identify species of Leishmania and the source of blood meals. A total of 2515 individual sand flies of 43 species were collected and identified, Trichophoromyia auraensis (839; 33.35%), Trichophoromyia spp. (537; 21.35%) and Evandromyia saulensis (187; 7.43%) were more abundant in the rural area (S = 41 species) than in the urban forest. No significant differences were found in species richness between forest and peridomestic environments in the rural area (H = 0.04; P > 0.05), but a larger number of species was found in the forest. Leishmania DNA was sequenced in 13 samples, confirming the presence of L. (V.) braziliensis in Th. auraensis (n = 1), Ev. saulensis (n = 2), Ev. walkeri (n = 1), Ps. llanosmartinsi (n = 1), Pi. nevesi (n = 2), Ps. davisi (n = 1), Ps. ayrozai (n = 1), Pa. aragaoi (n = 1), Ny. antunesi (n = 1) and Ev. infraspinosa (n

  1. Improvement of the green fluorescent protein reporter system in Leishmania spp. for the in vitro and in vivo screening of antileishmanial drugs.

    Science.gov (United States)

    Pulido, Sergio A; Muñoz, Diana L; Restrepo, Adriana M; Mesa, Carol V; Alzate, Juan F; Vélez, Iván D; Robledo, Sara M

    2012-04-01

    Development of new therapeutic approaches for leishmaniasis treatment requires new high throughput screening methodologies for the antileishmanial activity of the new compounds both in vitro and in vivo. Reporter genes as the GFP have become one of the most promissory and widely used tools for drug screening in several models, since it offers live imaging, high sensibility, specificity and flexibility; additionally, the use of GFP as a reporter gene in screening assays eliminates all the drawbacks presented in conventional assays and also those technical problems found using other reporter genes. The utility of the GFP as a reporter gene in drug screening assays with Leishmania parasites depends on the homogeneity and stability of the GFP transfected strains. Stable expression of the GFP in the Old World Leishmania species has been demonstrated using integration vectors; however, no reports exist yet about the success of this methodology in the New World species. Here we report the generation of New World Leishmania strains expressing the GFP protein from an integration vector, which replaces one copy of the 18S RNA in the chromosome with the GFP coding sequence by homologous recombination. We also prove that the expression of the integrated GFP is stable and homogeneous in the transfected parasites after months in culture without selective pressure or during its use in hamster infection assays. The fluorescent strains are useful for in vitro, ex vivo and in vivo drug screening assays since no considerable variations in virulence or infectivity where seen attributable to the genetic manipulation during both in vitro and in vivo infection experiments. The platform described here for drug testing assays based on the use of stable fluorescent Leishmania strains coupled to flow cytometry and fluorescent microscopy is more sensitive, more specific and faster than conventional assays used normally for the evaluation of compounds with potential antileishmanial activity

  2. Evaluación de la Dúplex PCR para el diagnóstico simultáneo de Mycobacterium spp. y Brucella spp. en bovinos

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    Ariel Escobar

    2013-01-01

    Full Text Available La tuberculosis y la brucelosis siguen siendo causas importantes de morbilidad y mortalidad en muchos países, para la detección de ambas enfermedades se requieren de herramientas eficientes y sensibles para efectivizar el diagnóstico. Este trabajo fue diri gido a evaluar y comparar la técnica de Dúplex PCR frente a la PCR anidada, para la detección de Brucella spp. (BR y Mycobacterium spp. (TB. Se utilizó un total de 100 muestras de tejidos a partir de nódulos linfáticos traqueo bronquiales, pulmón de bovi nos y, aislados bacterianos de TB y BR como controles positivos. Diez combinaciones de iniciadores dirigidos a flanquear un segmento de la secuencia 16S ARNr (BR y el gen antígeno MPB70 (TB fueron evaluados, el mejor resultado para la Dúplex PCR se logró con los iniciadores Bru - 2F/Bru - 2R para BR y para TB, Tub - 1F/Tub - N - R. Los productos de amplificación fueron de 225 y 230 - pb respectivamente. A fin de potencializar los resultados de la técnica propuesta, todas las muestras fueron inicialmente analizadas y comparadas entre la PCR simple y la PCR anidada (Kappa, k = 0,85 y, la concordancia entre los resultados obtenidos con la PCR anidada y la Dúplex PCR (k = 0,88, para las dos bacterias fue muy buena.

  3. Single and concomitant experimental infectionsby Endotrypanum spp. and Leishmania (Viannia guyanensis (Kinetoplastida: Trypanosomatidae in the Neotropical sand fly Lutzomyia longipalpis (Diptera: Psychodidae

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    André F Barbosa

    2006-12-01

    Full Text Available Lutzomyia longipalpis females received single and mixed infections with Endotrypanum and Leishmania. Two biological parameters were analyzed: the percentage of infected females and the distribution of flagellates in the gut of the females. The principal comparisons were performed between (1 two strains of Endotrypanum, (2 cloned versus primary sample of one strain of Endotrypanum, (3 Endotrypanum versus Leishmania guyanensis, and (4 the pattern of flagellates behaviour by optical microscopy in females with single or mixed infection versus the identification of parasites isolated from digestive tracts by isoenzyme electrophoresis. Flagellates of Endotrypanum showed distinct patterns of infection suggesting that there is variation between and within strains. The distribution of Endotrypanum and L. guyanensis differed significantly in relation to the colonization of the stomodeal valve. In co-infection with L. guyanensis, a large number of flagellates were seen to be plentifully infecting the stomodeal valve in significantly more specimens than in females infected by Endotrypanum only. However, the electrophoretic profiles of isoenzymes of parasites recovered from all co-infected specimens corresponded to Endotrypanum. This suggests that the mere correlation sand fly infection-biochemical analysis of isolates may induce parasitological incorrect consideration.

  4. Aislamiento microbiológico de Salmonella spp. y herramientas moleculares para su detección

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    Jose Gonzalez Pedraza

    2014-01-01

    Full Text Available Salmonella spp, es uno de los principales agentes causales de intoxicaciones alimentarías a nivel mundial, coloniza a la mayoría de los animales y el ser humano. No es detectable en muestras que tienen un bajo número de células y los métodos tradicionales para su aislamiento tienen baja especificidad, baja sensibilidad y consumen mucho tiempo. Esta revisión presenta un análisis sobre los métodos para la detección de Salmonella spp. y se profundiza en los estudios moleculares encaminados al diagnóstico de este microorganismo de importancia en salud pública. En los últimos años se han desarrollado diferentes protocolos utilizando métodos moleculares para la detección de Salmonella spp. a partir de muestras clínicas y alimentos. Los costos para la detección molecular de Salmonella spp. son elevados en com- paración con los métodos tradicionales, pero la alta sensibilidad y especificidad que ofrece la PCR, los beneficios al sector salud al lograr un diagnóstico rápido y preciso, la relación costo beneficio que otorga al sector productivo permitiendo liberar productos alimenticios al mercado con mayor rapidez, justifican la implementación de estas técnicas. La revisión de las ventajas y desventajas de los métodos microbiológicos tradicionales y moleculares para detectar Salmonella spp. en diferentes matrices, permite establecer la mejor estrategia a seguir en la detección y diagnóstico de microorganismos de difícil aislamiento. Dada la complejidad de las diferentes metodologías que existen para la detección de Salmonella spp, dichas técnicas serán presentadas en forma independiente.

  5. Anti-Leptospira spp. antibodies in captive animals in Paraíba State

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    Arthur Brasil

    2013-12-01

    Full Text Available The aim of this survey was to verify the occurrence of anti-Leptospira spp. antibodies in captive animals in the Parque Zoobotânico Arruda Câmara, João Pessoa, Paraíba State, Northeastern Brazil. Blood samples were collected from 49 animals: 26 mammals of the species Sapajus libidinosus, Cebus flavius, Saimiri sciureu, Coendu sp., Pseudalopex vetulus, Leopardus pardalis, Leopardus tigrinus, Galactitis vitata, Eira barbara, Nasua nasua, Tayassu tajacu and Ratus norvegicus; 10 birds of the species Penelope jacucaca, Pavo cristatus, Anodorhynchus hyacinthinus, Ara chlorothpterus, Pionites leucogaster, Polyborus plancus, Geranoaetus melanoleucus and Urubitinga urubitinga; and 13 reptiles of the species Caiman latirostris, Paleosuchus trigonatus, Caiman crocodilus, Tupinabis merinae, Tupinambis teguixin, Boa constrictor, Corallus hortulanus, Python molurus, Bufocephala vanderhaegei, Geochelone denticulata and Geochelone carboraria. Sera were examined by the microscopic agglutination teste (MAT using 24 serovars as antigens and cut-off point of 1:100. One ocelot (Leopardo pardalis presented positive reaction for the Icterohaemorrhagiae serovar with titer of 100, however, it did not show any clinical sign of the infection. Sinantropic rodents are the main reservoirs of this serovar, which suggests the need of maintenance and continuous evaluation of rodent control programs.

  6. Prevalence of Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in apparently healthy and CVBD-suspect dogs in Portugal - a national serological study

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    Cardoso Luís

    2012-03-01

    Full Text Available Abstract Background Canine vector-borne diseases (CVBDs are caused by a wide range of pathogens transmitted to dogs by arthropods including ticks and insects. Many CVBD-agents are of zoonotic concern, with dogs potentially serving as reservoirs and sentinels for human infections. The present study aimed at assessing the seroprevalence of infection with or exposure to Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in dogs in Portugal. Methods Based on 120 veterinary medical centres from all the regions of mainland and insular Portugal, 557 apparently healthy and 628 CVBD-suspect dogs were sampled. Serum, plasma or whole blood was tested for qualitative detection of D. immitis antigen and antibodies to E. canis, B. burgdorferi s. l., Anaplasma spp. and L. infantum with two commercial in-clinic enzyme-linked immunosorbent assay kits. Odds ratios (OR were calculated by logistic regression analysis to identify independent risk factors of exposure to the vector-borne agents. Results Total positivity levels to D. immitis, E. canis, B. burgdorferi, Anaplasma spp., L. infantum, one or more agents and mixed agents were 3.6%, 4.1%, 0.2%, 4.5%, 4.3%, 14.0% and 2.0% in the healthy group, and 8.9%, 16.4%, 0.5%, 9.2%, 25.2%, 46.3% and 11.6% in the clinically suspect group, respectively. Non-use of ectoparasiticides was a risk factor for positivity to one or more agents both in the apparently healthy (OR = 2.1 and CVBD-suspect (OR = 1.5 dogs. Seropositivity to L. infantum (OR = 7.6, E. canis (OR = 4.1 and D. immitis (OR = 2.4 were identified as risk factors for the presence of clinical signs compatible with CVBDs. Positivity to mixed agents was not found to be a risk factor for disease. Conclusions Dogs in Portugal are at risk of becoming infected with vector-borne pathogens, some of which are of zoonotic concern. CVBDs should be considered by practitioners and prophylactic measures must be put in

  7. An Insight into the proteome of Crithidia fasciculata choanomastigotes as a comparative approach to axenic growth, peanut lectin agglutination and differentiation of Leishmania spp. promastigotes.

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    Pedro J Alcolea

    Full Text Available The life cycle of the trypanosomatid Crithidia fasciculata is monogenetic, as the unique hosts of these parasites are different species of culicids. The comparison of these non-pathogenic microorganisms evolutionary close to other species of trypanosomatids that develop digenetic life cycles and cause chronic severe sickness to millions of people worldwide is of outstanding interest. A ground-breaking analysis of differential protein abundance in Crithidia fasciculata is reported herein. The comparison of the outcome with previous gene expression profiling studies developed in the related human pathogens of the genus Leishmania has revealed substantial differences between the motile stages of these closely related organisms in abundance of proteins involved in catabolism, redox homeostasis, intracellular signalling, and gene expression regulation. As L. major and L. infantum agglutinate with peanut lectin and non-agglutinating parasites are more infective, the agglutination properties were evaluated in C. fasciculata. The result is that choanomastigotes are able to agglutinate with peanut lectin and a non-agglutinating subpopulation can be also isolated. As a difference with L. infantum, the non-agglutinating subpopulation over-expresses the whole machinery for maintenance of redox homeostasis and the translation factors eIF5a, EF1α and EF2, what suggests a relationship between the lack of agglutination and a differentiation process.

  8. Natural Leishmania (Viannia) spp. infections in phlebotomine sand flies (Diptera: Psychodidae) from the Brazilian Amazon region reveal new putative transmission cycles of American cutaneous leishmaniasis

    Science.gov (United States)

    de Souza, Adelson Alcimar Almeida; dos Santos, Thiago Vasconcelos; Jennings, Yara Lúcia Lins; Ishikawa, Edna Aoba Yassui; Barata, Iorlando da Rocha; Silva, Maria das Graças Soares; Lima, José Aprígio Nunes; Shaw, Jeffrey; Lainson, Ralph; Silveira, Fernando Tobias

    2016-01-01

    In Amazonian Brazil the etiological agents of American cutaneous leishmaniasis (ACL) belong to at least seven Leishmania species but little is known about the putative phlebotomine sand fly vectors in different biomes. In 2002–2003 a survey of the phlebotomine fauna was undertaken in the “Floresta Nacional do Tapajós”, Belterra municipality, in the lower Amazon region, western Pará State, Brazil, where we recently confirmed the presence of a putative hybrid parasite, L. (V.) guyanensis × L. (V.) shawi shawi. Sand flies were collected from Centers for Disease Control (CDC) light traps, Shannon traps and by aspiration on tree bases. Females were dissected and attempts to isolate any flagellate infections were made by inoculating homogenized midguts into Difco B45 medium. Isolates were characterized by monoclonal antibodies and isoenzyme electrophoresis. A total of 9,704 sand flies, belonging to 68 species or subspecies, were collected. Infections were found in the following sand flies: L. (V.) naiffi with Psychodopygus hirsutus hirsutus (1) and Ps. davisi (2); and L. (V.) shawi shawi with Nyssomyia whitmani (3) and Lutzomyia gomezi (1). These results provide strong evidence of new putative transmission cycles for L. (V.) naiffi and L. (V.) s. shawi. PMID:27235194

  9. Genetic diversity of the mitochondrial cytochrome b gene in Lutzomyia spp., with special reference to Lutzomyia peruensis, a main vector of Leishmania (Viannia) peruviana in the Peruvian Andes.

    Science.gov (United States)

    Yamamoto, Kento; Cáceres, Abraham G; Gomez, Eduardo A; Mimori, Tatsuyuki; Iwata, Hiroyuki; Korenaga, Masataka; Sakurai, Tatsuya; Katakura, Ken; Hashiguchi, Yoshihisa; Kato, Hirotomo

    2013-05-01

    The genetic divergence caused by genetic drift and/or selection is suggested to affect the vectorial capacity and insecticide susceptibility of sand flies, as well as other arthropods. In the present study, cytochrome b (cyt b) gene sequences were determined in 13 species circulating in Peru to establish a basis for analysis of the genetic structure, and the intraspecific genetic diversity was assessed in the Lutzomyia (Lu.) peruensis, a main vector species of Leishmania (Viannia) peruviana in Peruvian Andes. Analysis of intraspecific genetic diversity in the cyt b gene sequences from 36 Lu. peruensis identified 3 highly polymorphic sites in the middle region of the gene. Haplotype and gene network analyses were performed on the cyt b gene sequences of 130 Lu. peruensis in 9 Andean areas from 3 Departments (Ancash, Lima and La Libertad). The results showed that the populations of La Libertad were highly polymorphic and that their haplotypes were distinct from those of Ancash and Lima, where dominant haplotypes were observed, suggesting that a population bottleneck may have occurred in Ancash and Lima, but not in La Libertad. The present study indicated that the middle region of the cyt b gene is useful for the analysis of genetic structure in sand fly populations. Copyright © 2013 Elsevier B.V. All rights reserved.

  10. Effect of Leishmania spp infection on the survival, life expectancy, fecundity and fertility of Lutzomyia longipalpiss.l. and Lutzomyia pseudolongipalpis

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    Irma Fatima Agrela

    2015-08-01

    Full Text Available We evaluated the effects of Leishmaniaspp infection on several population parameters of Lutzomyia longipalpis sensu lato andLutzomyia pseudolongipalpis, vectors of visceral leishmaniasis in Venezuela, under experimental conditions during the first post-feeding period. Females of both species were allowed to feed and engorge on a suspension of fresh washed human red blood cells in foetal calf serum. These blood cells were either non-infected or infected with one of the fourLeishmaniaspp strains and were offered through a chicken skin membrane. The longevity, life expectancy and the fecundity of uninfected flies were similar in both species, but the fertility was significantly lower in uninfected Lu. longipalpis females. In all cases, the infection of Lu. longipalpis and Lu. pseudolongipalpis by the Leishmaniastrains resulted in significant detrimental effects, which exerted a fitness cost expressed by reduced survival and life expectancy, as well as decreased fertility and fecundity compared with the control groups. Nevertheless, differences in these parameters were observed between these vector species depending on whether they were infected with the autochthonous Venezuelan Leishmania infantum strain (NESA or the Brazilian reference strain (PP75. The experimental data obtained agree with field data on the natural infection of these vector species and the significance of this scenario is discussed.

  11. 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

  12. In vitro anti-leishmania evaluation of nickel complexes with a triazolopyrimidine derivative against Leishmania infantum and Leishmania braziliensis.

    Science.gov (United States)

    Ramírez-Macías, Inmaculada; Maldonado, Carmen R; Marín, Clotilde; Olmo, Francisco; Gutiérrez-Sánchez, Ramón; Rosales, María J; Quirós, Miguel; Salas, Juan M; Sánchez-Moreno, Manuel

    2012-07-01

    Studies on the anti-proliferative activity in vitro of seven ternary nickel (II) complexes with a triazolopyrimidine derivative and different aliphatic or aromatic amines as auxiliary ligands against promastigote and amastigote forms of Leishmania infantum and Leishmania braziliensis have been carried out. These compounds are not toxic for the host cells and two of them are effective at lower concentrations than the reference drug used in the present study (Glucantime). In general, the in vitro growth rate of Leishmania spp. was reduced, its capacity to infect cells was negatively affected and the multiplication of the amastigotes decreased. Ultrastructural analysis and metabolism excretion studies were executed in order to propose a possible mechanism for the action of the assayed compounds. Our results show that the potential mechanism is at the level of organelles membranes, either by direct action on the microtubules or by their disorganization, leading to vacuolization, degradation and ultimately cell death. Copyright © 2012 Elsevier Inc. All rights reserved.

  13. PCR associated with hybridization with DNA radioactive probes for diagnosis of asymptomatic infection caused by Leishmania Chagasi; PCR associado a hibridizacao com sondas radioativas de DNA para a identificacao de infeccao subclinica causada por Leishmania Chagasi

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, Antero Silva Ribeiro de [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN), Belo Horizonte, MG (Brazil); Moreno, Elizabeth Castro [Fundacao Nacional de Saude, Belo Horizonte, MG (Brazil). Coordenacao Regional de Minas Gerais; Gomes, Rosangela Fatima; Melo, Maria Norma de; Carneiro, Mariangela [Minas Gerais Univ., Belo Horizonte, MG (Brazil). Dept. de Parasitologia; Fernandes, Octavio [Fundacao Inst. Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ (Brazil). Dept. de Medicina Tropical

    2002-07-01

    Detection systems for diagnosis of leishmaniasis based on PCR are very promising due to their sensitivity and specificity. Secondary detection by specific radioactive DNA probes, able to type the PCR amplified products, increase the specificity and raise about tem-fold the sensitivity of the assay. The aim of this work was evaluate PCR and hybridization as a tool to identify Leishmania (Leishmania) chagasi (the specie that cause the visceral leishmaniasis in Brazil) infection in asymptomatic persons living in a endemic area. Material and Methods: A group of 226 asymptomatic individuals, living in General Carneiro (MG), was selected. Blood samples were harvested and the DNA extracted from the mononucleate cells. PCR was performed using primers addressed to the kinetoplast DNA minicircles. This protocol gives a positive reaction for all Leishmania species. The amplified products were further hybridized with cloned L.chagasi minicircles labeled with {sup 32} P. Results: were identified 111 samples PCR positive, 2 of them hybridization negative and 133 samples hybridization positive, 24 of them PCR negative. The occurrence of samples with hybridization positive and PCR negative was expected since hybridization, with DNA probes labeled with {sup 32} P, increase the sensitivity of the assay. The samples that presented positive PCR and negative hybridization were probably due the presence of other Leishmania species, likely L. (V.) braziliensis (that produce tegumentary leishmaniasis in the region), since L. (L.) chagasi cloned minicircles were used as hybridization probe. We conclude that this procedure is a valuable tool to access subclinical L. (L.) chagasi infections in epidemiological studies. (author)

  14. Presencia de Salmonella spp. en un área del Caribe colombiano: un riesgo para la salud pública.

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    Johnny Durango

    2004-03-01

    Full Text Available Salmonella está asociada frecuentemente con la enfermedad diarreica aguda; la bacteria se propaga principalmente por la ingestión de alimentos o de aguas contaminadas o por personas infectadas que manipulan los alimentos. En Colombia no se conoce la distribución de los serotipos de Salmonella en los alimentos. El objetivo de este estudio fue el de establecer la frecuencia de Salmonella spp. en alimentos del Caribe colombiano. Se analizaron 636 muestras de alimentos obtenidas en ventas de comidas rápidas callejeras y en plazas de mercados de Barranquilla (n=245, Montería (n=222, Sincelejo (n=87 y Cartagena (n=82. El aislamiento de la bacteria se realizó por el método convencional de la Food and Drug Administration (FDA de los Estados Unidos. Se inocularon 25 g de cada muestra de alimento en 225 ml de caldo de preenriquecimiento; posteriormente, se inoculó 1 ml a los caldos de enriquecimiento y, finalmente, se sembraron en medios de cultivo selectivos para Salmonella. Las colonias sospechosas se identificaron por pruebas bioquímicas y serológicas convencionales para Salmonella. Se aislaron 47 (7,4% Salmonella spp. Del total de muestras de carne de res, 9,3% fueron positivas para Salmonella spp., 12,6% de chorizo, 7,9% de queso, 5,2% de carne de cerdo, 1,6% de pollo y 10,5% de arepa de huevo. Los principales serotipos encontrados fueron S. Anatum (26%, S. Newport (13%, S. Typhimurium (9%, S. Gaminara (9% y S. Uganda (9%. Este estudio permitió establecer la distribución de los serotipos de Salmonella spp. en alimentos de la Costa Atlántica colombiana. No se observaron diferencias estadísticas significativas entre los estratos 1 y 4, ni entre los estratos 2 y 3 (p>0,05, pero sí entre los estratos 1, 2 y 3 (p

  15. Antraquinona e surfactante para otimizaçao do processo Kraft com Pinus Spp.

    OpenAIRE

    Mocelin, Ezequiel Zatoni

    2013-01-01

    A madeira de Pinus é atualmente a matéria-prima mais utilizada na obtenção de fibras longas para papel, juntamente com o processo Kraft pela sua versatilidade e por produzir fibras com boa resistência. As atuais perspectivas florestais apontam para uma escassez da madeira de Pinus para os diversos fins, seja para madeira processada, seja para celulose e papel. O processo Kraft tem como desvantagem o baixo rendimento em celulose, devido à solubilização não somente da lignina como também parte ...

  16. Fenotipado radicular para la resistencia a la sequía en Solanum spp

    OpenAIRE

    ATUCHA ZAMKOVA, ANASTASIA-AINHOA

    2015-01-01

    [ES] La sequía es un gran problema para la agricultura, ya que destruye o merma considerablemente las cosechas. Por eso la mejora genética de las plantas para la tolerancia a sequía es una necesidad acuciante. Para mejorar las plantas genéticamente, es necesario tener herramientas de fenotipado adecuadas para poder identificar las plantas con las características deseadas. La berenjena cultivada es una planta que es bastante tolerante a la sequía, y posee una gran variedad de especies relac...

  17. Ocorrência de Mollicutes e Ureaplasma spp. em surto de doença reprodutiva em rebanho bovino no Estado da Paraíba

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    Sandra B. dos Santos

    2013-03-01

    Full Text Available Em março de 2012 foi diagnosticado um surto de doença reprodutiva em rebanho bovino no Estado da Paraíba, Brasil. Foram examinadas 32 vacas e dois touros da raça Girolando. As vacas apresentaram sinais de doença reprodutiva como repetição de cio, vulvovaginite granular, infertilidade e abortos. As amostras de suabes vaginais e prepuciais foram colhidas e submetidas a isolamento bacteriano e PCR. As reações da PCR para Mollicutes e Ureaplasma spp. foram realizadas com os iniciadores MGSO-GPO3 e UGP'F-UGP'R, respectivamente. Na Nested PCR para Ureaplasma diversum, os iniciadores usados foram UD1, UD2, UD3 e UD4. Para isolamento bacteriano, as amostras foram diluídas de 10-1 até 10-5, semeadas em meio "UB", líquido e placa, sendo incubadas por até 21 dias a 37ºC em jarra de microaerofilia. A frequência de Mollicutes detectada na PCR foi de 65,6% e para Ureaplasma spp. foi de 50,0%, enquanto que para U. diversum foi de 15,6%. No isolamento a frequência de Mollicutes foi de 57,1% e para Ureaplasma spp. foi de 28,6%. No ágar "UB" foi visualizado o crescimento misto de Mycoplasma spp. e Ureaplasma spp. em seis amostras. Foi confirmado o envolvimento de micro-organismos da Classe Mollicutes em surto de doença reprodutiva em vacas no sertão paraibano.

  18. Superoxide Dismutase (SOD Enzyme Activity Assay in Fasciola spp. Para-sites and Liver Tissue Extract

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    M Assady

    2011-09-01

    Full Text Available Background: The purpose of this comparative study was to detect superoxide dismutase (SOD activities in Fasciola hepatica, F. gigantica parasites, infected and healthy liver tissues in order to determine of species effects and liver infection on SODs activity level.Methods: Fasciola spp. parasites and sheep liver tissues (healthy and infected liver tissues, 10 samples for each, were collected, homogenized and investigated for protein measurement, protein detection and SOD enzyme activity assay. Protein concentration was measured by Bradford method and SODs band protein was detected on SDS-PAGE. SODs activity was determined by iodonitrotetrazolium chloride, INT, and xanthine substrates. Independent samples t-test was conducted for analysis of SODs activities difference.Results: Protein concentration means were detected for F. hepatica 1.3 mg/ ml, F. gigantica 2.9 mg/ml, healthy liver tissue 5.5 mg/ml and infected liver tissue 1.6 mg/ml (with similar weight sample mass. Specific enzyme activities in the samples were obtained 0.58, 0.57, 0.51, 1.43 U/mg for F. hepatica, F. gigantica, healthy liver and infected liver respectively. Gel electrophoresis of Fasciola spp. and sheep liver tissue extracts revealed a band protein with MW of 60 kDa. The statistical analysis revealed significant difference between SOD activities of Fasciola species and also between SOD activity of liver tissues (P<.05.Conclusion: Fasciola species and liver infection are effective causes on SOD enzyme activity level.

  19. Cell migration induced by Leishmania (Leishmania) amazonensis, Leishmania (Leishmania) major and Leishmania (Viannia) braziliensis into the peritoneal cavity of BALB/c mice

    OpenAIRE

    DT Wakimoto; KV Gaspareto; TGV Silveira; MVC Lonardoni; SMA Aristides

    2010-01-01

    In American cutaneous leishmaniasis, the initial infection phase is characterized by recruitment of neutrophils and monocytes. The migration of these cells in response to the presence of Leishmania in the peritoneum of affected animals remains unclear. The objective of this study was to investigate cell migration to the peritoneum of BALB/c mice after infection with Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis and Leishmania (Leishmania) major. Initially, Leishmania ...

  20. Cristalización y predicción de la estructura tridimensional de la proteína homóloga del receptor activado para la quinasa c (lack de leishmania

    Directory of Open Access Journals (Sweden)

    Edwin Patiño González

    2014-10-01

    Full Text Available Los parásitos del género Leishmania son los causantes de la enfermedad conocida como Leishmaniasis. Esta enfermedad es endémica en 98 países. Veinte especies de Leishmania sp han sido descritas como patógenos en humanos y varias de ellas presentan manifestaciones clínicas diferentes. No se dispone de vacuna, a pesar del considerable esfuerzo de muchos grupos de investigación. Las alternativas para descubrir nuevos medicamentos están basadas en el diseño de compuestos que interaccionen con blancos específicos, principalmente, proteínas encargadas de procesos metabólicos o celulares del patógeno, e.g. la parasitación de las células del huésped vertebrado. La eficiente parasitación del huésped vertebrado por Leishmania depende de la expresión de diferentes proteínas, incluyendo la proteína Lack. Parásitos deficientes de Lack no sobreviven internalizados en las células de los vertebrados. Este artículo presenta las condiciones de renaturalización, purificación y cristalización de la proteína Lack del patógeno humano Leishmania (Viannia panamensis. Además, los resultados de modelación estructural de esta proteína muestran una conformación proteica similar a un ventilador organizado en 7 aspas, cada una compuesta de 4 hojas β. La estructura de la proteína Lack resultó similar a la proteína asociada a ribosoma RACK1 de Trypanosoma brucei y Saccharomyces cerevisiae, y a la de otros eucariotas. Las características estructurales de la proteína Lack podrían ser usadas para la exploración de nuevos fármacos.

  1. Natural infection of bats with Leishmania in Ethiopia.

    Science.gov (United States)

    Kassahun, Aysheshm; Sadlova, Jovana; Benda, Petr; Kostalova, Tatiana; Warburg, Alon; Hailu, Asrat; Baneth, Gad; Volf, Petr; Votypka, Jan

    2015-10-01

    The leishmaniases, a group of diseases with a worldwide-distribution, are caused by different species of Leishmania parasites. Both cutaneous and visceral leishmaniasis remain important public health problems in Ethiopia. Epidemiological cycles of these protozoans involve various sand fly (Diptera: Psychodidae) vectors and mammalian hosts, including humans. In recent years, Leishmania infections in bats have been reported in the New World countries endemic to leishmaniasis. The aim of this study was to survey natural Leishmania infection in bats collected from various regions of Ethiopia. Total DNA was isolated from spleens of 163 bats belonging to 23 species and 18 genera. Leishmania infection was detected by real-time (RT) PCR targeting a kinetoplast (k) DNA and internal transcribed spacer one (ITS1) gene of the parasite. Detection was confirmed by sequencing of the PCR products. Leishmania kDNA was detected in eight (4.9%) bats; four of them had been captured in the Aba-Roba and Awash-Methara regions that are endemic for leishmaniasis, while the other four specimens originated from non-endemic localities of Metu, Bedele and Masha. Leishmania isolates from two bats were confirmed by ITS1 PCR to be Leishmania tropica and Leishmania major, isolated from two individual bats, Cardioderma cor and Nycteris hispida, respectively. These results represent the first confirmed observation of natural infection of bats with the Old World Leishmania. Hence, bats should be considered putative hosts of Leishmania spp. affecting humans with a significant role in the transmission. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  2. Canine leishmaniosis caused by Leishmania major and Leishmania tropica: comparative findings and serology.

    Science.gov (United States)

    Baneth, Gad; Yasur-Landau, Daniel; Gilad, Matan; Nachum-Biala, Yaarit

    2017-03-13

    Infection and clinical disease associated with Leishmania major and Leishmania tropica, two common agents of human cutaneous leishmaniosis, have rarely been reported in dogs. This study describes dogs infected with these Leishmania spp. prevalent in the Middle East and North Africa, and compares the serological response of dogs infected with Leishmania infantum, L. major or L. tropica to whole promastigote antigen enzyme-linked immunosorbent assay (ELISA) of each species and to rK39 dipstick. Leishmania major infection in a 5-month-old male dog was associated with alopecic and ulcerative periocular and limb skin lesions which responded to allopurinol treatment. Infection was detected by skin and blood polymerase chain reaction (PCR) and confirmed by DNA sequencing but the dog was seronegative. Leishmania tropica infection was detected in a 3-month-old female dog co-infected with Babesia vogeli and Anaplasma platys and with no skin lesions. PCR and DNA sequencing of the blood and parasite culture were positive for L. tropica. Sera from 11 dogs infected with L. infantum, L. major or L. tropica were reactive with all three Leishmania spp. antigens except for sera from a dog with L. major infection. No significant differences were found between reactivity of dog sera to the antigen of the infecting species, or to the other Leishmania spp. antigens. Sera from dogs infected with L. infantum and L. tropica were positive with the rK39 antigen kit, while dogs with L. major infection were seronegative. Skin lesions in L. major infected dogs from this study and previous reports (n = 2) were ulcerative and located on the muzzle, feet and foot pads and not associated with generalized lymphadenomegaly and splenomegaly. In previous L. tropica infections, skin lesions were proliferative mucocutaneous in young dogs (n = 2), or associated with widespread dermatitis, lymphadenomegaly and splenomegaly in older dogs with similarity to L. infantum infection (n = 2). This

  3. PCR para la confirmación de transmisión experimental de Leishmania chagasi a hámster sano por picadura de Lutzomyia longipalpis (Diptera: Psychodidae.

    Directory of Open Access Journals (Sweden)

    Olga L. Cabrera

    2003-06-01

    Full Text Available Se evaluó la efectividad de la PCR como herramienta en la detección de la transmisión experimental de Leishmania chagasi a hámster, Mesocricetus auratus, por picadura del insecto vector. Dos pares de hámsteres sanos y anestesiados fueron colocados en jaulas que contenían hembras de Lutzomyia longipalpis. Previamente, las hembras se infectaron experimentalmente con Leishmania chagasi y la infección se confirmó por disección en una submuestra. A los 37 y 51 días después de la exposición a los insectos infectados, las biopsias de hígado y bazo de cada hámster se sometieron a examen directo al microscopio, histopatología y PCR. El ADN se extrajo con Chelex 100®; en la amplificación se utilizó un par de iniciadores específicos para la región conservada de los minicírculos del ADN de Leishmania. El producto amplificado se separó en geles de agarosa y se visualizó bajo luz UV. En tres de las cuatro biopsias se observó una banda de 120 pares de bases, aproximadamente, correspondiente al tamaño esperado de la fracción del minicírculo. La técnica de PCR fue el único método que detectó la presencia del parásito. Estos resultados demostraron que la sensibilidad de la PCR acelera los procesos de incriminación vectorial de las especies vectoras de leishmaniasis.

  4. Potencial de pseudomonas spp. fluorescentes para biocontrole de alternaria ricini em mamoneira Potential of fluorescent pseudomonas spp. For biological control of alternaria ricini on castorbean

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    Francisco de A.G. da Silva

    1998-06-01

    Full Text Available The potential of fluorescent Pseudomonas spp. to control Alternaria leaf spot on castorbean, caused by Alternaria ricini, was studied under greenhouse conditions. Two periods for antagonist applications were tested: 48h before and simultaneously to the pathogen inoculation. Among the antagonists tested JA4 and BJ22 were the most effectives showing disease severity reduction of 20.9% and 17.8% respectively, when applied simultaneously. The effect of Pseudomonas spp. on the micelial growth and sporulation was also studied throughout three different methods (funel, streak and celophane. Inhibition of micelial growth and sporulation was observed. There was no correlation between in vitro and in vivo data. Antibiosis was showed as a mode of action for Pseudomonas spp. in relation to Alternaria ricini. Ultrastructural studies confirmed the inhibition of spore germination by the bacteria.

  5. Estudo de validação comparativo entre as técnicas Elisa e RIFI para diagnosticar Leishmania sp em cães errantes apreendidos no município de Campos dos Goytacazes, Estado do Rio de Janeiro Comparative validation study between the ELISA and RIFI techniques for diagnosing Leishmania sp in stray dogs caught in the municipality of Campos de Goytacazes, State of Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    Mariane Pinto Fernandes Távora

    2007-08-01

    Full Text Available Foi realizada uma pesquisa objetivando-se verificar a eficácia do teste ELISA, para detecção de anticorpos contra Leishmania sp em cães, comparando-o com o RIFI, padrão em humanos, e investigar a situação sorológica desta zoonose na microrregião. Os testes tiveram uma concordância de 97,6%, classificada como forte.A survey was carried out aiming to verify the ELISA test effectiveness for detecting antibodies against Leishmania sp in dogs, comparing with RIFI human pattern and for investigating sorological zoonosis situation in the microregion. An accordance about 97.6% considered strong was reported.

  6. Utilización de Eucalyptus spp. Alternativas de plantaciones uruguayas para pulpa Kraft

    Directory of Open Access Journals (Sweden)

    Javier Doldán

    2011-05-01

    Full Text Available Las plantaciones de Eucalyptus globulus han mostrado desiguales tasas de crecimiento en diferentes regiones de Uruguay. Esto ha motivado la búsqueda de otros orígenes de semilla y especies para la producción de pulpa de celulosa. Propiedades papeleras de las pulpas blanqueadas (ECF de especies alternativas llevan a intuir que podrían ser atractivas para mercados de pulpa de fibra corta. En este trabajo se realiza un comparativo del E. grandis, E. dunnii, E. maidenii y E. globulus (procedencia de semilla “Jeeralang” con el E. globulus predominante en Uruguay. Se discute el potencial de estas maderas como base para una mezcla en cocción, basándose en el análisis de propiedades físicas (densidad aparente básica, propiedades pulpables (rendimiento, carga de álcali activo en cocción Kraft y consumo de madera y propiedades papeleras. En trabajos previos se han encontrado diferencias significativas entre el comportamiento pulpable del Eucalyptus globulus y Eucalyptus maidenii, sugiriendo que nosería recomendable mezclar estas especies. La misma conclusión se podría extender a las especies de Eucalyptus estudiadas. Sin embargo, teniendo en cuenta los similares requerimientos en las cargas de álcali activo, la mezcla entre especies alternativas podría ser aplicada.AbstractEucalyptus globulus plantations have shown different growth rates in different sites in Uruguay. This fact has triggered the search for other pulp wood species and seed provenance. Paper making properties of ECF bleached pulps of alternative speciessuggest that these species could be perfectly used as hardwood bleached pulp raw materials. This study intends to compare alternativeUruguayan pulpwood species E. grandis, E. dunnii, E. maidenii and “Jeeralang” a seed provenance of E. globulus to the E. globulus most widely cultivated in the country. Physical properties of wood (Basic Density, Kraft pulping performance (pulp yield, active alkali and wood consumption

  7. PRESENCIA DE Wolbachia y Leishmania EN UNA POBLACION DE Lutzomyia evansi PRESENTE EN LA COSTA CARIBE DE COLOMBIA

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    Rafael J. Vivero-Gómez

    2016-07-01

    Full Text Available Lutzomyia evansi es importante en salud pública por su participación en la trasmisión de la leishmaniasis visceral y cutánea en la costa caribe de Colombia. Diversos estudios se han desarrollado sobre la poblaciones naturales de Lutzomyia evansi, sin embargo pocos estudios han explorado en profundidad la detección de microorganismos simbióticos (ej. Wolbachia y de manera simultánea la presencia de Leishmania sp.. El endosimbionte Wolbachia ha sido propuesto en la actualidad como control biológico de insectos vectores de diversas enfermedades tropicales. En el presente estudio el ADN de tres especies del género Lutzomyia colectadas en el municipio de Ovejas (Departamento de Sucre fue evaluado para detectar la infección natural por la bacteria Wolbachia y la presencia de parásitos del género Leishmania. El ADN total de 176 individuos adultos y 34 inmaduros (larvas y pupas de Lu. evansi, fue utilizado para evaluar la detección de Wolbachia mediante amplificación por PCR del gen WSP (Proteína Mayor de la Superficie de Wolbachia y la infección por Leishmania mediante amplificación por PCR de segmentos de los genes HPSN70 (Proteína de Choque Térmico. Se encontró un grupo de machos infectado de forma natural por Wolbachia y nueve grupos de hembras con infección natural por Leishmania, todos pertenecientes a Lutzomyia evansi. El análisis filogenético de la secuencia del gen WSP de Wolbachia indica la ubicación de la cepa detectada dentro del supergrupo B (haplogrupo wLeva y su relación con haplotipos previamente reportados de Lutzomyia evansi y Lutzomya dubitans. Una región de 418 pb del gen HSP-70N fue secuenciada y mostró similaridad con secuencias de Leishmania luego de realizar el análisis en BlastN. Se confirma la presencia de Wolbachia en poblaciones silvestres de machos de L. evansi y la infección natural por Leishmania spp. en hembras de la misma especie cuya infección por Wolbachia resulto negativa.

  8. A atividade da azitromicina contra a Leishmania (Viannia) braziliensis e a Leishmania (Leishmania) amazonensis no modelo golden hamster

    OpenAIRE

    Sinagra, Ángel; Luna, Concepción; Abraham, David; Iannella, Maria del Carmen; Riarte, Adelina; Krolewiecki, Alejandro J.

    2007-01-01

    New therapeutic alternatives against leishmaniasis remain a priority. The activity of azithromycin against Leishmania (Leishmania) major has been previously demonstrated. Different responses among species of Leishmania make species-specific drug screening necessary. The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis was evaluated in golden hamsters infected through footpad injections of metacyclic promastigotes, and compared with unt...

  9. The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis in the golden hamster model

    OpenAIRE

    Sinagra,Ángel; Luna,Concepción; Abraham,David; Iannella,Maria del Carmen; Riarte,Adelina; Krolewiecki,Alejandro J.

    2007-01-01

    New therapeutic alternatives against leishmaniasis remain a priority. The activity of azithromycin against Leishmania (Leishmania) major has been previously demonstrated. Different responses among species of Leishmania make species-specific drug screening necessary. The activity of azithromycin against Leishmania (Viannia) braziliensis and Leishmania (Leishmania) amazonensis was evaluated in golden hamsters infected through footpad injections of metacyclic promastigotes, and compared with unt...

  10. Sensibilidade a cobre, estreptomicina e oxitetraciclina em Xanthomonas spp. associadas à mancha-bacteriana do tomate para processamento industrial

    Directory of Open Access Journals (Sweden)

    Quezado-Duval Alice Maria

    2003-01-01

    Full Text Available Apesar de amplamente empregados em lavouras de tomate para processamento industrial no Brasil, fungicidas cúpricos e antibióticos registrados para uso agrícola nem sempre resultam em controle eficiente das bacterioses que afetam a cultura. O aparecimento de estirpes resistentes é uma das causas dessa baixa eficiência. Avaliou-se, in vitro, a sensibilidade a cobre, estreptomicina e oxitetraciclina de 389 isolados de Xanthomonas spp. associadas à mancha-bacteriana do tomateiro, sendo 92 de X. axonopodis pv. vesicatoria (60 do grupo "A"/raça T1 e 32 do "C"/raça T3, 93 de X. vesicatoria (grupo "B"/raça T2 e 204 de X. gardneri (grupo "D"/raça T2. Os isolados foram obtidos de plantas doentes em campos comerciais de tomate para processamento industrial nos estados de Goiás, Minas Gerais, Pernambuco e Bahia, nos anos de 1995 a 1998 e em 2000. Alíquotas de 5 ml de suspensões bacterianas foram depositadas em meio Nutriente-Ágar suplementado com sulfato de cobre, nas concentrações de 50 e 200 µg/ml; sulfato de estreptomicina, a 25 e 200 µg/ml e cloridrato de oxitetraciclina, a 25 µg/ml. Nenhum isolado foi resistente a oxitetraciclina, como também nenhum foi resistente ao cobre na concentração de 200 µg/ml do sulfato de cobre. No entanto, houve diferença entre isolados quanto à sensibilidade ao sulfato de cobre na concentração de 50 µg/ml e ao sulfato de estreptomicina nas duas concentrações empregadas. As freqüências de isolados de X. gardneri, X. axonopodis pv. vesicatoria (grupos "A" e "C" e X. vesicatoria resistentes à estreptomicina (25 µg/ml do produto usado foram, respectivamente, 98%, 38% e 2%, ao passo que, ao cobre, foram, respectivamente, 48%, 4% e 74%. Todos os isolados do grupo "C" foram sensíveis à estreptomicina e 97% sensíveis ao cobre.

  11. Ocorrência de Mollicutes e Ureaplasma spp. em surto de doença reprodutiva em rebanho bovino no Estado da Paraíba Occurrence of Mollicutes and Ureaplasma spp. in outbreak of reproductive disease in cattle herds, State of Paraíba, Brazil

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    Sandra B. dos Santos

    2013-03-01

    Full Text Available Em março de 2012 foi diagnosticado um surto de doença reprodutiva em rebanho bovino no Estado da Paraíba, Brasil. Foram examinadas 32 vacas e dois touros da raça Girolando. As vacas apresentaram sinais de doença reprodutiva como repetição de cio, vulvovaginite granular, infertilidade e abortos. As amostras de suabes vaginais e prepuciais foram colhidas e submetidas a isolamento bacteriano e PCR. As reações da PCR para Mollicutes e Ureaplasma spp. foram realizadas com os iniciadores MGSO-GPO3 e UGP'F-UGP'R, respectivamente. Na Nested PCR para Ureaplasma diversum, os iniciadores usados foram UD1, UD2, UD3 e UD4. Para isolamento bacteriano, as amostras foram diluídas de 10-1 até 10-5, semeadas em meio "UB", líquido e placa, sendo incubadas por até 21 dias a 37ºC em jarra de microaerofilia. A frequência de Mollicutes detectada na PCR foi de 65,6% e para Ureaplasma spp. foi de 50,0%, enquanto que para U. diversum foi de 15,6%. No isolamento a frequência de Mollicutes foi de 57,1% e para Ureaplasma spp. foi de 28,6%. No ágar "UB" foi visualizado o crescimento misto de Mycoplasma spp. e Ureaplasma spp. em seis amostras. Foi confirmado o envolvimento de micro-organismos da Classe Mollicutes em surto de doença reprodutiva em vacas no sertão paraibano.In March of 2012 was investigated a reproductive disease outbreak in cattle herds from Paraíba State, Brazil. Were examined 32 cows and two bulls Giroland breed. The cows showed signs and symptoms of reproductive failure such as repeat breeding, granular vulvovaginitis, infertility and abortions. Vaginal and preputial mucous samples were collected for analysis by PCR and isolation. The PCR reactions for Mollicutes and Ureaplasma spp. were realized with primers MGSO and GPO3, and UGP'F and UGP'R respectively. The nested PCR assay for Ureaplasma diversum was realized with primers UD1, UD2, UD3 and UD4. For bacteriologic isolation, obtained samples were diluted up to 10-1 at 10-5, inoculated

  12. Toxicidade para lagartas de Phthorimaea operculella (Zell. dos aleloquímicos 2-tridecanona e 2-undecanona presentes em tomateiro (Lycopersicon spp. Toxicity of 2-tridecanone and 2-undecanone allelochemicals present in tomato (Lycopersicon spp. for Phthorimaea operculella (Zell. larvae

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    M.U. Ventura

    1995-12-01

    Full Text Available Avaliou-se a toxicidade para Phthorimaea operculella (Zell. dos aleloquímicos 2-trídecanona e 2-undecanona, presentes nos exsudatos glandulares dos tricomas de Lycopersicon spp. Determinou-se a toxicidade dos exsudatos de L. hirsutum f. glabratum PI134417 (DL50, do 2-tridecanona (dose letal: DL50 e concentração letal: CL50 e do 2-undecanona (CL50 para lagartas de primeiro ínstar desse inseto. A DL50 foi de 5,54 glândulas e 775,53 ng por lagarta, respectivamente. Em relação à concentração letal, o 2-tridecanona foi mais tóxico do que o 2-undecanona. A toxicidade (CL50 do 2-tridecanona aumentou com a elevação da temperatura de 27 para 32°C e para o 2-undecanona ocorreu o inverso. Concluiu-se que estes compostos podem ter grande importância na resistência do tomateiro para esta praga.This research deals with the toxicity of 2-tridecanone and 2-undecanone allelochemicals, present in the grandular trichomes of Lycopersicon spp., to Phthorimaea operculella (Zell.. The toxicity of the exsudates of L. hirsutum f. glabratum PI 134117 (LD50, of the 2-tridecanone (lethal dose: LD50and lethal concentration: LC50 and 2-undecanone (LC50 to first instar larvae of this pest were determined. The LD50value was 5.54 glands and 775.53 ng per larvae, respectively. With respect to the lethal concentration, the 2-tridecanone was more toxic than the 2-undecanone. The toxicity (CL50 of the 2-tridecanone increased with the elevation of the temperature from 27 to 32°C, but decreased with the 2-undecanone. It was concluded that these compounds can have great importance in the resistance of the tomato plant to this pest.

  13. The Genome Sequence of Leishmania (Leishmania) amazonensis: Functional Annotation and Extended Analysis of Gene Models

    Science.gov (United States)

    Real, Fernando; Vidal, Ramon Oliveira; Carazzolle, Marcelo Falsarella; Mondego, Jorge Maurício Costa; Costa, Gustavo Gilson Lacerda; Herai, Roberto Hirochi; Würtele, Martin; de Carvalho, Lucas Miguel; e Ferreira, Renata Carmona; Mortara, Renato Arruda; Barbiéri, Clara Lucia; Mieczkowski, Piotr; da Silveira, José Franco; Briones, Marcelo Ribeiro da Silva; Pereira, Gonçalo Amarante Guimarães; Bahia, Diana

    2013-01-01

    We present the sequencing and annotation of the Leishmania (Leishmania) amazonensis genome, an etiological agent of human cutaneous leishmaniasis in the Amazon region of Brazil. L. (L.) amazonensis shares features with Leishmania (L.) mexicana but also exhibits unique characteristics regarding geographical distribution and clinical manifestations of cutaneous lesions (e.g. borderline disseminated cutaneous leishmaniasis). Predicted genes were scored for orthologous gene families and conserved domains in comparison with other human pathogenic Leishmania spp. Carboxypeptidase, aminotransferase, and 3′-nucleotidase genes and ATPase, thioredoxin, and chaperone-related domains were represented more abundantly in L. (L.) amazonensis and L. (L.) mexicana species. Phylogenetic analysis revealed that these two species share groups of amastin surface proteins unique to the genus that could be related to specific features of disease outcomes and host cell interactions. Additionally, we describe a hypothetical hybrid interactome of potentially secreted L. (L.) amazonensis proteins and host proteins under the assumption that parasite factors mimic their mammalian counterparts. The model predicts an interaction between an L. (L.) amazonensis heat-shock protein and mammalian Toll-like receptor 9, which is implicated in important immune responses such as cytokine and nitric oxide production. The analysis presented here represents valuable information for future studies of leishmaniasis pathogenicity and treatment. PMID:23857904

  14. Comparación de seis métodos coproscópicos para el diagnóstico del cromista Blastocystis spp | Comparison of six coproscopics methods for the diagnosis of the chromista Blastocystis spp

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    Milagros Figueroa Lara,

    2017-10-01

    Full Text Available Blastocystis spp. es uno de los parásitos intestinales zoonóticos de mayor prevalencia y distribución mundial. Está caracterizado por una gran variabilidad genética, lo que dificulta su estudio, generando controversias en cuanto a características morfológicas, ciclo vital, criterios diagnósticos y rol patógeno. Se evaluaron seis técnicas de laboratorio para la identificación de Blastocystis spp. en 391 muestras fecales de pacientes de ambos sexos y diferentes edades que asistieron al Laboratorio Clínico Universitario (LCU y a los laboratorios de emergencia y general del Servicio Autónomo del Hospital Universitario Antonio Patricio de Alcalá (SAHUAPA, Cumaná, estado Sucre, Venezuela, durante mayo-junio de 2013. Se consideró el examen directo como estándar de oro para la detección del cromista, y como pruebas de comparación los métodos de concentración: sedimentación espontánea en tubo (SET y Ritchie modificado, y las coloraciones: tinta china modificada, safranina-azul de metileno y May-Grünwald-Giemsa. Se obtuvo una prevalencia de parasitosis intestinal de 32,74% con el examen directo, con mayor frecuencia de Blastocystis spp. (17,39%. Al comparar los métodos de concentración y coloración, se pudo evidenciar que las técnicas que ofrecieron mejores porcentajes de sensibilidad fueron la tinción de May-Grünwald-Giemsa (94,12%, seguido de la tinta china modificada (92,65%; por último, la utilización de safranina-azul de metileno (76,47%, en cuanto a los porcentajes de especificidad superaron el (99,00% con índices Kappa excelentes (> 0,75. Los métodos de concentración resultaron con porcentajes de sensibilidad relativamente bajos, mientras que la especificidad se mantuvo por encima de (99,00%, razón por la cual los métodos de tinción tinta china y May-Grünwald-Giemsa pueden ser incluidos como técnicas complementarias para la detección y estudio morfológico de Blastocystis spp.

  15. Experimental treatment with sodium stibogluconate of hamsters infected with Leishmania (Leishmania) chagasi and Leishmania (Leishmania) amazonensis Tratamento experimental com stibogluconato de sódio em hamsters infectados com Leishmania (Leishmania) chagasi e Leishmania (Leishmania) amazonensis

    OpenAIRE

    Elizabeth M. de Figueiredo; Jaime Costa e Silva; Reginaldo P. Brazil

    1999-01-01

    The present paper reports the experimental treatment of hamsters infected with Leishmania chagasi and Leishmania amazonensis with sodium stibogluconate (20mg/kg/day x 20 days). Only with L. chagasi did the treatment result in the complete elimination of parasites from the spleen. However, no parasitological cure was achieved in hamsters infected with L. amazonensis.O presente trabalho é um relato do tratamento experimental de hamsters infectado com Leishmania chagasi e Leishmania amazonensis ...

  16. Factores de riesgo para la transmisión de leishmaniasis cutánea en niños de 0 a 5 años en un área endémica de Leishmania (Viannia braziliensis Risk factors for cutaneous leishmaniasis transmission in children aged 0 to 5 years in an endemic area of Leishmania (Viannia braziliensis

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    Julia Ampuero

    2005-02-01

    Full Text Available Fue realizado un estudio caso-control pareado en Corte de Pedra, Bahía, Brasil, área endémica de Leishmania (Viannia braziliensis para evaluar los diferentes factores de riesgo asociados a la presencia de leishmaniasis cutánea en niños de 0 a 5 años. Fueron considerados como casos, los niños con prueba de leishmanina positiva y que presentaban en el examen físico, una o más lesiones clínicas, activas o cicatrizales, compatibles con leishmaniasis cutánea. Fueron seleccionados 40 casos y 71 controles que fueron pareados por edad y área de residencia. La presencia de algún otro miembro de la familia con antecedente de leishmaniasis cutánea durante el año anterior a la aparición de la enfermedad en el niño demostró ser un importante factor de riesgo (MÔR MH = 17,75; IC95%: 4,08-77,25. No se encontraron evidencias de asociación con otros factores, como hábitos del niño dentro y fuera de casa, características de la vivienda y del peridomicilio, presencia de vectores y animales como probables reservorios. Estos hallazgos apoyan la hipótesis que el ser humano podría comportarse como un posible reservorio y servir de fuente de contagio para este grupo de edad.With the purpose of identifying risk factors for cutaneous leishmaniasis transmission in children from 0 to 5 years, a matched case-control study was carried out in Corte de Pedra, Bahia, Brazil, an endemic area of Leishmania (Viannia braziliensis. Children with a positive leishmanin skin test and one or more active lesions or scars consistent with cutaneous leishmaniasis were defined as cases. Forty cases and 71 controls were selected and matched by age and place of residence. The presence of a family member with a history of cutaneous leishmaniasis in the year prior to the appearance of the disease in the child was found to be an important risk factor (MÔR MH = 17.75; 95%CI: 4.08-77.25. No evidence of association between the disease and other risk factors was found, such

  17. Could Phlebotomus mascittii play a role as a natural vector for Leishmania infantum? New data.

    Science.gov (United States)

    Obwaller, Adelheid G; Karakus, Mehmet; Poeppl, Wolfgang; Töz, Seray; Özbel, Yusuf; Aspöck, Horst; Walochnik, Julia

    2016-08-19

    The occurrence of phlebotomine sand flies in Central Europe was questioned until they were recorded for the first time in Germany in 1999, and ten years later also in Austria. The aim of this study was to investigate sand flies collected in Austria for their carrier status of Leishmania spp. From 2012 to 2013 field studies were conducted in eastern Austria. Altogether, 22 individuals of sand flies were found, all morphologically identified as Phlebotomus (Transphlebotomus) mascittii Grassi, 1908. Twelve non-engorged female specimens with no visible remnants of a blood meal in their bodies were individually investigated for Leishmania spp. by ITS-1 real-time PCR. One out of these was positive for Leishmania, identified as Leishmania infantum by DNA sequencing. This finding suggests that L. infantum is not excreted by P. mascittii and possibly can establish an infection within P. mascittii. Interestingly, an asymptomatic dog living on the farm where this sand fly had been caught was also Leishmania-positive. This study provides new data on the suspected vector capacity of P. mascittii, being the northernmost sand fly species in Europe and in most central European regions the only sand fly species found. Proven vector capacity of P. mascittii for Leishmania spp. would be of significant medico-veterinary importance, not only with respect to expanding sand fly populations in Central Europe related to global warming, but also in the light of globalization and increasing movements of humans.

  18. 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

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    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.

  19. Gluconeogenesis in Leishmania mexicana

    Science.gov (United States)

    Rodriguez-Contreras, Dayana; Hamilton, Nicklas

    2014-01-01

    Gluconeogenesis is an active pathway in Leishmania amastigotes and is essential for their survival within the mammalian cells. However, our knowledge about this pathway in trypanosomatids is very limited. We investigated the role of glycerol kinase (GK), phosphoenolpyruvate carboxykinase (PEPCK), and pyruvate phosphate dikinase (PPDK) in gluconeogenesis by generating the respective Leishmania mexicana Δgk, Δpepck, and Δppdk null mutants. Our results demonstrated that indeed GK, PEPCK, and PPDK are key players in the gluconeogenesis pathway in Leishmania, although stage-specific differences in their contribution to this pathway were found. GK participates in the entry of glycerol in promastigotes and amastigotes; PEPCK participates in the entry of aspartate in promastigotes, and PPDK is involved in the entry of alanine in amastigotes. Furthermore, the majority of alanine enters into the pathway via decarboxylation of pyruvate in promastigotes, whereas pathway redundancy is suggested for the entry of aspartate in amastigotes. Interestingly, we also found that l-lactate, an abundant glucogenic precursor in mammals, was used by Leishmania amastigotes to synthesize mannogen, entering the pathway through PPDK. On the basis of these new results, we propose a revision in the current model of gluconeogenesis in Leishmania, emphasizing the differences between amastigotes and promastigotes. This work underlines the importance of studying the trypanosomatid intracellular life cycle stages to gain a better understanding of the pathologies caused in humans. PMID:25288791

  20. Cultivo de Penicillium spp. em resíduos da colheita de soja para produção de celulase, protease e amilase

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    Joice Raísa Barbosa Cunha

    Full Text Available RESUMO O objetivo deste estudo foi avaliar a produção de enzimas amilolíticas, celulolíticas e proteolíticas pela linhagem Penicillium spp. LEMI A8221 cultivada em estado sólido em resíduos da colheita de soja, ao longo de quatro dias, em diferentes condições de pH (5,0 e 6,0, temperatura (30 e 35°C e concentrações de substrato (70 e 90% p/v. As atividades máximas obtidas para α-amilase, β-amilase, CMCase e protease foram de 0,20; 0,13; 0,65 e 147 U.mg-1, respectivamente. As condições de fermentação influenciaram a atividade das enzimas, sendo a concentração de substrato, a variável mais significativa para o processo. O tempo de fermentação exerceu efeito apenas para as atividades de amilase sacarificante e CMCase, sendo registrados os menores valores de atividade para essas enzimas, nas primeiras 24 e 48 horas de fermentação, respectivamente. Conclui-se que o isolado Penicillium spp. LEMI A8221 pode ser considerado promissor agente biológico, com aplicação industrial, e o resíduo de soja apresentou-se como fonte de carbono alternativa, no cultivo em estado sólido, para produção de enzimas por esta linhagem microbiana.

  1. Effect of ionizing radiation on the morphology, physiology and growth of Leishmania ssp

    International Nuclear Information System (INIS)

    Bonetti, Franco C.; Spencer, Patrick J.; Nascimento, Nanci do; Junior A, Heitor F.

    2000-01-01

    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)

  2. Histogram score contributes for reliability of DNA content estimatives in Brachiaria spp Notas do histograma contribuem para a confiabilidade das estimativas do conteúdo de DNA de Brachiaria spp

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    Ana Luiza de Oliveira Timbó

    2012-12-01

    Full Text Available Flow cytometry allows to estimate the DNA content of a large number of plants quickly. However, inadequate protocols can compromise the reliability of these estimates leading to variations in the values of DNA content the same species. The objective of this study was to propose an efficient protocol to estimate the DNA content of Brachiaria spp. genotypes with different ploidy levels using flow cytometry. We evaluated four genotypes (B. ruziziensis diploid and artificially tetraploidized; a tetraploid B. brizantha and a natural triploid hybrid, three buffer solutions (MgSO4, Galbraith and Tris-HCl and three species as internal reference standards (Raphanus sativus, Solanum lycopersicum e Pisum sativum. The variables measured were: histogram score (1-5, coefficient of variation and estimation of DNA content. The best combination for the analysis of Brachiaria spp. DNA content was the use of MgSO4 buffer with R. sativus as a internal reference standard. Genome sizes expressed in picograms of DNA are presented for all genotypes and the importance of the histogram score on the results reliability of DNA content analyses were discussed.A citometria de fluxo permite estimar o conteúdo de DNA de um grande número de plantas rapidamente. No entanto, protocolos inadequados podem comprometer a confiabilidade dessas estimativas, levando a variações nos valores de conteúdo de DNA para uma mesma espécie. Neste trabalho, objetivou-se propor um protocolo eficiente para a estimativa do conteúdo de DNA de genótipos de Brachiaria spp. com diferentes níveis de ploidia, utilizando a citometria de fluxo. Foram avaliados quatro genótipos (B. ruziziensis, diploide e tetraploidizada artificialmente; B. brizantha tetraploide e um híbrido natural triploide, 3 soluções tampões (MgSO4, Galbraith e Tris-HCl e três espécies como padrões de referência interno (Raphanus sativus, Solanum lycopersicum e Pisum sativum. As variáveis mensuradas foram: nota do

  3. Utilidad del modelo animal ratón-pulmón para evaluar la virulencia de posibles cepas vacunales de Shigella spp

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    Olga M. Martínez

    2008-04-01

    Full Text Available Shigella flexneri y Shigella sonnei, como cualquier otra especie del género Shigella, se sitúan entre los principales agentes etiológicos de las enfermedades diarreicas agudas, sobre todo aquellas que ocurren en los países en vías de desarrollo, aunque por la baja dosis infectante de este enteropatógeno no se excluyen los países desarrollados. Esta situación conlleva a la elaboración de vacunas para prevenir esta enfermedad y la necesidad de un modelo animal que pruebe la eficacia protectora e inmunogénica de posibles candidatos vacunales contra la shigellosis, situación que ha motivado numerosos estudios por la dificultad de demostrar la enteropatía intestinal en los monos y humanos. Lo anteriormente expuesto, más la capacidad de Shigella spp para mostrar resistencia a los antimicrobianos, motivó la realización de este trabajo. En el mismo se constató la utilidad del modelo animal ratón-pulmón para evaluar la virulencia de candidatos vacunales a partir de este microorganismo. Se utilizó la técnica de inoculación intranasal con una concentración entre 107 y 109 UFC de cepas de Shigella flexneri y Shigella sonnei. Por todos los resultados obtenidos con el modelo animal ratón-pulmón se concluyó que este modelo puede ser eficiente para los estudios preclínicos de cualquier candidato vacunal a partir de Shigella spp.

  4. Antifungal property of chili, shallot and garlic extracts against pathogenic fungi, Phomopsis spp., isolated from infected leaves of para rubber (Hevea brasiliensis Muell. Arg.

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    Patcharawan Sittisart

    2017-12-01

    Full Text Available The aim of this study was to evaluate the antifungal property of extracts of chili, shallot and garlic (local varieties in Sisaket, Thailand against pathogenic fungi, Phomopsis spp., which were isolated from infected leaves of para rubber (Hevea brasiliensis Muell. Arg.. Seven isolates of Phomopsis spp. namely Phomopsis sp. SSK1.1, SSK1.2, SSK3.1, SSK4.1, SSK5.1, SSK5.2 and SSK7.1 were identified on the basis of morphological characteristics. Fresh plants were extracted with water to obtain crude extracts and their antifungal properties were tested on potato dextrose agar (PDA media. The study demonstrated that increasing the concentrations (20%, 40%, 60% or 80% of the chili extract exhibited a dependent increase in the inhibitory level on mycelial growth of Phomopsis spp. SSK3.1, SSK4.1 and SSK5.2. The inhibitory level on mycelial growth of shallot extract also increased in a dose-dependent manner in all isolates of Phomopsis. The garlic extract had significant inhibition on the growth of all isolates with complete inhibition at 80% concentration. The highest levels of percentage inhibition of mycelial growth were with garlic extract followed by shallot and chili extracts, respectively. The study also showed that these plant extracts contained some polyphenols (apigenin, gallic acid, catechin, quercetin, kaempferol and tannic acid which are well-known compounds possessing antifungal activity. Therefore, it is possible that the antifungal properties of these plant extracts were partly due to these polyphenols or unknown active compounds which could not be analyzed in this study. Collectively, these results suggest that local varieties of both shallot and garlic possess strong antifungal properties. Keywords: Chili, Garlic, Para rubber, Phomopsis, Shallot

  5. Comportamiento experimental del Sporothrix schenckii y la Leishmania mexicana en el hamster Experimental behavior of Sporothrix schenckii and Leishmania mexicana in hamsters

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    Luz Angela González de Polanía

    1990-10-01

    Full Text Available La descripción macroscópica del proceso de patogénesis en hamsters inoculados subcutáneamente en nariz con Sporothrix schenckii ó Leishmania mexicana spp. proporcionó bases para diferenciar estos dos microorganismos en un modelo animal utilizado comunmente para estudiarlos. Observaciones secuenciales durante 150 días permitieron afirmar que en las infecciones causadas por estos patógenos se presentaron edema y eritema como signos primarios, seguidos de alopecia, necrosis y ulceración. La producción de pus fué una característica distintiva para el S. schenckii. Estos signos clínicos se observaron más temprano en la esporotricosis que en la infección por L. mexicana, mostrando diferencias estadísticas significantes en días promedio de aparición. El presente trabajo muestra que las lesiones producidas tanto por el S. schenckii como la L. mexicana en este modelo experimental comparten signos clínicos, pero el tiempo de aparición de los mismos y su frecuencia relativa permiten diferenciarlas. Las condiciones de inoculación como: cepa de los microorganismos, dosis del inóculo, sitio y vía de inoculación, deben tenerse presentes en la evaluación de su comportamiento experimental.The macroscopic description of the pathogenic process of Sporothrix schenckii and Leishmania mexicana spp in hamsters inoculated subcutaneously in the nose provided bases for the differentiation of the behavior of these two microorganisms in a model frequently utilized for their study. Sequential observations over 150 days demonstrated that infections caused by these pathogens results initially in edema and erythema followed by loss of hair, necrosis and ulceration. The pus production was a characteristic presented only by S. schenckii. These clinical signs were observed earlier in sporotrichosis than in L. mexicana infection. Differences in the mean day of appearance were statistically significant. The lesions produced by S. schenckii and L. mexicana

  6. 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

  7. Deception and Manipulation: the arms of Leishmania, a successful parasite

    Directory of Open Access Journals (Sweden)

    Pedro eCecílio

    2014-10-01

    Full Text Available 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.

  8. Sistemática molecular de algunas especies de Lutzomyia spp del grupo verrucarum (Theodor 1965. Búsqueda de marcadores moleculares para la caracterización de especies

    Directory of Open Access Journals (Sweden)

    I. D. Vélez

    2000-02-01

    Full Text Available

    En el nuevo mundo la Leishmaniosis es transmitida por dípteros del género Lutzomyia frança. Este género presenta una amplia diversidad morfológica por lo que ha sido dividida en subgéneros, grupos y series de especies. Se ha propuesto que la reciente especiación de algunos grupos fue debida a cambios climáticos en el pasado, especialmente en el pleistoceno (hace 1.8 millones de años, los cuales sirvieron para aislar poblaciones coespecíficas en diferentes refugios durante períodos secos. El grupo verrucarum es uno de los agrupamientos subgenéricos en el que se incluyen 28 especies. Con base en la morfología de la genitalia de machos se ha subdividido este grupo en las series serrana, verrucarum y towsendi. De estas especies L. evansi se ha incriminado como vector alternativo de Leishmania (Leishmania infantum chagasi en un foco de Leishmaniosis visceral al norte de Colombia. Otras especies se han incriminado en la transmisión de Leishmania (Viannia, agente causante de Leishmaniosis cutánea.

    Debido a la gran controversia sobre la ubicación sistemática de muchos insectos de importancia médica, surgió un macroproyecto del cual hace parte este estudio y que tiene como propósito aportar al conocimiento sobre las relaciones filogenéticas entre especies del género Lutzomyia. Las especies del grupo verrucarum tienen un interés particular debido a las frecuentes dificultades en su determinación taxonómica basada en caracteres morfológicos. Este estudio tiene como objetivo particular realizar un análisis cladístico entre algunas especies alopátricas (distanciadas geográficamente presentes en Colombia, usando secuencias del gen que codifica para la unidad cuatro de la NADH deshidrogenasa mitocondrial

  9. Real-time PCR for Leishmania species identification: Evaluation and comparison with classical techniques.

    Science.gov (United States)

    de Morais, Rayana Carla Silva; da Costa Oliveira, Cintia Nascimento; de Albuquerque, Suênia da Cunha Gonçalves; Mendonça Trajano Silva, Lays Adrianne; Pessoa-E-Silva, Rômulo; Alves da Cruz, Heidi Lacerda; de Brito, Maria Edileuza Felinto; de Paiva Cavalcanti, Milena

    2016-06-01

    Cutaneous leishmaniasis (CL) is a parasitic disease caused by various Leishmania species. Several studies have shown that real time quantitative PCR (qPCR) can be used for Leishmania spp. identification by analyzing the melting temperature (Tm). Thus, the aim of this study was to evaluate the viability of qPCR for differentiating eight closely related Leishmania species that cause the same clinical form of the disease and to compare the results with classical techniques. qPCR assays for standardizing the Tm using reference strains were performed. After the CL diagnosis on blood samples of domestic animals, positive samples were analyzed by their Tm and qPCR products were purified and sequenced. Ten human samples previously characterized by Multilocus Enzyme Electrophoresis (MLEE) were also analyzed by Tm. A Restriction Fragment Length Polymorphism (RFLP) assay, a reference test, was also standardized, by using the reference strains. Through standardization of Tm for Leishmania spp., two Tm ranges were created for analysis: 1 (Tm = 78-79.99 °C) included Leishmania (V.) braziliensis, Leishmania (V.) panamensis, Leishmania (V.) lainsoni, Leishmania (V.) guyanensis and Leishmania (V.) shawi; and 2 (Tm = 80-82.2 °C) included Leishmania (V.) naiffi, Leishmania (L.) amazonensis and Leishmania (L.) mexicana. A total of 223 positive blood samples were analyzed, with 58 included in range 1 and 165 in range 2. L. (V.) braziliensis, L. (V.) panamensis and L. (V.) guyanensis were identified by sequencing, while L. (V.) braziliensis, L. (L.) mexicana and L. (V.) panamensis were identified by RFLP analysis. Ten human samples previously characterized by Multilocus Enzyme Electrophoresis (MLEE) were also analyzed by qPCR Tm analysis; five were classified in range 1 and five in range 2. A concordance of 80% was calculated between qPCR and the gold-standard (MLEE) with no significant difference between the methods (p = 0.6499); a similar result was observed for sequencing

  10. Método de difusión con discos para la determinación de sensibilidad a fluconazol en aislamientos de Candida spp Disk diffusion method for fluconazole susceptibility testing of Candida spp. isolates

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    L. Rodero

    2006-09-01

    Full Text Available Se estudiaron 1193 aislamientos clínicos para estandarizar y evaluar un método de difusión con discos de fluconazol de lectura visual, que permita detectar levaduras sensibles al antifúngico. Las especies analizadas fueron: Candida albicans (n=584, Candida parapsilosis (n=196, Candida tropicalis (n=200, Candida glabrata (n=113, Candida krusei (n=50, Candida spp. y otras levaduras oportunistas (n=50. Los discos fueron manufacturados en el INEI-ANLIS "Dr. Carlos G. Malbrán". Se midieron los halos de inhibición del crecimiento producidos por fluconazol y la concentración inhibitoria mínima (CIM por el método de referencia M27-A2 modificado por EUCAST. Se establecieron los valores de corte del método de difusión en: ≥16 mm para levaduras sensibles a fluconazol (CIM ≤ 8 µg/ml, entre 9 y 15 mm para sensibles dependientes de la dosis (CIM = 16-32 mg/ml y ≤ 8 mm para resistentes (CIM ≥ 64 µg/ml. El método de difusión tuvo 94,7% de concordancia con el de referencia, con 0,2% de errores very major y 0,3% de errores major. La reproducibilidad inter e intralaboratorio fue muy buena. Para detectar aislamientos sensibles a fluconazol, este método resulta confiable y de bajo costo; sin embargo, es conveniente que los aislamientos con halos ≤ 15 mm sean reevaluados por el método de referencia.In order to standardize and evaluate a disk diffusion method with visual reading to detect in vitro fluconazole susceptibility of yeast, 1193 clinical isolates were tested. These included 584 Candida albicans, 196 Candida parapsilosis, 200 Candida tropicalis, 113 Candida glabrata, 50 Candida krusei and 50 Candida spp. and other opportunistic yeasts. The disks were manufactured in the INEI-ANLIS "Dr. Carlos G. Malbrán". The disk diffusion method results were compared to MIC results obtained by the reference CLSI M27-A2 broth microdilution method modified by EUCAST. The interpretative breakpoints for in vitro susceptibility testing of fluconazole

  11. Preparo de Itens de Ensaio de Proficiência em Matriz Queijo para a Pesquisa de Salmonella spp. | Preparation of Proficienc Test Items for Salmonella spp. Detection in Cheese Matrix

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    Juliana de Castro Beltrão da Costa

    2015-08-01

    Full Text Available A participação de laboratórios analíticos em ensaio de proficiência permite a verificação da confiabilidade dos resultados gerados nas análises de controle da qualidade. O objetivo deste estudo foi produzir um lote de itens de ensaio (IE a ser utilizado em ensaio de proficiência (EP para pesquisa de Salmonella spp. em matriz queijo. Queijo Minas Frescal (QMF Ultrafiltrado foi utilizado como matriz e fortificado com uma cepa de Salmonella Enteritidis. A trealose foi utilizada como crioprotetor e a liofilização como técnica de preservação. O lote foi avaliado quanto a verificação do vácuo, teste da homogeneidade, estudo da estabilidade em longo prazo nas temperaturas de -70oC (referência e -20oC (armazenamento e em curto prazo nas temperaturas de 4, 25 e 35ºC (transporte. O lote apresentou presença de vácuo em 95,6% dos frascos e foi considerado suficientemente homogêneo. Os IE apresentaram estabilidade a -70ºC superior a 360 dias e a -20ºC superior a 160 dias. Os IE demonstraram ser estáveis por até seis dias nas temperaturas de 4 e 25ºC, mas não a 35ºC. Conclui-se que a metodologia utilizada foi satisfatória para produção de IE para o ensaio de pesquisa de Salmonella spp. em matriz queijo. ----------------------------------------------------------------------------------------------- The participation of analytical laboratories in proficiency testing allows the verification of the reliability of results in analysis of quality control. The aim of this study was to produce a batch of test items (TI to be used in a proficiency assay for Salmonella spp. research in a cheese matrix. Ultrafiltered Minas cheese was used as the matrix and spiked with a strain of Salmonella Enteritidis. Trehalose was used as cryoprotectant, and freeze-drying was used as the preservation technique. The batch was evaluated for vacuum verification, homogeneity study, and long-term stability testing at temperatures of -70ºC (reference

  12. Increased transmission potential of Leishmania major/Leishmania infantum hybrids

    OpenAIRE

    Volf, Petr; Benkova, Ivana; Myskova, Jitka; Sadlova, Jovana; Campino, Lenea; Ravel, Christophe

    2007-01-01

    Development of Leishmania infantum/Leishmania major hybrids was studied in two sand fly species. In Phlebotomus papatasi, which supported development of L. major but not L. infantum, the hybrids produced heavy late-stage infections with high numbers of metacyclic promastigotes. In the permissive vector Lutzomyia longipalpis, all Leishmania strains included in this study developed well. Hybrids were found to express L. major lipophosphoglycan, apparently enabling them to survive in P. papatasi...

  13. CD95 (FAS) and CD178 (FASL) induce the apoptosis of CD4+ and CD8+ cells isolated from the peripheral blood and spleen of dogs naturally infected with Leishmania spp.

    Science.gov (United States)

    Silva, Kathlenn Liezbeth Oliveira; Melo, Larissa Martins; Perosso, Juliana; Oliveira, Bruna Brito; Santos, Paulo Sérgio Patto Dos; Eugênio, Flávia de Rezende; Lima, Valéria Marçal Felix de

    2013-11-08

    Infected dogs are urban reservoirs of Leishmania chagasi, which is a causative agent of visceral leishmaniasis (VL). Dogs exhibit immune suppression during the course of this disease, and lymphocyte apoptosis is involved in this process. To investigate apoptosis and the expression levels of FAS-FAS-associated death domain protein (CD95 or APO-1), FASL-FAS ligand protein (CD178), and TRAIL-TNF-related apoptosis-inducing ligand (CD253) receptors in peripheral blood mononuclear cells and spleen leukocytes from 38 symptomatic dogs with moderate VL and 25 healthy dogs were evaluated by flow cytometry. The apoptosis rate of blood and splenic CD4+ and CD8+ cells was higher in infected dogs than in healthy dogs. The expression levels of FAS and FASL in blood and splenic CD4+ cells were lower in infected dogs than in healthy dogs. FAS expression in CD8+ cells was higher in infected dogs than in healthy dogs; in contrast, FASL expression was lower in infected dogs. The expression of the TRAIL receptor increased only in splenic CD8+ cells from infected dogs. The FAS and FAS-L blocking antibodies confirmed the importance of these receptors in apoptosis. Our results enhance the current understanding of the immune response in dogs infected with L. chagasi, facilitating the future development of therapeutic interventions to reduce lymphocyte depletion. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Phylogenetic position of Leishmania isolates from Khyber Pakhtunkhwa province of Pakistan.

    Science.gov (United States)

    Khan, Nazma Habib; Messenger, Louisa A; Wahid, Sobia; Sutherland, Colin J

    2016-08-01

    Several species of the genus Leishmania are causative agents of cutaneous leishmaniasis in Pakistan. This study aimed to determine phylogenetic placement of Leishmania species causing cutaneous leishmaniasis in Khyber Pakhtunkhwa province, Pakistan (34 Leishmania tropica, 3 Leishmania infantum), in-relation to species from other geographical areas using gene sequences encoding cytochrome b (cytb) and internal transcribed spacer 2 (its2). Based on cytochrome b sequence analysis, L. tropica strains from Pakistan and other geographical regions were differentiated into two genotype groups, A and B. Within the province, five distinct L. tropica genotypes were recognized; two in group A, three in group B. Two L. infantum isolates from the province were closely associated with both Afro-Eurasian and American species of the Leishmania donovani complex, including Leishmania chagasi, L. infantum and L. donovani from Sudan and Ethiopia; while a third L. infantum isolate could not be differentiated from visceralizing Kenyan and Indian L. donovani. We observed apposite phylogenetic placement of CL-causing L. tropica and L. infantum from Khyber Pakhtunkhwa. Affinities ascribed to Leishmania spp. From the region are valuable in tracing potential importation of leishmaniasis. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  15. Definición de las condiciones de temperatura y almacenamiento adecuadas en la detección de ADN de Leishmania por PCR en flebotominos.

    Directory of Open Access Journals (Sweden)

    Olga L. Cabrera

    2002-09-01

    Full Text Available Para estudios epidemiológicos y programas de control de las leishmaniasis es importante la determinación taxonómica del insecto vector y del agente etiológico causante de esta enfermedad. Por su eficacia y sensibilidad, la utilización de técnicas moleculares, como la reacción en cadena de la polimerasa (PCR ha permitido avances en entomología médica. La metodología tradicional usada para la búsqueda de infección en los flebótomos es un método dispendioso que requiere mucho tiempo y capacitación para emitir un diagnóstico acertado. En el presente trabajo se evaluaron las condiciones y prácticas adecuadas para el almacenamiento de flebótomos con el propósito de aplicar la PCR. Hembras de Lutzomyia longipalpis de la colonia del Instituto Nacional de Salud se infectaron experimentalmente con una cepa de Leishmania chagasi del valle alto del río Magdalena (Quipile, Cundinamarca. Los insectos infectados se preservaron en tres soluciones: etanol al 100%, etanol al 70% y en buffer Tris- EDTA (TE; submuestras de cada una se almacenaron a -80 °C, -20 °C y a temperatura ambiente. Para determinar los porcentajes de infección, algunas de las hembras se disecaron para buscar las formas flageladas al microscopio. La extracción del KADN se realizó con Chelex 100. Para la amplificación se utilizaron los iniciadores OL1 y OL2 de Leishmania con electroforesis en geles de agarosa al 1%. En cada una de las condiciones descritas se logró la amplificación de un fragmento de »120 pb, correspondiente a Leishmania spp. Estos resultados muestran la ventaja de incorporar como técnica de rutina la PCR para detectar flebótomos infectados de zonas endémicas de leishmaniasis visceral. Por costo-efectividad, se concluyó que las muestras entomológicas para estudios de incriminación vectorial utilizando la PCR, se pueden preservar a temperatura ambiente en etanol al 70%.

  16. Estudo do momento ótimo de reforma para povoamentos de Eucalyptus spp - o caso da redução dos custos.

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    Álvaro Nogueira de Souza

    2010-08-01

    Full Text Available Os objetivos deste trabalho foram: a propor e verificar a eficiência de um modelo matemático que permita estudar os efeitos do progresso tecnológico (redução dos custos sobre a época ótima de substituir povoamentos de Eucalyptus spp; b determinar a rotação florestal de um povoamento de Eucalyptus spp; c verificar qual seria o momento de substituir se as receitas estivessem constantes desde o início dos incentivos fiscais, ou seja, na década de 60, e, d projetar o momento de substituir o povoamento no futuro, considerando os custos decrescentes e a produtividade constante a valores de hoje. Utilizou-se a Função Gompertz para o cálculo da produção de madeira. O critério utilizado para a avaliação econômica da viabilidade dos projetos foi o Valor Presente Líquido. O modelo proposto permitiu calcular o efeito do progresso tecnológico nos custos no período entre 1960 e 2000 e projetar os valores dos custos para o período entre 2000 e 2040. Concluiu-se que: a a rotação, com valores atuais, encontra-se, em 7 anos, mantidas as receitas da década de 60; b o momento ótimo de reforma estaria em seis cortes; c o modelo mostrou-se eficiente para estimativas em um horizonte de até 40 anos; o estudo da cadeia de substituição mostrou que as épocas ótimas de reforma caíram ao longo dos anos, de seis cortes na década de 60, três na década de 80 e dois atualmente; a tendência é permanecer nos dois cortes passando a três com um pequeno aumento na produtividade das talhadias.

  17. Actuaciones para el control y erradicación del teosinte (Zea mays spp.) en Aragón

    OpenAIRE

    Pardo Sanclemente, Gabriel; Fuertes, S.; Betrán, E.; Cirujeda Ranzenberger, Alicia

    2015-01-01

    En el verano de 2014 se tuvo constancia de la presencia de teosinte (Zea mays spp.) como mala hierba en campos de maíz de Aragón y en menos medida en Cataluña. Se conoce por teosinte a un grupo de especies y subespecies del género Zea, la mayoría de ellas de la misma especie que el maíz y originarias de Méjico. El maíz actual procede de antiguos teosintes mejorados, tras miles de años de selección. Debido al potencial infestante y competidor de esta especie, la Dirección General de Alimentaci...

  18. Variação fenotípica para caracteres silviculturais em populações de Aspidosperma spp. sem estrutura de progênies. Genetic variation for silvicultural traits in populations of Aspidosperma spp. without progeny arrays.

    Directory of Open Access Journals (Sweden)

    Camila Regina Silva Baleroni RECCO

    2016-06-01

    Full Text Available O objetivo deste estudo foi estimar os parâmetros genéticos para os caracteres silviculturais em Aspidosperma polyneuron Müll.Arg. e Aspidosperma cylindrocarpon Müll.Arg., para fins de conservação ex situ e uso em programas de recuperação ambiental. A coleta de sementes foi feita em árvores de polinização aberta no município de Andradina-SP e o teste experimental foi instalado na Fazenda de Ensino e Pesquisa (FEPE/UNESP, em 1997, com as espécies consorciadas com Inga marginata Willd. (ingá feijão em linhas alternadas. O delineamento experimental utilizado foi o de blocos casualizados completos com dois tratamentos (Aspidosperma spp., 32 repetições com parcelas lineares e 4 plantas por parcela. Aos 15 anos de idade foram mensurados diâmetro à altura do peito – DAP, altura de total – ALT e sobrevivência – SOB e os parâmetros genéticos foram estimados pelo programa Selegen. A espécie cylindrocarpon apresentou maior desenvolvimento nos caracteres de crescimento (DAP e ALT quando comparada com A. polyneuron. A taxa de sobrevivência foi de 71% demostrando boa adaptabilidade para ambas as espécies ao local de implantação do experimento. A acurácia foi alta, sugerindo maior confiabilidade das análises estatísticas. A herdabilidade foi moderada, indicando considerável controle genético nas espécies quando se observa que o coeficiente de determinação dentro de parcela foi menor que 10% para ALT e DAP, mostrando que as parcelas lineares são homogêneas, ou seja, o delineamento experimental adotado foi eficiente. Portanto, a ausência do uso de estrutura de progênies nas populações de Aspidosperma spp. estudadas não impossibilitou que fossem obtidas informações relevantes sobre as populações naturais de Aspidosperma, porém vale ressaltar que é de suma importância que novas coletas de sementes sejam realizadas para um estudo com base em teste de progênies. This study aimed to estimate genetic parameters

  19. Occurrence of anti-Brucella abortus and anti-Leptospira spp. antibodies in buffaloes from Paraíba state, Northeastern Brazil

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    Arthur Willian de Lima Brasil

    2015-07-01

    Full Text Available The objective of this study was to determine the frequency of animals that tested positive for brucellosis and leptospirosis and the risk factors for these diseases in the State of Paraíba. A total of 136 buffaloes from 14 herds were examined. For brucellosis, we used the buffered acidified plate antigen (BAPA test as screening method and 2-mercaptoethanol as confirmatory test. For leptospirosis, we conducted a microscopic agglutination test (MAT, with a cut-off point of 1:100. Of the animals examined, two [1.5%; 95% CI = 0.4%–5.2%] were positive for brucellosis, and 38 (27.9%; 95% CI = 21.1%–36.0% were positive for leptospirosis. The brucellosis-positive animals were from two (14.3% herds, while nine (64.3% herds had leptospirosis-seropositive animals. The more frequent Leptospira spp. serotypes were Bratislava, Pomona, and Canicola. We concluded that leptospirosis was widespread in buffaloes in the state of Paraíba and suggested that breeding alongside horses and pigs might be na important factor in the spread of leptospirosis-positive animals. The presence of brucellosispositive animals indicated the possibility of negative-impacting measures on disease control in bovines, and it is therefore recommended that greater attention be given to these animals for brucellosis control.

  20. Frações de zinco em solo arenoso e suas relações com disponibilidade para Cynodon spp cv. Tifton-85 Zinc fractions in a sandy soil and its relations with availability to Cynodon spp cv. Tifton-85

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    E. M. André

    2003-06-01

    Full Text Available Para avaliar um esquema de fracionamento de zinco em Argissolo arenoso e suas relações com a disponibilidade de Zn para Cynodon spp cv. Tifton-85, realizou-se um experimento em casa de vegetação, em esquema fatorial 5 x 2 x 2 (5 doses de Zn, 2 doses de calcário e 2 épocas de amostragem, em delineamento inteiramente casualizado, com três repetições. As doses de Zn foram de 0; 2,5; 5,0; 7,5 e 10 mg dm-3; metade dos vasos não recebeu calagem (V = 42 % e metade recebeu a calagem com vistas em elevar o índice de saturação por bases a 70 %. As épocas de amostragem foram 30 e 150 dias após aplicação de Zn, respectivamente, antes do plantio e depois do 3º corte de Tifton-85. A aplicação de Zn resultou em aumento significativo do elemento nas frações: trocável, óxidos de Mn, matéria orgânica e óxidos de Fe. O Zn ligado aos óxidos de Mn aumentou significativamente com a calagem. Após 150 dias de experimentação, houve diminuição do Zn trocável, ligado aos óxidos de Mn, à matéria orgânica e aos óxidos de Fe, e aumento na fração residual. A distribuição de Zn nas frações do solo foi: residual > óxidos de Fe > óxidos de Mn > trocável > matéria orgânica. As relações entre as características estudadas mostraram que tanto o Zn-DTPA quanto o Zn trocável, ligado à M.O. e aos óxidos de Mn, foram eficientes para representar o Zn absorvido pela planta.To evaluate a fractionation scheme for Zn in a sandy soil (Ultisol and its relation with Zn availability for Cynodon spp cv. Tifton-85, an experiment was carried out in greenhouse, with a factorial scheme (5 doses of zinc, 2 doses of lime and 2 times of sampling, in a completely randomized block design, with three replications. The doses of Zn were: 0; 2.5; 5.0; 7.5 and 10 mg dm-3; half of the pots did not received liming (V = 42% and, in the other half, liming was done to increase the base saturation index to 70%. The sampling times were 30 and 150 days after Zn

  1. Avaliação sorológica para detecção de anticorpos anti-Leishmania em cães e gatos no bairro de Santa Rita de Cássia, Município de Barra Mansa, Estado do Rio de Janeiro Serological evaluation for detection of anti-Leishmania antibodies in dogs and cats in the district of Santa Rita de Cássia, municipality of Barra Mansa, State of Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    Fabiano Borges Figueiredo

    2009-04-01

    Full Text Available A partir de um caso canino de leishmaniose tegumentar americana na localidade de Santa Rita de Cássia, município de Barra Mansa, Rio de Janeiro, foi realizado um inquérito sorológico em 177 cães e 43 gatos. Na avaliação das amostras de soros caninos, 10% foram positivos na reação de imunofluorescência indireta e 10,7% no ensaio imunoenzimático. Entre as amostras de soros felinos testados, nenhum animal foi positivo na reação de imunofluorescência indireta e apenas um (2,4% felino apresentou reação positiva ao ensaio imunoenzimático. A detecção de Leishmania braziliensis, autóctone em Barra Mansa, faz um alerta para a instalação de um possível foco de leishmaniose tegumentar americana nessa região.From a canine case of american cutaneous leishmaniasis in the locality of Santa Rita de Cássia, municipality of Barra Mansa, Rio de Janeiro, a serological survey was performed on 177 dogs and 43 cats. Evaluation of the canine serum samples showed that 10% had a positive reaction in the indirect immunofluorescence test and 10.7% in the enzyme-linked immunosorbent assay. Among the feline serum samples tested, none of the animals had a positive reaction in the indirect immunofluorescence test and only one (2.4% showed a positive reaction in the enzyme-linked immunosorbent assay. The detection of an autochthonous case of Leishmania braziliensis in Barra Mansa gives warning that a focus of american cutaneous leishmaniasis is possibly becoming established in this region.

  2. Leishmania (V.) braziliensis infecting bats from Pantanal wetland, Brazil: First records for Platyrrhinus lineatus and Artibeus planirostris.

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    de Castro Ferreira, Eduardo; Pereira, Agnes Antônio Sampaio; Silveira, Maurício; Margonari, Carina; Marcon, Glaucia Elisete Barbosa; de Oliveira França, Adriana; Castro, Ludiele Souza; Bordignon, Marcelo Oscar; Fischer, Erich; Tomas, Walfrido Moraes; Dorval, Maria Elizabeth Cavalheiros; Gontijo, Célia Maria Ferreira

    2017-08-01

    In the New World genus Leishmania parasites are etiological agents of neglected zoonoses known as leishmaniasis. Its epidemiology is very complex due to the participation of several species of sand fly vectors and mammalian hosts, and man is an accidental host. Control is very difficult because of the different epidemiological patterns of transmission observed. Studies about Leishmania spp. infection in bats are so scarce, which represents a large gap in knowledge about the role of these animals in the transmission cycle of these pathogens, especially when considering that Chiroptera is one of the most abundant and diverse orders among mammals. Leishmaniasis in Mato Grosso do Sul, Brazil are remarkably frequent, probably due to the abundance of its regional mastofauna. The recent record of L. braziliensis in bats from this state indicates the need to clarify the role of these mammals in the transmission cycle. In this study we evaluated the presence of Leishmania parasites in the skin of different species of bats, using PCR directed to Leishmania spp. kDNA for screening followed by PCR/RFLP analysis of the hsp70 gene for the identification of parasite species. Leishmania species identification was confirmed by PCR directed to the G6PD gene of L. braziliensis, followed by sequencing of the PCR product. Samples from 47 bats were processed, of which in three specimens (6.38%) was detected the presence of Leishmania sp. kDNA. PCR/RFLP and sequencing identified the species involved in the infection as L. braziliensis in all of them. This is the first report of Leishmania braziliensis in bats from Pantanal ecosystem and the first record of this species in Platyrrhinus lineatus and Artibeus planirostris, bats with a wide distribution in South America. These results reinforce the need to deepen the knowledge about the possibility of bats act as reservoirs of Leishmania spp. especially considering their ability of dispersion and occupation of anthropic environments

  3. Thrichomys laurentius (Rodentia; Echimyidae as a putative reservoir of Leishmania infantum and L. braziliensis: patterns of experimental infection.

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    André Luiz Rodrigues Roque

    Full Text Available The importance of the genus Thrichomys in the retention of infection and transmission of Leishmania species is supported by previous studies that describe an ancient interaction between caviomorphs and trypanosomatids and report the natural infection of Thrichomys spp. Moreover, these rodents are widely dispersed in Brazil and recognized as important hosts of other tripanosomatids. Our main purpose was to evaluate the putative role of Thrichomys laurentius in the retention of infection and amplification of the transmission cycle of Leishmania infantum and L. braziliensis. Male and female T. laurentius (n = 24 born in captivity were evaluated for the retention of infection with these Leishmania species and followed up by parasitological, serological, hematological, biochemical, histological, and molecular assays for 3, 6, 9, or 12 months post infection (mpi. T. laurentius showed its competence as maintenance host for the two inoculated Leishmania species. Four aspects should be highlighted: (i re-isolation of parasites 12 mpi; (ii the low parasitic burden displayed by T. laurentius tissues; (iii the early onset and maintenance of humoral response, and (iv the similar pattern of infection by the two Leishmania species. Both Leishmania species demonstrated the ability to invade and maintain itself in viscera and skin of T. laurentius, and no rodent displayed any lesion, histological changes, or clinical evidence of infection. We also wish to point out the irrelevance of the adjective dermotropic or viscerotropic to qualify L. braziliensis and L. infantum, respectively, when these species are hosted by nonhuman hosts. Our data suggest that T. laurentius may act at least as a maintenance host of both tested Leishmania species since it maintained long-lasting infections. Moreover, it cannot be discarded that Leishmania spp. infection in free-ranging T. laurentius could result in higher parasite burden due the more stressing conditions in the wild

  4. Molecular detection of Leishmania infantum and Leishmania tropica in rodent species from endemic cutaneous leishmaniasis areas in Morocco.

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    Echchakery, Mohamed; Chicharro, Carmen; Boussaa, Samia; Nieto, Javier; Carrillo, Eugenia; Sheila, Ortega; Moreno, Javier; Boumezzough, Ali

    2017-10-02

    Leishmaniasis remains a major public health problem in African nations, including Morocco, where little is known about the vertebrate reservoirs involved in the causal parasites' transmission cycles. The present study investigates the role of rodent species as potential reservoirs of Leishmania spp. in central Morocco, where both L. tropica and L. infantum have been reported. Rodents were caught from 22 sites in central Morocco, by using Sherman metal traps, and identified morphologically. For each specimen, genomic DNA was extracted from different tissues using the Speed Tools DNA extraction Kit. Then, samples were PCR-analyzed, targeting the SSU rRNA gene to detect Leishmania spp. DNA, followed by amplification of the internal transcribed spacer 1 (ITS1) and its sequencing to identify the species. A total of 197 rodents belonging to ten species were captured and identified: Rattus rattus (40.61%), Mus musculus (25.38%), Apodemus sylvaticus (8.63%), Mus spretus (7.11%), Meriones shawi (5.58%), Rattus norvegicus (4.57%), Meriones libycus (3.05%), Mastomys erythroleucus (2.03%), Gerbillus campestris (2.03%) and Lemniscomys barbarus (1.01%). Molecular analysis revealed the presence of Leishmania species in 18 specimens: six R. rattus (out of 80 captured; 7.5%), 11 M. musculus (out of 50 captured; 22%), and one R. norvegicus (out of 9 captured; 11.11%). To the best of our knowledge, L. infantum and L. tropica were identified in rodent species for the first time in Morocco. These findings suggest that rodent species may be involved in L. infantum and L. tropica transmission cycles in this country but that further studies are needed to confirm their role as reservoirs of Leishmania species in Morocco.

  5. Innate Immunity against Leishmania Infections

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    Gurung, Prajwal; Kanneganti, Thirumala-Devi

    2015-01-01

    Leishmaniasis is a major health problem that affects more than 300 million people throughout the world. The morbidity associated with the disease causes serious economic burden in Leishmania endemic regions. Despite the morbidity and economic burden associated with Leishmaniasis, this disease rarely gets noticed and is still categorized under neglected tropical diseases. The lack of research combined with the ability of Leishmania to evade immune recognition has rendered our efforts to design therapeutic treatments or vaccines challenging. Herein, we review the literature on Leishmania from innate immune perspective and discuss potential problems as well as solutions and future directions that could aid in identifying novel therapeutic targets to eliminate this parasite. PMID:26249747

  6. Bioprospecção de isolados de Trichoderma spp. para o controle de Rhizoctonia solani na produção de mudas de pepino Bioprospection of Trichoderma spp. isolates to control Rhizoctonia solani on cucumber seedling production

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    Cleusa Maria Mantovanello Lucon

    2009-03-01

    Full Text Available O objetivo deste trabalho foi selecionar e identificar isolados de Trichoderma spp. para o controle do tombamento causado por Rhizoctonia solani (AG-4 em plântulas de pepino (Cucumis sativus L., além de avaliar o efeito de concentrações crescentes e de combinações dos isolados mais eficientes no controle da doença. Os experimentos foram conduzidos em casa de vegetação, com 490 isolados. O tombamento das mudas foi avaliado uma semana após a aplicação à base das plântulas de substrato infestado com antagonista (1% e patógeno (1%. Os doze isolados que proporcionaram mais de 85% de redução da doença foram testados em concentrações crescentes para o controle do patógeno (1%: 0,5, 1, 2, 3 e 4%. Também foi avaliado o efeito das combinações dos cinco isolados mais promissores. Os isolados mais efetivos foram identificados pelo sequenciamento da região espaçadores internos transcritos (ITS do DNA ribossômico. Dos 490 isolados testados 44 (9% reduziram o tombamento. As concentrações de antagonistas superiores a 2% foram as mais efetivas no controle da doença. Apenas duas combinações resultaram no aumento do controle da doença. Os isolados mais efetivos foram identificados como T. hamatum (IB08, IB30, IB60, T. harzianum (IB34, IB35, T. atroviride (IB13, T. spirale (IB16, IB24 e T. asperellum (IB44. Não foi possível a identificação da espécie de três isolados.The objective of this work was to select and identify Trichoderma spp. isolates for the control of Rhizoctonia solani (AG-4 damping-off on cucumber (Cucumis sativus L. seedlings, as well as to evaluate the effects of increasing concentrations and different combinations of the most efficient isolates in the disease control. The experiments were carried out in a greenhouse with 490 isolates. The disease on cucumber seedlings was evaluated one week after the application of a commercial substrate infested with both antagonist (1% and pathogen (1% to the seedlings

  7. LeishCyc: a biochemical pathways database for Leishmania major

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    Doyle Maria A

    2009-06-01

    Full Text Available Abstract Background Leishmania spp. are sandfly transmitted protozoan parasites that cause a spectrum of diseases in more than 12 million people worldwide. Much research is now focusing on how these parasites adapt to the distinct nutrient environments they encounter in the digestive tract of the sandfly vector and the phagolysosome compartment of mammalian macrophages. While data mining and annotation of the genomes of three Leishmania species has provided an initial inventory of predicted metabolic components and associated pathways, resources for integrating this information into metabolic networks and incorporating data from transcript, protein, and metabolite profiling studies is currently lacking. The development of a reliable, expertly curated, and widely available model of Leishmania metabolic networks is required to facilitate systems analysis, as well as discovery and prioritization of new drug targets for this important human pathogen. Description The LeishCyc database was initially built from the genome sequence of Leishmania major (v5.2, based on the annotation published by the Wellcome Trust Sanger Institute. LeishCyc was manually curated to remove errors, correct automated predictions, and add information from the literature. The ongoing curation is based on public sources, literature searches, and our own experimental and bioinformatics studies. In a number of instances we have improved on the original genome annotation, and, in some ambiguous cases, collected relevant information from the literature in order to help clarify gene or protein annotation in the future. All genes in LeishCyc are linked to the corresponding entry in GeneDB (Wellcome Trust Sanger Institute. Conclusion The LeishCyc database describes Leishmania major genes, gene products, metabolites, their relationships and biochemical organization into metabolic pathways. LeishCyc provides a systematic approach to organizing the evolving information about Leishmania

  8. Comparación de los métodos de inmunoensayo enzimático automatizado (VIDAS y PCR para la detección de Salmonella spp. en expendios de la ciudad de Santa Marta (Colombia

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    Libardo Acosta

    2013-01-01

    Full Text Available Objetivo: comparar los métodos de inmunoensayo enzimático automatizado VIDAS, (Vi- tek Immunodiagnostic Assay System, y el análisis molecular por Reacción en Cadena de la Polimerasa (PCR para detectar Salmonella spp en expendios de la ciudad de Santa Marta (Colombia. Materiales y métodos: Previamente se reportó la estandarización de la técnica de PCR para la detección de Salmonella spp. en un lapso de 12 horas. En colaboración con el Labo- ratorio de Salud Pública del departamento del Magdalena se procedió a tomar un total de 65 muestras de alimentos para análisis de vigilancia rutinaria, discriminados así: carne de res: 14 (21.5 %, embutidos: 18 (27.7 %, pollo: 7(10.8 %, pescado: 3(4.6 %, harinas: 13 (20 %, lácteos: 5 (7.7 % salsas: 4(6.2 % y ensalada: 1(1.5 %, en la ciudad de Santa Marta entre septiembre y noviembre de 2010. Las 65 muestras fueron sometidas a análisis microbiológico para la determinación de Salmonella spp. en el Laboratorio de Salud Pú- blica del departamento del Magdalena siguiendo el protocolo del VIDAS. Una alícuota de 1 mL del preenriquecimiento no selectivo fue enviada refrigerada al laboratorio de biología molecular de la Universidad Cooperativa de Colombia para ser analizada por PCR. Resultados: Los resultados muestran que los alimentos analizados en el Laboratorio de Salud Pública del departamento del Magdalena con VIDAS presentaron Salmonella spp. únicamente en cárnicos: 5/65 (7.7 %, mientras que esas mismas muestras analizadas por PCR mostraban la presencia de Salmonella spp. en 36/65 (55.4 %. Conclusiones: Los resultados indican que la PCR puede ser aplicada para realizar vigilan- cia epidemiológica y obtener resultados cualitativos confiables en menor tiempo.

  9. Dietas a base de forraje tradicional y nopal (Opuntia spp. enriquecido con proteínas para alimentar cabras

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    Arnoldo Flores-Hernández

    2017-01-01

    Full Text Available El objetivo fue determinar la composición química de las dietas y su efecto en la ganancia de peso vivo, consumo y rechazo de alimento. El estudio se realizó mediante un diseño completamente azar con tres repeticiones y como factor principal tres dietas cuyos componentes fueron : dieta 1 40% heno de alfalfa + 60% heno de sorgo, dieta 2 40% nopal seco natural + 60 % heno de sorgo y dieta 3 40% nopal seco enriquecido co n proteínas + 60% heno de sorgo . Los resultados mostraron que la dieta de nopal enriquecido con proteínas y la dieta de nopal seco natural poseen una calidad nutritiva mayor, pero las condiciones de secado y/o almacenamiento tuvieron efecto en la calidad de l os componentes de las dietas evaluadas. La ganancia de peso vivo fue mayor en cabras alimentadas con la dieta de nopal - E , pero el porcentaje de alimento rechazado (menor palatabili dad fue mayor para esta dieta. La ganancia de peso vivo fue menor en alfalfa y nopal seco natural , respectivamente . La ganancia de peso y el tiempo de alimentación mostraron correlación positiva y significativa ; sin embargo, se debe determinar el tiempo óptimo de alimentación y evaluar el costo - beneficio de las dietas utilizadas.

  10. Disseminated Leishmaniasis Caused by Leishmania Tropica in a Puppy from Karaj, Central Iran

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    M Mohebali

    2011-06-01

    Full Text Available A 5-month old puppy with muco-cutaneous lesions in the chin, around lips and eyes was exam­ined physically and microscopically for leishmaniasis. Muco-cutaneous lesions containing a large num­ber of amastigotes of Leishmania spp. were observed. Amastigotes were also detected in liver and spleen of the puppy. The animal was positive with Dipstick rK39 kit and high level of anti-Leishmania antibodies was detected by direct agglutination test (DAT. DNA, Using PCR-RFLP technique extracted from cultured Leishmania promastigotes and L. tropica was identified. This is the first report of concurrent mucosal and visceral involvement of L. tropica in a puppy from Iran.

  11. The proliferation potential of promastigotes of the main Leishmania species of the old world in NNN culture medium prepared using blood of four different mammals.

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    Ladopoulos, Theodoros; Ntais, Pantelis; Tsirigotakis, Nikolaos; Dokianakis, Emmanouil; Antoniou, Maria

    2015-10-01

    The efficacy of the in vitro cultivation of promastigotes of four Leishmania spp. was tested in the biphasic Novy-MacNeal-Nicolle (NNN) medium prepared using blood from different animals (horse, donkey, goat and sheep). The aim was to test which NNN preparation gave the best yield in the shortest time for different parasite species, in order to obtain a large crop of promastigotes for experimental work and for antigen preparation. Promastigotes of Leishmania infantum, Leishmania donovani, Leishmania tropica and Leishmania major, the four main parasite species occurring in the old world, were defrosted from -80 °C and placed, at equal numbers, in the 4 different NNN preparations. At the end of the 7th day, the NNN medium using horse blood produced the greatest number of promastigotes for all Leishmania spp. tested, whilst goat blood proved the poorest medium, providing culture results only for L. infantum. This finding may be explained by the fact that Leishmania is a nicotinamide adenine dinucleotide (NAD) auxotroph and horse erythrocytes support NAD-dependent microorganisms. Copyright © 2015 Elsevier Inc. All rights reserved.

  12. Evaluación del efecto del ácido nalidíxico, ampicilina, kanamicina, penicilina G y polimixina B en los cultivos de promastigotes de leishmania

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    Sofía Duque Beltrán

    1992-06-01

    Full Text Available Se evaluó el efecto de diferentes concentraciones de ácido nalidíxico, ampicilina, kanarnicina, penicilina G y polimixina B, sobre la población de promastigotes de Leishmania braziliensis braziliensis , Leishmania donovani chagasi y Leishmania mexicana amazonensis in vitro. La penicilina G y la ampicilina se pueden utilizar hasta concentraciones de 1000 ug/ml y 500 ug/ml respectivamente en cultivo de promastigotes de cualquier cepa de Leishmania sin que éstos se afecten. La polimixina B disminuye la población de prosmastigotes por lo cual es preferible no usarse en cultivos de Leishmania. El ácido nalidíxico y kanamicina pueden ser utilizados in vitro pero teniéndose en cuenta la especie de Leishmania y la concentración de anfimicrobiano recomendado para la misma.

  13. 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

  14. Leishmania major infection in a dog with cutaneous manifestations.

    Science.gov (United States)

    Baneth, Gad; Nachum-Biala, Yaarit; Shabat Simon, Maytal; Brenner, Ori; Gaier, Sarit; Rojas, Alicia; Yasur-Landau, Daniel

    2016-05-10

    Leishmania major is a main cause of cutaneous leishmaniasis in humans in an area that stretches from India through Central Asia, the Middle East, to North and West Africa. In Israel, it is a common infection of humans with rodents as the reservoir hosts and Phlebotomus papatasi as its sand fly vector. A 6 months old spayed female mixed breed dog was referred to the Hebrew University Veterinary Teaching Hospital with a large ulcerative dermal lesion on the muzzle, and lesions in the foot pads and left hind leg. Histopathology of a skin biopsy found chronic lymphohistiocytic dermatitis with the presence of Leishmania spp. amastigotes in the muzzle. Physical examination indicated that the dog was overall in a good clinical condition and the main findings were the skin lesions and enlarged prescapular lymph nodes. Complete blood count and serum biochemistry profile were within reference ranges. Serology by ELISA was positive for Leishmania spp. and PCR of the prescapular lymph node was positive by an ITS1 region PCR-high resolution melt analysis. However, the melt curve and subsequent DNA sequencing indicated that infection was caused by L. major and not L. infantum, which is the main causative agent of canine leishmaniosis in the Mediterranean region. DNA was extracted from the paraffin embedded muzzle biopsy and PCR with sequencing also indicated L. major. The dog's young age and the absence of hyperglobulinemia and anemia were not typical of L. infantum infection. The dog was treated with allopurinol and the skin lesions improved and later disappeared when the dog was re-evaluated. This is the first molecularly-confirmed case of L. major infection in a dog. Two previous reports of L. major in dogs originated from Saudi-Arabia and Egypt in 1985 and 1987 were confirmed by enzymatic biochemical techniques. Serology for L. infantum was positive probably due to the well documented serological cross-reactivity between Leishmania spp. Although dogs and wild carnivores are

  15. Novel selective inhibitor of Leishmania (Leishmania) amazonensis arginase.

    Science.gov (United States)

    da Silva, Edson R; Boechat, Nubia; Pinheiro, Luiz C S; Bastos, Monica M; Costa, Carolina C P; Bartholomeu, Juliana C; da Costa, Talita H

    2015-11-01

    Arginase is a glycosomal enzyme in Leishmania that is involved in polyamine and trypanothione biosynthesis. The central role of arginase in Leishmania (Leishmania) amazonensis was demonstrated by the generation of two mutants: one with an arginase lacking the glycosomal addressing signal and one in which the arginase-coding gene was knocked out. Both of these mutants exhibited decreased infectivity. Thus, arginase seems to be a potential drug target for Leishmania treatment. In an attempt to search for arginase inhibitors, 29 derivatives of the [1,2,4]triazolo[1,5-a]pyrimidine system were tested against Leishmania (Leishmania) amazonensis arginase in vitro. The [1,2,4]triazolo[1,5-a]pyrimidine scaffold containing R1  = CF3 exhibited greater activity against the arginase rather than when the substituent R1  = CH3 in the 2-position. The novel compound 2-(5-methyl-2-(trifluoromethyl)-[1,2,4]triazolo[1,5-a]pyrimidin-7-yl)hydrazinecarbothioamide (30) was the most potent, inhibiting arginase by a non-competitive mechanism, with the Ki and IC50 values for arginase inhibition estimated to be 17 ± 1 μm and 16.5 ± 0.5 μm, respectively. These results can guide the development of new drugs against leishmaniasis based on [1,2,4]triazolo[1,5-a]pyrimidine derivatives targeting the arginase enzyme. © 2015 John Wiley & Sons A/S.

  16. Changes to cholesterol trafficking in macrophages by Leishmania parasites infection.

    Science.gov (United States)

    Semini, Geo; Paape, Daniel; Paterou, Athina; Schroeder, Juliane; Barrios-Llerena, Martin; Aebischer, Toni

    2017-08-01

    Leishmania spp. are protozoan parasites that are transmitted by sandfly vectors during blood sucking to vertebrate hosts and cause a spectrum of diseases called leishmaniases. It has been demonstrated that host cholesterol plays an important role during Leishmania infection. Nevertheless, little is known about the intracellular distribution of this lipid early after internalization of the parasite. Here, pulse-chase experiments with radiolabeled cholesteryl esterified to fatty acids bound to low-density lipoproteins indicated that retention of this source of cholesterol is increased in parasite-containing subcellular fractions, while uptake is unaffected. This is correlated with a reduction or absence of detectable NPC1 (Niemann-Pick disease, type C1), a protein responsible for cholesterol efflux from endocytic compartments, in the Leishmania mexicana habitat and infected cells. Filipin staining revealed a halo around parasites within parasitophorous vacuoles (PV) likely representing free cholesterol accumulation. Labeling of host cell membranous cholesterol by fluorescent cholesterol species before infection revealed that this pool is also trafficked to the PV but becomes incorporated into the parasites' membranes and seems not to contribute to the halo detected by filipin. This cholesterol sequestration happened early after infection and was functionally significant as it correlated with the upregulation of mRNA-encoding proteins required for cholesterol biosynthesis. Thus, sequestration of cholesterol by Leishmania amastigotes early after infection provides a basis to understand perturbation of cholesterol-dependent processes in macrophages that were shown previously by others to be necessary for their proper function in innate and adaptive immune responses. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  17. APROVECHAMIENTO DE PULPA Y CÁSCARA DE PLÁTANO (Musa paradisiaca spp PARA LA OBTENCIÓN DE MALTODEXTRINA

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    DIANA VICTORIA MELO-SABOGAL

    Full Text Available Los subproductos de las cadenas agroindustriales constituyen alrededor del 90% de la parte del cultivo, por lo cual, establecer alternativas de aprovechamiento puede ser importantes para los productores. En este trabajo se evaluó el efecto del tratamiento enzimático sobre almidón obtenido de pulpa y cáscara de plátano (Musa paradisiaca spp. y se estableció el efecto de la temperatura de secado en almidón hidrolizado sobre propiedades químicas, físicas y reológicas. Se cuantificó la actividad de agua, el contenido de humedad y el contenido de dextrosa equivalente, azúcares reductores presentes en la maltodextrina por cromatografía líquida de alta eficacia; además se determinaron parámetros de color y reológicos. La hidrólisis enzimática presentó efecto significativo sobre la cantidad de dextrosa equivalente, permitió la obtención de maltotriosa, maltosa y glucosa, compuestos propios de maltodextrina, además, tuvo efecto sobre los parámetros reológicos, evidenciándose el cambio de la estructura del almidón. La temperatura de 130°C fue la que tuvo mayor efecto sobre las variables de actividad de agua, contenido de humedad y cambio de color. Lo anterior demuestra que es posible la obtención de maltodextrinas a partir de pulpa y cáscara de plátano por vía enzimática y secado por atomización a temperatura de 130°C.

  18. Naturally infected Lutzomyia sand flies in a Leishmania-endemic area of Brazil.

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    Carvalho, Gustavo M L; Andrade Filho, Jose D; Falcao, Alda L; Rocha Lima, Ana C V M; Gontijo, Celia M F

    2008-06-01

    In Brazil, Leishmania transmission involves several species of phlebotomine sand flies that are closely associated with different parasites and reservoirs, giving rise to different transmission cycles. The present study focused on naturally infected phlebotomines originating from Santa Luzia, a municipality near Belo Horizonte, capital of the Brazilian state of Minas Gerais, in which leishmaniasis are endemic. Systematic and non systematic approaches,involving the use of light traps and direct aspiration from resting sites, respectively, were used to collect females and flies. Identification of the captured insects and determination of natural infection by Leishmania spp. were performed using both conventional dissection methods and polymerase chain reaction (PCR). The dissection of 102 sand flies allowed five species of Lutzomyia to be identified, although no flagellate parasite forms were observed.In addition, 211 sand flies were identified, were separated according to species, and were combined into 11 pools of up to 20 individuals each. PCR analyses showed that two of these pools were infected with Leishmania:one pool of Lu. whitmani was infected with Le. (Viannia) spp. and another of Lu. cortelezzii was infected with Le. chagasi. This suggests that Lu. whitmani may be a possible vector of Leishmania in the study area, and more work needs to be performed to assess the role of Lu. cortelezzii as a vector.

  19. Leishmania (Viannia) braziliensis infection in wild small mammals in ecotourism area of Brazil.

    Science.gov (United States)

    Tonelli, Gabriel Barbosa; Tanure, Aline; Rego, Felipe Dutra; Carvalho, Gustavo Mayr de Lima; Stumpp, Rodolfo; Ássimos, Gabriela Ribeiro; Campos, Aldenise Martins; Lima, Ana Cristina Viana Mariano da Rocha; Gontijo, Célia Maria Ferreira; Paz, Gustavo Fontes; Andrade Filho, José Dilermando

    2017-01-01

    Leishmaniases are parasitic diseases transmitted to mammalian hosts by sand fly vectors (Diptera: Psychodidae). Despite the increasing occurrence of visceral and cutaneous leishmaniasis cases in urban centers, their transmission still occur primarily in wild environments and may be associated with professional activities and recreation, such as ecotourism. The Reserva Particular do Patrimônio Natural Santuário do Caraça (RPPNSC) is one of the largest ecotourism attractions in the State of Minas Gerais, Brazil, and comprises an area of environmental preservation with 11,233 hectares presenting a transitional vegetation between Cerrado and Atlantic Forest. The present study describes the abundance of small mammals in RPPNSC, the isolation and identification of Leishmania in five wild animals. Small mammals were bimonthly trapped along 6 trails within the RPPNSC with 10 Tomahawk traps each. Two trails were located in peridomiciliary areas near tourist lodging facilities, and four trails were located at sites visited by tourists in forest areas. The most prevalent species were Akodon cursor, Cerradomys subflavus and Oligoryzomys nigripes. Six isolates of Leishmania were obtained from these animals and identified as Leishmania braziliensis through HSP70-PCR RFLP method. Leishmania spp. DNA was detected by kDNA-PCR method and isolated by biphasic culture. Studies point to some of the captured species as potential wild reservoirs of Leishmania, suggesting they may be involved in the transmission cycle in these wild environments.

  20. Direct detection of Leishmania from clinical samples.

    Science.gov (United States)

    Waitumbi, John N; Bast, Joshua; Nyakoe, Nancy; Magiri, Charles; Quintana, Miguel; Takhampunya, Ratree; Schuster, Anthony L; Van de Wyngaerde, Marshall T; McAvin, James C; Coleman, Russell E

    2017-01-01

    The ability to rapidly and accurately diagnose leishmaniasis is a military priority. Testing was conducted to evaluate diagnostic sensitivity and specificity of field-expedient Leishmania genus and visceral Leishmania specific dual-fluorogenic, hydrolysis probe (TaqMan), polymerase chain reaction assays previously established for use in vector surveillance. Blood samples of patients with confirmed visceral leishmaniasis and controls without the disease from Baringo District, Kenya, were tested. Leishmania genus assay sensitivity was 100% (14/14) and specificity was 84% (16/19). Visceral Leishmania assay sensitivity was 93% (13/14) and specificity 80% (4/5). Cutaneous leishmaniasis (CL) skin scrapes of patients from Honduras were also evaluated. Leishmania genus assay sensitivity was 100% (10/10). Visceral Leishmania assay specificity was 100% (10/10) from cutaneous leishmaniasis samples; no fluorescence above background was reported. These results show promise in a rapid, sensitive, and specific method for Leishmania direct detection from clinical samples.

  1. 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

    : 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.

  2. Detection of Leishmania amazonensis and Leishmania braziliensis in Culicoides (Diptera, Ceratopogonidae) in an endemic area of cutaneous leishmaniasis in the Brazilian Amazonia.

    Science.gov (United States)

    Rebêlo, José Manuel Macário; Rodrigues, Bruno Leite; Bandeira, Maria da Conceição Abreu; Moraes, Jorge Luiz Pinto; Fonteles, Raquel Silva; Pereira, Silma Regina Ferreira

    2016-12-01

    Biting midges in the genus Culicoides act as vectors of arboviruses throughout the world and as vectors of filariasis in Latin America, the Caribbean, and parts of Africa. Although Culicoides spp. are currently not considered to be vectors of Leishmania protozoa, the high abundance of biting midges in areas with active cutaneous leishmaniasis transmission points to the possibility of Culicoides infection by these pathogens. We used PCR to test captured Culicoides species for natural infection with Leishmania spp. We tested 450 Culicoides females, divided into 30 pools of 15 individuals each, as follows: nine pools of C. foxi (135 specimens), seven pools of C. filariferus (105), seven pools of C. insignis (105), five pools of C. ignacioi (75), and two pools of C. flavivenula (30). PCR confirmed the presence of Leishmania braziliensis DNA in C. ignacioi (0.14%), C. insignis (0.14%), and C. foxi (0.11); and Le. amazonensis DNA in C. filariferus (0.14%) and C. flavivenula (0.50%). We conclude that these Culicoides species can be naturally infected, but vector competence and transmission capability must be confirmed in future studies. Our results warrant further investigation into the role of these biting midge species in the leishmaniasis epidemiological cycle. © 2016 The Society for Vector Ecology.

  3. Molecular Detection of Leishmania DNA in Wild-Caught Phlebotomine Sand Flies (Diptera: Psychodidae) From a Cave in the State of Minas Gerais, Brazil.

    Science.gov (United States)

    Carvalho, G M L; Brazil, R P; Rêgo, F D; Ramos, M C N F; Zenóbio, A P L A; Andrade Filho, J D

    2017-01-01

    Leishmania spp. are distributed throughout the world, and different species are associated with varying degrees of disease severity. In Brazil, Leishmania transmission involves several species of phlebotomine sand flies that are closely associated with different parasites and reservoirs, and thereby giving rise to different transmission cycles. Infection occurs during the bloodmeals of sand flies obtained from a variety of wild and domestic animals, and sometimes from humans. The present study focused on detection of Leishmania DNA in phlebotomine sand flies from a cave in the state of Minas Gerais. Detection of Leishmania in female sand flies was performed with ITS1 PCR-RFLP (internal transcribed spacer 1) using HaeIII enzyme and genetic sequencing for SSUrRNA target. The survey of Leishmania DNA was carried out on 232 pools and the parasite DNA was detected in four: one pool of Lutzomyia cavernicola (Costa Lima, 1932), infected with Le. infantum (ITS1 PCR-RFLP), two pools of Evandromyia sallesi (Galvão & Coutinho, 1939), both infected with Leishmania braziliensis complex (SSUrRNA genetic sequencing analysis), and one pool of Sciopemyia sordellii (Shannon & Del Ponte, 1927), infected with subgenus Leishmania (SSUrRNA genetic sequencing analysis). The present study identified the species for Leishmania DNA detected in four pools of sand flies, all of which were captured inside the cave. These results represent the first molecular detection of Lu cavernicola with Le infantum DNA, Sc sordellii with subgenus Leishmania DNA, and Ev sallesi with Leishmania braziliensis complex DNA. The infection rate in females captured for this study was 0.17%. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  4. Desarrollo de sistema de información geográfica para gestión fitosanitaria en unidades productivas de caña de azúcar (Saccharum spp.)

    OpenAIRE

    Balmaseda, C; Romero, Madelin

    2012-01-01

    La fitoprotección del cultivo de la caña de azúcar (Saccharum spp.) requiere de la aplicación de modernas herramientas y procedimientos tecnológicos con la finalidad de mantener la vigilancia y facilitar el proceso de toma de decisiones en el control fitosanitario. Para ello se desarrolló un sistema de información geográfica (SIG) eligiendo como base cartográfica el catastro especializado de una empresa. El modelo conceptual desarrollado muestra las relaciones que existen entre variables cons...

  5. Desarrollo de un proceso a escala de laboratorio para el aislamiento, conservación y producción de cepas nativas de Monascus spp. a partir de la biodiversidad fúngica ecuatoriana

    OpenAIRE

    Ayala Pastaz, Klever Bayardo

    2017-01-01

    Desarrollar un proceso a escala de laboratorio para la conservación y producción de cepas nativas de Monascus purpureus, mediante bioprocesos que permita el máximo aprovechamiento del recurso natural fúngico y el manejo sostenible de la biodiversidad. Se evidencia un limitado conocimiento en el manejo y aprovechamiento sustentable de la especie Monascus spp. nativo ecuatoriano, tanto en su proceso de aislamiento, conservación y producción; así como del ciclo de vida en el medio natural y m...

  6. HIV aspartyl peptidase inhibitors interfere with cellular proliferation, ultrastructure and macrophage infection of Leishmania amazonensis.

    Directory of Open Access Journals (Sweden)

    Lívia O Santos

    Full Text Available BACKGROUND: Leishmania is the etiologic agent of leishmanisais, a protozoan disease whose pathogenic events are not well understood. Current therapy is suboptimal due to toxicity of the available therapeutic agents and the emergence of drug resistance. Compounding these problems is the increase in the number of cases of Leishmania-HIV coinfection, due to the overlap between the AIDS epidemic and leishmaniasis. METHODOLOGY/PRINCIPAL FINDINGS: In the present report, we have investigated the effect of HIV aspartyl peptidase inhibitors (PIs on the Leishmania amazonensis proliferation, ultrastructure, interaction with macrophage cells and expression of classical peptidases which are directly involved in the Leishmania pathogenesis. All the HIV PIs impaired parasite growth in a dose-dependent fashion, especially nelfinavir and lopinavir. HIV PIs treatment caused profound changes in the leishmania ultrastructure as shown by transmission electron microscopy, including cytoplasm shrinking, increase in the number of lipid inclusions and some cells presenting the nucleus closely wrapped by endoplasmic reticulum resembling an autophagic process, as well as chromatin condensation which is suggestive of apoptotic death. The hydrolysis of HIV peptidase substrate by L. amazonensis extract was inhibited by pepstatin and HIV PIs, suggesting that an aspartyl peptidase may be the intracellular target of the inhibitors. The treatment with HIV PIs of either the promastigote forms preceding the interaction with macrophage cells or the amastigote forms inside macrophages drastically reduced the association indexes. Despite all these beneficial effects, the HIV PIs induced an increase in the expression of cysteine peptidase b (cpb and the metallopeptidase gp63, two well-known virulence factors expressed by Leishmania spp. CONCLUSIONS/SIGNIFICANCE: In the face of leishmaniasis/HIV overlap, it is critical to further comprehend the sophisticated interplays among Leishmania

  7. The Gut Microbiome of the Vector Lutzomyia longipalpis Is Essential for Survival of Leishmania infantum.

    Science.gov (United States)

    Kelly, Patrick H; Bahr, Sarah M; Serafim, Tiago D; Ajami, Nadim J; Petrosino, Joseph F; Meneses, Claudio; Kirby, John R; Valenzuela, Jesus G; Kamhawi, Shaden; Wilson, Mary E

    2017-01-17

    The vector-borne disease leishmaniasis, caused by Leishmania species protozoa, is transmitted to humans by phlebotomine sand flies. Development of Leishmania to infective metacyclic promastigotes in the insect gut, a process termed metacyclogenesis, is an essential prerequisite for transmission. Based on the hypothesis that vector gut microbiota influence the development of virulent parasites, we sequenced midgut microbiomes in the sand fly Lutzomyia longipalpis with or without Leishmania infantum infection. Sucrose-fed sand flies contained a highly diverse, stable midgut microbiome. Blood feeding caused a decrease in microbial richness that eventually recovered. However, bacterial richness progressively decreased in L. infantum-infected sand flies. Acetobacteraceae spp. became dominant and numbers of Pseudomonadaceae spp. diminished coordinately as the parasite underwent metacyclogenesis and parasite numbers increased. Importantly, antibiotic-mediated perturbation of the midgut microbiome rendered sand flies unable to support parasite growth and metacyclogenesis. Together, these data suggest that the sand fly midgut microbiome is a critical factor for Leishmania growth and differentiation to its infective state prior to disease transmission. Leishmania infantum, a parasitic protozoan causing fatal visceral leishmaniasis, is transmitted to humans through the bite of the sand fly Lutzomyia longipalpis Development of the parasite to its virulent metacyclic state occurs in the sand fly gut. In this study, the microbiota within the Lu. longipalpis midgut was delineated by 16S ribosomal DNA (rDNA) sequencing, revealing a highly diverse community composition that lost diversity as parasites developed to their metacyclic state and increased in abundance in infected flies. Perturbing sand fly gut microbiota with an antibiotic cocktail, which alone had no effect on either the parasite or the fly, arrested both the development of virulent parasites and parasite expansion

  8. Persistence of Leishmania antigen in C57Bl/6j inbred mice infected with Leishmania (Leishmania amazonensis Persistência do antígeno da Leishmania no camundongo isogênico C57Bl/6j infectado com a Leishmania (Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    C. Vasconcellos

    1999-07-01

    Full Text Available PURPOSE. To develop an animal model for studying mucocutaneous leishmaniasis. METHODS. The hind footpad of C57Bl/6j inbred mice was experimentally infected with 10(7 Leishmania (Leishmania amazonensis promastigote and the skin was studied through light and electron transmission microscopy and immunohistochemistry (PAP techniques. RESULTS. There were morphological evidences of cellular immune mechanisms and hypersensitivity reaction after eight weeks of infection and metastasis and well shaped parasites at ultrastructural level by fifty-one weeks post infection. Relapse of infection with mucosa lesions occurred around the 50th week after inoculation. CONCLUSION. The use of this animal model in long term follow up could be an useful experimental model for human mucocutaneous leishmaniasis.OBJETIVO. Desenvolver um modelo experimental para o estudo da leishmaniose cutâneo-mucosa. MÉTODOS. O coxim plantar traseiro de camundongos isogênicos C57Bl/6j foi inoculado com 10(7 formas promastigotas da Leishmania (Leishmania amazonensis e a pele foi estudada através da microscopia óptica e eletrônica e de técnica imunohistoquímica (PAP. RESULTADOS. Ocorreram evidências morfológicas de mecanismos imunes mediados por células, concomitantemente ao de reação de hipersensibilidade, após a oitava semana de infecção e a presença de parasitas com ultraestrutura preservada na quinquagésima primeira semana após a infecção. Houve recidiva da infecção com surgimento de lesões mucosas por volta da 50a semana pós inoculação. CONCLUSÃO. Este modelo animal, com um período de tempo de seguimento prolongado, poderia ser empregado como modelo para o estudo experimental da leishmaniose cutâneo-mucosa.

  9. Purificación de dos proteínas de Leishmania Viannia panamensis y su utilización en el diagnóstico de casos de leishmaniosis cutánea de larga evolución y difícil diagnóstico parasitológico

    Directory of Open Access Journals (Sweden)

    Sonia Agudelo

    2001-04-01

    Full Text Available

    La respuesta inmune humoral inducida por Leishmania spp, se inicia desde la etapa temprana de la infección y se mantiene hasta cuando la mayoría de los parásitos son eliminados. Los isotipos, subclases y niveles de anticuerpos que evalúan esta respuesta dependen en gran medida de la forma clínica y el estado de evolución de la enfermedad. En casos de lesiones mucosas o cutáneas de larga evolución donde la posibilidad de encontrar parásitos es baja, toma gran importancia evaluar esta respuesta humoral. Sin embargo, las técnicas serológicas existentes para detectar anticuerpos antileishmania presentan baja sensibilidad y alta reacción cruzada con parásitos de Trypanosoma cruzi, surgiendo, como una alternativa para mejorar estas pruebas, el uso de antígenos proteicos purificados (1,2. Teniendo como base los resultados obtenidos por Agudelo y col (3, quienes demostraron que las fracciones proteicas de 36 y 50 kDa de Leishmania Viannia panamensis son altamente sensibles y
    100% específicas de Leishmania, nos proponemos purificar estas proteínas a fin de utilizarlas para mejorar la sensibilidad y especificidad de pruebas serológicas que sean económicas y de ejecución práctica en zonas endémicas de leishmaniosis.

     

  10. Canine antibody response to Phlebotomus perniciosus bites negatively correlates with the risk of Leishmania infantum transmission.

    Directory of Open Access Journals (Sweden)

    Michaela Vlkova

    2011-10-01

    Full Text Available BACKGROUND: Phlebotomine sand flies are blood-sucking insects that can transmit Leishmania parasites. Hosts bitten by sand flies develop an immune response against sand fly salivary antigens. Specific anti-saliva IgG indicate the exposure to the vector and may also help to estimate the risk of Leishmania spp. transmission. In this study, we examined the canine antibody response against the saliva of Phlebotomus perniciosus, the main vector of Leishmania infantum in the Mediterranean Basin, and characterized salivary antigens of this sand fly species. METHODOLOGY/PRINCIPAL FINDINGS: Sera of dogs bitten by P. perniciosus under experimental conditions and dogs naturally exposed to sand flies in a L. infantum focus were tested by ELISA for the presence of anti-P. perniciosus antibodies. Antibody levels positively correlated with the number of blood-fed P. perniciosus females. In naturally exposed dogs the increase of specific IgG, IgG1 and IgG2 was observed during sand fly season. Importantly, Leishmania-positive dogs revealed significantly lower anti-P. perniciosus IgG2 compared to Leishmania-negative ones. Major P. perniciosus antigens were identified by western blot and mass spectrometry as yellow proteins, apyrases and antigen 5-related proteins. CONCLUSIONS: Results suggest that monitoring canine antibody response to sand fly saliva in endemic foci could estimate the risk of L. infantum transmission. It may also help to control canine leishmaniasis by evaluating the effectiveness of anti-vector campaigns. Data from the field study where dogs from the Italian focus of L. infantum were naturally exposed to P. perniciosus bites indicates that the levels of anti-P. perniciosus saliva IgG2 negatively correlate with the risk of Leishmania transmission. Thus, specific IgG2 response is suggested as a risk marker of L. infantum transmission for dogs.

  11. Characterization of Leishmania Soluble Exo-Antigen

    National Research Council Canada - National Science Library

    Cui, Liwang

    2003-01-01

    .... Vaccine development is the ultimate solution for this problem. Our previous research indicates that Leishmania parasites secrete, excrete, or shed antigens into the medium during in vitro culture...

  12. Desarrollo de un bioproceso para la conservación de cepas nativas de Trichoderma spp., a partir de la biodiversidad fúngica ecuatoriana

    OpenAIRE

    Benavides Sotelo, Edmundo Napoleon

    2017-01-01

    Trichoderma spp., es un moho cosmopolita, que se encuentra de forma natural en plantas, rizosferas, sedimentos, entre otros sustratos. Dentro de sus principales aplicaciones se encuentra su uso como biocontrolador y/o biofertilizante en cultivos agrícolas, gracias a su actividad antagonista hacia fitopatógenos. Adicionalmente produce otra serie de compuestos de gran valor biotecnológico. Muchas industrias requieren de cepas puras de Trichoderma spp., siendo esta la razón de ser de los Centros...

  13. Arginase expression modulates nitric oxide production in Leishmania (Leishmania) amazonensis.

    Science.gov (United States)

    Acuña, Stephanie Maia; Aoki, Juliana Ide; Laranjeira-Silva, Maria Fernanda; Zampieri, Ricardo Andrade; Fernandes, Juliane Cristina Ribeiro; Muxel, Sandra Marcia; Floeter-Winter, Lucile Maria

    2017-01-01

    Arginase is an enzyme that converts L-arginine to urea and L-ornithine, an essential substrate for the polyamine pathway supporting Leishmania (Leishmania) amazonensis replication and its survival in the mammalian host. L-arginine is also the substrate of macrophage nitric oxide synthase 2 (NOS2) to produce nitric oxide (NO) that kills the parasite. This competition can define the fate of Leishmania infection. The transcriptomic profiling identified a family of oxidoreductases in L. (L.) amazonensis wild-type (La-WT) and L. (L.) amazonensis arginase knockout (La-arg-) promastigotes and axenic amastigotes. We highlighted the identification of an oxidoreductase that could act as nitric oxide synthase-like (NOS-like), due to the following evidences: conserved domain composition, the participation of NO production during the time course of promastigotes growth and during the axenic amastigotes differentiation, regulation dependence on arginase activity, as well as reduction of NO amount through the NOS activity inhibition. NO quantification was measured by DAF-FM labeling analysis in a flow cytometry. We described an arginase-dependent NOS-like activity in L. (L.) amazonensis and its role in the parasite growth. The increased detection of NO production in the mid-stationary and late-stationary growth phases of La-WT promastigotes could suggest that this production is an important factor to metacyclogenesis triggering. On the other hand, La-arg- showed an earlier increase in NO production compared to La-WT, suggesting that NO production can be arginase-dependent. Interestingly, La-WT and La-arg- axenic amastigotes produced higher levels of NO than those observed in promastigotes. As a conclusion, our work suggested that NOS-like is expressed in Leishmania in the stationary growth phase promastigotes and amastigotes, and could be correlated to metacyclogenesis and amastigotes growth in a dependent way to the internal pool of L-arginine and arginase activity.

  14. Detection of Leishmania RNA virus in Leishmania parasites.

    Directory of Open Access Journals (Sweden)

    Haroun Zangger

    Full Text Available Patients suffering from cutaneous leishmaniasis (CL caused by New World Leishmania (Viannia species are at high risk of developing mucosal (ML or disseminated cutaneous leishmaniasis (DCL. After the formation of a primary skin lesion at the site of the bite by a Leishmania-infected sand fly, the infection can disseminate to form secondary lesions. This metastatic phenotype causes significant morbidity and is often associated with a hyper-inflammatory immune response leading to the destruction of nasopharyngeal tissues in ML, and appearance of nodules or numerous ulcerated skin lesions in DCL. Recently, we connected this aggressive phenotype to the presence of Leishmania RNA virus (LRV in strains of L. guyanensis, showing that LRV is responsible for elevated parasitaemia, destructive hyper-inflammation and an overall exacerbation of the disease. Further studies of this relationship and the distribution of LRVs in other Leishmania strains and species would benefit from improved methods of viral detection and quantitation, especially ones not dependent on prior knowledge of the viral sequence as LRVs show significant evolutionary divergence.This study reports various techniques, among which, the use of an anti-dsRNA monoclonal antibody (J2 stands out for its specific and quantitative recognition of dsRNA in a sequence-independent fashion. Applications of J2 include immunofluorescence, ELISA and dot blot: techniques complementing an arsenal of other detection tools, such as nucleic acid purification and quantitative real-time-PCR. We evaluate each method as well as demonstrate a successful LRV detection by the J2 antibody in several parasite strains, a freshly isolated patient sample and lesion biopsies of infected mice.We propose that refinements of these methods could be transferred to the field for use as a diagnostic tool in detecting the presence of LRV, and potentially assessing the LRV-related risk of complications in cutaneous leishmaniasis.

  15. OBTENCIÓN DE UN ALIMENTO EXTRUIDO PARA TILAPIA ROJA (Oreochromis spp UTILIZANDO ENSILAJE BIOLÓGICO DE PESCADO A OBTENÇÃO DE UMA RAÇÃO EXTRUSADA PARA A TILÁPIA VERMELHA (Oreochromis spp UTILIZANDO SILAGEM DE PESCADO BIOLÓGICA OBTAINING AN EXTRUDED NOURISHMENT FOR RED TILAPIA (Oreochromis ssp USING FISH BIOLOGICAL SILAGE

    Directory of Open Access Journals (Sweden)

    JAIDER OSWALDO PANTOJA

    2011-12-01

    Full Text Available En la represa 'La Salvajina', municipio de Suárez (Cauca, la transformación de Tilapia roja (Oreochromis spp deja como subproductos mayoritarios cabezas, huesos, piel, escamas y vísceras, los cuales a través del proceso de ensilaje pueden brindar una alternativa proteica en la formulación de alimentos para peces. La presente investigación obtuvo pelets extruidos para tilapia roja en fase de engorde, sustituyendo parcialmente harina de pescado por ensilaje biológico. Las variables de respuesta del pelet (propiedades físicas, permitieron a través de un diseño factorial 23, evaluar los parámetros del proceso de extrusión (temperatura °C, velocidad de tornillo rpm y humedad %. Los resultados indicaron significancia por parte de factores e interacciones, siendo la humedad el factor de mayor incidencia. El punto medio (PM del diseño, formulado con 15% de ensilaje y procesado con niveles medios (125°C, 170 rpm, 28%, presentó un porcentaje superior al 90% de flotabilidad, además de resultados aceptables en el índice de expansión, densidad específica, índice de absorción de agua, estabilidad en agua y durabilidad; logrando condiciones favorables para la obtención de un alimento con propiedades físicas similares a las de un alimento comercial.Na barragem, 'A Salvajina', no municipio de Suárez (Cauca, a transformação da Tilápia Vermelha (Oreochromis spp, deixa como principais produtos: cabeças, ossos, pele, escamas e as vísceras, que através do processo de ensilagem podem constituir uma alternativa de proteína na formulação de rações para peixes. Esta pesquisa obteve aglomerados extrusados para tilápia vermelha na engorda, substituindo parcialmente a farinha de peixe por silagem biológica. As variáveis de resposta de pelet (propriedades físicas, permitiram através de um planejamento fatorial 23, avaliar os parâmetros de processo de extrusão (temperatura °C, velocidade do parafuso rpm e umidade %. Os resultados

  16. Phlebotomus sergenti in a Cutaneous Leishmaniasis Focus in Azilal Province (High Atlas, Morocco): Molecular Detection and Genotyping of Leishmania tropica, and Feeding Behavior

    Science.gov (United States)

    Ajaoud, Malika; Es-Sette, Nargys; Charrel, Rémi N; Laamrani-Idrissi, Abderahmane; Nhammi, Haddou; Riyad, Myriam; Lemrani, Meryem

    2015-01-01

    Background Phlebotomus (Paraphlebotomus) sergenti is at least one of the confirmed vectors for the transmission of cutaneous leishmaniasis caused by Leishmania tropica and distributed widely in Morocco. This form of leishmaniasis is considered largely as anthroponotic, although dogs were found infected with Leishmania tropica, suggestive of zoonosis in some rural areas. Methodology and Findings This survey aimed at (i) studying the presence of Leishmania in field caught Phlebotomus sergenti, (ii) investigating genetic diversity within Leishmania tropica and (iii) identifying the host-blood feeding preferences of Phlebotomus sergenti. A total of 4,407 sand flies were collected in three rural areas of Azilal province, using CDC miniature light traps. Samples collected were found to consist of 13 species: Phlebotomus spp. and 3 Sergentomyia spp. The most abundant species was Phlebotomus sergenti, accounting for 45.75 % of the total. 965 female Phlebotomus sergenti were screened for the presence of Leishmania by ITS1-PCR-RFLP, giving a positive rate of 5.7% (55/965), all being identified as Leishmania tropica. Nucleotide heterogeneity of PCR-amplified ITS1-5.8S rRNA gene-ITS2 was noted. Analyses of 31 sequences obtained segregated them into 16 haplotypes, of which 7 contain superimposed peaks at certain nucleotide positions, suggestive of heterozygosity. Phlebotomus sergenti collected were found to feed on a large variety of vertebrate hosts, as determined by Cytochrome b sequencing of the DNA from the blood meals of 64 engorged females. Conclusion Our findings supported the notion that Phlebotomus sergenti is the primary vector of Leishmania tropica in this focus, and that the latter is genetically very heterogeneous. Furthermore, our results might be suggestive of a certain level of heterozygosity in Leishmania tropica population. This finding, as well as the feeding of the vectors on different animals are of interest for further investigation. PMID:25826399

  17. Phlebotomus sergenti in a cutaneous leishmaniasis focus in Azilal province (High Atlas, Morocco: molecular detection and genotyping of Leishmania tropica, and feeding behavior.

    Directory of Open Access Journals (Sweden)

    Malika Ajaoud

    2015-03-01

    Full Text Available Phlebotomus (Paraphlebotomus sergenti is at least one of the confirmed vectors for the transmission of cutaneous leishmaniasis caused by Leishmania tropica and distributed widely in Morocco. This form of leishmaniasis is considered largely as anthroponotic, although dogs were found infected with Leishmania tropica, suggestive of zoonosis in some rural areas.This survey aimed at (i studying the presence of Leishmania in field caught Phlebotomus sergenti, (ii investigating genetic diversity within Leishmania tropica and (iii identifying the host-blood feeding preferences of Phlebotomus sergenti. A total of 4,407 sand flies were collected in three rural areas of Azilal province, using CDC miniature light traps. Samples collected were found to consist of 13 species: Phlebotomus spp. and 3 Sergentomyia spp. The most abundant species was Phlebotomus sergenti, accounting for 45.75 % of the total. 965 female Phlebotomus sergenti were screened for the presence of Leishmania by ITS1-PCR-RFLP, giving a positive rate of 5.7% (55/965, all being identified as Leishmania tropica. Nucleotide heterogeneity of PCR-amplified ITS1-5.8S rRNA gene-ITS2 was noted. Analyses of 31 sequences obtained segregated them into 16 haplotypes, of which 7 contain superimposed peaks at certain nucleotide positions, suggestive of heterozygosity. Phlebotomus sergenti collected were found to feed on a large variety of vertebrate hosts, as determined by Cytochrome b sequencing of the DNA from the blood meals of 64 engorged females.Our findings supported the notion that Phlebotomus sergenti is the primary vector of Leishmania tropica in this focus, and that the latter is genetically very heterogeneous. Furthermore, our results might be suggestive of a certain level of heterozygosity in Leishmania tropica population. This finding, as well as the feeding of the vectors on different animals are of interest for further investigation.

  18. Progress towards a Leishmania vaccine.

    Science.gov (United States)

    Tabbara, Khaled S

    2006-07-01

    Leishmaniasis is a vector-born protozoan disease. Approximately 12 million individuals are affected worldwide with an estimated annual incidence of 1.5-2 million. Two clinical manifestations are recognized, cutaneous, and visceral, both of which are common in the Middle East. In both forms, infection is chronic, with potential deformities, persistence following cure, and lifelong risk of reactivation. Attempts to develop an effective human Leishmania vaccine have not yet succeeded. Leishmanization, a crude form of live vaccination historically originated in this part of the world. Experimental vaccination has been extensively studied in model animals in the past 2 decades. In this review, major human killed vaccine trials are surveyed, and modern trends in Leishmania vaccine development, including subunit vaccines, naked DNA vaccines, and transmission blocking vaccines are explored. Recent findings of a link between persistence of live parasites, and maintenance of long-term immunity suggest live vaccination with attenuated strains, as a future vaccination strategy.

  19. Leishmania species identification using FTA card sampling directly from patients' cutaneous lesions in the state of Lara, Venezuela.

    Science.gov (United States)

    Kato, Hirotomo; Watanabe, Junko; Mendoza Nieto, Iraida; Korenaga, Masataka; Hashiguchi, Yoshihisa

    2011-10-01

    A molecular epidemiological study was performed using FTA card materials directly sampled from lesions of patients with cutaneous leishmaniasis (CL) in the state of Lara, Venezuela, where causative agents have been identified as Leishmania (Viannia) braziliensis and L. (Leishmania) venezuelensis in previous studies. Of the 17 patients diagnosed with CL, Leishmania spp. were successfully identified in 16 patients based on analysis of the cytochrome b gene and rRNA internal transcribed spacer sequences. Consistent with previous findings, seven of the patients were infected with L. (V.) braziliensis. However, parasites from the other nine patients were genetically identified as L. (L.) mexicana, which differed from results of previous enzymatic and antigenic analyses. These results strongly suggest that L. (L.) venezuelensis is a variant of L. (L.) mexicana and that the classification of L. (L.) venezuelensis should be reconsidered. Copyright © 2011 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.

  20. Lutzomyia migonei is a permissive vector competent for Leishmania infantum.

    Science.gov (United States)

    Guimarães, Vanessa Cristina Fitipaldi Veloso; Pruzinova, Katerina; Sadlova, Jovana; Volfova, Vera; Myskova, Jitka; Filho, Sinval Pinto Brandão; Volf, Petr

    2016-03-17

    Leishmania infantum is the most widespread etiological agent of visceral leishmaniasis (VL) in the world, with significant mortality rates in human cases. In Latin America, this parasite is primarily transmitted by Lutzomyia longipalpis, but the role of Lutzomyia migonei as a potential vector for this protozoan has been discussed. Laboratory and field investigations have contributed to this hypothesis; however, proof of the vector competence of L. migonei has not yet been provided. In this study, we evaluate for the first time the susceptibility of L. migonei to L. infantum. Females of laboratory-reared L. migonei were fed through a chick-skin membrane on rabbit blood containing L. infantum promastigotes, dissected at 1, 5 and 8 days post-infection (PI) and checked microscopically for the presence, intensity and localisation of Leishmania infections. In addition, morphometric analysis of L. infantum promastigotes was performed. High infection rates of both L. infantum strains tested were observed in L. migonei, with colonisation of the stomodeal valve already on day 5 PI. At the late-stage infection, most L. migonei females had their cardia and stomodeal valve colonised by high numbers of parasites, and no significant differences were found compared to the development in L. longipalpis. Metacyclic forms were found in all parasite-vector combinations since day 5 PI. We propose that Lutzomyia migonei belongs to sand fly species permissive to various Leishmania spp. Here we demonstrate that L. migonei is highly susceptible to the development of L. infantum. This, together with its known anthropophily, abundance in VL foci and natural infection by L. infantum, constitute important evidence that L. migonei is another vector of this parasite in Latin America.

  1. Dual Transcriptome Profiling of Leishmania-Infected Human Macrophages Reveals Distinct Reprogramming Signatures.

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    Fernandes, Maria Cecilia; Dillon, Laura A L; Belew, Ashton Trey; Bravo, Hector Corrada; Mosser, David M; El-Sayed, Najib M

    2016-05-10

    Macrophages are mononuclear phagocytes that constitute a first line of defense against pathogens. While lethal to many microbes, they are the primary host cells of Leishmania spp. parasites, the obligate intracellular pathogens that cause leishmaniasis. We conducted transcriptomic profiling of two Leishmania species and the human macrophage over the course of intracellular infection by using high-throughput RNA sequencing to characterize the global gene expression changes and reprogramming events that underlie the interactions between the pathogen and its host. A systematic exclusion of the generic effects of large-particle phagocytosis revealed a vigorous, parasite-specific response of the human macrophage early in the infection that was greatly tempered at later time points. An analogous temporal expression pattern was observed with the parasite, suggesting that much of the reprogramming that occurs as parasites transform into intracellular forms generally stabilizes shortly after entry. Following that, the parasite establishes an intracellular niche within macrophages, with minimal communication between the parasite and the host cell later during the infection. No significant difference was observed between parasite species transcriptomes or in the transcriptional response of macrophages infected with each species. Our comparative analysis of gene expression changes that occur as mouse and human macrophages are infected by Leishmania spp. points toward a general signature of the Leishmania-macrophage infectome. Little is known about the transcriptional changes that occur within mammalian cells harboring intracellular pathogens. This study characterizes the gene expression signatures of Leishmania spp. parasites and the coordinated response of infected human macrophages as the pathogen enters and persists within them. After accounting for the generic effects of large-particle phagocytosis, we observed a parasite-specific response of the human macrophages early in

  2. Identification of a RAC/AKT-like gene in Leishmania parasites as a putative therapeutic target in leishmaniasis.

    Science.gov (United States)

    Varela-M, Rubén E; Ochoa, Rodrigo; Muskus, Carlos E; Muro, Antonio; Mollinedo, Faustino

    2017-10-10

    Leishmaniasis is one of the world's most neglected diseases caused by at least 20 different species of the protozoan parasite Leishmania. Although new drugs have become recently available, current therapy for leishmaniasis is still unsatisfactory. A subgroup of serine/threonine protein kinases named as related to A and C protein kinases (RAC), or protein kinase B (PKB)/AKT, has been identified in several organisms including Trypanosoma cruzi parasites. PKB/AKT plays a critical role in mammalian cell signaling promoting cell survival and is a major drug target in cancer therapy. However, the role of protozoan parasitic PKB/AKT remains to be elucidated. We have found that anti-human AKT antibodies recognized a protein of about 57 kDa in Leishmania spp. parasites. Anti-human phospho-AKT(Thr308) antibodies identified a protein in extracts from Leishmania spp. that was upregulated following parasite exposure to stressful conditions, such as nutrient deprivation or heat shock. Incubation of AKT inhibitor X with Leishmania spp. promastigotes under stressful conditions or with Leishmania-infected macrophages led to parasite cell death. We have identified and cloned a novel gene from Leishmania donovani named Ld-RAC/AKT-like gene, encoding a 510-amino acid protein of approximately 57.6 kDa that shows a 26.5% identity with mammalian AKT1. Ld-RAC/AKT-like protein contains major mammalian PKB/AKT hallmarks, including the typical pleckstrin, protein kinase and AGC kinase domains. Unlike mammalian AKT that contains key phosphorylation sites at Thr308 and Ser473 in the activation loop and hydrophobic motif, respectively, Ld-RAC/AKT-like protein has a Thr residue in both motifs. By domain sequence comparison, we classified AKT proteins from different origins in four major subcategories that included different parasites. Our data suggest that Ld-RAC/AKT-like protein represents a Leishmania orthologue of mammalian AKT involved in parasite stress response and survival, and

  3. In-silico Leishmania Target Selectivity of Antiparasitic Terpenoids

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    Ifedayo Victor Ogungbe

    2013-07-01

    Full Text Available Neglected Tropical Diseases (NTDs, like leishmaniasis, are major causes of mortality in resource-limited countries. The mortality associated with these diseases is largely due to fragile healthcare systems, lack of access to medicines, and resistance by the parasites to the few available drugs. Many antiparasitic plant-derived isoprenoids have been reported, and many of them have good in vitro activity against various forms of Leishmania spp. In this work, potential Leishmania biochemical targets of antiparasitic isoprenoids were studied in silico. Antiparasitic monoterpenoids selectively docked to L. infantum nicotinamidase, L. major uridine diphosphate-glucose pyrophosphorylase and methionyl t-RNA synthetase. The two protein targets selectively targeted by germacranolide sesquiterpenoids were L. major methionyl t-RNA synthetase and dihydroorotate dehydrogenase. Diterpenoids generally favored docking to L. mexicana glycerol-3-phosphate dehydrogenase. Limonoids also showed some selectivity for L. mexicana glycerol-3-phosphate dehydrogenase and L. major dihydroorotate dehydrogenase while withanolides docked more selectively with L. major uridine diphosphate-glucose pyrophosphorylase. The selectivity of the different classes of antiparasitic compounds for the protein targets considered in this work can be explored in fragment- and/or structure-based drug design towards the development of leads for new antileishmanial drugs.

  4. Metacyclic promastigotes of Leishmania amazonensis selection using gamma irradiation

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    Bonetti, Franco C.; Nascimento, Nanci do [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil). Centro de Biologia Molecular]. E-mail: fbonetti@usp.br; Andrade Junior, Heitor Franco de [Instituto de Medicina Tropical de Sao Paulo, SP (Brazil). Lab. de Protozoologia

    2005-07-01

    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 {sup 60}Co 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. (a0011uth.

  5. Metacyclic promastigotes of Leishmania amazonensis selection using gamma irradiation

    International Nuclear Information System (INIS)

    Bonetti, Franco C.; Nascimento, Nanci do; Andrade Junior, Heitor Franco de

    2005-01-01

    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 60 Co 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)

  6. Experimental mixed infection of Leishmania (Leishmania) amazonensis and Leishmania (L.) infantum in hamsters (Mesocricetus auratus).

    Science.gov (United States)

    DE Lima Celeste, Jordanna Luíza; Venuto Moura, Ana Paula; França-Silva, João Carlos; Matos DE Sousa, Gabriela; Oliveira Silva, Soraia; Norma Melo, Maria; Luiz Tafuri, Wagner; Carvalho Souza, Carolina; Monteiro DE Andrade, Hélida

    2017-08-01

    In South America, visceral leishmaniasis is frequently caused by Leishmania infantum and, at an unknown frequency, by Leishmania amazonensis. Therefore, mixed infections with these organisms are possible. Mixed infections might affect the clinical course, immune response, diagnosis, treatment and epidemiology of the disease. Here we describe the clinical course of mixed infections with L. amazonensis and L. infantum in a hamster model. We show that mixed infections are associated with more severe clinical disease than infection with L. amazonensis or L. infantum alone. In spleens with mixed infections, L. infantum outcompeted L. amazonensis in the tissue, but not in culture from tissue. We found increased levels of IgG in animals infected with L. infantum. Although more than 30 bands were revealed in a Western blot, the highest immunogenicity was observed with proteins having molecular masses of 95 and 90 kDa, whereas proteins with molecular masses of lower than 50 kDa were reactive frequently with serum from hamsters infected with L. amazonensis, and proteins with molecular masses of 80 and 70 kDa were reactive only with serum from hamsters infected with L. infantum. This finding has important implications regarding the biology of Leishmania and humoral immune responses to infections with these organisms.

  7. Evaluación de la selección H, mediante hermanos completos y su síntesis, para resistencia a Diatraea spp. de dos variedades de maíz

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    López John Jairo

    1986-12-01

    Full Text Available El criterio de selección fue el menor número de orificios de salida del insecto, en pares de plantas, hembra y macho, éstas, últimas también auto polinizadas. En infestación natural, se estudió el efecto del sistema de selección en el daño causado por el insecto, y en los caracteres correlacionados: rendimiento, días a floración femenina, prolificidad, altura de mazorca y estatura de planta. Se determinó el avance por ciclo de selección para cada uno de estos caracteres por el método de la regresión, tomando el porcentaje relativo de cada carácter con respecto a la variedad original, para cada uno de los niveles de síntesis en las dos variedades. Se obtuvieron materiales derivados en los que aparentemente se fueron acumulando genes para otros caracteres relacionados con rendimiento.An evaluation of the H selection, through full-sib in them and their synthesis, was carry out, for resistance to Diatraea spp., in two corn varieties (ETO and DIACOL V. 351. The judgment of selection was the less number of holes from the insect exit, in pairs of plants, male and female, these last ones self pollennized. Under natural infestation, the effect of the selection system was studied in the damage caused by the insect, and in the correlate yield character, the femenine floration days, prolificity, ear corn height and plant stature. The advance was determined by the selection cycle of each one ot these characters under the regretion method, taking a relative percentage of each character in respect to the original variety, for each one of the synthesis level. Derived materials were obtained in which genes for the resistance to Diatraea spp., apparently were acumulated, and indirectly genes were qathered for other characters related to the yielding.

  8. Leishmania amazonensis DNA in wild females of Lutzomyia 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.

  9. DISEÑO Y PRODUCCIÓN DE UNA HERRAMIENTA MOLECULAR PARA EL ESTUDIO DEL N-TERMINAL DE LA NICOTINAMIDA MONONUCLEÓTIDO ADENILIL TRANSFERASA (NMNAT EN Leishmania braziliensis

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    Jesus Castaño

    2012-12-01

    Full Text Available Leishmania braziliensis es un parásitoprotozoario causante de la mayor partede casos de leishmaniasis cutánea enal menos quince países del continenteamericano. La Organización Mundial dela Salud (OMS ha reportado que cercade doce millones de personas están infectadasen el mundo y que este númeroaumenta cada año. Debido al delicadoproblema de salud pública derivado dela prevalencia de esta enfermedad sehace necesario el estudio del metabolismode este parásito. En tal sentido se haestudiado la proteína NMNAT de esteparásito, la cual es una enzima centraldel metabolismo de todos los organismosal estar encargada de la síntesisdel NAD+, un importante cofactor enreacciones redox de procesos centralesdel metabolismo celular. En la NMNATde L. braziliensis se ha encontrado unasecuencia de 44 aminoácidos en el extremoN-terminal carente de homologíacon la proteína del hospedero. En esteestudio se produjeron anticuerpos IgGespecíficos contra esta secuencia, utilizandocomo antígenos péptidos quecontuvieran la secuencia mencionada.Los anticuerpos obtenidos mostraron unreconocimiento de la NMNAT recombinantede L. braziliensis mediante ensayopor western blot.

  10. A preliminary parasitological survey of hepatozoon spp. Infection in dogs in mashhad, iran.

    Science.gov (United States)

    Amoli, Aa Rahmani; Khoshnegah, J; Razmi, Ghr

    2012-01-01

    We attempted to determine the prevalence of Hepatozoon spp. infection in Mashhad, northeast of Iran, via blood smear parasitology. The prevalence was investigated by examination of blood smear parasitology, using blood samples collected from 254 dogs (51 strays and 203 privately owned-dogs). Two stray dogs (2/51; 3.92%) and two privately-owned dogs (2/203; 0.98%) were infected with Hepatozoon spp. Therefore, as per blood smear parasitology, the prevalence of Hepatozoon spp. infection was 1.57% (4/254). Sixteen out of 254 dogs (6.29%) were infested with ticks; all of which were Rhipicephalus sanguineus. One of the dogs infected with Hepatozoon spp. exhibited ticks at the time of examination. Concurrent infection with Ehrlichia canis and Leishmania infantum was not detected in the four Hepatozoon spp. infected dogs. This is the first epidemiological study on the prevalence of Hepatozoon spp. infection in dogs in Iran.

  11. The use of successive milking as a procedure for the elimination of infection on due to Prototheca spp in cases of bovine clinical mastitis. (A case report / Utilização de ordenhas sucessivas como procedimento para eliminação de infecção por prototheca spp de casos de mastite clínica bovina (relato de caso

    Directory of Open Access Journals (Sweden)

    Márcia Kiyoe Shimada

    2000-01-01

    Full Text Available This work relates the case of two animals (Black and White Holstein from a bovine milking herd, from north of Paraná State, in which mastitis due Prototheca spp was diagnosed. As a procedure for the infection elimination, it was opted for the use of successive milking performed six times daily, during eight days. The animals were followed up through mycological cultures from milk Samples after 15, 30 and 60 days from the beginning of the procedure. The clinical signs regressed after three days and the mycologicai cultures were negative after the 15th day.O presente trabalho relata um caso de dois animais HPB de um rebanho bovino leiteiro, da região norte do Estado do Paraná, dos quais diagnosticou-se mastite clínica por Prototheca spp. Como procedimento para eliminação da infecção optou-se peta utilização de ordenhas sucessivas praticadas seis vezes diariamente durante oito dias. Os animais foram acompanhados através de culturas micológicas das amostras de leite após 15, 30 e 60 dias do inicio do procedimento. Os sinais clínicos regrediram após três dias, e a partir do 15° dia as culturas foram negativas.

  12. Avaliação de Matrizes de Carne Bovina na Produção de Itens de Ensaio de Proficiência para Pesquisa de Salmonella spp.*

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    Marcelo Luiz Lima Brandão

    2014-01-01

    Full Text Available Objetivo: Avaliar três tipos de matrizes cárneas (autoclavada a 121ºC/15 min, cozida por 20 min e enlatada na produção de itens de ensaio (IE contendo Salmonella spp., a serem utilizados em um Ensaio de Proficiência (EP, Material e Métodos: O lote de IE foi preparado utilizando a técnica de liofilização e a trealose como crioprotetor e avaliados quando a sua homogeneidade e estabilidade, Resultados: Os lotes produzidos com carne cozida e com carne enlatada não se apresentaram suficientemente homogêneos, Os IE produzidos em matriz carne autoclavada foram considerados suficientemente homogêneos e estáveis à temperatura de ≤-70ºC durante o período de quatro meses, No estudo de estabilidade em curta duração, os IE apresentaram-se suficientemente estáveis nas temperaturas de -20, 4 e 25ºC durante três dias, Conclusão: A carne autoclavada a 121ºC/15 min foi considerada uma matriz satisfatória para produção de IE contendo Salmonella spp, aplicáveis a um EP.

  13. Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin.

    Science.gov (United States)

    Prestes, Suzane Ribeiro; Guerra, Jorge Augusto de Oliveira; Romero, Gustavo Adolfo Sierra; Magalhaes, Laylah Kelre Costa; Santana, Rosa Amelia Gonçalves; Maciel, Marcel Gonçalves; Custódio, Ana; Barbosa, Maria das Graças Vale; Silveira, Henrique

    2015-01-01

    In the Americas, mucosal leishmaniasis is primarily associated with infection by Leishmania (Viannia) braziliensis. However, Leishmania (Viannia) guyanensis is another important cause of this disease in the Brazilian Amazon. In this study, we aimed at detecting Leishmaniadeoxyribonucleic acid (DNA) within paraffin-embedded fragments of mucosal tissues, and characterizing the infecting parasite species. We evaluated samples collected from 114 patients treated at a reference center in the Brazilian Amazon by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses. Direct examination of biopsy imprints detected parasites in 10 of the 114 samples, while evaluation of hematoxylin and eosin-stained slides detected amastigotes in an additional 17 samples. Meanwhile, 31/114 samples (27.2%) were positive for Leishmania spp. kinetoplast deoxyribonucleic acid (kDNA) by PCR analysis. Of these, 17 (54.8%) yielded amplification of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identification. Six of the samples were identified as L. (V.) braziliensis, while the remaining 11 were identified as L. (V.) guyanensis. The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffin-embedded tissues. Moreover, our findings confirm that L. (V.) guyanensisis a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.

  14. Detection and molecular identification of leishmania RNA virus (LRV) in Iranian Leishmania species.

    Science.gov (United States)

    Hajjaran, Homa; Mahdi, Maryam; Mohebali, Mehdi; Samimi-Rad, Katayoun; Ataei-Pirkooh, Angila; Kazemi-Rad, Elham; Naddaf, Saied Reza; Raoofian, Reza

    2016-12-01

    Leishmania RNA virus (LRV) was first detected in members of the subgenus Leishmania (Viannia), and later, the virulence and metastasis of the New World species were attributed to this virus. The data on the presence of LRV in Old World species are confined to Leishmania major and a few Leishmania aethiopica isolates. The aim of this study was to survey the presence of LRV in various Iranian Leishmania species originating from patients and animal reservoir hosts. Genomic nucleic acids were extracted from 50 cultured isolates belonging to the species Leishmania major, Leishmania tropica, and Leishmania infantum. A partial sequence of the viral RNA-dependent RNA polymerase (RdRp) gene was amplified, sequenced and compared with appropriate sequences from the GenBank database. We detected the virus in two parasite specimens: an isolate of L. infantum derived from a visceral leishmaniasis (VL) patient who was unresponsive to meglumine antimoniate treatment, and an L. major isolate originating from a great gerbil, Rhombomys opimus. The Iranian LRV sequences showed the highest similarities to an Old World L. major LRV2 and were genetically distant from LRV1 isolates detected in New World Leishmania parasites. We could not attribute treatment failure in VL patient to the presence of LRV due to the limited number of specimens analyzed. Further studies with inclusion of more clinical samples are required to elucidate the potential role of LRVs in pathogenesis or treatment failure of Old World leishmaniasis.

  15. Presencia de Giardia lamblia y Cryptosporidium spp. en aguas residuales depuradas reutilizadas para riego agrícola en la isla de Tenerife, España. Efectos del transporte a larga distancia sobre la calidad del agua reutilizada

    Directory of Open Access Journals (Sweden)

    N. Abreu Acosta

    2002-06-01

    Full Text Available El empleo de aguas residuales depuradas para riego agrícola es la alternativa de reutilización con más relevancia hoy día, siempre y cuando se desarrolle con garantías sanitarias y medioambientales.Giardia lamblia y Cryptosporidium spp. son dos protozoos patógenos entéricos de amplia distribución ambiental, frecuentes en hábitats acuáticos. La investigación y detección de estos parásitos en aguas ha adquirido importancia en los últimos años debido a que poseen formas de dispersión resistentes a los tratamientos habituales, aplicados tanto en procesos de potabilización como de depuración, y a que han sido clasificados como patógenos emergentes causantes de importantes brotes de transmisión hídrica.En este trabajo se estudia la presencia de quistes de Giardia lamblia y ooquistes de Cryptosporidium spp. en el agua residual depurada de la ciudad de Santa Cruz de Tenerife que es transportada hasta el sur de la isla de Tenerife para su reutilización en agricultura. Asimismo se investiga el efecto del transporte, el almacenamiento y el tratamiento avanzado sobre la concentración de quistes y ooquistes a través del sistema, y la existencia de relaciones con otros parámetros bacteriológicos y físico-químicos.Los resultados obtenidos ponen de manifiesto comportamientos variables de los contenidos en quistes y ooquistes frente a los tratamientos aplicados, y el efecto depurador que el transporte a larga distancia, parece tener sobre el agua residual depurada.No se han encontrado relaciones entre las concentraciones de quistes y ooquistes en el agua residual depurada y el contenido de los indicadores tradicionales de contaminación fecal.

  16. Distribution and identification of sand flies naturally infected with Leishmania from the Southeastern Peruvian Amazon.

    Science.gov (United States)

    Zorrilla, Victor; De Los Santos, Maxy B; Espada, Liz; Santos, Rocío Del Pilar; Fernandez, Roberto; Urquia, Albino; Stoops, Craig A; Ballard, Sarah-Blythe; Lescano, Andres G; Vásquez, Gissella M; Valdivia, Hugo O

    2017-11-01

    Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian and military populations that are frequently exposed in endemic settings. The Peruvian region of Madre de Dios located near the border with Brazil is one of the most endemic CL regions in South America with more than 4,451 reported cases between 2010 and 2015 according to the Peruvian epidemiology directorate. However, little is known regarding the diversity and distribution of sand fly vectors in this region. In this study, we aimed to characterize the sand fly fauna in this endemic setting and identify sand fly species naturally infected with Leishmania possibly involved in pathogen transmission. Sand fly collections were carried out during 2014 and 2015 in the communities of Flor de Acre, Villa Primavera, Mavila and Arca Pacahuara using CDC light traps and Shannon traps. Collected specimens were identified and non-blood-fed females were selected for Leishmania infection screening using kinetoplastid DNA-PCR (kDNA-PCR) and nested Real time PCR for species identification. A total of 10,897 phlebotomines belonging to the genus Lutzomyia (58 species) and Brumptomyia (2 species) were collected. Our study confirmed the widespread distribution and abundance of Lutzomyia (Trichophoromyia) spp. (24%), Lu. whitmani (19.4%) and Lu. yucumensis (15.8%) in the region. Analysis of Shannon diversity index indicates variability in sand fly composition across sites with Villa Primavera presenting the highest sand fly diversity and abundance. Leishmania screening by kDNA-PCR resulted in 45 positive pools collected from Flor de Acre (34 pools), Mavila (10 pools) and Arca Pacahuara (1 pool) and included 14 species: Lu. yucumensis, Lu. aragoi, Lu. sallesi, Lu. sherlocki, Lu. shawi, Lu. walkeri, Lu nevesi, Lu. migonei, Lu. davisi, Lu. carrerai, Lu. hirsuta, Lu. (Trichophoromyia) spp., Lu. llanosmartinsi and Lu. whitmani. Lutzomyia sherlocki, Lu. walkeri and Lu. llanosmartinsi had the

  17. Distribution and identification of sand flies naturally infected with Leishmania from the Southeastern Peruvian Amazon.

    Directory of Open Access Journals (Sweden)

    Victor Zorrilla

    2017-11-01

    Full Text Available Cutaneous leishmaniasis (CL is an important health problem in the New World affecting civilian and military populations that are frequently exposed in endemic settings. The Peruvian region of Madre de Dios located near the border with Brazil is one of the most endemic CL regions in South America with more than 4,451 reported cases between 2010 and 2015 according to the Peruvian epidemiology directorate. However, little is known regarding the diversity and distribution of sand fly vectors in this region. In this study, we aimed to characterize the sand fly fauna in this endemic setting and identify sand fly species naturally infected with Leishmania possibly involved in pathogen transmission.Sand fly collections were carried out during 2014 and 2015 in the communities of Flor de Acre, Villa Primavera, Mavila and Arca Pacahuara using CDC light traps and Shannon traps. Collected specimens were identified and non-blood-fed females were selected for Leishmania infection screening using kinetoplastid DNA-PCR (kDNA-PCR and nested Real time PCR for species identification.A total of 10,897 phlebotomines belonging to the genus Lutzomyia (58 species and Brumptomyia (2 species were collected. Our study confirmed the widespread distribution and abundance of Lutzomyia (Trichophoromyia spp. (24%, Lu. whitmani (19.4% and Lu. yucumensis (15.8% in the region. Analysis of Shannon diversity index indicates variability in sand fly composition across sites with Villa Primavera presenting the highest sand fly diversity and abundance. Leishmania screening by kDNA-PCR resulted in 45 positive pools collected from Flor de Acre (34 pools, Mavila (10 pools and Arca Pacahuara (1 pool and included 14 species: Lu. yucumensis, Lu. aragoi, Lu. sallesi, Lu. sherlocki, Lu. shawi, Lu. walkeri, Lu nevesi, Lu. migonei, Lu. davisi, Lu. carrerai, Lu. hirsuta, Lu. (Trichophoromyia spp., Lu. llanosmartinsi and Lu. whitmani. Lutzomyia sherlocki, Lu. walkeri and Lu. llanosmartinsi had the

  18. Distribution and identification of sand flies naturally infected with Leishmania from the Southeastern Peruvian Amazon

    Science.gov (United States)

    Zorrilla, Victor; De Los Santos, Maxy B.; Espada, Liz; Santos, Rocío del Pilar; Fernandez, Roberto; Urquia, Albino; Stoops, Craig A.; Ballard, Sarah-Blythe; Lescano, Andres G.; Vásquez, Gissella M.; Valdivia, Hugo O.

    2017-01-01

    Background Cutaneous leishmaniasis (CL) is an important health problem in the New World affecting civilian and military populations that are frequently exposed in endemic settings. The Peruvian region of Madre de Dios located near the border with Brazil is one of the most endemic CL regions in South America with more than 4,451 reported cases between 2010 and 2015 according to the Peruvian epidemiology directorate. However, little is known regarding the diversity and distribution of sand fly vectors in this region. In this study, we aimed to characterize the sand fly fauna in this endemic setting and identify sand fly species naturally infected with Leishmania possibly involved in pathogen transmission. Methods Sand fly collections were carried out during 2014 and 2015 in the communities of Flor de Acre, Villa Primavera, Mavila and Arca Pacahuara using CDC light traps and Shannon traps. Collected specimens were identified and non-blood-fed females were selected for Leishmania infection screening using kinetoplastid DNA-PCR (kDNA-PCR) and nested Real time PCR for species identification. Results A total of 10,897 phlebotomines belonging to the genus Lutzomyia (58 species) and Brumptomyia (2 species) were collected. Our study confirmed the widespread distribution and abundance of Lutzomyia (Trichophoromyia) spp. (24%), Lu. whitmani (19.4%) and Lu. yucumensis (15.8%) in the region. Analysis of Shannon diversity index indicates variability in sand fly composition across sites with Villa Primavera presenting the highest sand fly diversity and abundance. Leishmania screening by kDNA-PCR resulted in 45 positive pools collected from Flor de Acre (34 pools), Mavila (10 pools) and Arca Pacahuara (1 pool) and included 14 species: Lu. yucumensis, Lu. aragoi, Lu. sallesi, Lu. sherlocki, Lu. shawi, Lu. walkeri, Lu nevesi, Lu. migonei, Lu. davisi, Lu. carrerai, Lu. hirsuta, Lu. (Trichophoromyia) spp., Lu. llanosmartinsi and Lu. whitmani. Lutzomyia sherlocki, Lu. walkeri and Lu

  19. Aplicaciones de la embriogénesis somática para la propagación y mejora de Quercus ilex L. y Pinus spp.

    OpenAIRE

    Blasco Carlos, Miquel

    2013-01-01

    Esta tesis está enfocada a la aplicación de la embriogénesis somática para la conservación y mejora de Quercus ilex L., Pinus pinaster Aiton y Pinus pinea L., especies forestales representativas de los bosques de la Península Ibérica. La propagación clonal mediante embriogénesis somática (ES) es la herramienta más adecuada para la propagación de genotipos de élite y para la implementación de la llamada silvicultura multivarietal. Además los cultivos embriogénicos son explantos adecuados para ...

  20. Presence of anti-Leishmania (Viannia braziliensis antibodies in blood donors in the West-Central region of the State of Paraná, Brazil

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    Lais de Souza Braga

    2015-10-01

    Full Text Available ABSTRACTINTRODUCTION:Serological screening in blood banks does not include all transmittable diseases. American cutaneous leishmaniasis (ACL has a high detection rate in the municipalities of the State of Paraná.METHODS:This study analyzed the presence of anti- Leishmania braziliensisantibodies in 176 blood donors who live in these endemic areas. The variables were analyzed with the χ2 test and Stata 9.1 software. RESULTS: Twenty (11.4% samples were positive for the presence of anti- L. braziliensisantibodies. CONCLUSIONS: The high percentage of donors with anti- Leishmania spp. antibodies indicates the need to study the risk of ACL transmission through blood donors.

  1. Determinação da dose de radiação gama para reduzir a população de Salmonella spp em carne de frango Determination of gamma radiation doses to reduce Salmonella spp in chiken meat

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    A.F. Santos

    2003-08-01

    Full Text Available O consumo de carne de frango contaminada com Salmonella é uma causa importante de salmonelose em todo o mundo. Essa doença transmitida por alimentos, é um problema de saúde pública e causa de perdas econômicas substanciais. O processo de irradiação é um método eficiente de conservação de alimentos por reduzir o número de microrganismos patogênicos e deteriorantes, sem que as características organolépticas e nutricionais do alimento sejam alteradas significativamente. Os objetivos desta pesquisa foram determinar o valor D10 de Salmonella Typhimurium ATCC 14028, inoculada em sobrecoxas de frango, e recomendar uma dose de radiação para ser aplicada a esse alimento a fim de torná-lo seguro do ponto de vista microbiológico. O valor D10 foi calculado a partir da curva de sobreviventes dessa bactéria em sobrecoxas de frango, após terem sido expostas às doses de 0kGy; 0,1kGy; 0,2kGy; 0,3kGy; 0,5kGy; 0,6kGy; 0,7kGy e 0,8kGy. O valor D10 variou de 0,241kGy a 0,480kGy. Considerando o maior valor D10 e o maior nível de contaminação de Salmonella spp encontrado em sobrecoxas de frango - 0,4NMP/g - adquiridas em feiras livres da cidade de São Paulo, a dose de radiação gama recomendada para garantir a segurança do produto em relação à presença de Salmonella spp é de 3,8kGy.The consumption of chicken meat contaminated with Salmonella spp is an important cause of salmonellosis worldwide. This food-borne disease is a public health problem and causes substantial economical loss. Irradiation process is an effective method for food preservation because it causes no significant change in organoleptic and nutritional food characteristics and destroys pathogenic and spoilage microorganisms. D10 values were calculated through the number of survivors for S. Typhimurium ATCC 14028, inoculated in chicken thighs, after being irradiated with different doses of gamma radiation - 0.0kGy; 0.1kGy; 0.2kGy; 0.3kGy; 0.5kGy; 0.7kGy and 0.8kGy . D10

  2. Amplified DNAs in laboratory stocks of Leishmania tarentolae: extrachromosomal circles structurally and functionally similar to the inverted-H-region amplification of methotrexate-resistant Leishmania major

    International Nuclear Information System (INIS)

    Petrillo-Peixoto, M.L.; Beverley, S.M.

    1988-01-01

    We describe the structure of amplified DNA that was discovered in two laboratory stocks of the protozoan parasite Leishmania tarentolae. Restriction mapping and molecular cloning revealed that a region of 42 kilobases was amplified 8- to 30-fold in these lines. Southern blot analyses of digested DNAs or chromosomes separated by pulsed-field electrophoresis showed that the amplified DNA corresponded to the H region, a locus defined originally by its amplification in methotrexate-resistant Leishmania major. Similarities between the amplified DNA of the two species included (i) extensive cross-hybridization; (ii) approximate conservation of sequence order; (iii) extrachromosomal localization; (iv) an overall inverted, head-to-head configuration as a circular 140-kilobase tetrameric molecule; (v) two regions of DNA sequence rearrangement, each of which was closely associated with the two centers of the inverted repeats; (vi) association with methotrexate resistance; and (vii) phenotypically conservative amplification, in which the wild-type chromosomal arrangement was retained without apparent modification. Our data showed that amplified DNA mediating drug resistance arose in unselected L. tarentolae, although the pressures leading to apparently spontaneous amplification and maintenance of the H region are not known. The simple structure and limited extent of DNA amplified in these and other Leishmania lines suggests that the study of gene amplification in Leishmania spp. offers an attractive model system for the study of amplification in cultured mammalian cells and tumors. We also introduced a method for measuring the size of large circular DNAs, using gamma-irradiation to introduce limited double-strand breaks followed by sizing of the linear DNAs by pulsed-field electrophoresis

  3. Phylogenomic reconstruction supports supercontinent origins for Leishmania.

    Science.gov (United States)

    Harkins, Kelly M; Schwartz, Rachel S; Cartwright, Reed A; Stone, Anne C

    2016-03-01

    Leishmania, a genus of parasites transmitted to human hosts and mammalian/reptilian reservoirs by an insect vector, is the causative agent of the human disease complex leishmaniasis. The evolutionary relationships within the genus Leishmania and its origins are the source of ongoing debate, reflected in conflicting phylogenetic and biogeographic reconstructions. This study employs a recently described bioinformatics method, SISRS, to identify over 200,000 informative sites across the genome from newly sequenced and publicly available Leishmania data. This dataset is used to reconstruct the evolutionary relationships of this genus. Additionally, we constructed a large multi-gene dataset, using it to reconstruct the phylogeny and estimate divergence dates for species. We conclude that the genus Leishmania evolved at least 90-100 million years ago, supporting a modified version of the Multiple Origins hypothesis that we call the Supercontinent hypothesis. According to this scenario, separate Leishmania clades emerged prior to, and during, the breakup of Gondwana. Additionally, we confirm that reptile-infecting Leishmania are derived from mammalian forms and that the species that infect porcupines and sloths form a clade long separated from other species. Finally, we firmly place the guinea-pig infecting species, Leishmaniaenriettii, the globally dispersed Leishmaniasiamensis, and the newly identified Australian species from a kangaroo, as sibling species whose distribution arises from the ancient connection between Australia, Antarctica, and South America. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Aplicación de las pruebas de PCR convencional simple y múltiple para la identificación de aislamientos de Leptospira spp. en Colombia Application of conventional and multiplex PCR assays for identification of isolates of Leptospira spp. in Colombia

    Directory of Open Access Journals (Sweden)

    Natali Moreno

    2010-12-01

    Full Text Available Debido a las dificultadas asociadas con la identificación serológica de aislamientos de Leptospira ssp, se genera gran interés en la pruebas moleculares por su poder discriminatorio, reproducibilidad y fácil interpretación. Objetivo. Aplicar y validar la prueba de PCR convencional, usando dos pares de iniciadores descritos previamente y dirigidos a los genes lipL32 (PCR simple y secY/flaB (PCR múltiple, con el fin de evaluar su aplicación para identificar especies patógenas y saprófitas de Leptospira spp. Materiales y métodos. Para la estandarización de las pruebas de PCR se usó 22 cepas de referencia internacional y 12 aislamientos colombianos. Se determinó el nivel de detección de cada pareja de iniciadores, su especificidad frente a otros microorganismos causantes de enfermedades endémicas en Colombia y su capacidad de identificar especies dentro del grupo de Leptospira. Resultados. El límite de detección de la PCR simple lipL32 fue una dilución 1:10000 y para la PCR múltiple secY/flaB fue una dilución 1:100 para el gen secY y 1:1000 para flaB. La especificidad de todos los iniciadores fue de 100%. La PCR simple lipL32, mostró amplificado específico para 21/22 cepas de referencia mientras que la PCR multiple secY/flaB lo fue para 18/22 cepas. De los 12 aislamientos colombianos, siete fueron positivos por PCR lipL32 y seis lo fueron por PCR secY/flaB. Conclusiones. Los resultados más consistentes fueron obtenidos con la PCR simple lipL32 tanto en límite de detección, especificidad y utilidad para la identificación de Leptospira spp, por lo que esta prueba es aplicable a la identificación molecular de aislamientos patógenos de Leptospira spp de diversas fuentes.Serological identification of Leptospira ssp isolates is difficult to achieve. Thus, molecular testing may be of great interest thanks to its high discrimination power, reproducibility and easy interpretation. Objective. To implement and validate conventional

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

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    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

  6. Leishmania, Babesia and Ehrlichia in urban pet dogs: co-infection or cross-reaction in serological methods?

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    Felipe da Silva Krawczak

    2015-02-01

    Full Text Available INTRODUCTION: The present study was designed to assess the occurrence of co-infection or cross-reaction in the serological techniques used for detecting the anti-Leishmania spp., -Babesia canis vogeli and -Ehrlichia canis antibodies in urban dogs from an area endemic to these parasites. METHODS: The serum samples from dogs were tested for the Babesia canis vogeli strain Belo Horizonte antigen and Ehrlichia canis strain São Paulo by immunofluorescence antibody test (IFAT and by anti-Leishmania immunoglobulin G (IgG antibody detection to assess Leishmania infection. We used the following four commercial kits for canine visceral leishmaniasis: ELISA, IFAT, Dual Path Platform (DPP (Bio Manguinhos(r/FIOCRUZ/MS and a rK39 RDT (Kalazar Detect Canine Rapid Test; Inbios. RESULTS : Of 96 serum samples submitted to serological assays, 4 (4.2% were positive for Leishmania as determined by ELISA; 12 (12.5%, by IFAT; 14 (14.6% by rK39 RDT; and 20 (20.8%, by DPP. Antibodies against Ehrlichia and Babesia were detected in 23/96 (23.9% and 30/96 (31.2% samples, respectively. No significant association was identified between the results of tests for detecting Babesia or Ehrlichia and those for detecting Leishmania (p-value>0.05. CONCLUSIONS: In the present study, we demonstrated co-infection with Ehrlichia or Babesia and Leishmania in dogs from Minas Gerais (Brazil; we also found that the serological tests that were used did not cross-react.

  7. Vaccine development against Leishmania donovani

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    Amrita eDas

    2012-05-01

    Full Text Available Visceral leishmaniasis (VL caused by Leishmania donovani and Leishmania infantum/ chagasi represents the second most challenging infectious disease worldwide, affecting nearly 500,000 people and 60,000 deaths annually. Zoonotic VL (ZVL caused by L. infantum is re-emergent canid zoonoses which represents a complex epidemiological cycle in New world where domestic dogs serve as reservoir host responsible for potentially fatal human infection where dog culling is the only control measure for eliminating reservoir host. Lifelong immunity in human against reinfection has motivated several attempts in developing prophylactic vaccines against the disease but very few have progressed beyond experimental stage. Absence of any licensed vaccine along with high toxicity and increasing resistance to the current chemotherapeutic drugs has further complicated the situation in endemic regions of the world. Advances in vaccinology, including recombinant proteins, novel antigen-delivery systems/adjuvants, heterologous prime-boost regimens and strategies for intracellular antigen presentation, have contributed to recent advances in vaccine development against VL. Attempts to develop an effective vaccine for use in domestic dogs in areas of canine VL should be pursued for preventing human infection. Studies in animal models and human patients have revealed the pathogenic mechanisms of disease progression and features of protective immunity. This review will summarize the accumulated knowledge about pathogenesis, immune response and prerequisites for protective immunity against human VL. Authors will discuss promising vaccine targets, their developmental status and future prospects in a quest for rational vaccine development against VL. In addition, several challenges such as safety issues, a renewed and coordinated commitment to basic research, preclinical studies and trial design will be addressed to overcome the problems faced in developing effective vaccines

  8. Evaluación de la curva de crecimiento, cuatro sustratos, dos métodos de esporulación, tres portadores inertes para Trichoderma spp. y su comportamiento frente a ocho fungicidas y cuatro hongos patógenos en laboratorio.

    OpenAIRE

    Erazo Sandoval, Norma Soledad

    2001-01-01

    Como un método alternativo en el combate de enfermedades causadas por patógenos del suelo y de la filósfera, se consideró a Trichoderma spp por sus múltiples cualidades que lo han convertido en el candidato ideal en el manejo de agroecosistemas. Sin embargo, para que sea fácilmente disponible y aceptado, es necesario contar con formulaciones apropiadas, por lo que esta investigación tuvo como objetivo la Determinación de la curva de crecimiento y esporulación de Trichoderma spp.. Se utili...

  9. High density of Leishmania major and rarity of other mammals' Leishmania in zoonotic cutaneous leishmaniasis foci, Iran.

    Science.gov (United States)

    Bordbar, Ali; Parvizi, Parviz

    2014-03-01

    Only Leishmania major is well known as a causative agent of zoonotic cutaneous leishmaniasis (ZCL) in Iran. Our objective was to find Leishmania parasites circulating in reservoir hosts, sand flies and human simultaneously. Sand flies, rodents and prepared smears of humans were sampled. DNA of Leishmania parasites was extracted, and two fragments of ITS-rDNA gene amplified by PCR. RFLP and sequencing were employed to identify Leishmania parasites. Leishmania major and L. turanica were identified unequivocally by targeting and sequencing ITS-rDNA from humans, rodents and sand flies. The new Leishmania species close to gerbilli (GenBank Accession Nos. EF413076; EF413087) was discovered only in sand flies. Based on parasite detection of ITS-rDNA in main and potential reservoir hosts and vectors and humans, we conclude that at least two Leishmania species are common in the Turkmen Sahra ZCL focus. Phylogenetic analysis proved that the new Leishmania is closely related to Leishmania mammal parasites (Leishmania major, Leishmania turanica, Leishmania gerbilli). Its role as a principal agent of ZCL is unknown because it was found only in sand flies. Our findings shed new light on the transmission cycles of several Leishmania parasites in sand flies, reservoir hosts and humans. © 2014 John Wiley & Sons Ltd.

  10. 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

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    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.

  11. Produção de farinha de banana verde (Musa spp. para aplicação em pão de trigo integral

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    Bruna Andina Andrade

    2017-11-01

    Full Text Available Resumo A farinha de banana verde (FBV é uma alternativa viável para o aproveitamento dos frutos e possui alto teor de amido resistente (AR, que apresenta papel fisiológico similar ao das fibras alimentares. O objetivo deste trabalho foi produzir FBV de dois genótipos de bananeira (Prata e Caturra, além de selecionar aquele com maior teor de AR, para posterior substituição de parte da farinha de trigo integral por FBV, em uma formulação de pão de fôrma integral. Para a produção das farinhas, os frutos foram descascados, cortados em rodelas e estas foram submersas em uma solução de ácido ascórbico e ácido cítrico, sendo então distribuídas em bandejas e secas a 50 °C, por 7 horas, e moídas. As farinhas foram submetidas às análises de composição proximal e determinação do teor de AR. Posteriormente, foram processados pães integrais com a substituição de 10%, 15% e 20% de farinha de trigo integral pela FBV que obteve o maior teor de AR. Os pães controle e aquele que apresentou maior teor de AR e boas características tecnológicas, como volume específico, dureza e cor, foram submetidos aos testes sensoriais de aceitação e intenção de compra. O teor de AR encontrado para as FBV Prata e Caturra foram 24,1% e 13,7%, respectivamente. O pão com substituição de 15% de farinha de trigo integral por FBV foi escolhido para a análise sensorial. As formulações controle e com 15% de FBV atingiram Índices de Aceitação Sensorial de 88,7% e 82,1%, respectivamente. A atitude dos avaliadores foi positiva, segundo a qual 56% dos avaliadores “comprariam frequentemente” ou “comprariam sempre” o pão com 15% de FBV, não tendo sido observada rejeição para nenhuma das formulações.

  12. Members of a large retroposon family are determinants of post-transcriptional gene expression in Leishmania.

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    Frédéric Bringaud

    2007-09-01

    Full Text Available Trypanosomatids are unicellular protists that include the human pathogens Leishmania spp. (leishmaniasis, Trypanosoma brucei (sleeping sickness, and Trypanosoma cruzi (Chagas disease. Analysis of their recently completed genomes confirmed the presence of non-long-terminal repeat retrotransposons, also called retroposons. Using the 79-bp signature sequence common to all trypanosomatid retroposons as bait, we identified in the Leishmania major genome two new large families of small elements--LmSIDER1 (785 copies and LmSIDER2 (1,073 copies--that fulfill all the characteristics of extinct trypanosomatid retroposons. LmSIDERs are approximately 70 times more abundant in L. major compared to T. brucei and are found almost exclusively within the 3'-untranslated regions (3'UTRs of L. major mRNAs. We provide experimental evidence that LmSIDER2 act as mRNA instability elements and that LmSIDER2-containing mRNAs are generally expressed at lower levels compared to the non-LmSIDER2 mRNAs. The considerable expansion of LmSIDERs within 3'UTRs in an organism lacking transcriptional control and their role in regulating mRNA stability indicate that Leishmania have probably recycled these short retroposons to globally modulate the expression of a number of genes. To our knowledge, this is the first example in eukaryotes of the domestication and expansion of a family of mobile elements that have evolved to fulfill a critical cellular function.

  13. Comparative study of bacteremias caused by Enterococcus spp. with and without high-level resistance to gentamicin. The Grupo Andaluz para el estudio de las Enfermedades Infecciosas.

    Science.gov (United States)

    Caballero-Granado, F J; Cisneros, J M; Luque, R; Torres-Tortosa, M; Gamboa, F; Díez, F; Villanueva, J L; Pérez-Cano, R; Pasquau, J; Merino, D; Menchero, A; Mora, D; López-Ruz, M A; Vergara, A

    1998-02-01

    A prospective, multicenter study was carried out over a period of 10 months. All patients with clinically significant bacteremia caused by Enterococcus spp. were included. The epidemiological, microbiological, clinical, and prognostic features and the relationship of these features to the presence of high-level resistance to gentamicin (HLRG) were studied. Ninety-three patients with enterococcal bacteremia were included, and 31 of these cases were caused by HLRG (33%). The multivariate analysis selected chronic renal failure, intensive care unit stay, previous use of antimicrobial agents, and Enterococcus faecalis species as the independent risk factors that influenced the development of HLRG. The strains with HLRG showed lower levels of susceptibility to penicillin and ciprofloxacin. Clinical features (except for chronic renal failure) were similar in both groups of patients. HLRG did not influence the prognosis for patients with enterococcal bacteremia in terms of either the crude mortality rate (29% for patients with bacteremia caused by enterococci with HLRG and 28% for patients not infected with strains with HLRG) or the hospital stay after the acquisition of enterococcal bacteremia. Hemodynamic compromise, inappropriate antimicrobial therapy, and mechanical ventilation were revealed in the multivariate analysis to be the independent risk factors for mortality. Prolonged hospitalization was associated with the nosocomial acquisition of bacteremia and polymicrobial infections.

  14. Screening for Inhibitors of Essential Leishmania Glucose Transporters

    Science.gov (United States)

    2013-07-01

    Leishmania Glucose Transporters PRINCIPAL INVESTIGATOR: Scott M. Landfear, Ph.D. CONTRACTING ORGANIZATION: Oregon Health & Science...COVERED 1 July 2009- 30 June 2013 4. TITLE AND SUBTITLE 5a. CONTRACT NUMBER Screening for Inhibitors of Essential Leishmania Glucose Transporters 5b...The objective of this project was to identify compounds that selectively inhibit the essential Leishmania glucose transporters and could hence serve

  15. Resistência de porta-enxertos para pessegueiro e ameixeira aos nematóides causadores de galhas (Meloidogyne spp. Resistance of rootstock for peach tree and plum to root-knot nematodes (Meloidogyne spp.

    Directory of Open Access Journals (Sweden)

    José Carlos Fachinello

    2000-03-01

    Full Text Available O trabalho foi realizado com o objetivo de comparar o comportamento de cinco diferentes porta-enxertos para frutas de caroço em frente a duas espécies de nematóides do gênero Meloidogyne (Meloidogyne javanica e Meloidogyne incognita. O cultivar GF 677 foi obtido a partir do cultivo in vitro e os demais porta-enxertos a partir de sementes. Aos dois meses, as plântulas foram repicadas e inoculadas com uma mistura de Meloidogyne javanica e M. incognita aos 30, 60 e 70 dias após o plantio em canteiros incorporando-se 0,2kg de solo altamente infestado com os nematóides ao redor de cada planta. Ao final do experimento, avaliaram-se o desenvolvimento das plantas e o grau de infecção de cada cultivar. O grau de resistência dos cultivares foi estimado a partir do índice de galhas, obtido através de uma escala de grau ou nota, a qual varia de 0 a 5, em função do número de galhas ou ootecas. Os resultados obtidos demonstraram que o cultivar Okinawa não apresentou galhas no sistema radicular e na análise de crescimento foi superior aos demais cultivares. Os cultivares R-15-2 e Aldrighi foram considerados resistentes aos fitonematóides por apresentarem pequeno número de galhas no sistema radicular. Já o cultivar GF 677 apresentou maior número de galhas no sistema radicular, chegando a 126 galhas/g de raiz.The work was carried out with the objective of comparing the response of five different stone fruit rootstocks to two nematode species (Meloidogyne javanica and Meloidogyne incognita. The cultivar GF677, which was obtained from in vitro cultivation and the others rootstocks, were obtained from seedlings. Two month old plants were transplanted and inoculated with a mixture of both nematode species at 30, 60, 70 days after planting, by incorporating 0.2kg of highly infested soil around each cultivar. The development of the roststocks were evaluated at the end of the experiment. Number of galls, ranging from 0 to 5 was used to score the

  16. USO DE MÉTODOS MULTIVARIANTES PARA LA AGRUPACIÓN DE AISLAMIENTOS DE Colletotrichum spp. CON BASE EN CARACTERÍSTICAS MORFOLÓGICAS Y CULTURALES USE OF MULTIVARIATE METHODS FOR GROUPING STRAINS OF Colletotrichum spp. BASED ON CULTURAL AND MORPHOLOGICAL CHARACTERS

    Directory of Open Access Journals (Sweden)

    Guillermo Correa Londoño

    2007-06-01

    Full Text Available Se evaluaron 15 variables morfológicas y culturales, con mezcla de variables cuantitativas y cualitativas, a 58 aislamientos de Colletotrichum spp. con cuatro repeticiones. Se generó una agrupación guiada por el Análisis de Coordenadas Principales (ACOP, basado en el coeficiente de similitud de Gower. Las dos primeras dimensiones del ACOP permitieron la perfecta discriminación de tres grandes grupos: la primera dimensión separaba a Colletotrichum gloeosporioides y su teleomorfo Glomerella cingulata de C. dematium; la segunda dimensión permitía separar a C. gloeosporioides de su teleomorfo G. cingulata. Mediante el análisis simultáneo de los tres primeros ejes factoriales, se detectaron 21 grupos menores. Para la caracterización de los grupos se usó la representación Biplot de la tabla de contingencia de los grupos y las variables. Se trabajó con las frecuencias relativas al tamaño de los grupos para evitar la alta influencia que de otra forma tendrían los grupos más numerosos sobre las configuraciones Biplot.Fifteen cultural and morphological characteristics, with a mix of quantitative and qualitative variables, were evaluated on 58 strains of Colletotrichum spp. repeated four times. Using Principal Coordinates Analysis (PCOA, based on Gower’s similarity coefficient, a grouping was obtained. The two first dimensions of PCOA allowed the perfect discrimination of three big groups: the first dimension separated Colletotrichum gloeosporioides and its teleomorph Glomerella cingulata from C. dematium; the second dimension discriminated between C. gloeosporioides and its teleomorph G. cingulata. By analyzing simultaneously the first three factorial axis, 21 minor groups were detected. In order to characterize the groups, a Biplot representation of the groups and variables contingency table was used. Relative frequencies to the size of the groups were used to avoid the high influence that otherwise the more numerous groups would have

  17. Arginase activity in pathogenic and non-pathogenic species of Leishmania parasites.

    Science.gov (United States)

    Badirzadeh, Alireza; Taheri, Tahereh; Taslimi, Yasaman; Abdossamadi, Zahra; Heidari-Kharaji, Maryam; Gholami, Elham; Sedaghat, Baharehsadat; Niyyati, Maryam; Rafati, Sima

    2017-07-01

    Proliferation of Leishmania (L.) parasites depends on polyamine availability, which can be generated by the L-arginine catabolism and the enzymatic activity of arginase (ARG) of the parasites and of the mammalian hosts. In the present study, we characterized and compared the arginase (arg) genes from pathogenic L. major and L. tropica and from non-pathogenic L. tarentolae. We quantified the level of the ARG activity in promastigotes and macrophages infected with pathogenic L. major and L. tropica and non-pathogenic L. tarentolae amastigotes. The ARG's amino acid sequences of the pathogenic and non-pathogenic Leishmania demonstrated virtually 98.6% and 88% identities with the reference L. major Friedlin ARG. Higher ARG activity was observed in all pathogenic promastigotes as compared to non-pathogenic L. tarentolae. In vitro infection of human macrophage cell line (THP1) with pathogenic and non-pathogenic Leishmania spp. resulted in increased ARG activities in the infected macrophages. The ARG activities present in vivo were assessed in susceptible BALB/c and resistant C57BL/6 mice infected with L. major, L. tropica and L. tarentolae. We demonstrated that during the development of the infection, ARG is induced in both strains of mice infected with pathogenic Leishmania. However, in L. major infected BALB/c mice, the induction of ARG and parasite load increased simultaneously according to the time course of infection, whereas in C57BL/6 mice, the enzyme is upregulated solely during the period of footpad swelling. In L. tropica infected mice, the footpads' swellings were slow to develop and demonstrated minimal cutaneous pathology and ARG activity. In contrast, ARG activity was undetectable in mice inoculated with the non-pathogenic L. tarentolae. Our data suggest that infection by Leishmania parasites can increase ARG activity of the host and provides essential polyamines for parasite salvage and its replication. Moreover, the ARG of Leishmania is vital for parasite

  18. Leishmania Hijacks Myeloid Cells for Immune Escape

    Directory of Open Access Journals (Sweden)

    María Martínez-López

    2018-05-01

    Full Text Available Protozoan parasites of the Leishmania genus are the causative agents of leishmaniasis, a group of neglected tropical diseases whose clinical manifestations vary depending on the infectious Leishmania species but also on host factors. Recognition of the parasite by host myeloid immune cells is a key to trigger an effective Leishmania-specific immunity. However, the parasite is able to persist in host myeloid cells by evading, delaying and manipulating host immunity in order to escape host resistance and ensure its transmission. Neutrophils are first in infiltrating infection sites and could act either favoring or protecting against infection, depending on factors such as the genetic background of the host or the parasite species. Macrophages are the main host cells where the parasites grow and divide. However, macrophages are also the main effector population involved in parasite clearance. Parasite elimination by macrophages requires the priming and development of an effector Th1 adaptive immunity driven by specific subtypes of dendritic cells. Herein, we will provide a comprehensive outline of how myeloid cells regulate innate and adaptive immunity against Leishmania, and the mechanisms used by the parasites to promote their evasion and sabotage. Understanding the interactions between Leishmania and the host myeloid cells may lead to the development of new therapeutic approaches and improved vaccination to leishmaniases, an important worldwide health problem in which current therapeutic or preventive approaches are limited.

  19. Cryopreservation of Leishmania Species in Manisa Province.

    Science.gov (United States)

    Çavuş, İbrahim; Ocak, Fulya; Kaya, Tuğba; Özbilgin, Ahmet

    2017-09-01

    It was aimed to assess the success of the cryopreservation process which is carried out in order to preserve the genetic material and the virulence of the Leishmania species that are an important health problem in our region. Leishmania tropica, L. infantum, L. major, and L. donovani strains in Novy-MacNeal-Nicolle (NNN) medium in MCBU were used. Promastigotes cultured in the NNN medium were transferred to RPMI 1640 medium; promastigotes in the logarithmic phase were washed three times with PBS, and 15% dimethylsulfoxide (DMSO) was added. Leishmania species were transferred to 12 separate tubes. The tubes were stored at -86°C for one night by placing them in Coolcell boxes. The tubes were transferred into a liquid nitrogen tank. One cryotube per Leishmania strain is thawed monthly and cultured in NNN medium. For the duration of study it was observed that each Leishmania isolate preserved 60-65% of their viability and entered the logarithmic phase on the 7th day following the inoculation in the NNN medium. Abnormalities in the structures and movements of the promastigotes were not observed in microscopic examinations. The following conclusions were made: cryopreservation is important for studies planned related to leishmaniasis and cryopreservation with DMSO is successful.

  20. Leishmania infections: Molecular targets and diagnosis.

    Science.gov (United States)

    Akhoundi, Mohammad; Downing, Tim; Votýpka, Jan; Kuhls, Katrin; Lukeš, Julius; Cannet, Arnaud; Ravel, Christophe; Marty, Pierre; Delaunay, Pascal; Kasbari, Mohamed; Granouillac, Bruno; Gradoni, Luigi; Sereno, Denis

    2017-10-01

    Progress in the diagnosis of leishmaniases depends on the development of effective methods and the discovery of suitable biomarkers. We propose firstly an update classification of Leishmania species and their synonymies. We demonstrate a global map highlighting the geography of known endemic Leishmania species pathogenic to humans. We summarize a complete list of techniques currently in use and discuss their advantages and limitations. The available data highlights the benefits of molecular markers in terms of their sensitivity and specificity to quantify variation from the subgeneric level to species complexes, (sub) species within complexes, and individual populations and infection foci. Each DNA-based detection method is supplied with a comprehensive description of markers and primers and proposal for a classification based on the role of each target and primer in the detection, identification and quantification of leishmaniasis infection. We outline a genome-wide map of genes informative for diagnosis that have been used for Leishmania genotyping. Furthermore, we propose a classification method based on the suitability of well-studied molecular markers for typing the 21 known Leishmania species pathogenic to humans. This can be applied to newly discovered species and to hybrid strains originating from inter-species crosses. Developing more effective and sensitive diagnostic methods and biomarkers is vital for enhancing Leishmania infection control programs. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  1. Antileishmanial activity and tubulin polymerization inhibition of podophyllotoxin derivatives on Leishmania infantum

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    José Miguel Escudero-Martínez

    2017-12-01

    Full Text Available Leishmania microtubules play an important role not only in cell division, but also in keeping the shape of the parasite and motility of its free-living stages. Microtubules result from the self-assembly of alpha and beta tubulins, two phylogenetically conserved and very abundant eukaryotic proteins in kinetoplastids. The colchicine binding domain has inspired the discovery and development of several drugs currently in clinical use against parasites. However, this domain is less conserved in kinetoplastids and may be selectively targeted by new compounds. This report shows the antileishmanial effect of several series of compounds (53, derived from podophyllotoxin (a natural cyclolignan isolated from rhizomes of Podophyllum spp. and podophyllic aldehyde, on a transgenic, fluorescence-emitting strain of Leishmania infantum. These compounds were tested on both promastigotes and amastigote-infected mouse splenocytes, and in mammalian – mouse non-infected splenocytes and liver HepG2 cells – in order to determine selective indexes of the drugs. Results obtained with podophyllotoxin derivatives showed that the hydroxyl group at position C-7α was a structural requisite to kill the parasites. On regards podophyllic aldehyde, derivatives with C9-aldehyde group integrated into a bicyclic heterostructure displayed more potent antileishmanial effects and were relatively safe for host cells. Docking studies of podophyllotoxin and podophyllic aldehyde derivatives showed that these compounds share a similar pattern of interaction at the colchicine site of Leishmania tubulin, thus pointing to a common mechanism of action. However, the results obtained suggested that despite tubulin is a remarkable target against leishmaniasis, there is a poor correlation between inhibition of tubulin polymerization and antileishmanial effect of many of the compounds tested, fact that points to alternative pathways to kill the parasites. Keywords: Leishmania, Tubulin, DNA

  2. Molecular and serological detection of tick-borne pathogens in dogs from an area endemic for Leishmania infantumin Mato Grosso do Sul, Brazil

    Directory of Open Access Journals (Sweden)

    Keyla Carstens Marques de Sousa

    Full Text Available Tick-borne pathogens affect a wide range of vertebrate hosts. To identify tick-borne pathogens among dogs from Campo Grande, MS, Brazil testing seropositive for Leishmania infantum (syn. L. chagasi, a serological and molecular study was conducted to detectEhrlichia canis, Anaplasma platys and Babesia vogeli in 60 serum and spleen samples. A confirmatory diagnosis ofL. infantum based on serological and molecular assays was also performed, as was sequence alignment and phylogenetic analysis to assess the identity of the parasite species infecting these animals. IgG antibodies toEhrlichia spp., B. vogeli and L. infantum were found, respectively, in 39 (65%, 49 (81.6% and 60 (100% of the sampled dogs. Twenty-seven (45%, fifty-four (90%, fifty-three (88.3%, two (3.3% and one (1.6% dog were positive, respectively, forE. canis, Leishmania spp., Leishmania donovani complex, Babesia sp. and Anaplasma sp. in PCR assays. After sequencing, the amplicons showed 99% of identity with E. canis, B. vogeli, A. platys andLeishmania chagasi isolates. The findings of this study indicate that L. infantum-seropositive dogs from Campo Grande are exposed to multiple tick-borne pathogens, which should therefore be included in the differential diagnosis of dogs with clinical suspicion of leishmaniasis.

  3. 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)

    Bonetti, Franco Claudio

    2006-01-01

    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 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)

  4. Diagnóstico ecológico para la conservación de Jatropha spp. (Euphorbiaceae y sus hábitats, en la Reserva Tehuacán-Cuicatlán, México

    Directory of Open Access Journals (Sweden)

    Nancy Hernández-Nicolás

    2015-03-01

    Full Text Available Jatropha spp., es un importante recurso fitogenético usado como alimento, medicina y biocombustible. En este estudio verificamos la identidad taxonómica de las especies de Jatropha para la Reserva de la Biosfera Tehuacán-Cuicatlán, y las unidades ambientales (UA que ocupan. Evaluamos el estado de conservación de sus hábitats y la vulnerabilidad de las poblaciones de Jatropha. Los sitios de muestreo seleccionados en la Reserva fueron 15. El trabajo taxonómico estuvo basado en especímenes, descripciones originales y material tipo de herbarios y aquellos en línea. Las UA fueron clasificadas mediante variables biofísicas y el software gvSIG. Los atributos ecológicos fueron determinados con análisis cuantitativo por el método de cuadrado centrado en un punto; el disturbio fue estimado a través de indicadores de sitio, y el estado de conservación de las poblaciones de Jatropha con el Método para la Evaluación de Riesgo de Extinción de Plantas en México (MER. Jatropha frecuentemente dominó la fisonomía de las comunidades vegetales. La distribución actual de las especies de Jatropha en la Reserva estuvo principalmente determinada por las variables altitud, temperatura y precipitación. Las especies confirmadas fueron: Jatropha ciliata Sessé ex Cerv., Jatropha neopauciflora Pax, Jatropha oaxacana J. Jiménez Ram. & R. Torres, Jatropha rufescens Brandegee, y Jatropha rzedowskii J. Jiménez Ram., las cuales estuvieron distribuidas en cuatro de las seis UA definidas. J. neopauciflora y J. rzedowskii son las especies más ampliamente distribuidas; estas últimas especies concurrieron en cuatro, J. oaxacana en dos, mientras que J. rufescens y J. ciliata en una UA, siendo ésta la especie más restringida. La riqueza de géneros en las comunidades asociadas varió entre 16 y 42. Los máximos y mínimos Índices de Valores de Importancia fueron observados en San Nicolás Tepoxtitlan para J. neopauciflora (53.75% y J. rzedowskii (1.50%. El

  5. First evidence of autochthonous cases of Leishmania (Leishmania) infantum in horse (Equus caballus) in the Americas and mixed infection of Leishmania infantum and Leishmania (Viannia) braziliensis.

    Science.gov (United States)

    Soares, Isabel R; Silva, Soraia O; Moreira, Filipe Moraghi; Prado, Luan Gavião; Fantini, Priscila; Maranhão, Renata de Pino Albuquerque; da Silva Filho, José Monteiro; Melo, Maria Norma; Palhares, Maristela S

    2013-11-08

    This study reports the first evidence of infection by Leishmania infantum in Equus caballus in Americas and the first mixed infection of L. infantum/Leishmania braziliensis on this mammalian species in the world. The diagnoses was based on presence of parasites in lesions and bone marrow aspirates, their identification by using specific primers for L. infantum and L. braziliensis complexes and also serological methods IFAT and ELISA. The analysis of the PCR products suggested mixed infection in three animals. Further studies involving equine leishmaniasis are carrying out in order to clarify the dynamic of Leishmania sp. in this mammalian specie and their role in the transmission of those parasites in urban endemic area of Belo Horizonte, Minas Gerais State, Brazil. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Mapa del proteoma de parásitos de Leishmania Viannia a partir de electroforesis de geles de poliacramida en dos dimensiones y otras técnicas asociadas

    OpenAIRE

    Rafael Góngora; Nathalie Acestor; Manfredo Quadroni; Nicolas Fasel; Nancy G. Saravia; John Walker

    2003-01-01

    En este estudio demostramos el potencial de la electroforesis en dos dimensiones (2DE) como herramienta para la caracterización del proteoma de Leishmania (expresión proteica complementaria del genoma). Los proteomas por 2DE en el rango neutro (pH 5-7) de extractos solubles de promastigotes de Leishmania (Viannia) guyanensis y Leishmania (Viannia) panamensis fueron reproducibles usando tampón de lisis de urea y nonidet P-40. Con la tinción de azul de Coomassie y nitrato de plata se detectaron...

  7. Avaliação de genótipos de Mentha arvensis, Mentha x piperita e Mentha spp. para a produção de mentol

    OpenAIRE

    Deschamps,Cícero; Monteiro,Rodrigo; Machado,Marília P; Scheer,Agnes de P; Cocco,Lílian; Yamamoto,Carlos

    2013-01-01

    O gênero Mentha possui 18 espécies e diversos quimiotipos com diferenças inter e intra-específicas. Apesar da grande variabilidade fitoquímica, o maior interesse econômico na produção de óleo essencial de menta deve-se à obtenção do constituinte mentol. Objetivou-se avaliar 19 genótipos de três espécies de Mentha para a produção de mentol. O experimento foi conduzido a campo com delineamento experimental em blocos casualizados, sendo avaliado o desenvolvimento vegetativo, a produtividade e a ...

  8. Nuclear DNA replication and repair in parasites of the genus Leishmania: Exploiting differences to develop innovative therapeutic approaches.

    Science.gov (United States)

    Uzcanga, Graciela; Lara, Eliana; Gutiérrez, Fernanda; Beaty, Doyle; Beske, Timo; Teran, Rommy; Navarro, Juan-Carlos; Pasero, Philippe; Benítez, Washington; Poveda, Ana

    2017-03-01

    Leishmaniasis is a common tropical disease that affects mainly poor people in underdeveloped and developing countries. This largely neglected infection is caused by Leishmania spp, a parasite from the Trypanosomatidae family. This parasitic disease has different clinical manifestations, ranging from localized cutaneous to more harmful visceral forms. The main limitations of the current treatments are their high cost, toxicity, lack of specificity, and long duration. Efforts to improve treatments are necessary to deal with this infectious disease. Many approved drugs to combat diseases as diverse as cancer, bacterial, or viral infections take advantage of specific features of the causing agent or of the disease. Recent evidence indicates that the specific characteristics of the Trypanosomatidae replication and repair machineries could be used as possible targets for the development of new treatments. Here, we review in detail the molecular mechanisms of DNA replication and repair regulation in trypanosomatids of the genus Leishmania and the drugs that could be useful against this disease.

  9. Innate Immunity to Leishmania Infection: Within Phagocytes

    Directory of Open Access Journals (Sweden)

    Marcela Freitas Lopes

    2014-01-01

    Full Text Available Infection by Leishmania takes place in the context of inflammation and tissue repair. Besides tissue resident macrophages, inflammatory macrophages and neutrophils are recruited to the infection site and serve both as host cells and as effectors against infection. Recent studies suggest additional important roles for monocytes and dendritic cells. This paper addresses recent experimental findings regarding the regulation of Leishmania major infection by these major phagocyte populations. In addition, the role of IL-4 on dendritic cells and monocytes is discussed.

  10. Xenodiagnostico con Lutzomyia youngi en casos venezolanos de leishmaniasis cutaned por Leishmania braziliensis Xenodiagnosis with Lutzomyia youngi in Venezuelan cases of cutaneous leishmaniais due to Leishmania braziliensis

    Directory of Open Access Journals (Sweden)

    Elina Rojas

    1989-03-01

    Full Text Available Xenodiagnósticos con Lutzomya yungi aplicados en los bordes de las úlceras de pacientes infectados con Leishmania braziliensis antes y después del tratamiento con 10 dosis de antimonial pentavalente y un aminoglicósido, evidencian la condición reservoria de leishmanias del enfermo, para flebótomos endofágicos y la utilidad de un tratamiento específico-temprano que no solamente conduce a la curación clínica, sino a la eliminación del riesgo de una eventual transmisión intradomiciliar por insectos que pican dentro del domicilio durante la noche.Eight patients infected with Leishmania braziliensis were used for xenodiagnosis with Lutzomtyia youngi, before and after specific antileishmanial treatment with "glucantime" and "gabbromycin". All of them infected sandflies fed on the borders of the skin lesions before the treatment, suggesting that infected persons might act as reservoirs of infection for an indoor-bitting sandfly species. The negative results obtained by xenodiagnosis carried out after specific treatment of the same individuals indicated cure of the patients, and a reduction of risk for further intradomiciliary transmission.

  11. AVALIAÇÃO DA TERAPIA FOTODINÂMICA COM AZUL DE METILENO EM Leishmania major e Leishmania braziliensis: ESTUDO in vitro

    Directory of Open Access Journals (Sweden)

    Danielle El Atra Coelho

    2017-03-01

    Full Text Available A Leishmaníase é uma doença crônica causada pelo protozoário do gênero Leishmania, cujo tratamento é agressivo. A Terapia Fotodinâmica (TFD é uma alternativa promissora que combina luz, fotossensibilizador (FS e oxigênio molecular, para causar a morte celular. O objetivo desse trabalho foi avaliar, in vitro, a ação da TFD com Azul de metileno (AM em promastigotas de Leishmania, por teste de MTT, curva de crescimento e morfologia do parasito. O teste de MTT demonstrou alteração de ambas as espécies após interação com o AM no escuro e após TFD. A análise das curvas demonstrou que a TFD influenciou o crescimento das espécies. A análise morfológica revelou que o AM no escuro não causou alterações expressivas como a TFD, sendo a cepa de L. braziliensis mais afetada que a cepa de L. major. Pode-se concluir que a TFD com AM foi promissora contra promastigotas de Leishmania, pois foi capaz de diminuir o crescimento e alterar a morfologia dos parasitos em cultura.

  12. Infection by Mycoplasma spp., feline immunodeficiency virus and feline leukemia virus in cats from an area endemic for visceral leishmaniasis.

    Science.gov (United States)

    Marcondes, Mary; Hirata, Karina Y; Vides, Juliana P; Sobrinho, Ludmila S V; Azevedo, Jaqueline S; Vieira, Thállitha S W J; Vieira, Rafael F C

    2018-03-20

    Visceral leishmaniasis (VL) has been increasingly recognized in cats living in areas endemic for the disease. Co-infection with Leishmania infantum and other infectious agents is well established in dogs. However, for cats, data on co-infections with L. infantum and other infectious agents are still sparse. The aim of this study was to identify the prevalence of vector-borne pathogens, Mycoplasma spp., feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) in cats from an area endemic for VL in southeastern Brazil. Of the 90 cats, eight (8.9%) were infected with Mycoplasma spp., five (5.5%) were FIV- positive and one (1.1%) was FeLV-positive. Co-infection with L. infantum and at least one other infectious agent was found in 9/50 (18.0%; CI: 8.6-31.4%) cats. In Group 1 (cats infected naturally by L. infantum), 4/50 (8.0%) cats were positive for FIV, 4/50 (8%) for Mycoplasma spp. and 1/50 (2.0%) was co-infected with FeLV and Mycoplasma spp. In Group 2 (cats non-infected with L. infantum), 2/40 (5.0%) cats were infected with Mycoplasma spp. and 1/40 (2.5%) was co-infected with FIV and Mycoplasma spp. All cats were negative for Ehrlichia spp., Babesia spp. and Anaplasma platys. A low prevalence of co-infection in Leishmania-infected and non-infected cats was found. Co-infections with Leishmania and vector-borne diseases in cats are not common in this area endemic for VL in Brazil.

  13. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Seleção e caracterização molecular de isolados de Bacillus thuringiensis para o controle de Spodoptera spp.

    Directory of Open Access Journals (Sweden)

    Kelly Christiane Constanski

    2015-08-01

    Full Text Available Resumo:O objetivo deste trabalho foi selecionar e caracterizar molecularmente isolados de Bacillus thuringiensistóxicos a Spodoptera eridaniaeS. frugiperda. Trinta e quatro isolados foram submetidos ao bioensaio, dos quais três foram selecionados e usados para a estimativa da CL50. Os isolados selecionados não diferiram da linhagem padrão HD-1. Na caracterização molecular, identificou-se a presença dos genes cry1 e cry2, nos isolados BR37 e BR94, e dos genes cry4A, cry4B, cry10, cry11 e cyt1 no isolado BR58, o que confirmou o perfil proteico obtido de 130, 70 e 65 kDa. Foram identificados cristais bipiramidais e esféricos. O isolado BR58, apesar de não conter os genes relacionados à toxicidade a Lepidoptera, causa mortalidade em ambas as espécies

  16. Effects of nitro-heterocyclic derivatives against Leishmania (Leishmania) infantum promastigotes and intracellular amastigotes.

    Science.gov (United States)

    Petri e Silva, Simone Carolina Soares; Palace-Berl, Fanny; Tavares, Leoberto Costa; Soares, Sandra Regina Castro; Lindoso, José Angelo Lauletta

    2016-04-01

    Leishmaniasis is an overlooked tropical disease affecting approximately 1 million people in several countries. Clinical manifestation depends on the interaction between Leishmania and the host's immune response. Currently available treatment options for leishmaniasis are limited and induce severe side effects. In this research, we tested nitro-heterocyclic compounds (BSF series) as a new alternative against Leishmania. Its activity was measured in Leishmania (Leishmania) infantum promastigotes and intracellular amastigotes using MTT colorimetric assay. Additionally, we assessed the phosphatidylserine exposure by promastigotes, measured by flow cytometry, as well as nitric oxide production, measured by Griess' method. The nitro-heterocyclic compounds (BSF series) showed activity against L. (L.) infantum promastigotes, inducting the phosphatidylserine exposition by promastigotes, decreasing intracellular amastigotes and increasing oxide nitric production. The selectivity index was more prominent to Leishmania than to macrophages. Compared to amphotericin b, our compounds presented higher IC50, however the selectivity index was more specific to parasite than to amphotericin b. In conclusion, these nitro-heterocyclic compounds showed to be promising as an anti-Leishmania drug, in in vitro studies. Copyright © 2016 Elsevier Inc. All rights reserved.

  17. Leaf Surface Scanning Electron Microscopy of 16 Mulberry Genotypes (Morus spp. with Respect to their Feeding Value in Silkworm (Bombyx mori L. Rearing Microscopía Electrónica de Barrido de la Superficie Foliar de 16 Genotipos de Morus spp. en Relación a su Valor Alimenticio para Crianza del Gusano de la Seda (Bombyx mori L.

    Directory of Open Access Journals (Sweden)

    B.K Singhal

    2010-06-01

    Full Text Available Mulberry (Morus spp. is the only silkworm (Bombyx mori L. food plant. In Indian sub tropics, S-146 is the only popular and ruling mulberry genotype for silkworm rearing. As a result, mulberry leaf availability is always the limiting factor, and therefore, sub tropics are contributing less than 1% of the country’s total silk production compared with more than 60% under tropical conditions. Besides climatic conditions, this is due to a very limited number of mulberry genotypes available in this region for silkworm rearing. However, in the mean time, 15 mulberry genotypes viz. ‘Tr-10’,‘Chinese White’,‘K-2’,‘Sujanpur Local’,‘BC2-59’,‘S-1635’,‘C-1730’,‘Mandalaya’,‘S-30’‘(Vishala,‘RFS-175’,‘Anantha’,‘C-2016’,‘C-2017’,‘S-41’ and‘V-1’ were also introduced in the sub tropics, but remained unexplored. In sericulture, leaf surface is also an important parameter for, both, the silkworm’s acceptability of not having any feeding impediment and the mulberry improvement programs. The objective of this study was to explore the possibilities of using these 16 mulberry genotypes for their leaf surface characteristics by scanning electron microscopy and using them for sericulture. Based on leaf yield, stomatal size, stomatal number per unit of area and trichomes and idioblasts length, these genotypes were grouped into different categories. The mulberry genotype ‘Mandalaya’, in addition to the ruling genotype ‘S-146’ excelled because of their higher leaf yield and desired leaf surface characteristics. Furthermore, the genotypes ‘K-2’, ‘S-41’ and ‘Sujanpur Local’ are also suggested to develop high yield mulberry genotypes in the Indian sub tropics.La morera (Morus spp. es la única planta de alimento para el gusano de la seda (Bombyx mori L.. En los sub-trópicos de la India, ‘S-146’ es el único genotipo popular y predominante de morera para criarlo. Como resultado, la

  18. 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)

  19. 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)

  20. First Cases of Cutaneous Leishmaniasis Caused by Leishmania (Viannia) naiffi Infection in Surinam

    NARCIS (Netherlands)

    van Thiel, Pieter-Paul A. M.; van Gool, Tom; Kager, Piet A.; Bart, Aldert

    2010-01-01

    Cutaneous leishmaniasis in Surinam is generally caused by infection by Leishmania guyanensis. We report three cases of infection with Leishmania (Viannia) naiffi, a Leishmania species not described from Surinam before. Treatment with pentamidine proved to be effective

  1. Involvement of Leishmania donovani major surface glycoprotein ...

    Indian Academy of Sciences (India)

    The major surface glycoprotein gp63 of the kinetoplastid protozoal parasite Leishmania is implicated as a ligand mediating uptake of the parasite into, and survival within, the host macrophage. By expressing gp63 antisense RNA from an episomal vector in L. donovani promastigotes, gp63-deficient transfectants were ...

  2. Evaluación de los cebadores TS3 e ITS4 para la detección de infecciones por Candida spp. en muestras de flujo vaginal y humor acuoso Evaluation of ITS3 and ITS4 primers for detection of Candida spp. Candida spp. infections from vaginal samples and aqueous humor

    Directory of Open Access Journals (Sweden)

    Juan Felipe Ramirez

    2008-06-01

    Full Text Available Antecedentes. Los métodos convencionales para a identificación de levaduras del género Candida requieren mucho tiempo, necesitan cantidad importante de la muestra y, en muchos casos, es necesario realizar cultivo. Objetivos. Evaluar un par de cebadores para la identificación de levaduras del género Candida y demostrar su potencial aplicación para el diagnóstico de infecciones intraoculares. Materiales y métodos. Se utilizaron 17 cepas de referencia, 29 aislamientos clínicos de flujo vaginal, 7 muestras de humor acuoso y una de humor vítreo. Evaluamos tres métodos para el rompimiento de la pared celular: congelación descongelación, sonicación, y la enzima liticasa. Para la purificación del ácido nucleico, se utilizó en los tres casos el estuche comercial Wizard Genomics; en la reacción de PCR, el estuche comercial Go Taq Green Master Mix y los iniciadores ITS3 e ITS4 a una concentración de 0,5 µM. Resultados. De los tres métodos el que mejor resultados ofreció fue el uso de enzima liticasa más el estuche comercial. Con los iniciadores ITS3 e ITS4 fue posible identificar las levaduras únicamente a nivel de género y no se presentó reacción cruzada con otros microorganismos comúnmente encontrados en diferentes muestras de tejido humano. La sensibilidad fue de 100 fg. Se lograron identificar por PCR todos los aislamientos a partir de flujo vaginal y de una muestra de humor acuso. Para las demás muestras de humor acuoso el diagnóstico fue para otros agentes causales, entre ellos, Toxoplasma sp. Conclusión. Consideramos que ésta es una metodología adecuada, la cual permite identificar levaduras del género Candida con gran sensibilidad y reproducibilidad.Background. Conventional methods for Candida identification are time consuming, require high amounts of the sample and in several cases the culture is mandatory. Aims. To evaluate two primers for the identification of Candida and to demonstrate their application for

  3. Commensal Staphylococcus spp., Acinetobacter spp. and ...

    African Journals Online (AJOL)

    ANTHONY

    2012-07-31

    Jul 31, 2012 ... Intermittent assessment of resistance genes in the ecosystem should be ..... among resistant Acinetobacter spp. isolated from integrated fish .... independent studies on the emerging phylogenetic view of bacterial .... Functional.

  4. 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"

    Directory of Open Access Journals (Sweden)

    Raquel S. Pacheco

    1987-12-01

    Full Text Available 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 subespécie de Leishmania por exometabólitos de outra subespécie, um fenômeno ainda não notificado, é sugerido em nossas recentes observações em experimentos de clonagem celular com Leishmania mexicana mexicana e Leishmania mexicana amazonensis. Os clones foram identificados usando a técnica de análise de esquizodemas. O fenômeno observado é claramente relevante em estudos de isolamento parasitário, metabolismo, imunidade cruzada e quimioterapia.

  5. Serosurvey of Rickettsia spp. in dogs and humans from an endemic area for Brazilian spotted fever in the State of São Paulo, Brazil Sorologia para Rickettsia spp. em cães e humanos de uma área endêmica para febre maculosa brasileira no Estado de São Paulo, Brasil

    Directory of Open Access Journals (Sweden)

    Adriano Pinter

    2008-02-01

    Full Text Available The present study provides a rickettsial serosurvey in 25 dogs and 35 humans in an endemic area for Brazilian spotted fever in the State of São Paulo, where the tick Amblyomma aureolatum is the main vector. Testing canine and human sera by indirect immunofluorescence against four Rickettsia antigens (R. rickettsii, R. parkeri, R. felis and R. bellii showed that 16 (64% of canine sera and 1 (2.8% of human sera reacted to at least one of these rickettsial antigens with titers ³ 64. Seven canine sera and the single reactive human serum showed titers to R. rickettsii at least four times those of any of the other three antigens. The antibody titers in these 7 animals and 1 human were attributed to stimulation by R. rickettsii infection. No positive canine or human serum was attributed to stimulation by R. parkeri, R. felis, or R. bellii. Our serological results showed that dogs are important sentinels for the presence of R. rickettsii in areas where the tick A. aureolatum is the main vector of Brazilian spotted fever.Este estudo avaliou a ocorrência de anticorpos anti-Rickettsia em 25 cães e 35 humanos, em uma área endêmica para a febre maculosa brasileira no Estado de São Paulo, onde o principal vetor é o carrapato Amblyomma aureolatum. Soros dos cães e humanos foram testados pela técnica de imunofluorescência indireta contra quatro antígenos de riquétsias (R. rickettsii, R. parkeri, R. felis, R. bellii, mostrando que soros de 16 (64% cães e 1 (2,8% humano reagiram com títulos ³ 64 para pelo menos um dos antígenos de riquétsias. Sete soros caninos e o único soro humano reativo demonstraram títulos para R. rickettsii no mínimo quatro vezes maior do que aqueles para os outros antígenos de riquétsias. Os títulos de anticorpos nesses cães e um humano foram considerados homólogos a R. rickettsii, enquanto que nenhum soro de cão ou humano foi considerado reativamente homólogo para R. parkeri, R. felis ou R. bellii. Os

  6. Seroprevalence rates of antibodies against Leishmania infantum and other protozoan and rickettsial parasites in dogs Soroprevalência de anticorpos contra Leishmania infantum e outras espécies de protozoários e rickettsia em cães

    Directory of Open Access Journals (Sweden)

    Silvana de Cássia Paulan

    2013-03-01

    Full Text Available Canine visceral leishmaniasis (CVL is caused by the protozoan Leishmania infantum, which infects dogs and humans in many regions of Brazil. The present study involved an indirect fluorescent antibody test (IFAT to analyze L. infantum, Ehrlichia spp., Babesia canis, Toxoplasma gondii and Neospora caninum infection rates in serum samples from 93 dogs in a rural settlement in Ilha Solteira, SP, Brazil. The seroprevalence rates of anti-L. infantum, anti-Ehrlichia, anti-B. canis, anti-T. gondii and anti-N. caninum antibodies were 37.6%, 75.3%, 72%, 47.3% and 6.4%, respectively. In addition to IFAT, direct microscopic examination of popliteal lymph node aspirates revealed 26.9% of CVL positive dogs. Serological tests revealed that 17.2% of the dogs were seropositive for a single parasite, 29% for two parasites, 33% for three, 16.1% for four, and 1.1% for five parasites, while 3.2% were seronegative for five parasites. The presence of antibodies against these parasites in serum samples from dogs confirmed their exposure to these parasites in this rural area. Because of the potential zoonotic risk of these diseases, mainly leishmaniasis, ehrlichiosis and toxoplasmosis, special attention should focus on programs for the improvement of diagnostic assays and control measures against these parasites.Leishmaniose Visceral Canina (LVC é causada pelo protozoário Leishmania infantum, podendo infectar cães e humanos em várias regiões do Brasil. O presente estudo teve por objetivo realizar a reação de imunofluorescência indireta (RIFI para analisar os índices de infecção parasitária para L. infantum, Ehrlichia spp., Babesia canis, Toxoplasma gondii e Neospora caninum, em 93 amostras de soro de cães de um assentamento rural no município de Ilha Solteira, SP, Brasil. A taxa de soroprevalência de cães com anticorpos anti-L. infantum, anti-Ehrlichia, anti-B. canis, anti-T. gondii e anti-N. caninum foi de 37,6%, 75,3%, 72%, 47,3% e 6

  7. 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.

  8. Carfentrazone-ethyl aplicado em pós-emergência para o controle de Ipomea spp. e Commelina benghalensis na cultura da cana-de-açúcar Carfentrazone-ethyl applied in post-emergence to control Ipomoea spp. and Commelina benghalensis in sugarcane crop

    Directory of Open Access Journals (Sweden)

    P.J. Christoffoleti

    2006-01-01

    Full Text Available Este trabalho teve como objetivos determinar a eficácia e seletividade do carfentrazone-ethyl no controle de plantas daninhas na cultura da cana-de-açúcar e avaliar curvas de dose-resposta desse herbicida em Ipomoea spp. e Commelina benghalensis. Desenvolveu-se um ensaio de campo, em pós-emergência das plantas daninhas e da cultura (cv. RB72-454. O delineamento experimental utilizado foi do tipo blocos ao acaso, com os seguintes tratamentos: carfentrazone a 5, 10, 20, 30 e 50 g ha-1 e metribuzin a 2.400 g ha-1. As plantas daninhas que infestavam a área eram Ipomoea nil, I. grandifolia, I. quamoclit, Momordica charantia e C. benghalensis. As avaliações de controle visual e fitotoxicidade foram realizadas aos 15, 30 e 45 dias após a aplicação dos herbicidas. Para elaboração das curvas de dose-resposta, um experimento foi instalado em casa de vegetação, com as plantas daninhas I. nil, I. hederifolia, I. grandifolia, I. quamoclit e C. benghalensis. As doses do herbicida carfentrazone foram: 10D, 4D, 2D, D, 0,5D, 0,25D, 0,125D, 0,1D e 0,01D, em que D = 20 g ha¹. Os dados foram analisados a partir de curvas de dose-resposta de controle visual e massa fresca. No experimento de campo, observou-se excelente controle das plantas daninhas, sendo a dose de 50 g ha-1 de carfentrazone a mais eficiente. Nenhum tratamento causou danos à cana-de-açúcar. O experimento de casa de vegetação permitiu concluir que a ordem decrescente de sensibilidade das espécies de corda-de-viola ao carfentrazone-ethyl é: I. hederifolia > I. quamoclit > I. nil > I. grandifolia; e que C. benghalensis é controlada pelo carfentrazone a partir da dose de 5 g ha-1.This work aimed to evaluate the efficacy and selectivity of the herbicide carfentrazone-ethyl applied in post-emergence to control weeds in sugarcane crop and to evaluate dose-response curves of this herbicide in four species of Ipomoea spp. and Commelina benghalensis. Field experiment was carried out in

  9. Adenine phosphoribosyltransferase-deficient Leishmania donovani

    International Nuclear Information System (INIS)

    Kaur, K.; Iovannisci, D.M.; Ullman, B.

    1986-01-01

    To elucidate the relative roles of two routes for adenine salvage, the authors use biochemical genetic approaches to isolate clonal strains of Leishmania donovani promasatigotes genetically deficient in APRTase activity. The studies suggest that the metabolic rate of adenine in these organisms is initiated by deamination. The radiolabel incorporation experiments and biochemical experiments are described in which the rate of uptake of radiolabelled purine nucleobases (C 14) was determined. Results are presented

  10. Aislamiento de tres cepas de leishmania

    Directory of Open Access Journals (Sweden)

    Florentino Rey

    1942-08-01

    Full Text Available Con el ánimo de estudiar el problema parasitológico de la Leishmaniasis cutánea en Colombia, emprendimos algunas experiencias al respecto. Este es un estudio de aislamiento de tres cepas de Leishmania obtenidas de tres enfermos de distintas regiones del país, Investigadores colombianos que se han ocupado con anterioridad de este problema, obtuvieron siempre resultados negativos. Comunicaciones posteriores informarán sobre estudios experimentales de las cepas mencionadas.

  11. Leishmania infantum and Leishmania braziliensis: differences and similarities to evade the innate immune system

    Directory of Open Access Journals (Sweden)

    Sarah Athayde Couto Falcão

    2016-08-01

    Full Text Available Visceral Leishmaniasis is a severe form of the disease, caused by Leishmania infantum in the New World. Patients present an anergic immune response that favors parasite establishment and spreading through tissues like bone marrow and liver. On the other hand, Leishmania braziliensis causes localized cutaneous lesions, which can be self healing in some individuals. Interactions between host and parasite are essential to understand disease pathogenesis and progression. In this context, dendritic cells (DCs act as essential bridges that connect innate and adaptive immune responses. In this way, the aim of this study was to compare the effects of these two Leishmania species, in some aspects of human dendritic cells biology to better understanding of the evasion mechanisms of Leishmania from host innate immune response. To do so, DCs were obtained from monocytes from whole peripheral blood’s healthy volunteers donors and infected with L. infantum or L. braziliensis for 24 hours. We observed similar rates of infection (around 40% as well as parasite burden for both Leishmania species. Concerning surface molecules, we observed that both parasites induced CD86 expression when DCs were infected for 24h. On the other hand, we detected a lower surface expression of CD209 in the presence of both L. braziliensis and L. infantum, but only the last one promoted the survival of dendritic cells after 24 hours. Therefore, DCs infected by both Leishmania species showed a higher expression of CD86 and a decrease of CD209 expression, suggesting that both enter DCs through CD209 molecule. However, only L. infantum had the ability to inhibit DC apoptotic death, as an evasion mechanism that enables its spreading to organs like bone marrow and liver. Lastly, L. braziliensis was more silent parasite, once it did not inhibit DC apoptosis in our in vitro model.

  12. Leishmania infantum Asparagine Synthetase A Is Dispensable for Parasites Survival and Infectivity.

    Science.gov (United States)

    Faria, Joana; Loureiro, Inês; Santarém, Nuno; Macedo-Ribeiro, Sandra; Tavares, Joana; Cordeiro-da-Silva, Anabela

    2016-01-01

    A growing interest in asparagine (Asn) metabolism has currently been observed in cancer and infection fields. Asparagine synthetase (AS) is responsible for the conversion of aspartate into Asn in an ATP-dependent manner, using ammonia or glutamine as a nitrogen source. There are two structurally distinct AS: the strictly ammonia dependent, type A, and the type B, which preferably uses glutamine. Absent in humans and present in trypanosomatids, AS-A was worthy of exploring as a potential drug target candidate. Appealingly, it was reported that AS-A was essential in Leishmania donovani, making it a promising drug target. In the work herein we demonstrate that Leishmania infantum AS-A, similarly to Trypanosoma spp. and L. donovani, is able to use both ammonia and glutamine as nitrogen donors. Moreover, we have successfully generated LiASA null mutants by targeted gene replacement in L. infantum, and these parasites do not display any significant growth or infectivity defect. Indeed, a severe impairment of in vitro growth was only observed when null mutants were cultured in asparagine limiting conditions. Altogether our results demonstrate that despite being important under asparagine limitation, LiAS-A is not essential for parasite survival, growth or infectivity in normal in vitro and in vivo conditions. Therefore we exclude AS-A as a suitable drug target against L. infantum parasites.

  13. Leishmania infantum Asparagine Synthetase A Is Dispensable for Parasites Survival and Infectivity.

    Directory of Open Access Journals (Sweden)

    Joana Faria

    2016-01-01

    Full Text Available A growing interest in asparagine (Asn metabolism has currently been observed in cancer and infection fields. Asparagine synthetase (AS is responsible for the conversion of aspartate into Asn in an ATP-dependent manner, using ammonia or glutamine as a nitrogen source. There are two structurally distinct AS: the strictly ammonia dependent, type A, and the type B, which preferably uses glutamine. Absent in humans and present in trypanosomatids, AS-A was worthy of exploring as a potential drug target candidate. Appealingly, it was reported that AS-A was essential in Leishmania donovani, making it a promising drug target. In the work herein we demonstrate that Leishmania infantum AS-A, similarly to Trypanosoma spp. and L. donovani, is able to use both ammonia and glutamine as nitrogen donors. Moreover, we have successfully generated LiASA null mutants by targeted gene replacement in L. infantum, and these parasites do not display any significant growth or infectivity defect. Indeed, a severe impairment of in vitro growth was only observed when null mutants were cultured in asparagine limiting conditions. Altogether our results demonstrate that despite being important under asparagine limitation, LiAS-A is not essential for parasite survival, growth or infectivity in normal in vitro and in vivo conditions. Therefore we exclude AS-A as a suitable drug target against L. infantum parasites.

  14. Little evidence of seasonal variation of natural infection by Leishmania infantum in dogs in Spain.

    Science.gov (United States)

    Fernández-Bellon, H; Solano-Gallego, L; Rodríguez-Cortés, A; Ferrer, L; Gallego, M; Alberola, J; Ramis, A

    2008-08-01

    Leishmania infantum, the etiological agent of canine leishmaniosis in the Mediterranean region, is vectored by Phlebotomus spp sandflies, which are active during the warmer months of the year. In order to determine whether seasonality in transmission induces seasonal changes in the prevalence of infection by L. infantum and of parasite-specific immune response, two groups of dogs, one in February (n=37) and another in October (n=42), were studied. Clinical signs compatible with leishmaniosis, as well as presence of microscopic skin lesions in the muzzle were recorded for all dogs. Assays were also performed for detection of L. infantum parasites in muzzle skin samples (PCR, immunohistochemistry and culture), specific serum antibodies (ELISA), and specific lymphocyte proliferation and interferon-gamma production. Although prevalence of non-specific clinical signs increased significantly after the sandfly season, this was not the case for Leishmania-specific markers: positivity by PCR (24% vs. 21%) or immunohistochemistry (3% vs. 2%) of muzzle skin samples, as well as lymphocyte proliferation (59% vs. 50%) or interferon-gamma production (21% vs. 27%) were similar in February and in October. Only prevalence of positive specific antibody titers increased noticeably in October (8% vs. 20%), although this was not statistically significant. Overall, the sandfly season did not have a marked impact on the prevalence L. infantum infection or parasite-specific immune responses analyzed in this study.

  15. Eupomatenoid-5 Isolated from Leaves of Piper regnellii Induces Apoptosis in Leishmania amazonensis

    Directory of Open Access Journals (Sweden)

    Francielle Pelegrin Garcia

    2013-01-01

    Full Text Available Leishmania spp. are protozoa responsible for leishmaniasis, a neglected disease that kills up to 50,000 people every year. Current therapies mainly rely on antimonial drugs that are inadequate because of their poor efficacy and safety and increased drug resistance. An urgent need exists to find new and more affordable drugs. Our previous study demonstrated the antileishmanial activity of eupomatenoid-5, a neolignan obtained from leaves of Piper regnellii var. pallescens. The aim of the present study was to clarify the mode of action of eupomatenoid-5 against L. amazonensis. We used biochemical and morphological techniques and demonstrated that eupomatenoid-5 induced cell death in L. amazonensis promastigotes, sharing some phenotypic features observed in metazoan apoptosis, including increased reactive oxygen species production, hypopolarization of mitochondrial potential, phosphatidylserine exposure, decreased cell volume, and G0/G1 phase cell cycle arrest.

  16. Screening For Inhibitors Of Essential Leishmania Glucose Transporters

    Science.gov (United States)

    2011-07-01

    parasite life cycle and, unlike he amastigote form that lives inside mammalian macrophages, s viable provided that an alternative energy source such as pro...glucose transporters havebeenvalidated asnewdrug targets for proto- zoan parasites including Plasmodium falciparum, Leishmania mexicana and Trypanosoma...such as Leishmania species, Trypanosoma rucei, and Plasmodium falciparum, the causative agents of leish- aniasis, African sleeping sickness, and malaria

  17. Erythema exsudativum multiforme after a Leishmania skin test

    NARCIS (Netherlands)

    Wind, Bas S.; Guimarães, Luiz H.; Machado, Paulo R. L.

    2014-01-01

    A 45-year-old otherwise healthy male from an endemic region for Leishmania braziliensis infection in Bahia, Brazil, presented with three erosive hemorrhagic infiltrated plaques on the left shin accompanied with lymphadenopathy in the groin since one month. A Leishmania skin test performed on the

  18. Detecção de DNA de Leishmania braziliensis em pacientes de leishmaniose tegumentar americana Detección de DNA de Leishmania braziliensis en pacientes de leishmaniose tegumentaria americana Detection of Leishmania braziliensis DNA in American tegumentary leishmaniasis patients

    Directory of Open Access Journals (Sweden)

    Leila Martins

    2010-06-01

    Full Text Available Foi realizado diagnóstico para leishmaniose tegumentar americana a partir de sangue de pacientes residentes em dois municípios endêmicos do estado de Pernambuco. O DNA de 119 amostras de sangue foi extraído e submetido a reação em cadeia da polimerase. Utilizaram-se primers do minicírculo do DNA do cinetoplasto (kDNA de Leishmania braziliensis, circulante em Pernambuco, cuja seqüência-alvo gera um fragmento de 750 pares de bases. No total 58 (48,7% indivíduos apresentaram amplificação positiva e 61 (51,3% negativa. Das amostras positivas para a PCR, 37 (≅ 64% pertenciam a indivíduos tratados e sem lesão. Conclui-se que a técnica de PCR é eficaz para identificar o DNA de leishmânia em material de biópsias e em sangue venoso.Fue realizado diagnóstico para leishmaniosis tegumentaria americana a partir de sangre de pacientes residentes en dos municipios endémicos del estado de Pernambuco (Noreste de Brasil. El DNA de 119 muestras de sangre fue extraído y sometido a la reacción en cadena de la polimerasa. Se utilizaron primers del minicírculo del DNA del cinetoplasto (kDNA de Leishmania braziliensis, circulante en Pernambuco, cuya secuencia blanco genera un fragmento de 750 pares de bases. En total 58 (48,7% individuos presentaron amplificación positiva y 61 (51,3% negativa. De las muestras positivas para la PCR, 37 (≅64% pertenecían a individuos tratados y sin lesión. Se concluyó que la técnica de la PCR es eficaz para identificar el DNA de Leishmania en material de biopsias y en sangre venosa.Diagnostic tests for American tegumentary leishmaniasis were performed on blood samples of patients living in two endemic municipalities in the state of Pernambuco, Northeastern Brazil. DNA was extracted from 119 samples and used as template for polymerase chain reaction (PCR analysis. The tests used primers specific for the kinetoplast mini-circle DNA (kDNA of Leishmania braziliensis, a species circulating in Pernambuco, which

  19. UTILIZAÇÃO DE RESÍDUOS DA MADEIRA DE Pinus spp. COMO SUBSTITUIÇÃO AO AGREGADO MIÚDO NA PRODUÇÃO DE BLOCOS DE CONCRETO PARA ALVENARIA ESTRUTURAL

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    Adauto José Miranda de Lima

    2014-01-01

    Full Text Available This research was developed with the objective of evaluating the potential use of residues of the production of Pinus spp. sawnwood, as substitution to the fine aggregate of the concrete used for the production of blocks for structural masonry. The evaluation of the physical characteristics (density, porosity and water absorption and mechanics (compression and tension strength of the concrete was evaluated in agreement with the ABNT normalization. Substitutions of the fine aggregate were tested by percentages of 0, 20, 40, 60, 80 and 100% of residue of Pinus spp. in natura, Pinus spp. in natura and with addition of 4,5% CaCl 2 .2H 2 O and residue of Pinus spp. pre-treated with extraction in cold water for 48 hours (AF-48H. The compositions were initially used for the production, in laboratory, of cylindrical specimens of 50 mm diameter, by the use of a molding system for vibrate-compression and later, for producing blocks, with optimized compositions, in vibrate-press commercial machine. Satisfactory results were obtained with substitutions of up to 50% of the mineral fine aggregate for the residue of Pinus spp.

  20. A diagnostic assay based on variable intergenic region distinguishes between Leishmania donovani and Leishmania infantum

    Czech Academy of Sciences Publication Activity Database

    Chocholová, Eva; Jirků, Milan; Lukeš, Julius

    2008-01-01

    Roč. 55, č. 1 (2008), s. 75-78 ISSN 0015-5683 R&D Projects: GA MŠk LC07032; GA MŠk 2B06129 Institutional research plan: CEZ:AV0Z60220518 Keywords : Leishmania * assay * diagnosis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.307, year: 2008

  1. 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.

  2. Molecular detection of Leishmania infection due to Leishmania major and Leishmania turanica in the vectors and reservoir host in Iran.

    Science.gov (United States)

    Rassi, Yavar; Oshaghi, Mohammad Ali; Azani, Sadegh Mohammadi; Abaie, Mohammad Reza; Rafizadeh, Sina; Mohebai, Mehdi; Mohtarami, Fatemeh; Zeinali, Mohammad kazem

    2011-02-01

    An epidemiological study was carried out on the vectors and reservoirs of cutaneous leishmaniasis in rural areas of Damghan district, Semnan province, central Iran, during 2008-2009. Totally, 6110 sand flies were collected using sticky papers and were subjected to molecular methods for detection of Leishmania parasite. Phlebotomus papatasi Scopoli was the common species in outdoor and indoor resting places. Polymerase chain reaction technique showed that 24 out of 218 P. papatasi (11%) and 4 out of 62 Phlebotomus caucasicus Marzinovskyi (6.5%) were positive for parasites Leishmania major Yakimoff and Schokhor. Twenty-one rodent reservoir hosts captured using Sherman traps were identified as Rhombomys opimus Lichtenstein (95%) and Meriones libycus Lichtenstein (5%). Microscopic investigation on blood smear of the animals for amastigote parasites revealed 8 (40%) rodents infected with R. opimus. L. major infection in these animals was then confirmed by polymerase chain reaction against internal transcribed spacer ribosomal DNA (rDNA) loci of the parasite followed by restriction fragment length polymorphism. Further, sequence analysis of 297 bp of ITS1-rDNA loci revealed the presence of L. major and Leishmania turanica in P. papatasi, and L. major in R. opimus. This is the first molecular report of L. major infection in both vectors (P. papatasi and P. caucasicus) and reservoir host (R. opimus) in this region. The results indicated that P. papatas was the primary vector of the disease and circulating the parasite between human and reservoirs, and P. caucasicus could be considered as a secondary vector. Further, our study showed that R. opimus is the most important host reservoir for maintenance of the parasite source in the area.

  3. 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

    Directory of Open Access Journals (Sweden)

    Yeda L. Nogueira

    2006-12-01

    élula 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.

  4. Detection and characterization of Leishmania (Leishmania and Leishmania (Viannia by SYBR green-based real-time PCR and high resolution melt analysis targeting kinetoplast minicircle DNA.

    Directory of Open Access Journals (Sweden)

    Marcello Ceccarelli

    Full Text Available Leishmaniasis is a neglected disease with a broad clinical spectrum which includes asymptomatic infection. A thorough diagnosis, able to distinguish and quantify Leishmania parasites in a clinical sample, constitutes a key step in choosing an appropriate therapy, making an accurate prognosis and performing epidemiological studies. Several molecular techniques have been shown to be effective in the diagnosis of leishmaniasis. In particular, a number of PCR methods have been developed on various target DNA sequences including kinetoplast minicircle constant regions. The first aim of this study was to develop a SYBR green-based qPCR assay for Leishmania (Leishmania infantum detection and quantification, using kinetoplast minicircle constant region as target. To this end, two assays were compared: the first used previously published primer pairs (qPCR1, whereas the second used a nested primer pairs generating a shorter PCR product (qPCR2. The second aim of this study was to evaluate the possibility to discriminate among subgenera Leishmania (Leishmania and Leishmania (Viannia using the qPCR2 assay followed by melting or High Resolution Melt (HRM analysis. Both assays used in this study showed good sensitivity and specificity, and a good correlation with standard IFAT methods in 62 canine clinical samples. However, the qPCR2 assay allowed to discriminate between Leishmania (Leishmania and Leishmania (Viannia subgenera through melting or HRM analysis. In addition to developing assays, we investigated the number and genetic variability of kinetoplast minicircles in the Leishmania (L. infantum WHO international reference strain (MHOM/TN/80/IPT1, highlighting the presence of minicircle subclasses and sequence heterogeneity. Specifically, the kinetoplast minicircle number per cell was estimated to be 26,566±1,192, while the subclass of minicircles amplifiable by qPCR2 was estimated to be 1,263±115. This heterogeneity, also observed in canine clinical

  5. due to Klebsiella spp.

    African Journals Online (AJOL)

    Primary acute conununity-acquired pneumonia was unconunon. ... organisms such as Klebsiella spp. are frequent causes of nosocomial infections."o The larrer ... Accepted 11 Sep 1992. recognised cause of community-acquired Gram-nega-.

  6. 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.

  7. Arginine and Polyamines Fate in Leishmania Infection

    Science.gov (United States)

    Muxel, Sandra M.; Aoki, Juliana I.; Fernandes, Juliane C. R.; Laranjeira-Silva, Maria F.; Zampieri, Ricardo A.; Acuña, Stephanie M.; Müller, Karl E.; Vanderlinde, Rubia H.; Floeter-Winter, Lucile M.

    2018-01-01

    Leishmania is a protozoan parasite that alternates its life cycle between the sand fly and the mammalian host macrophages, involving several environmental changes. The parasite responds to these changes by promoting a rapid metabolic adaptation through cellular signaling modifications that lead to transcriptional and post-transcriptional gene expression regulation and morphological modifications. Molecular approaches such as gene expression regulation, next-generation sequencing (NGS), microRNA (miRNA) expression profiling, in cell Western blot analyses and enzymatic activity profiling, have been used to characterize the infection of murine BALB/c and C57BL/6 macrophages, as well as the human monocytic cell-lineage THP-1, with Leishmania amazonensis wild type (La-WT) or arginase knockout (La-arg-). These models are being used to elucidate physiological roles of arginine and polyamines pathways and the importance of arginase for the establishment of the infection. In this review, we will describe the main aspects of Leishmania-host interaction, focusing on the arginine and polyamines pathways and pointing to possible targets to be used for prognosis and/or in the control of the infection. The parasite enzymes, arginase and nitric oxide synthase-like, have essential roles in the parasite survival and in the maintenance of infection. On the other hand, in mammalian macrophages, defense mechanisms are activated inducing alterations in the mRNA, miRNA and enzymatic profiles that lead to the control of infection. Furthermore, the genetic background of both parasite and host are also important to define the fate of infection. PMID:29379478

  8. 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.

    Directory of Open Access Journals (Sweden)

    Jarina Pena DaMata

    Full Text Available 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.

  9. Native rodent species are unlikely sources of infection for Leishmania (Viannia braziliensis along the Transoceanic Highway in Madre de Dios, Peru.

    Directory of Open Access Journals (Sweden)

    Lisa A Shender

    Full Text Available An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In Peru's Amazon region, the department of Madre de Dios (MDD rises above the rest of the country in terms of the annual incidence rates of human leishmaniasis. Leishmania (Viannia braziliensis is the species most frequently responsible for the form of disease that results in tissue destruction of the nose and mouth. However, essentially nothing is known regarding the reservoirs of this vector-borne, zoonotic parasite in MDD. Wild rodents have been suspected, or proven, to be reservoirs of several Leishmania spp. in various ecosystems and countries. Additionally, people who live or work in forested terrain, especially those who are not regionally local and whose immune systems are thus naïve to the parasite, are at most risk for contracting L. (V. braziliensis. Hence, the objective of this study was to collect tissues from wild rodents captured at several study sites along the Amazonian segment of the newly constructed Transoceanic Highway and to use molecular laboratory techniques to analyze samples for the presence of Leishmania parasites. Liver tissues were tested via polymerase chain reaction from a total of 217 rodents; bone marrow and skin biopsies (ear and tail were also tested from a subset of these same animals. The most numerous rodent species captured and tested were Oligoryzomys microtis (40.7%, Hylaeamys perenensis (15.7%, and Proechimys spp. (12%. All samples were negative for Leishmania, implying that although incidental infections may occur, these abundant rodent species are unlikely to serve as primary reservoirs of L. (V. braziliensis along the Transoceanic Highway in MDD. Therefore, although these rodent species may persist and even thrive in moderately altered landscapes, we did not find any evidence to suggest they pose a risk for L. (V. braziliensis transmission to human inhabitants in this highly prevalent region.

  10. Native rodent species are unlikely sources of infection for Leishmania (Viannia) braziliensis along the Transoceanic Highway in Madre de Dios, Peru.

    Science.gov (United States)

    Shender, Lisa A; De Los Santos, Maxy; Montgomery, Joel M; Conrad, Patricia A; Ghersi, Bruno M; Razuri, Hugo; Lescano, Andres G; Mazet, Jonna A K

    2014-01-01

    An estimated 2.3 million disability-adjusted life years are lost globally from leishmaniasis. In Peru's Amazon region, the department of Madre de Dios (MDD) rises above the rest of the country in terms of the annual incidence rates of human leishmaniasis. Leishmania (Viannia) braziliensis is the species most frequently responsible for the form of disease that results in tissue destruction of the nose and mouth. However, essentially nothing is known regarding the reservoirs of this vector-borne, zoonotic parasite in MDD. Wild rodents have been suspected, or proven, to be reservoirs of several Leishmania spp. in various ecosystems and countries. Additionally, people who live or work in forested terrain, especially those who are not regionally local and whose immune systems are thus naïve to the parasite, are at most risk for contracting L. (V.) braziliensis. Hence, the objective of this study was to collect tissues from wild rodents captured at several study sites along the Amazonian segment of the newly constructed Transoceanic Highway and to use molecular laboratory techniques to analyze samples for the presence of Leishmania parasites. Liver tissues were tested via polymerase chain reaction from a total of 217 rodents; bone marrow and skin biopsies (ear and tail) were also tested from a subset of these same animals. The most numerous rodent species captured and tested were Oligoryzomys microtis (40.7%), Hylaeamys perenensis (15.7%), and Proechimys spp. (12%). All samples were negative for Leishmania, implying that although incidental infections may occur, these abundant rodent species are unlikely to serve as primary reservoirs of L. (V.) braziliensis along the Transoceanic Highway in MDD. Therefore, although these rodent species may persist and even thrive in moderately altered landscapes, we did not find any evidence to suggest they pose a risk for L. (V.) braziliensis transmission to human inhabitants in this highly prevalent region.

  11. Arginase activity of Leishmania isolated from patients with cutaneous leishmaniasis.

    Science.gov (United States)

    Badirzadeh, A; Taheri, T; Abedi-Astaneh, F; Taslimi, Y; Abdossamadi, Z; Montakhab-Yeganeh, H; Aghashahi, M; Niyyati, M; Rafati, S

    2017-09-01

    Cutaneous leishmaniasis (CL) is one of the most important vector-borne parasitic diseases, highly endemic in Iran, and its prevalence is increasing all over the country. Arginase (ARG) activity in isolated Leishmania parasites from CL patients is yet to be explored. This study aimed to compare the ARG activity of isolated Leishmania promastigotes from CL patients with a standard strain of Leishmania major and its influences on the disease pathogenesis. We recruited 16 confirmed CL patients from Qom Province, in central Iran; after detection of Leishmania species using PCR-RFLP, we assessed the levels of ARG in the isolated promastigotes and determined the parasites' growth rate. Only L. major was identified from CL patients. The level of ARG activity in the isolated Leishmania promastigotes from CL patients was significantly higher than that obtained from the standard strain of L. major. No significant correlations between ARG activity and lesion size, number or duration were observed; in contrast, a significant negative correlation was seen between ARG level and Leishmania' growth rate. The obtained results suggest that increased ARG expression and activity in the isolated Leishmania promastigotes might contribute to the higher parasite infectivity and play a major role in the pathogenicity of the CL. © 2017 John Wiley & Sons Ltd.

  12. Molecular identification of Leishmania species in Taybad district, Iran

    Directory of Open Access Journals (Sweden)

    Salehi Ghodratollah

    2014-09-01

    Full Text Available Objective: To identify Leishmania species in patients with cutaneous leishmaniasis in the city of Taybad in Razavi Khorasan Province from April 2012 to March 2013. Methods: Among 52 persons who referred to Health Center of Taybad with suspected skin lesions, stained slide smears of 35 patients showed positive result for Leishmania. Also polymerase chain reaction assay performed using specific kDNA primers. Data of patients were analyzed with SPSS. Results: Of 35 positive smears for Leishmania, 21 (60% belonged to males and 14 (40% belonged to females. Polymerase chain reaction bands were observed in all 35 samples of which 31 (88.6% samples showed Leishmania tropica and 4 (11.4% showed Leishmania major. The highest infected age group was 11-20 years old. Conclusions: Both anthroponotic cutaneous leishmaniasis and zoonotic cutaneous leishmaniasis are present in Taybad. Leishmania tropica is the dominant causative species for anthroponotic cutaneous leishmaniasis. Further study is recommended to discover probable reservoir and vector for Leishmania major in Taybad.

  13. Leptospira spp. y leptospirosis humana

    Directory of Open Access Journals (Sweden)

    Claudia M. Romero-Vivas

    2016-01-01

    Full Text Available La leptospirosis, la enfermedad bacteriana zoonótica y emergente más importante en el mundo, es causada por las especies patógenas de Leptospira spp. Han sido descritas veinte especies de Leptospira spp.; se ha determinado la secuencia del ADN genómico de algunas cepas patógenas; la función de la mayoría de los genes involucrados en su patogénesis permanece desconocida. La leptospirosis humana presenta un rango de síntomas que van desde una fiebre indiferenciada hasta una ictericia, hemorragia, fallas renales y pulmonares severas. La administración temprana e intravenosa de penicilina G es requerida para reducir las tasas de mortalidad, pero los antibióticos pueden no ser efectivos en la enfermedad pulmonar severa. En las Américas, las áreas de alto riesgo son Brasil, Centroamérica y el Caribe. En Colombia se han realizado pocos estudios. La prueba serológica de oro, la microaglutinación, tiene alta sesibilidad y especificidad cuando se usan baterías de serovariedades locales, pero es serogrupo específica. Las vacunas generan respuestas específicas para la serovariedad usada, pero no previenen la infección o trasmisión. Problemas en el diagnóstico de laboratorio de la leptospirosis conllevan a un subregistro en el número de casos; altas tasas de mortalidad asociadas a fallas renal y pulmonar son resultado de las dificultades en el manejo de los casos.

  14. Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis.

    Science.gov (United States)

    Nemati, Sara; Fazaeli, Asghar; Hajjaran, Homa; Khamesipour, Ali; Anbaran, Mohsen Falahati; Bozorgomid, Arezoo; Zarei, Fatah

    2017-08-01

    Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

  15. Accuracy of qPCR for quantifying Leishmania kDNA in different skin layers of patients with American tegumentary leishmaniasis.

    Science.gov (United States)

    Sevilha-Santos, L; Dos Santos Júnior, A C M; Medeiros-Silva, V; Bergmann, J O; da Silva, E F; Segato, L F; Arabi, A Y M; de Paula, N A; Sampaio, R N R; Lima, B D; Gomes, C M

    2018-05-03

    Superficial swab sampling of American tegumentary leishmaniasis (ATL) lesions shows higher amounts of Leishmania than those from biopsy. Subcutaneous involvement is also important in ATL, but parasite quantification according to lesion depth has not been evaluated. We aim to present the best depth at which sampling should be performed for molecular exams of ATL. Patients with a clinical presentation compatible with ATL were allocated to ATL and control groups. Qualitative and quantitative qPCR assays were performed using SYBR Green and primers amplifying the kDNA minicircle of Leishmania spp. in different skin layers, including the epidermis, the superior dermis, the inferior dermis, and the hypodermis. Fifty-nine patients were included in this study, including 40 who had been diagnosed with ATL and 19 controls. The number of parasites was greater in samples of the epidermis and superior dermis (159.1 × 10 6 , range 4.0-781.7, and 75.4 × 10 6 , range 8.0-244.5, mean Leishmania parasite equivalents per μg of tissue DNA, respectively) than those in samples of the inferior dermis and hypodermis (54.6, range 8.0-256.6, and 16.8 × 10 6 , range 8.0-24.1, mean Leishmania parasite equivalents per μg of tissue DNA, respectively). The best diagnostic accuracy was achieved in the superior dermis (77.9%) and was significantly greater than that in the hypodermis (63.3%; p 0.039). We conclude that superficial sampling can retrieve a greater quantity of parasites. Future studies of the role of transepidermal elimination as a mechanism of host defence in ATL must be performed as there is a considerable quantity of Leishmania kDNA in the epidermis. Copyright © 2018 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  16. Presencia de Giardia lamblia y Cryptosporidium spp. en aguas residuales depuradas reutilizadas para riego agrícola en la isla de Tenerife, España. Efectos del transporte a larga distancia sobre la calidad del agua reutilizada Giardia lamblia and Cryptosporidium spp. Presence in treated wastewater reutilised for irrigation in Tenerife island, Spain. Long-distance transport effects in the reutilised water quality

    Directory of Open Access Journals (Sweden)

    Escolástico Aguiar González

    2002-06-01

    Full Text Available El empleo de aguas residuales depuradas para riego agrícola es la alternativa de reutilización con más relevancia hoy día, siempre y cuando se desarrolle con garantías sanitarias y medioambientales.Giardia lamblia y Cryptosporidium spp. son dos protozoos patógenos entéricos de amplia distribución ambiental, frecuentes en hábitats acuáticos. La investigación y detección de estos parásitos en aguas ha adquirido importancia en los últimos años debido a que poseen formas de dispersión resistentes a los tratamientos habituales, aplicados tanto en procesos de potabilización como de depuración, y a que han sido clasificados como patógenos emergentes causantes de importantes brotes de transmisión hídrica.En este trabajo se estudia la presencia de quistes de Giardia lamblia y ooquistes de Cryptosporidium spp. en el agua residual depurada de la ciudad de Santa Cruz de Tenerife que es transportada hasta el sur de la isla de Tenerife para su reutilización en agricultura. Asimismo se investiga el efecto del transporte, el almacenamiento y el tratamiento avanzado sobre la concentración de quistes y ooquistes a través del sistema, y la existencia de relaciones con otros parámetros bacteriológicos y físico-químicos.Los resultados obtenidos ponen de manifiesto comportamientos variables de los contenidos en quistes y ooquistes frente a los tratamientos aplicados, y el efecto depurador que el transporte a larga distancia, parece tener sobre el agua residual depurada.No se han encontrado relaciones entre las concentraciones de quistes y ooquistes en el agua residual depurada y el contenido de los indicadores tradicionales de contaminación fecal.Treated wastewater use for irrigation is, nowadays, the more relevant reutilization alternative of wastewater, if it is developed with sanitary and environmental warranties.Giardia lamblia and Cryptosporidium spp. two wide distributed enteric pathogen protozoan, which are frequent in aquatic

  17. A Preliminary Parasitological Survey of Hepatozoon Spp. Infection in Dogs in Mashhad, Iran

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    J Khoshnegah

    2012-12-01

    Full Text Available Background: We attempted to determine the prevalence of Hepatozoon spp. infection in Mashhad, northeast of Iran, via blood smear parasitology.Methods: The prevalence was investigated by examination of blood smear parasitology, using blood samples collected from 254 dogs (51 strays and 203 privately owned-dogs.Results: Two stray dogs (2/51; 3.92% and two privately-owned dogs (2/203; 0.98% were infected with Hepatozoon spp. Therefore, as per blood smear parasitology, the prevalence of Hepatozoon spp. infection was 1.57% (4/254. Sixteen out of 254 dogs (6.29% were infested with ticks; all of which were Rhipicephalus sanguineus. One of the dogs infected with Hepatozoon spp. exhibited ticks at the time of examination. Concurrent infection with Ehrlichia canis and Leishmania infantum was not detected in the four Hepatozoon spp. infected dogs.Conclusion: This is the first epidemiological study on the prevalence of Hepatozoon spp. infection in dogs in Iran.

  18. Cyclic nucleotide specific phosphodiesterases of Leishmania major

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    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

  19. Vector Competence of Lutzomyia cruzi Naturally Demonstrated for Leishmania infantum and Suspected for Leishmania amazonensis.

    Science.gov (United States)

    de Oliveira, Everton Falcão; Oshiro, Elisa Teruya; Fernandes, Wagner Souza; Ferreira, Alda Maria Teixeira; de Oliveira, Alessandra Gutierrez; Galati, Eunice Aparecida Bianchi

    2017-01-11

    Corumbá city is one of the oldest visceral leishmaniasis-endemic foci in the state of Mato Grosso do Sul, Brazil, where the transmission of Leishmania infantum has been attributed to Lutzomyia cruzi Aiming at investigating the parameters of the vectorial capacity of Lu. cruzi for L. infantum, a project was undertaken in this city. Among these parameters, vector competence was investigated and the results obtained are reported herein. Of the 12 hamsters exposed to feed wild-caught female sandflies, two developed infection with L. infantum and surprisingly, one with Leishmania amazonensis In addition, hamsters with L. infantum infection were bitten only by females of Lu. cruzi, whereas the hamster infected with L. amazonensis was bitten by 124 Lu. cruzi females and one of Evandromyia corumbaensis Although there is a strong suspicion regarding the competence of Lu. cruzi in transmitting L. amazonensis naturally, it was not demonstrated. © The American Society of Tropical Medicine and Hygiene.

  20. Desarrollo y evaluación de una prueba de reacción en cadena de la polimerasa (PCR, utilizando la secuencia del gen hilA para diagnóstico de fiebre entérica por Salmonella spp.

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    Miryan Margot Sánchez

    2004-06-01

    Full Text Available El diagnóstico de fiebre entérica por Salmonella spp. se basa en el aislamiento de la bacteria en hemocultivos el cual consume tiempo, no siempre está disponible y tiene poca utilidad en pacientes con tratamiento antibiótico previo. Por consiguiente, se hace necesario el desarrollo de una prueba rápida, sensible y específica para el diagnóstico de fiebre entérica. Salmonella spp. utiliza el gen hilA (componente de la isla de patogenicidad I para invadir células epiteliales y producir infección. Al usar la secuencia de este gen se diseñó una prueba de PCR para detectar la bacteria en sangre y se evaluó su sensibilidad, especificidad, valor predictivo positivo y valor predictivo negativo, utilizando la metodología prueba de una prueba. La prueba de oro fue el hemocultivo. Se estudiaron 34 individuos con sintomatología de fiebre entérica con aislamiento de Salmonella serotipo Typhi en hemocultivos; 35 individuos con sepsis por otros bacilos Gram negativos aislados de hemocultivo (Klebsiella pneumoniae, 9; Serratia marcescens, 5; Escherichia coli, 4; Pseudomonas aeruginosa, 9; Providencia alcalifaciens, 4, y Enterobacter cloacae, 4 y 150 muestras de sangre de voluntarios asintomáticos. La sensibilidad, especificidad, valor pronóstico positivo y valor pronóstico negativo de la PCR fue del 100%. El número mínimo de UFC/ml que la PCR detecta en sangre es de 10.

  1. Cutaneous leishmaniasis with lymphadenopathy due to Leishmania donovani

    NARCIS (Netherlands)

    Faber, W. R.; Wonders, J.; Jensema, A. J.; Chocholova, E.; Kager, P. A.

    2009-01-01

    Summary We describe a case of cutaneous leishmaniasis with lymphadenopathy due to Leishmania donovani, which was successfully treated with oral miltefosine. Given the increased prevalence of travelling, patients presenting with lymph-node enlargement should have leishmaniasis included in the

  2. Novel Leishmania and Malaria Potassium Channels: Candidate Therapeutic Targets

    National Research Council Canada - National Science Library

    McDonald, Thomas V

    2005-01-01

    .... major and T. cruzi). Using a combination of cultured mammalian cells and Xenopus oocytes for heterologous expression we have evidence that 2 channels from malaria [PFK1 & PFK22] and Leishmania [LMK1 & LMK2] generate K+...

  3. Efficacy of Recombinant Canine Distemper Virus Expressing Leishmania Antigen against Leishmania Challenge in Dogs.

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    Ryuichi Miura

    Full Text Available Canine distemper virus (CDV vaccination confers long-term protection against CDV reinfection. To investigate the utility of CDV as a polyvalent vaccine vector for Leishmania, we generated recombinant CDVs, based on an avirulent Yanaka strain, that expressed Leishmania antigens: LACK, TSA, or LmSTI1 (rCDV-LACK, rCDV-TSA, and rCDV-LmSTI1, respectively. Dogs immunized with rCDV-LACK were protected against challenge with lethal doses of virulent CDV, in the same way as the parental Yanaka strain. To evaluate the protective effects of the recombinant CDVs against cutaneous leishmaniasis in dogs, dogs were immunized with one recombinant CDV or a cocktail of three recombinant CDVs, before intradermal challenge (in the ears with infective-stage promastigotes of Leishmania major. Unvaccinated dogs showed increased nodules with ulcer formation after 3 weeks, whereas dogs immunized with rCDV-LACK showed markedly smaller nodules without ulceration. Although the rCDV-TSA- and rCDV-LmSTI1-immunized dogs showed little protection against L. major, the cocktail of three recombinant CDVs more effectively suppressed the progression of nodule formation than immunization with rCDV-LACK alone. These results indicate that recombinant CDV is suitable for use as a polyvalent live attenuated vaccine for protection against both CDV and L. major infections in dogs.

  4. Efficacy of Recombinant Canine Distemper Virus Expressing Leishmania Antigen against Leishmania Challenge in Dogs.

    Science.gov (United States)

    Miura, Ryuichi; Kooriyama, Takanori; Yoneda, Misako; Takenaka, Akiko; Doki, Miho; Goto, Yasuyuki; Sanjoba, Chizu; Endo, Yasuyuki; Fujiyuki, Tomoko; Sugai, Akihiro; Tsukiyama-Kohara, Kyoko; Matsumoto, Yoshitsugu; Sato, Hiroki; Kai, Chieko

    2015-01-01

    Canine distemper virus (CDV) vaccination confers long-term protection against CDV reinfection. To investigate the utility of CDV as a polyvalent vaccine vector for Leishmania, we generated recombinant CDVs, based on an avirulent Yanaka strain, that expressed Leishmania antigens: LACK, TSA, or LmSTI1 (rCDV-LACK, rCDV-TSA, and rCDV-LmSTI1, respectively). Dogs immunized with rCDV-LACK were protected against challenge with lethal doses of virulent CDV, in the same way as the parental Yanaka strain. To evaluate the protective effects of the recombinant CDVs against cutaneous leishmaniasis in dogs, dogs were immunized with one recombinant CDV or a cocktail of three recombinant CDVs, before intradermal challenge (in the ears) with infective-stage promastigotes of Leishmania major. Unvaccinated dogs showed increased nodules with ulcer formation after 3 weeks, whereas dogs immunized with rCDV-LACK showed markedly smaller nodules without ulceration. Although the rCDV-TSA- and rCDV-LmSTI1-immunized dogs showed little protection against L. major, the cocktail of three recombinant CDVs more effectively suppressed the progression of nodule formation than immunization with rCDV-LACK alone. These results indicate that recombinant CDV is suitable for use as a polyvalent live attenuated vaccine for protection against both CDV and L. major infections in dogs.

  5. Natural canine infection by Leishmania infantum and Leishmania amazonensis and their implications for disease control

    Directory of Open Access Journals (Sweden)

    Letícia da Cruz Sanches

    Full Text Available Abstract Leishmaniasis is a major public health problem worldwide. Because Leishmania can adapt to new hosts or vectors, knowledge concerning the current etiological agent in dogs is important in endemic areas. This study aimed to identify the Leishmania species detected in 103 samples of peripheral blood from dogs that were naturally infected with these protozoa. The diagnosis of leishmaniasis was determined through parasitological examination, the indirect enzyme-linked immunosorbent assay (ELISA and the polymerase chain reaction (PCR. The Leishmania species were identified by means of PCR-restriction fragment length polymorphism (PCR-RFLP. The samples were subjected to PCR using oligonucleotide primers that amplify the intergenic region ITS1 of the rRNA gene in order to identify the species. The amplified DNA was digested using the restriction enzyme HaeIII. A restriction profile identical to L. amazonensis was shown in 77/103 samples and the profile was similar to L. infantum in 17/103. However, a mixed profile was shown in 9/103 samples, which impeded species identification. In conclusion, the infection in these dogs was predominantly due to L. amazonensis, thus indicating that diagnosing of cases of canine leishmaniasis needs to be reexamined, since the causative agent identified is not restricted to L. infantum.

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

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    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.

  7. Evaluación de la respuesta de isotipos de inmunoglobulina especifica a Leishmania en leishmaniasis tegumentaria Americana Evaluation of specific immunoglobin isotype response to Leishmania in American tegumentary leishmaniasis

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    Maricel Labrada

    1989-09-01

    Full Text Available Con el fin de determinar las clases de anticuerpo producido contra el parásito y la cinética de los mismos en relación a la evolución de la infección, se estudiaron los sueros de 133 pacientes infectados con Leishmania del complejo braziliensis. Se utilizó la prueba de inmunofluorescencia indirecta y amastigotas de L. mexicana amazonensis como antígeno. En los sueros obtenidos al momento de consultar para el diagnóstico se encontró IgM en 54 de los sueros absorbidos con Straphylococcus aureus Cowan I y en 5 de los no absorbidos. La IgM sólo se encontro en los sueros de pacientes con tiempo devolución de las lesiones Leishmania-specific immunoglobulin subclass response was evaluated in 133 patients infected with Leishmania braziliensis. The indirect immunofluorescent antibody test (IFAT was employed with amastigotes of L. mexicana amazonensis as antigen. Among the 133 sera obtained at consultation fo rdiagnosis of active lesions, IgM was detected in 54 following absorption with Staphylococcus aureus Cowan strain I, and in 5 sera prior to absorption. IgM reactive with Leishmania antigen was only found in sera from patients whose lesions had envolved over past two months or less. Leishmania-specific IgG was detected in all sera prior to absorption. Sera obtained at the time of recurrence of after complete healing of lesions presented only specific IgC. The combined use of the Montenegro skin test and specific IgM increased the sensitivity of immunodiagnostic methods in patients with lesions of less than 2 months duration. Normal control volunteers were negative for specific IgM and unreactive to Montenegro skin testin. Among 16 patients with non-leishmanial lesion, 3 with sporotrichosis showed IgG reactive with Leishmania; none, including 4 with lesions of less than two months duration, showed specific IgM. We conclude that in patients infected with L. brasiliensis the presence of specific IgG and IgM is associated with the time of

  8. Uji Infeksi Mycosphaerella spp Terhadap Bibit Eucalyptus spp

    OpenAIRE

    Lidya Morita Sondang

    2009-01-01

    Tujuan penelitian ini adalah untuk mengetahui tingkat ketahanan 2 klon Eucalyptus spp yaitu Eucalyptus grandis x Eucalyptus pellita dan Eucalyptus grandis x Eucalyptus urophylla terhadap Mycosphaerella spp serta mengetahui virulensi Mycospaherella spp pada 2 kelas umur (2 dan 3 bulan) pada tanaman Eucalyptus spp. Penelitian ini dilaksanakan dengan pengambilan sampel bibit tanaman Eucalyptus grandis x Eucalyptus pellita dan Eucalyptus grandis x Eucalyptus urophylla dari pembibitan PT.Toba Pulp...

  9. Calcein+/PI- as an early apoptotic feature in Leishmania.

    Science.gov (United States)

    Basmaciyan, Louise; Azas, Nadine; Casanova, Magali

    2017-01-01

    Although leishmaniases are responsible for high morbidity and mortality all over the world, no really satisfying treatment exists. Furthermore, the corresponding parasite Leishmania undergoes a very characteristic form of programmed cell death. Indeed, different stimuli can induce morphological and biochemical apoptotic-like features. However, the key proteins involved in mammal apoptosis, such as caspases and death receptors, are not encoded in the genome of this parasite. Currently, little is known about Leishmania apoptosis, notably owing to the lack of specific tools for programmed cell death analysis in these parasites. Furthermore, there is a need for a better understanding of Leishmania programmed cell death in order (i) to better understand the role of apoptosis in unicellular organisms, (ii) to better understand apoptosis in general through the study of an ancestral eukaryote, and (iii) to identify new therapeutic targets against leishmaniases. To advance understanding of apoptosis in Leishmania, in this study we developed a new tool based on the quantification of calcein and propidium iodide by flow cytometry. This double labeling can be employed to distinguish early apoptosis, late apoptosis and necrosis in Leishmania live cells with a very simple and rapid assay. This paper should, therefore, be of interest for people working on Leishmania and related parasites.

  10. Calcein+/PI- as an early apoptotic feature in Leishmania.

    Directory of Open Access Journals (Sweden)

    Louise Basmaciyan

    Full Text Available Although leishmaniases are responsible for high morbidity and mortality all over the world, no really satisfying treatment exists. Furthermore, the corresponding parasite Leishmania undergoes a very characteristic form of programmed cell death. Indeed, different stimuli can induce morphological and biochemical apoptotic-like features. However, the key proteins involved in mammal apoptosis, such as caspases and death receptors, are not encoded in the genome of this parasite. Currently, little is known about Leishmania apoptosis, notably owing to the lack of specific tools for programmed cell death analysis in these parasites. Furthermore, there is a need for a better understanding of Leishmania programmed cell death in order (i to better understand the role of apoptosis in unicellular organisms, (ii to better understand apoptosis in general through the study of an ancestral eukaryote, and (iii to identify new therapeutic targets against leishmaniases. To advance understanding of apoptosis in Leishmania, in this study we developed a new tool based on the quantification of calcein and propidium iodide by flow cytometry. This double labeling can be employed to distinguish early apoptosis, late apoptosis and necrosis in Leishmania live cells with a very simple and rapid assay. This paper should, therefore, be of interest for people working on Leishmania and related parasites.

  11. 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.

  12. 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

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    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.

  13. Quantificação da IL-10 e do INF-γ em cães com ou sem sinais clínicos de infecção com Leishmania (Leishmania chagasi

    Directory of Open Access Journals (Sweden)

    Carla J.R.M. do Rosário

    Full Text Available RESUMO: As leishmanioses compreendem um complexo de doenças causadas por parasitos intracelulares obrigatórios pertencentes ao gênero Leishmania. Consideradas como importante problema de saúde pública, sendo os cães domésticos os principais responsáveis pela manutenção da cadeia epidemiológica da doença, estima-se que mais da metade dos cães infectados não manifestam sinais clínicos da enfermidade. Avaliou-se o perfil de IL-10 e INF- γ de cães naturalmente infectados com Leishmania (Leishmania chagasi no município de São Luís-MA. Foram coletadas 50 amostras, sendo 20 de animais positivos e sintomáticos para Leishmaniose Visceral Canina (LVC, 20 de animais positivos e assintomáticos e 10 de animais sabidamente negativos para a LVC. As amostras foram analisadas pelo teste imunocromatográfico rápido Dual Path Platform (DPP/Biomanguinhos® e pelo ELISA (EIE/Biomanguinhos® indireto para detecção de anticorpos anti-Leishmania. Após as confirmações dos testes, foi realizado o ELISA de captura para quantificação das citocinas IL-10 e INF-γ através do kit Milliplex MAP. Houve diferença estatística entre os grupos, observando um aumento de IL-10 em soros de cães sintomáticos para LVC, comparado com o grupo de animais assintomáticos, sugerindo que animais com essa expressão de IL-10 podem estar associados à susceptibilidade a doença. Assim como o aumento dos níveis de INF-γ observados em cães assintomáticos, comparado com o grupo de cães sintomáticos, poderiam estar relacionados à cronicidade da doença.

  14. Asymptomatic dogs are highly competent to transmit Leishmania (Leishmania) infantum chagasi to the natural vector.

    Science.gov (United States)

    Laurenti, Márcia Dalastra; Rossi, Claudio Nazaretian; da Matta, Vânia Lúcia Ribeiro; Tomokane, Thaise Yumie; Corbett, Carlos Eduardo Pereira; Secundino, Nágila Francinete Costa; Pimenta, Paulo Filemon Paulocci; Marcondes, Mary

    2013-09-23

    We evaluated the ability of dogs naturally infected with Leishmania (Leishmania) infantum chagasi to transfer the parasite to the vector and the factors associated with transmission. Thirty-eight infected dogs were confirmed to be infected by direct observation of Leishmania in lymph node smears. Dogs were grouped according to external clinical signs and laboratory data into symptomatic (n=24) and asymptomatic (n=14) animals. All dogs were sedated and submitted to xenodiagnosis with F1-laboratory-reared Lutzomyia longipalpis. After blood digestion, sand flies were dissected and examined for the presence of promastigotes. Following canine euthanasia, fragments of skin, lymph nodes, and spleen were collected and processed using immunohistochemistry to evaluate tissue parasitism. Specific antibodies were detected using an enzyme-linked immunosorbent assay. Antibody levels were found to be higher in symptomatic dogs compared to asymptomatic dogs (p=0.0396). Both groups presented amastigotes in lymph nodes, while skin parasitism was observed in only 58.3% of symptomatic and in 35.7% of asymptomatic dogs. Parasites were visualized in the spleens of 66.7% and 71.4% of symptomatic and asymptomatic dogs, respectively. Parasite load varied from mild to intense, and was not significantly different between groups. All asymptomatic dogs except for one (93%) were competent to transmit Leishmania to the vector, including eight (61.5%) without skin parasitism. Sixteen symptomatic animals (67%) infected sand flies; six (37.5%) showed no amastigotes in the skin. Skin parasitism was not crucial for the ability to infect Lutzomyia longipalpis but the presence of Leishmania in lymph nodes was significantly related to a positive xenodiagnosis. Additionally, a higher proportion of infected vectors that fed on asymptomatic dogs was observed (p=0.0494). Clinical severity was inversely correlated with the infection rate of sand flies (p=0.027) and was directly correlated with antibody

  15. Leishmaniasis cutánea zosteriforme causada por Leishmania (Viannia panamensis y Leishmania (Viannia braziliensis: reporte de tres casos

    Directory of Open Access Journals (Sweden)

    Camilo Andrés Morales

    2014-09-01

    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.

  16. Leishmania (Viannia) guyanensis in tegumentary leishmaniasis.

    Science.gov (United States)

    Borges, Arissa Felipe; Gomes, Rodrigo Saar; Ribeiro-Dias, Fátima

    2018-06-01

    Leishmania (Viannia) guyanensis is a causal agent of American tegumentary leishmaniasis (ATL). This protozoan has been poorly investigated; however, it can cause different clinical forms of ATL, ranging from a single cutaneous lesion to severe lesions that can lead to destruction of the nasopharyngeal mucosa. L. (V.) guyanensis and the disease caused by this species can present unique aspects revealing the need to better characterize this parasite species to improve our knowledge of the immunopathological mechanisms and treatment options for ATL. The mechanisms by which some patients develop a more severe form of ATL remain unclear. It is known that the host immune profile and parasite factors may influence the clinical manifestations of the disease. Besides intrinsic parasite factors, Leishmaniavirus RNA 1 (LRV1) infecting L. guyanensis can contribute to ATL immunopathogenesis. In this review, general aspects of L. guyanensis infection in humans and mouse models are presented.

  17. 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%.

  18. Seroepidemiology and molecular diversity of Leishmania donovani complex in Georgia.

    Science.gov (United States)

    Babuadze, Giorgi; Farlow, Jason; de Koning, Harry P; Carrillo, Eugenia; Chakhunashvili, Giorgi; Murskvaladze, Mari; Kekelidze, Merab; Karseladze, Irakli; Kokaia, Nora; Kalandadze, Irine; Tsereteli, David; Markhvashvili, Ivane; Sidamonidze, Ketevan; Chanturia, Gvantsa; Adeishvili, Ekaterine; Imnadze, Paata

    2016-05-13

    Leishmaniasis includes multiple clinical syndromes, most notably visceral, cutaneous, and mucosal forms. Visceral leishmaniasis (VL), also known as kala-azar, is a potentially fatal disease endemic to large parts of Africa and Asia, and in South-Eastern Europe (Greece, Turkey, Georgia). Visceral leishmaniasis is a parasitic zoonosis caused by species of the L. donovani complex. In the classical epidemiological model the main reservoir for VL are canines. The study included a cohort of 513 individuals of both genders (190 males and 323 females) from the ages of 1 to 70 years that were screened in ten villages across two districts in Kakheti using the Kalazar Detect™ rK39 rapid diagnostic test. The phylogenetic diversity patterns of local strains, based on the rDNA internal transcribed spacer (ITS) sequences, were assessed for samples obtained from patients with suspected L. donovani infection, from canine reservoirs and from Phlebotomus sand flies obtained from different geographical areas of Georgia and from Azerbaijan. Out of a total of 600 domestic dog blood samples 95 (15.8 %) were positive by rK39 rapid diagnostic tests. For symptomatic domestic dogs, the testing of conjunctival swabs or bone marrow aspirates revealed a higher VL incidence in Kvareli District (Kvareli; 19.4 %, n = 329) compared with that observed for Sagarejo District (Sagarejo; 11.4 %, n = 271). A total of 231 sand flies of both genders were collected during the 2-month period; of the 114 females, 1.75 % were PCR positive for the presence of Leishmania spp. VL infection rates remain high in both canines and humans in Georgia, with disease in several known natural foci. The genetic relationships derived from rDNA internal transcribed spacer (ITS) sequence comparisons identified genetic subgroups, revealing preliminary insights into the genetic structure of L. donovani complex members currently circulating in the South Caucasus and demonstrates the utility of ITS-based genotyping

  19. Physalis angulata induces death of promastigotes and amastigotes of Leishmania (Leishmania) amazonensis via the generation of reactive oxygen species.

    Science.gov (United States)

    Da Silva, B J M; Da Silva, R R P; Rodrigues, A P D; Farias, L H S; Do Nascimento, J L M; Silva, E O

    2016-03-01

    Leishmaniasis are a neglected group of emerging diseases that have been found in 98 countries and are caused by protozoa of the genus Leishmania. The therapy for leishmaniasis causes several side effects and leads to drug-resistant strains. Natural products from plants have exhibited activities against Leishmania in various experimental models. Physalis angulata is a widely used plant in popular medicine, and in the literature it has well-documented leishmanicidal activity. However, its mechanism of action is still unknown. Thus, this study aims to evaluate the mechanism driving the leishmanicidal activity of an aqueous extract of P. angulata root (AEPa). AEPa was effective against both promastigotes and intracellular amastigote forms of Leishmania amazonensis. This effect was mediated by an increase of reactive oxygen species (ROS), but not of nitric oxide (NO). The increased production of ROS induces cell death by phenotypes seems by apoptosis cell death in Leishmania, but not autophagy or necrosis. In addition, morphological analysis of macrophages showed that AEPa induced a high number of cytoplasmic projections, increased the volume of cytoplasm and number of vacuoles, caused cytoskeleton alterations and resulted in high spreading ability. AEPa also promoted superoxide anion (O2(-)) production in both uninfected macrophages and those infected with Leishmania. Therefore, these results revealed that AEPa causes cell death by phenotypes seems by apoptosis cell death in L. amazonensis and modulates macrophage activation through morphofunctional alterations and O2(-) generation to induce Leishmania death. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Quantification of anti-Leishmania antibodies in saliva of dogs.

    Science.gov (United States)

    Cantos-Barreda, Ana; Escribano, Damián; Bernal, Luis J; Cerón, José J; Martínez-Subiela, Silvia

    2017-08-15

    Detection of serum anti-Leishmania antibodies by quantitative or qualitative techniques has been the most used method to diagnose Canine Leishmaniosis (CanL). Nevertheless, saliva may represent an alternative to blood because it is easy to collect, painless and non-invasive in comparison with serum. In this study, two time-resolved immunofluorometric assays (TR-IFMAs) for quantification of anti-Leishmania IgG2 and IgA antibodies in saliva were developed and validated and their ability to distinguish Leishmania-seronegative from seropositive dogs was evaluated. The analytical study was performed by evaluation of assay precision, sensitivity and accuracy. In addition, serum from 48 dogs (21 Leishmania-seropositive and 27 Leishmania-seronegative) were analyzed by TR-IFMAs. The assays were precise, with an intra- and inter-assay coefficients of variation lower than 11%, and showed high level of accuracy, as determined by linearity under dilution (R 2 =0.99) and recovery tests (>88.60%). Anti-Leishmania IgG2 antibodies in saliva were significantly higher in the seropositive group compared with the seronegative (pLeishmania IgA antibodies between both groups were observed. Furthermore, TR-IFMA for quantification of anti-Leishmania IgG2 antibodies in saliva showed higher differences between seropositive and seronegative dogs than the commercial assay used in serum. In conclusion, TR-IFMAs developed may be used to quantify anti-Leishmania IgG2 and IgA antibodies in canine saliva with an adequate precision, analytical sensitivity and accuracy. Quantification of anti-Leishmania IgG2 antibodies in saliva could be potentially used to evaluate the humoral response in CanL. However, IgA in saliva seemed not to have diagnostic value for this disease. For future studies, it would be desirable to evaluate the ability of the IgG2 assay to detect dogs with subclinical disease or with low antibody titers in serum and also to study the antibodies behaviour in saliva during the

  1. First evidence of intraclonal genetic exchange in trypanosomatids using two Leishmania infantum fluorescent transgenic clones.

    Directory of Open Access Journals (Sweden)

    Estefanía Calvo-Álvarez

    2014-09-01

    Full Text Available The mode of reproduction in Leishmania spp has been argued to be essentially clonal. However, recent data (genetic analysis of populations and co-infections in sand flies have proposed the existence of a non-obligate sexual cycle in the extracellular stage of the parasite within the sand fly vector. In this article we propose the existence of intraclonal genetic exchange in the natural vector of Leishmania infantum.We have developed transgenic L. infantum lines expressing drug resistance markers linked to green and red fluorescent reporters. We hypothesized whether those cells with identical genotype can recognize each other and mate. Both types of markers were successfully exchanged within the sand fly midgut of the natural vector Phlebotomus perniciosus when individuals from these species were fed with a mixture of parental clones. Using the yellow phenotype and drug resistance markers, we provide evidence for genetic exchange in L. infantum. The hybrid progeny appeared to be triploid based on DNA content analysis. The hybrid clone analyzed was stable throughout the complete parasite life cycle. The progress of infections by the hybrid clone in BALB/c mice caused a reduction in parasite loads in both spleen and liver, and provided weight values similar to those obtained with uninfected mice. Spleen arginase activity was also significantly reduced relative to parental strains.A L. infantum hybrid lineage was obtained from intraclonal genetic exchange within the midgut of the natural vector, suggesting the ability of this parasite to recognize the same genotype and mate. The yellow hybrid progeny is stable throughout the whole parasite life cycle but with a slower virulence, which correlates well with the lower arginase activity detected both in vitro and in vivo infections.

  2. Aislamiento de genes diferencialmente expresados en amastigotes de Leishmania (Viannia panamensis

    Directory of Open Access Journals (Sweden)

    Róbinson Ramírez

    2001-04-01

    Full Text Available

    En Colombia cada año se diagnostican 6.500 casos nuevos de
    leishmaniasis cutánea y Leishmania (Viannia panamensis es la responsable del 95% de los casos. Durante su ciclo de vida, el parásito reside en el insecto vector como promastigote y se diferencia a amastigote después de ser inoculado en el hospedero mamífero. La transformación de promastigote a amastigote implica cambios morfológicos y bioquímicos que tienen lugar en el fagolisosoma del macrófago, y se caracterizan, entre otros, por la expresión de enzimas con actividad óptima a pH ácido además del cambio de los glicoconjugados de membrana. La supervivencia del parásito está asociada con su capacidad para inhibir funciones efectoras en el macrófago infectado permitiéndole al amastigote
    su replicación; sin embargo, poco se conoce acerca de los productos génicos expresados diferencialmente en este estadio por lo que nos propusimos con este trabajo aislar genes diferencialmente expresados en amastigotes de Leishmania (Viannia panamensis.

     

     

  3. Influência da idade na geração de modelos de espectroscopia NIR, para predição de propriedades da madeira de Eucalyptus spp.

    Directory of Open Access Journals (Sweden)

    Flaviana Reis Milagres

    2013-12-01

    Full Text Available O objetivo deste trabalho foi o desenvolvimento de modelos NIRS para predizer as propriedades físicas e químicas de madeiras de Eucalyptus, com idades variando de 3 a 7 anos. Foram utilizadas 136 amostras e realizadas análises laboratoriais de densidade básica, teores de lignina e extrativos. Amostras de cavacos foram moídas em moinho tipo ciclone, para coleta de espectros NIR. Os modelos foram desenvolvidos por regressão de mínimos quadrados parciais (PLS, sendo testadas transformações matemáticas. Para validação dos modelos, foram utilizados grupos de amostras externas com diferentes idades, que não participaram no desenvolvimento do modelo. Os modelos desenvolvidos resultaram em consideráveis coeficientes de determinação para densidade básica (R² 0,84 a 0,89 e lignina (R² 0,72 a 0,88, mas os melhores valores foram encontrados para o teor de extrativos (R² 0,92 a 0,98. Pelos valores de relação de desempenho de desvio (RPD, verificou-se que amostras com idades diferentes das utilizadas no desenvolvimento dos modelos não apresentaram boas predições, exceto para densidade básica cujos modelos desenvolvidos especificamente para as idades de 3 e 7 anos, predisseram satisfatoriamente amostras com idades entre 4 e 6 anos.

  4. Application of riboprinting for the identification of isolates of cutaneous Leishmania spp

    NARCIS (Netherlands)

    Nimri, L. F.; Schallig, H. D. F. H.

    2003-01-01

    Riboprinting is one of several molecular methods that can generate comparative data independently of the complexity of the organism's morphology. Restriction fragment length polymorphism (RFLP) profiles derived from digestion of polymerase chain reaction (PCR) products of the ribosomal 18S from

  5. Purification of a Fe-SOD excreted by Leishmania braziliensis for specific antibodies detection in Mexican human sera: Cutting-edge the knowledge

    Directory of Open Access Journals (Sweden)

    Silvia Stefania Longoni

    2016-06-01

    Full Text Available Clinical diagnosis of leishmaniasis is highly complex, presenting a wide range of clinical manifestations, sometimes non-specific, and thus the epidemiological study and diagnostic need specific molecular markers for each Leishmania species. Leishmania spp. posses different Fe-SOD isoforms, one of which is excreted into the external milieu and, presenting immunogenic characteristics, is a very reliable molecular marker. Superoxide dismutases (SODs are antioxidant metal-enzymes responsible for the dismutation of superoxide ion into hydrogen peroxide and molecular oxygen, and it is considered an important virulence factor. In this manuscript we have purified the iron(Fe-SOD excreted by Leishmania braziliensis using ion-exchange and molecular-sieve chromatography and we have studied it as an antigen in serodiagnostic analyses in ELISA and Western blot techniques, testing 213 human sera from Mexico. Indeed, L. braziliensis Fe-SODe has been purified 123.26 times with a specific activity of about 893.66 U/mg of protein. Applying the purified enzymes in serological tests we found 17.84% sera positive. We have demonstrated that the purified enzyme is more sensitive than the non-purified ones and we also demonstrated, for the first time, the presence of antibodies against L. braziliensis, not the main species in the country, in human population from Hidalgo and Nuevo Leon States.

  6. A calmodulin-like protein (LCALA) is a new Leishmania amazonensis candidate for telomere end-binding protein.

    Science.gov (United States)

    Morea, Edna G O; Viviescas, Maria Alejandra; Fernandes, Carlos A H; Matioli, Fabio F; Lira, Cristina B B; Fernandez, Maribel F; Moraes, Barbara S; da Silva, Marcelo S; Storti, Camila B; Fontes, Marcos R M; Cano, Maria Isabel N

    2017-11-01

    Leishmania spp. telomeres are composed of 5'-TTAGGG-3' repeats associated with proteins. We have previously identified LaRbp38 and LaRPA-1 as proteins that bind the G-rich telomeric strand. At that time, we had also partially characterized a protein: DNA complex, named LaGT1, but we could not identify its protein component. Using protein-DNA interaction and competition assays, we confirmed that LaGT1 is highly specific to the G-rich telomeric single-stranded DNA. Three protein bands, with LaGT1 activity, were isolated from affinity-purified protein extracts in-gel digested, and sequenced de novo using mass spectrometry analysis. In silico analysis of the digested peptide identified them as a putative calmodulin with sequences identical to the T. cruzi calmodulin. In the Leishmania genome, the calmodulin ortholog is present in three identical copies. We cloned and sequenced one of the gene copies, named it LCalA, and obtained the recombinant protein. Multiple sequence alignment and molecular modeling showed that LCalA shares homology to most eukaryotes calmodulin. In addition, we demonstrated that LCalA is nuclear, partially co-localizes with telomeres and binds in vivo the G-rich telomeric strand. Recombinant LCalA can bind specifically and with relative affinity to the G-rich telomeric single-strand and to a 3'G-overhang, and DNA binding is calcium dependent. We have described a novel candidate component of Leishmania telomeres, LCalA, a nuclear calmodulin that binds the G-rich telomeric strand with high specificity and relative affinity, in a calcium-dependent manner. LCalA is the first reported calmodulin that binds in vivo telomeric DNA. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Leishmania major methionine sulfoxide reductase A is required for resistance to oxidative stress and efficient replication in macrophages.

    Directory of Open Access Journals (Sweden)

    Fiona M Sansom

    Full Text Available Leishmania are protozoan parasites that proliferate within the phagolysome of mammalian macrophages. While a number of anti-oxidant systems in these parasites have been shown to protect against endogenous as well as host-generated reactive oxygen species, the potential role of enzymes involved in the repair of oxidatively damaged proteins remains uncharacterized. The Leishmania spp genomes encode a single putative methionine sulfoxide reductase (MsrA that could have a role in reducing oxidized free and proteinogenic methionine residues. A GFP-fusion of L. major MsrA was shown to have a cytoplasmic localization by immunofluorescence microscopy and subcellular fractionation. An L. major msrA null mutant, generated by targeted replacement of both chromosomal allelles, was viable in rich medium but was unable to reduce exogenous methionine sulfoxide when cultivated in the presence of this amino acid, indicating that msrA encodes a functional MsrA. The ΔmsrA mutant exhibited increased sensitivity to H(2O(2 compared to wild type parasites and was unable to proliferate normally in macrophages. Wild type sensitivity to H(2O(2 and infectivity in macrophages was restored by complementation of the mutant with a plasmid encoding MsrA. Unexpectedly, the ΔmsrA mutant was able to induce normal lesions in susceptible BALB/c indicating that this protein is not essential for pathogenesis in vivo. Our results suggest that Leishmania MsrA contributes to the anti-oxidative defences of these parasites, but that complementary oxidative defence mechansims are up-regulated in lesion amastigotes.

  8. Experimental Acquisition, Development, and Transmission of Leishmania tropica by Phlebotomus duboscqi

    Science.gov (United States)

    2013-01-01

    ac ta t ropica Experimental acquisition, development, and transmission of Leishmania tropica by Phlebotomus duboscqi Hanafi A. Hanafi, El...August 2012 Accepted 2 September 2012 Available online 10 September 2012 Keywords: Phlebotomus duboscqi Leishmania tropica Transmission Vector...competency a b s t r a c t We report experimental infection and transmission of Leishmania tropica (Wright), by the blood-feeding sand

  9. 46-kDa protein located in the flagellar pocket of Leishmania ...

    Indian Academy of Sciences (India)

    NII

    Cloning and expression of endocytic Rab GTPases from Leishmania. Fractionation of early compartment from. Leishmania containing endocytic probes. Rab7:WT. Rab5: WT. Localization of Rab5 and Rab7 in Leishmania. Phase. LTR. Merge. Rab7-GFP. Rab5-GFP. LTR. Phase. Merge. Rab5-GFP. Rab7-GFP. Lysosomes.

  10. Restrição do 16S-23S DNAr intergênico para avaliação da diversidade de Azospirillum amazonense isolado de Brachiaria spp. Restriction of 16S-23S intergenic rDNA for diversity evaluation of Azospirillum amazonense isolated from different Brachiaria spp.

    Directory of Open Access Journals (Sweden)

    Fábio Bueno dos Reis Junior

    2006-03-01

    Full Text Available O objetivo deste trabalho foi avaliar a diversidade intra-específica de isolados de Azospirillum amazonense e estabelecer a possível influência de diferentes espécies de Brachiaria ssp. e diferentes condições edafoclimáticas. A caracterização da diversidade desses isolados foi conduzida, utilizando-se a análise de restrição da região intergênica 16S-23S DNAr. As estirpes estudadas separaram-se em dois grupos, definidos a 56% de similaridade. As espécies de Brachiaria ssp. influenciaram a diversidade de estirpes. A maioria dos isolados oriundos de B. decumbens e B. brizantha está inserida no primeiro grupo, enquanto os oriundos de B. humidicola concentram-se no segundo grupo.The aim of this work was to study the intra-specific diversity of Azospirillum amazonense isolates and to establish possible influences of different Brachiaria spp. and edaphoclimatic conditions. The characterization of the diversity among the isolates of A. amazonense studied was conducted using restriction analysis of the 16S-23S rDNA intergenic spacer region. The evaluated strains were separated in two groups, defined at 56% of similarity. Brachiaria spp. showed effects on strain diversity. Most part of the isolates from B. decumbens and B. brizantha are inserted in the first group, while B. humidicola isolates concentrate in the second group.

  11. Potencial terapéutico del aceite esencial de Chenopodium ambrosioides y algunos de sus componentes frente a Leishmania

    OpenAIRE

    Monzote Fidalgo, Lianet

    2010-01-01

    La leishmaniosis es una enfermedad causada por parásitos protozoos del género Leishmania, clasificada por la Organización Mundial de la Salud como una de las infecciones tropicales más importantes. Los fármacos disponibles son tóxicos, costosos y la resistencia parasitaria ha emergido. En las áreas endémicas la población depende fundamentalmente de la medicina natural para el tratamiento de la leishmaniosis. En este trabajo, se evaluó la actividad del aceite esencial extraído de Chenopodium a...

  12. Manipulación genética y el estudio del parásito protozoario Leishmania.

    Directory of Open Access Journals (Sweden)

    Tania M. Cortázar

    2004-12-01

    Full Text Available Durante los últimos 15 años se ha dado paso al entendimiento de muchos aspectos de la genómica funcional de Leishmania gracias a los avances en la metodología de transfección de ADN dentro de la célula de este protozoario, la eliminación y la complementación de genes por medio de recombinación homóloga y las estrategias para la selección de células transfectadas. Estos acercamientos tienen el potencial de brindar información sobre la expresión génica y la función de las proteínas en el contexto del parásito intacto. Dado que el genoma de Leishmania muestra una carencia acentuada de los factores conocidos de iniciación de la transcripción y que la expresión génica está regulada casi completamente a nivel postranscripcional (a través del empalme de los ARNm y los mecanismos que involucran el procesamiento diferencial de la región no traducida 3' del ARNm (3?UTR, la transfección génica representa una herramienta útil para la identificación y el análisis funcional de los genes de interés así como de los mecanismos que dirigen su regulación. El desarrollo de los sistemas de manipulación genética también ha abierto nuevos horizontes para la identificación de genes esenciales involucrados en la virulencia, la supervivencia intracelular y la resistencia a drogas de Leishmania, así como para la validación de proteínas específicas del parásito como nuevos blancos quimio e inmunoterapéuticos. En esta revisión presentamos los avances más recientes en el campo de la manipulación genética en Leishmania, los cuales permiten análisis estructurales, funcionales y de fenotipo, por medio de la eliminación y complementación génica a través de la transfección transitoria o permanente de genes en este parásito.

  13. Actividad contra Leishmania sp. (Kinetoplastida: Trypanosomatidae) de plantas en una Reserva Biológica de Costa Rica

    OpenAIRE

    Misael Chinchilla-Carmona; Idalia Valerio-Campos; Ronald Sánchez-Porras; Vanessa Bagnarello-Madrigal; Laura Martínez-Esquivel; Antonieta González-Paniagua; Javier Alpizar-Cordero; Maribel Cordero-Villalobos; Daniela Rodríguez-Chaves

    2014-01-01

    La leishmaniosis es una enfermedad muy importante para el ser humano pero su tratamiento es bastante difícil. Por esta razón muchos investigadores han venido buscando plantas que contengan componentes químicos activos contra esta parasitosis. En Costa Rica no se tienen estudios al respecto y por eso el objetivo de este estudio fue la búsqueda de componentes activos contra Leishmania sp. en plantas de Costa Rica; 67 especies de la Reserva Biológica Alberto Manuel Brenes (REBAMB) fueron selecci...

  14. Phlebotomine sandflies (Diptera, Phlebotomidae) of Lanzarote Island (Canary Islands, Spain): Ecological survey and evaluation of the risk of Leishmania transmission.

    Science.gov (United States)

    Morillas-Márquez, Francisco; Díaz-Sáez, Victoriano; Morillas-Mancilla, María Jesús; Corpas-López, Victoriano; Merino-Espinosa, Gemma; Gijón-Robles, Patricia; Martín-Sánchez, Joaquina

    2017-04-01

    Phlebotomine sandflies are natural vectors of Leishmania spp. and their expansion throughout has been evidenced in the last few years due to the global warming and changes in human behavior, worsening leishmaniasis problem. However, phlebotomine sandflies have been captured in small numbers on the Canary Islands, particularly on the island of Lanzarote, where only one limited survey was carried out almost thirty years ago. The proximity of this island to Morocco, in addition to the high number of tourists, sometimes accompanied by their dogs, from leishmaniasis endemic regions, highlights the importance of studying the sandfly fauna on this island in order to determine the transmission risk of leishmaniasis Thirty-eight sampling sites spread across the island were studied, and ecological features were gathered to identify the ecological traits associated to the presence of sandflies. Only 85 sandfly specimens were captured (1.18/m 2 ) with the following species distribution: Sergentomyia minuta (0.15 specimens/m 2 ), which was reported for the first time on this island, and S. fallax (1.03/m 2 ). Sandfly captured were achieved in only 7 out of 38 stations. No specimen of the Phlebotomus genus was captured and given that none of the species captured has been demonstrated vectors of human pathogenic Leishmania and considering that they were captured in low frequency and density, it can be concluded that the current leishmaniasis transmission risk is null. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. Neutrophils reduce the parasite burden in Leishmania (Leishmania amazonensis-infected macrophages.

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    Erico Vinícius de Souza Carmo

    2010-11-01

    Full Text Available Studies on the role of neutrophils in Leishmania infection were mainly performed with L. (L major, whereas less information is available for L. (L amazonensis. Previous results from our laboratory showed a large infiltrate of neutrophils in the site of infection in a mouse strain resistant to L. (L. amazonensis (C3H/HePas. In contrast, the susceptible strain (BALB/c displayed a predominance of macrophages harboring a high number of amastigotes and very few neutrophils. These findings led us to investigate the interaction of inflammatory neutrophils with L. (L. amazonensis-infected macrophages in vitro.Mouse peritoneal macrophages infected with L. (L. amazonensis were co-cultured with inflammatory neutrophils, and after four days, the infection was quantified microscopically. Data are representative of three experiments with similar results. The main findings were 1 intracellular parasites were efficiently destroyed in the co-cultures; 2 the leishmanicidal effect was similar when cells were obtained from mouse strains resistant (C3H/HePas or susceptible (BALB/c to L. (L. amazonensis; 3 parasite destruction did not require contact between infected macrophages and neutrophils; 4 tumor necrosis factor alpha (TNF-α, neutrophil elastase and platelet activating factor (PAF were involved with the leishmanicidal activity, and 5 destruction of the parasites did not depend on generation of oxygen or nitrogen radicals, indicating that parasite clearance did not involve the classical pathway of macrophage activation by TNF-α, as reported for other Leishmania species.The present results provide evidence that neutrophils in concert with macrophages play a previously unrecognized leishmanicidal effect on L. (L. amazonensis. We believe these findings may help to understand the mechanisms involved in innate immunity in cutaneous infection by this Leishmania species.

  16. 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.

  17. Phylogenetic analysis of HSP70 and cyt b gene sequences for Chinese Leishmania isolates and ultrastructural characteristics of Chinese Leishmania sp.

    Science.gov (United States)

    Yuan, Dongmei; Qin, Hanxiao; Zhang, Jianguo; Liao, Lin; Chen, Qiwei; Chen, Dali; Chen, Jianping

    2017-02-01

    Leishmaniasis is a worldwide epidemic disease caused by the genus Leishmania, which is still endemic in the west and northwest areas of China. Some viewpoints of the traditional taxonomy of Chinese Leishmania have been challenged by recent phylogenetic researches based on different molecular markers. However, the taxonomic positions and phylogenetic relationships of Chinese Leishmania isolates remain controversial, which need for more data and further analysis. In this study, the heat shock protein 70 (HSP70) gene and cytochrome b (cyt b) gene were used for phylogenetic analysis of Chinese Leishmania isolates from patients, dogs, gerbils, and sand flies in different geographic origins. Besides, for the interesting Leishmania sp. in China, the ultrastructure of three Chinese Leishmania sp. strains (MHOM/CN/90/SC10H2, SD, GL) were observed by transmission electron microscopy. Bayesian trees from HSP70 and cyt b congruently indicated that the 14 Chinese Leishmania isolates belong to three Leishmania species including L. donovani complex, L. gerbilli, and L. (Sauroleishmania) sp. Their identity further confirmed that the undescribed Leishmania species causing visceral Leishmaniasis (VL) in China is closely related to L. tarentolae. The phylogenetic results from HSP70 also suggested the classification of subspecies within L. donovani complex: KXG-918, KXG-927, KXG-Liu, KXG-Xu, 9044, SC6, and KXG-65 belong to L. donovani; Cy, WenChuan, and 801 were proposed to be L. infantum. Through transmission electron microscopy, unexpectedly, the Golgi apparatus were not observed in SC10H2, SD, and GL, which was similar to previous reports of reptilian Leishmania. The statistical analysis of microtubule counts separated SC10H2, SD, and GL as one group from any other reference strain (L. donovani MHOM/IN/80/DD8; L. tropica MHOM/SU/74/K27; L. gerbilli MRHO/CN/60/GERBILLI). The ultrastructural characteristics of Leishmania sp. partly lend support to the phylogenetic inference that

  18. Natural hybrid of Leishmania infantum/L. donovani: development in Phlebotomus tobbi, P. perniciosus and Lutzomyia longipalpis and comparison with non-hybrid strains differing in tissue tropism.

    Science.gov (United States)

    Seblova, Veronika; Myskova, Jitka; Hlavacova, Jana; Votypka, Jan; Antoniou, Maria; Volf, Petr

    2015-11-25

    from Cyprus, however, in all other parameters studied, including localization of infection and colonization of stomodeal valve, dermotropic and viscerotropic strains were not significantly different. We showed high susceptibility of P. tobbi to various Leishmania spp. This, together with the presence of O-glycosylated midgut proteins in their midguts demonstrate that P. tobbi is a permissive vector. Two dermotropic and three viscerotropic strains from the L. donovani complex developed late-stage infections in natural L. infantum vectors, P. perniciosus and L. longipalpis and none of the parameters studied seem to be linked with different tropism of parasites in the vertebrate host.

  19. Mechanism of ascaridole activation in Leishmania.

    Science.gov (United States)

    Geroldinger, Gerald; Tonner, Matthias; Hettegger, Hubert; Bacher, Markus; Monzote, Lianet; Walter, Martin; Staniek, Katrin; Rosenau, Thomas; Gille, Lars

    2017-05-15

    Endoperoxides (EP) are an emerging class of drugs which have potential in antiparasitic therapy, but also in other fields. For malaria therapy the EP artemisinin (Art) and its derivatives are successfully used. We have shown in the past that the EP ascaridole (Asc) is useful for the treatment of cutaneous leishmaniasis in a mouse model. Biomimetic experiments suggested that these drugs need activation in the respective target pathogens to exert their function. In spite of this idea, direct activation of EP to radicals inside cells has never been demonstrated. Therefore, this study was initiated to explore the activation of Asc in biomimetic systems and inside Leishmania in comparison to Art. Using electron paramagnetic resonance spectroscopy (EPR) in combination with spin-trapping we identified the secondary alkyl radical intermediates arising from reduction by Fe 2+ in cell-free systems. Combined GC/NMR analysis confirmed the loss of isopropyl residues from Asc during this process as intermediates. This activation of Asc was stimulated by low molecular Fe 2+ complexes or alternatively by hemin in conjunction with thiol reductants, such as cysteine (Cys). In Leishmania tarentolae promastigotes (LtP) as model for pathogenic forms of Leishmania carbon-centered radicals were identified in the presence of Asc by EPR spin-trapping. Both Asc and Art inhibited the viability in LtP with IC 50 values in the low micromolar range while IC 50 values for J774 macrophages were considerably higher. A similar structure without EP bridge (1,4-cineole) resulted in no detectable radicals and possessed much less cytotoxicity in LtP and no selectivity for LtP compared to J774 cells. The Asc-derived radical formation in LtP was inhibited by the iron chelator deferoxamine (DFO), and stimulated by Cys (a suitable reductant for hemin). The IC 50 values for LtP viability in the presence of Asc or Art were increased significantly by the spin trap DMPO, while Cys and DFO increased only IC 50

  20. Dihydrotestosterone enhances growth and infectivity of Leishmania Mexicana.

    Science.gov (United States)

    Sánchez-García, L; Wilkins-Rodriguez, A; Salaiza-Suazo, N; Morales-Montor, J; Becker, I

    2018-03-01

    A strong sex-associated susceptibility towards Leishmania has been reported in males, yet little is known on the effect of hormones in Leishmania physiopathogenicity. Due to the enhanced susceptibility of males to Leishmania mexicana infections, we were interested in analysing the effect exerted by the main androgen produced in males (DHT) on L. mexicana promastigotes. Thus, the aim of this study was to assess the regulation exerted by dihydrotestosterone (DHT) on L. mexicana replication, infectivity, survival and development of tissue lesions. Experiments included growth curves of L. mexicana promastigotes incubated with different doses of DHT, their infection rate, intracellular survival and lesion development in BALB/c mice. Our data show that DHT significantly enhances parasite replication, infection rate and survival in bone marrow-derived macrophages (BMMФ). Promastigotes in the presence of DHT produced significantly larger lesions in BALB/c earlobes. These results suggest that DHT probably plays a critical role during L. mexicana infections, and the higher susceptibility of males possibly relates to benefits gained by the parasite from host-derived hormones. Our data shed new light on the physiopathology of Leishmania infections and are the first attempt to understand the direct interaction between Leishmania and androgens, particularly DHT. Understanding this trans-regulation process employed by parasites to exploit host molecules sheds new light on L. mexicana physiopathogenesis and opens a possible field for studies on drug development. © 2017 John Wiley & Sons Ltd.

  1. El análisis molecular y el inmunogénico sugieren la ausencia de las proteínas hidrofílicas de superficie en Leishmania (Viannia panamensis

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    Marcel Marín

    2008-09-01

    Full Text Available Introducción. Los dos subgéneros en los cuales se divide el género Leishmania: Viannia y Leishmania, presentan diferencias significativas en las manifestaciones clínicas que causan, en su comportamiento de crecimiento en cultivos in vitro, en sus características genéticas y en la expresión de varias proteínas, entre ellas las de la familia hidrofílica de superficie superficie. Objetivo. Caracterizar las proteínas hidrofílicas de superficie en Leishmania (Viannia panamensis. Materiales y métodos. Se amplificaron los genes hasp en L. (V. panamensis usando cebadores específicos para la especie Leishmania (Leishmania major. Los productos de la amplificación fueron clonados, secuenciados y analizados con herramientas bioinformáticas. Posteriormente, se realizó un análisis serológico por medio de ensayo inmunoabsorbente ligado a enzimas y Western blot para detectar la presencia de anticuerpos específicos contra las proteínas hidrofílicas recombinantes de superficie de L. (L. major en sueros de pacientes con leishmaniasis de zonas endémicas de Colombia. Resultados. Se encontró una copia de un pseudogen en L. (V. panamensis, el cual presentó una identidad del 60% con el gen haspa de L. (L. major. Sólo se encontraron anticuerpos contra las proteínas recombinantes de superficie hidrofílicas en sueros de pacientes con leishmaniasis visceral. Conclusión. Estos resultados sugieren que no existe ninguna copia de un gen funcional hasp en L. (V. panamensis, lo que indica una pérdida de la familia de genes en esta especie de Leishmania perteneciente al subgénero Viannia.

  2. Comparación de diferentes métodos para identificar las especies del género Proteus Comparison of different methods in order to identify Proteus spp

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    S. T. Castro

    2006-09-01

    Full Text Available Los objetivos de este trabajo fueron: a identificar a nivel de especie aislamientos de Proteus siguiendo la combinación de los esquemas de Farmer y O'Hara; b determinar la utilidad del sistema comercial API 20E y de un esquema reducido de pruebas (agar TSI y agar MIO: movilidad, indol y ornitina, comparar estos procedimientos con la metodología convencional y evaluar su sensibilidad y especificidad, y c evaluar la utilidad del perfil proteico en la identificación de las distintas especies. Se estudiaron 205 aislamientos de Proteus spp. aislados en el período comprendido entre enero de 1998 y setiembre de 2004, recuperados de distintos materiales clínicos correspondientes a pacientes hospitalizados y ambulatorios atendidos en el Hospital de Clínicas. Los organismos fueron identificados mediante la metodología convencional, por el sistema API 20E y con un esquema reducido de pruebas; 48 de ellos fueron sometidos a un SDS-PAGE. API 20E identificó 79 de 87 aislamientos de P. mirabilis (90,8%, 103/103 del complejo P. vulgaris y 15/15 de P. penneri. Ocho aislamientos identificados como Proteus spp. resultaron ser P. mirabilis, al incluir una prueba adicional (maltosa. En la identificación, el esquema reducido coincidió en un 100% con la metodología convencional. A diferencia del sistema API 20E, el esquema reducido alcanza la correcta identificación de todas las especies en laboratorios de baja complejidad, sin la necesidad de pruebas adicionales. El perfil proteico permitió la correcta diferenciación de las tres especies, independientemente de las diferentes atipias de P. mirabilis.The objectives were: a to identify Proteus strains to species level, following Farmer's and O'Hara's conventional biochemical reactions; b to evaluate the sensitivity and specificity of both the API 20E method and a schema of reduced reactions (TSI and MIO agar: motility, indole and ornithine comparing them with conventional methodology, and c to evaluate the

  3. 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.

  4. Determinación de la condición fisiológica de la semilla de Capsicum spp y efecto del método de secado para su almacenamiento.

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    Cardozo C. I.

    2004-04-01

    Full Text Available Semillas del género Capsicum se caracterizaron fisiológicamente. Basados en datos de viabilidad y germinación se estimó la tolerancia a la desecación y conservación por lo cual se estableció la condición ortodoxa de las semillas. Sin embargo, se destaca la importancia de la variable vigor, que permitió establecer diferencias entre las especies respecto de su longevidad y deducir que los contenidos de humedad de 10% y 7.5% y temperatura de 10°C admiten mayor capacidad de conservación. Los cuatro métodos de secado (secado artificial, sílica gel, sol y sombra y los dos sistemas de ambiente controlado no presentaron diferencias significativas en viabilidad. Se detectaron niveles moderados de latencia en las semillas que pudieron interferir en la interpretación de los resultados. Palabras claves: C. annuum, C. frutescens, C. baccatum, viabilidad, germinación. ABSTRACT Determination of physiological status of Capsicum spp seed and efect of the dry method for storage. Capsicum spp seeds was physiologycal characterizated. Based on data of viability and germination to estimate the tolerance to drying and conservation, the orthodox condition of the seeds could be established. Nevertheless, the importance of the variable vigor stands out, that allowed to establish differences between the species with respect to its longevity, It was settled down that 10 and 7.5% of humidity contents and temperature of 10°C allow greater capacity of conservation. The four methods of seed drying and the capacity of conservation under two different controlled environment systems were not significant differences. Moderate levels of the seed dormancy could be interfered the interpretation of the results. Keywords: C. annuum, C. frutescens, C. baccalum, viability, germination

  5. Rattus norvegicus (Rodentia: Muridae Infected by Leishmania (Leishmania infantum (syn. Le. chagasi in Brazil

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    Fabiana de Oliveira Lara-Silva

    2014-01-01

    Full Text Available In the present study we surveyed the fauna of phlebotomine sand flies and small mammals in peridomestic areas from a Brazilian municipality where the American cutaneous leishmaniasis (ACL is endemic. A total of 608 female phlebotomine sand flies were captured during nine months in 2009 and 2010. Seven different species were represented with 60% of them being Lutzomyia intermedia and Lu. whitmani, both incriminated vectors of ACL. Lu. longipalpis, a proven vector of visceral leishmaniasis (VL was also captured at high proportion (12.8%. Genomic DNA analysis of 136 species-specific pools of female sand flies followed by molecular genotyping showed the presence of Leishmania infantum DNA in two pools of Lu. longipalpis. The same Leishmania species was found in one blood sample from Rattus norvegicus among 119 blood and tissue samples analysed. This is the first report of Le. infantum in R. norvegicus in the Americas and suggests a possible role for this rodent species in the zoonotic cycle of VL. Our study coincided with the reemergence of VL in Governador Valadares.

  6. Curcumin overcomes the inhibitory effect of nitric oxide on Leishmania.

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    Chan, Marion Man-Ying; Adapala, Naga Suresh; Fong, Dunne

    2005-04-01

    Upon Leishmania infection, macrophages are activated to produce nitrogen and oxygen radicals simultaneously. It is well established that the infected host cells rely on nitric oxide (NO) as the major weapon against the intracellular parasite. In India where leishmaniasis is endemic, the spice turmeric is used prolifically in food and for insect bites. Curcumin, the active principle of turmeric, is a scavenger of NO. This report shows that curcumin protects promastigotes and amastigotes of the visceral species, Leishmania donovani, and promastigotes of the cutaneous species, L. major, against the actions of S-nitroso-N-acetyl-D,L-penicillamine (SNAP) and DETANONOate, which release NO, 3-morpholino-sydnonimine hydrochloride (SIN-1), which releases NO and superoxide, and peroxynitrite, which is formed from the reaction of NO with superoxide. Thus, curcumin, as an antioxidant, is capable of blocking the action of both NO and NO congeners on the Leishmania parasite.

  7. Leishmania carbon metabolism in the macrophage phagolysosome- feast or famine?

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    McConville, Malcolm J; Saunders, Eleanor C; Kloehn, Joachim; Dagley, Michael J

    2015-01-01

    A number of medically important microbial pathogens target and proliferate within macrophages and other phagocytic cells in their mammalian hosts. While the majority of these pathogens replicate within the host cell cytosol or non-hydrolytic vacuolar compartments, a few, including protists belonging to the genus Leishmania, proliferate long-term within mature lysosome compartments.  How these parasites achieve this feat remains poorly defined. In this review, we highlight recent studies that suggest that Leishmania virulence is intimately linked to programmed changes in the growth rate and carbon metabolism of the obligate intra-macrophage stages. We propose that activation of a slow growth and a stringent metabolic response confers resistance to multiple stresses (oxidative, temperature, pH), as well as both nutrient limitation and nutrient excess within this niche. These studies highlight the importance of metabolic processes as key virulence determinants in Leishmania.

  8. Molecular crosstalks in Leishmania-sandfly-host relationships

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    Volf P.

    2008-09-01

    Full Text Available Sandflies (Diptera: Phlebotominae are vectors of Leishmania parasites, causative agents of important human and animal diseases with diverse manifestations. This review summarizes present knowledge about the vectorial part of Leishmania life cycle and parasite transmission to the vertebrate host. Particularly, it focuses on molecules that determine the establishment of parasite infection in sandfly midgut. It describes the concept of specific versus permissive sandfly vectors, explains the epidemiological consequences of broad susceptibility of permissive sandflies and demonstrates that genetic exchange may positively affect Leishmania fitness in the vector. Last but not least, the review describes recent knowledge about circulating antibodies produced by hosts in response to sandfly bites. Studies on specificity and kinetics of antibody response revealed that anti-saliva IgG could be used as a marker of host exposure to sandflies, i.e. as a useful tool for evaluation of vector control.

  9. Propiedades antifúngicas diferenciadas de extractos vegetales como estrategia potencial para la prevención, manejo y control del "moho verde" (Trichoderma spp.). En el cultivo de los hongos comestibles (Lentinula, Pleurotus).

    OpenAIRE

    Reyes Quintanar, Claudia Katia

    2014-01-01

    En la actualidad, se utilizan agroquímicos como el benomil para el control de Trichoderma en el cultivo de hongos comestibles; sin embargo, su uso es cada vez más restringido por la sociedad debido a su potencial toxicidad y persistencia en el ambiente. Los alimentos producidos con productos naturales son cada vez más demandados, debido a esto, el objetivo del presente estudio fue el obtener extractos vegetales con propiedades antifúngicas diferenciadas para el control del “moho verde” (Trich...

  10. From genomes to vaccines: Leishmania as a model.

    Science.gov (United States)

    Almeida, Renata; Norrish, Alan; Levick, Mark; Vetrie, David; Freeman, Tom; Vilo, Jaak; Ivens, Alasdair; Lange, Uta; Stober, Carmel; McCann, Sharon; Blackwell, Jenefer M

    2002-01-01

    The 35 Mb genome of Leishmania should be sequenced by late 2002. It contains approximately 8500 genes that will probably translate into more than 10 000 proteins. In the laboratory we have been piloting strategies to try to harness the power of the genome-proteome for rapid screening of new vaccine candidate. To this end, microarray analysis of 1094 unique genes identified using an EST analysis of 2091 cDNA clones from spliced leader libraries prepared from different developmental stages of Leishmania has been employed. The plan was to identify amastigote-expressed genes that could be used in high-throughput DNA-vaccine screens to identify potential new vaccine candidates. Despite the lack of transcriptional regulation that polycistronic transcription in Leishmania dictates, the data provide evidence for a high level of post-transcriptional regulation of RNA abundance during the developmental cycle of promastigotes in culture and in lesion-derived amastigotes of Leishmania major. This has provided 147 candidates from the 1094 unique genes that are specifically upregulated in amastigotes and are being used in vaccine studies. Using DNA vaccination, it was demonstrated that pooling strategies can work to identify protective vaccines, but it was found that some potentially protective antigens are masked by other disease-exacerbatory antigens in the pool. A total of 100 new vaccine candidates are currently being tested separately and in pools to extend this analysis, and to facilitate retrospective bioinformatic analysis to develop predictive algorithms for sequences that constitute potentially protective antigens. We are also working with other members of the Leishmania Genome Network to determine whether RNA expression determined by microarray analyses parallels expression at the protein level. We believe we are making good progress in developing strategies that will allow rapid translation of the sequence of Leishmania into potential interventions for disease

  11. Leishmania (L.) mexicana Infected Bats in Mexico: Novel Potential Reservoirs

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    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

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

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    Miriam Berzunza-Cruz

    2015-01-01

    Full Text Available 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.

  13. Further support for a palaearctic origin of Leishmania

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    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.

  14. The past, present, and future of Leishmania genomics and transcriptomics

    Science.gov (United States)

    Cantacessi, Cinzia; Dantas-Torres, Filipe; Nolan, Matthew J.; Otranto, Domenico

    2015-01-01

    It has been nearly 10 years since the completion of the first entire genome sequence of a Leishmania parasite. Genomic and transcriptomic analyses have advanced our understanding of the biology of Leishmania, and shed new light on the complex interactions occurring within the parasite–host–vector triangle. Here, we review these advances and examine potential avenues for translation of these discoveries into treatment and control programs. In addition, we argue for a strong need to explore how disease in dogs relates to that in humans, and how an improved understanding in line with the ‘One Health’ concept may open new avenues for the control of these devastating diseases. PMID:25638444

  15. LR1: a candidate RNA virus of Leishmania.

    OpenAIRE

    Tarr, P I; Aline, R F; Smiley, B L; Scholler, J; Keithly, J; Stuart, K

    1988-01-01

    Although viruses are important biological agents and useful molecular tools, little is known about the viruses of parasites. We report here the discovery of a candidate for an RNA virus in a kinetoplastid parasite. This potential virus, which we term LR1, is present in the promastigote form of the human pathogen Leishmania braziliensis guyanensis CUMC1-1A but not in 11 other stocks of Leishmania that were examined nor in Trypanosoma brucei. The candidate viral RNA has a size of approximately ...

  16. IDENTIFICATION OF DIFFERENT FUSARIUM SPP. IN ALLIUM SPP. IN GERMANY.

    Science.gov (United States)

    Boehnke, B; Karlovsky, P; Pfohl, K; Gamliel, A; Isack, Y; Dehne, H W

    2015-01-01

    In 2013 Allium cepa bulbs from different fields in Northern and Southern Germany, seeds and sets from onion breeders were analysed for infestation with Fusarium species. The same investigation was done in 2014 with different edible Allium spp. from local markets. Different Fusarium spp. were isolated and identified by morphological characterisation. 24 different Fusarium spp. were identified. The diversity of Fusarium spp. and the intensity of infestation was higher on edible bulbs compared to the younger sets and seeds. The analysed onions and other edible Allium spp. from local markets showed also high contents of different Fusarium species. The most prevalent identified Fusarium sp. in the analysed Allium spp. in Germany was Fusarium oxysporum which can cause the Fusarium Basal Rot, followed by Fusarium solani. Fusarium proliferatum, which can cause the Fusarium Salmon Blotch in onions, could be detected in about half of the sampled onion fields and in approximately 10% of all analysed onions from fields. Also in the onion sets, on the surface of the seeds and in other edible Allium spp. F. proliferatum could be identified. Besides F. proliferatum, further mycotoxin producing Fusarium spp. like Fusarium equiseti or Fusarium tricinctum were identified. Other Fusarium spp. like Fusarium sporotrichioides and Fusarium poae were first described in Allium sp. in this study. The two most prevalent Fusarium spp. F. oxysporum and F. solani are able to produce mycotoxins like enniatins, fumonisins, moniliformin and T-2 toxins. Fusarium sp. like F. proliferatum, F. equiseti and F. tricinctum are able to produce additional toxins like beauvericins, zearalenone and diacetoscirpenol. This high number of Fusarium spp., which are able to produce a broad spectrum of different mycotoxins, could be a potential health risk for human beings and livestock.

  17. Prevalence and risk factors associated with Leishmania infection in Trang Province, southern Thailand.

    Science.gov (United States)

    Manomat, Jipada; Leelayoova, Saovanee; Bualert, Lertwut; Tan-Ariya, Peerapan; Siripattanapipong, Suradej; Mungthin, Mathirut; Naaglor, Tawee; Piyaraj, Phunlerd

    2017-11-01

    Autochthonous cutaneous and visceral leishmaniasis (VL) caused by Leishmania martiniquensis and Leishmania siamensis have been considered emerging infectious diseases in Thailand. The disease burden is significantly underestimated, especially the prevalence of Leishmania infection among HIV-positive patients. A cross-sectional study was conducted to determine the prevalence and risk factors associated with Leishmania infection among patients with HIV/AIDS living in Trang province, southern Thailand, between 2015 and 2016. Antibodies against Leishmania infection were assayed using the direct agglutination test (DAT). DNA of Leishmania was detected by ITS1-PCR using the buffy coat. Species of Leishmania were also identified. Of 724 participants, the prevalence of Leishmania infection was 25.1% (182/724) using either DAT or PCR assays. Seroprevalence of Leishmania infection was 18.5% (134/724), while Leishmania DNA detected by the PCR method was 8.4% (61/724). Of these, 24.9% (180/724) were asymptomatic, whereas 0.3% (2/724) were symptomatic VL and VL/CL (cutaneous leishmaniasis). At least five species were identified: L. siamensis, L. martiniquensis, L. donovani complex, L. lainsoni, and L. major. Multivariate analysis showed that CD4+ levels Leishmania infection. Those who were PCR positive for Leishmania DNA were significantly associated with a detectable viral load, whereas non-injection drug use (NIDU) and CD4+ levels Leishmania seropositivity. A magnitude of the prevalence of underreporting Leishmania infection among Thai patients with HIV was revealed in this study. Effective public health policy to prevent and control disease transmission is urgently needed.

  18. Distribución geográfica de especies de Leishmania aisladas de pacientes consultantes al Instituto Nacional de Dermatología Federico Lleras Acosta, E.S.E., 1995-2005

    Directory of Open Access Journals (Sweden)

    Clemencia Elena Ovalle

    2006-10-01

    Full Text Available Introducción. El mapa de la distribución de especies de Leishmania en Colombia no se ha actualizado desde hace siete años. Objetivo. Describir la distribución de las especies de Leishmania a partir de los aislamientos de pacientes consultantes al Instituto Nacional de Dermatología durante el período 1995 a 2005 Materiales y métodos. Se realizó un estudio descriptivo de distribución geográfica de las especies de Leishmania, a partir de los aislamientos obtenidos de 137 pacientes consultantes al Instituto Nacional de Dermatología Federico Lleras Acosta E. S. E, con diagnóstico confirmado de leishmaniasis. Se revisó la historia clínica obteniendo la información de forma clínica, sitio probable de infección, edad y género. Se tomaron y cultivaron muestras de las lesiones y los aislamientos se tipificaron por anticuerpos monoclonales, comparando el 10% de los resultados con los obtenidos por reacción en cadena de la polimerasa (PCR y patrones isoenzimáticos. Resultados. La frecuencia encontrada de los 137 aislamientos fue: Leishmania panamensis 74,45%; Leishmania braziliensis15,33%; Leishmania guyanensis 0,73%; Complejo Leishmania mexicana 3,65%, Leishmania mexicana 5,11% y el 0,73% restante correspondió a un aislamiento que no se pudo caracterizar por anticuerpos monoclonales. La distribución de L. braziliensis, L. panamensis y L. guyanensis fue concordante con lo reportado en estudios anteriores pero para las especies del complejo L. mexicana se encontraron procedencias no reportadas previamente: Caldas, Santander, Cundinamarca, Caquetá, Casanare, Cauca y Valle del Cauca. Conclusión. El complejo L. mexicana presenta una distribución más amplia de la reportada previamente. La utilidad de los monoclonales especie-específicos, los patrones isoenzimáticos y la PCR para L. mexicana y L. amazonensis fue limitada.

  19. Elaboración de harina de 3 variedades de plátano verde (musa spp) y su uso como materia prima para la planificación.

    OpenAIRE

    Jiménez Márquez, Erika

    2012-01-01

    En México y otros países, durante el manejo la cosecha, una fracción de los racimos de plátano se desprende y se pierde, generando problemas ambientales. Algunas alternativas para reducir esos desechos son la producción de harinas de plátano verde; pues, representan una fuente alternativa para la obtención de almidón, que actualmente se obtiene de maíz y papa. El almidón es uno de los nutrientes más importantes por su alto valor energético. Existen fracciones de almidón denominadas almidón re...

  20. Irrigação suplementar da cana-de-açúcar (Saccharum spp): um modelo de análise de decisão para o estado de Alagoas

    OpenAIRE

    Márcio Aurélio Lins dos Santos

    2005-01-01

    As usinas de açúcar e álcool no Estado de Alagoas têm grande importância econômica, para usar sua capacidade instalada torna-se necessário o uso de técnicas que aumentem sua produtividade e/ou diminuam os custos de produção da cana-deaçúcar. A irrigação suplementar apresenta-se como uma destas alternativas. A tecnologia de irrigação se tornou uma alternativa rentável para as condições do Estado de Alagoas. Com base na análise de viabilidade da tecnologia de irrigação em canasoca, a árvore de...

  1. Seroprevalence rates of antibodies againstLeishmania infantum and other protozoan and rickettsial parasites in dogs

    Directory of Open Access Journals (Sweden)

    Silvana de Cássia Paulan

    Full Text Available Canine visceral leishmaniasis (CVL is caused by the protozoan Leishmania infantum, which infects dogs and humans in many regions of Brazil. The present study involved an indirect fluorescent antibody test (IFAT to analyze L. infantum,Ehrlichia spp., Babesia canis,Toxoplasma gondii and Neospora caninuminfection rates in serum samples from 93 dogs in a rural settlement in Ilha Solteira, SP, Brazil. The seroprevalence rates of anti-L. infantum, anti-Ehrlichia, anti-B. canis, anti-T. gondii and anti-N. caninum antibodies were 37.6%, 75.3%, 72%, 47.3% and 6.4%, respectively. In addition to IFAT, direct microscopic examination of popliteal lymph node aspirates revealed 26.9% of CVL positive dogs. Serological tests revealed that 17.2% of the dogs were seropositive for a single parasite, 29% for two parasites, 33% for three, 16.1% for four, and 1.1% for five parasites, while 3.2% were seronegative for five parasites. The presence of antibodies against these parasites in serum samples from dogs confirmed their exposure to these parasites in this rural area. Because of the potential zoonotic risk of these diseases, mainly leishmaniasis, ehrlichiosis and toxoplasmosis, special attention should focus on programs for the improvement of diagnostic assays and control measures against these parasites.

  2. Excreción de promastigotos de Leishmania pifanoi por Lutzomyia youngi experimentalmente infectada

    Directory of Open Access Journals (Sweden)

    Elina Rojas

    1995-12-01

    Full Text Available Se describe el desarrollo poblacional promastigótico de Leishmania pifanoi en Lutzomyia youngi experimentalmente infectada y mantenida con sacarosa al 50% bajo condiciones constantes de temperatura y humedad. Se reconocen dos etapas para la diferenciación y el crecimiento de los parásitos entre las dos y ciento veinte horas postprandiales. Hasta 48 horas tiene lugar la diferenciación pleomórfica de amastigotos en promastigotos cortos, que se multiplican por división binaria hasta las 60 horas, cuando ocurre la ruptura de la membrana peritrófica. La segunda etapa tiene lugar entre las 72 y 96 horas cuando algunos parásitos migran hacia la válvula esofágica y los demás parásitos libres son excretados en gotitas fecales como promastigotos grandes y activos. Las primeras gotitas excretadas dan reacción positiva a glucosa o contienen cristales de urato. El exceso de promastigotos de la segunda fase de desarrollo es eliminado en las últimas excretas que dan reacción positiva con las pruebas Hemoscreen y Biuret para proteínas totales y también para glucosa, y constituyen el 82% del total de gotas excretadas. La excreción de parásitos por Lu. youngi es fase normal del desarrollo de L. pifanoi en un vector.

  3. Componentes antiinflamatórios na saliva do Lutzomyia longipalpis, vetor da Leishmania chagasi

    Directory of Open Access Journals (Sweden)

    Marta Chagas Monteiro

    2005-06-01

    Full Text Available A inoculação da saliva de vetores na pele do hospedeiro é importante tanto para a alimentação do inseto quanto para a transmissão e estabelecimento de várias infecções. Em leishmaniose, vários estudos demonstram que a saliva dos vetores Lutzomyia e Phlebotomus contém substâncias com atividades imunossupressoras, imunomodulatórias, vasodilatadoras, anti-plaquetárias e anticoagulantes. Os componentes salivares auxiliam a alimentação do inseto através do aumento do fluxo sanguíneo, assim como induzem a imunossupressão no hospedeiro, o que é fundamental para o estabelecimento da infecção por Leishmania. Neste trabalho foi observado que a saliva induz a produção de IL-10, citocina antiinflamatória, não alterando a produção de IFN-g , citocina próinflamatória, no foco da inflamação. Além disso, a saliva potencializa o edema induzido por carragenina.

  4. Excreción de promastigotos de Leishmania pifanoi por Lutzomyia youngi experimentalmente infectada

    Directory of Open Access Journals (Sweden)

    Rojas Elina

    1995-01-01

    Full Text Available Se describe el desarrollo poblacional promastigótico de Leishmania pifanoi en Lutzomyia youngi experimentalmente infectada y mantenida con sacarosa al 50% bajo condiciones constantes de temperatura y humedad. Se reconocen dos etapas para la diferenciación y el crecimiento de los parásitos entre las dos y ciento veinte horas postprandiales. Hasta 48 horas tiene lugar la diferenciación pleomórfica de amastigotos en promastigotos cortos, que se multiplican por división binaria hasta las 60 horas, cuando ocurre la ruptura de la membrana peritrófica. La segunda etapa tiene lugar entre las 72 y 96 horas cuando algunos parásitos migran hacia la válvula esofágica y los demás parásitos libres son excretados en gotitas fecales como promastigotos grandes y activos. Las primeras gotitas excretadas dan reacción positiva a glucosa o contienen cristales de urato. El exceso de promastigotos de la segunda fase de desarrollo es eliminado en las últimas excretas que dan reacción positiva con las pruebas Hemoscreen y Biuret para proteínas totales y también para glucosa, y constituyen el 82% del total de gotas excretadas. La excreción de parásitos por Lu. youngi es fase normal del desarrollo de L. pifanoi en un vector.

  5. Comparação entre método bioquímico e reação em cadeia de polimerase para identificação de Lactobacillus spp., isolados de aves Comparison between biochemical and polymerase chain reaction methods for the identification of Lactobacillus spp. isolated from chickens

    Directory of Open Access Journals (Sweden)

    M.R. Barros

    2009-04-01

    Full Text Available Lactobacilos foram isolados do inglúvio e cecos de reprodutoras pesadas e caracterizados como Gram-positivo, catalase negativo, produtores de gás em glicose e não produtores de H2S em triple sugar iron e pela fermentação de carboidratos. Utilizaram-se os iniciadores: Lac 1/23-10C para detecção de Lactobacillus acidophilus, L. crispatus, L. amylovorus, L. gasseri, L. helveticus e L. jensenii; Lac 2/LU-1' para L. acidophilus; Fer 3/Fer 4 para L. fermentum; Reu 1/Reu 2 para L. reuteri e Sal 1 e Sal 2 para L. salivarius. L. reuteri e L. salivarius foram identificados pela reação em cadeia de polimerase (PCR e pelo teste bioquímico, enquanto L. acidophilus, L. fermentum e Lactobacillus sp. somente pelo teste bioquímico. Os resultados obtidos na PCR foram mais precisos quando comparados aos obtidos com o método bioquímico, que demonstrou ser subjetivo devido às variações na fermentação de carboidratos, principalmente na diferenciação entre L. fermentum e L. reuteri.Lactobacilli were isolated from crops and ceca of broiler breeders and characterized by positive Gram staining, negative catalase test, production of gas from glucose, and negative for H2S production from triple sugar iron, and carbohydrates fermentation. Primers: Lac1/23-10C for detecting Lactobacillus acidophilus, L. crispatus, L. amylovorus, L. gasseri, L. helveticus, and L. jensenii; Lac2/LU-1' for L. acidophilus; Fer3/Fer4 for L. fermentum; Reu1/Reu2 for L. reuteri, and Sal1/Sal2 for L. salivarius were used. L. reuteri and L. salivarius were identified by both polymerase chain reaction (PCR and biochemical tests. However, L. acidophilus, L. fermentum, and Lactobacillus sp. were only identified by biochemical tests. PCR results were more precise, considering the variability of carbohydrate fermentation among the strains, especially for identifying L. fermentum and L. reuteri.

  6. Serological and molecular survey of Leishmania parasites in apparently healthy dogs in the West Bank, Palestine

    Directory of Open Access Journals (Sweden)

    Hamarsheh Omar

    2012-08-01

    Full Text Available Abstract Background Canine visceral leishmaniasis (CVL is caused by Leishmania infantum in all Mediterranean countries. The Leishmania parasite is transmitted by the bite of a corresponding sand fly vector and primarily maintained in nature by wild and domestic reservoirs, including dogs, foxes and jackals. Infected dogs are the primary reservoir host in endemic regions and are the most significant risk disposing humans to infection. The present study aimed at assessing the prevalence of infection with Leishmania and identification of Leishmania infantum in domestic dogs in the West Bank, Palestine. Methods The infection rate among domestic dogs collected from seven districts in the Palestinian West Bank was investigated by examination of parasites in culture from the buffy coat using serological and molecular methods; based on ELISA, internal transcribed spacer 1 (ITS1 and cysteine protease (CPB PCR. Results Out of 215 dogs examined for Leishmania, 36 (16.7% were positive in at least one method. Twenty three animals (11.5% were positive for Leishmania DNA, whereas, ELISA and culture revealed 16 (7.5%, and 4 (1.5% respectively. CPB-PCR on one of three culture-positive isolates revealed Leishmania infantum as the causative agent for Leishmania infection in dogs. Conclusions Our study showed that canine leishmania infection is prevalent with varying degrees in all the seven studied districts in Palestine despite the absence of human VL cases in 4 of these districts. The causative agent was confirmed to be Leishmania infantum.

  7. Studies on Stibanate unresponsive isolates of Leishmania donovani

    Indian Academy of Sciences (India)

    Unknown

    503. Keywords. Drug resistance; Leishmania donovani; sodium stibogluconate; visceral leishmaniasis ... cases to less than 5% in treated ones (Zuckerman and. Lainson 1977). Later on .... Effect of Stibanate on in vitro growth of Ag83 promastigotes. (A) Virulent .... In rural areas where health care facilities are not adequate ...

  8. Natural Sesquiterpene Lactones Induce Oxidative Stress in Leishmania mexicana

    NARCIS (Netherlands)

    Barrera, P.; Sulsen, V.P.; Lozano, E.; Rivera, M.; Beer, M.F.; Tonn, C.; Martino, V.S.; Sosa, M.A.

    2013-01-01

    Leishmaniasis is a worldwide parasitic disease, caused by monoflagellate parasites of the genus Leishmania. In the search for more effective agents against these parasites, the identification of molecular targets has been attempted to ensure the efficiency of drugs and to avoid collateral damages on

  9. Bibliography of Leishmania and Leishmanial Diseases. Volume 2

    Science.gov (United States)

    1980-08-01

    Leishmaniosi visceizile latente in adulto rivelatasi PHOTINOS, P. B, (1930A) Formes atypiques nel corso di una, Infezione tifoidea . N infiltries en... fiebre recurrente y con gran flagelle’s au cours des leishmanioses cantidad de leishmanias en sangre penl- ’cliniques et expegrinientales. Ann.ferlca

  10. 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

    Roč. 198, č. 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

  11. Generation of species-specific DNA probes for Leishmania aethiopica

    NARCIS (Netherlands)

    Laskay, T.; Kiessling, R.; Rinke deWit, T. F.; Wirth, D. F.

    1991-01-01

    We report here the cloning of kinetoplast DNA (kDNA) sequences from Leishmania aethiopica in order to develop a specific and sensitive method for the identification of the parasite. Analysis of the cloned kDNA sequences showed different taxonomic specificities demonstrating sequence diversity within

  12. Leishmania serology in the diagnosis of cutaneous leishmaniasis

    International Nuclear Information System (INIS)

    Mashood, A.H.; Malik, N.; Abbasi, S.

    2013-01-01

    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)

  13. Genetic Diversity in Natural Populations of New World Leishmania

    Directory of Open Access Journals (Sweden)

    Cupolillo Elisa

    1998-01-01

    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.

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

    Science.gov (United States)

    Dostálová, Anna; Volf, Petr

    2012-12-03

    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.

  15. Biosynthesis of silver nanoparticles by Leishmania tropica | Rahi ...

    African Journals Online (AJOL)

    A novel biosynthesis route for Silver Nanoparticles (Ag-NPs) was attempted in the present study using Leishmania tropica the causative agent of cutaneous leishmaniasis in different countries, particularly in Mediterranean region in Iraq. Silver nanoparticles were successfully synthesized from AgNO3 by reduction of ...

  16. Recognition of Leishmania antigens by T lymphocytes from nonexposed individuals

    DEFF Research Database (Denmark)

    Kemp, M; Hansen, M B; Theander, T G

    1992-01-01

    Crude antigen preparations of Leishmania promastigote sonicates were found to induce in vitro proliferation and gamma interferon production in peripheral blood mononuclear cells (PBMC) from individuals without known exposure to the parasite. The proliferating cells were mainly CD2-positive T cell...

  17. Activation of human T lymphocytes by Leishmania lipophosphoglycan

    DEFF Research Database (Denmark)

    Kemp, M; Theander, T G; Handman, E

    1991-01-01

    This study describes Leishmania antigen-induced activation of lymphocytes isolated from Kenyan donors, previously treated for visceral leishmaniasis, and from Danish and Kenyan controls. Peripheral blood mononuclear cells (PBMC) from cured Kala-Azar patients proliferated and produced Interferon......, the results suggest that human T lymphocytes can respond to glycolipid antigens....

  18. In vitro and in vivo activity of an organic tellurium compound on Leishmania (Leishmania chagasi.

    Directory of Open Access Journals (Sweden)

    Isabella Aparecida Salerno Pimentel

    Full Text Available Tellurium compounds have shown several biological properties and recently the leishmanicidal effect of one organotellurane was demonstrated. These findings led us to test the effect of the organotellurium compound RF07 on Leishmania (Leishmania chagasi, the agent of visceral leishmaniasis in Latin America. In vitro assays were performed in L. (L. chagasi-infected bone marrow derived macrophages treated with different concentrations of RF07. In in vivo experiments Golden hamsters were infected with L. (L. chagasi and injected intraperitoneally with RF07 whereas control animals received either Glucantime or PBS. The effect of RF07 on cathepsin B activity of L. (L. chagasi amastigotes was assayed spectrofluorometrically using fluorogenic substrates. The main findings were: 1 RF07 showed significant leishmanicidal activity against intracellular parasites at submicromolar concentrations (IC50 of 529.7±26.5 nM, and the drug displayed 10-fold less toxicity to macrophages (CC50 of 5,426±272.8 nM; 2 kinetics assays showed an increasing leishmanicidal action of RF07 at longer periods of treatment; 3 one month after intraperitoneal injection of RF07 L. (L. chagasi-infected hamsters showed a reduction of 99.6% of parasite burden when compared to controls that received PBS; 4 RF07 inhibited the cathepsin B activity of L. (L. chagasi amastigotes. The present results demonstrated that the tellurium compound RF07 is able to destroy L. (L. chagasi in vitro and in vivo at concentrations that are non toxic to the host. We believe these findings support further study of the potential of RF07 as a possible alternative for the chemotherapy of visceral leishmaniasis.

  19. The Genetic Relationship between Leishmania aethiopica and Leishmania tropica Revealed by Comparing Microsatellite Profiles.

    Science.gov (United States)

    Krayter, Lena; Schnur, Lionel F; Schönian, Gabriele

    2015-01-01

    Leishmania (Leishmania) aethiopica and L. (L.) tropica cause cutaneous leishmaniases and appear to be related. L. aethiopica is geographically restricted to Ethiopia and Kenya; L. tropica is widely dispersed from the Eastern Mediterranean, through the Middle East into eastern India and in north, east and south Africa. Their phylogenetic inter-relationship is only partially revealed. Some studies indicate a close relationship. Here, eight strains of L. aethiopica were characterized genetically and compared with 156 strains of L. tropica from most of the latter species' geographical range to discern the closeness. Twelve unlinked microsatellite markers previously used to genotype strains of L. tropica were successfully applied to the eight strains of L. aethiopica and their microsatellite profiles were compared to those of 156 strains of L. tropica from various geographical locations that were isolated from human cases of cutaneous and visceral leishmaniasis, hyraxes and sand fly vectors. All the microsatellite profiles were subjected to various analytical algorithms: Bayesian statistics, distance-based and factorial correspondence analysis, revealing: (i) the species L. aethiopica, though geographically restricted, is genetically very heterogeneous; (ii) the strains of L. aethiopica formed a distinct genetic cluster; and (iii) strains of L. aethiopica are closely related to strains of L. tropica and more so to the African ones, although, by factorial correspondence analysis, clearly separate from them. The successful application of the 12 microsatellite markers, originally considered species-specific for the species L. tropica, to strains of L. aethiopica confirmed the close relationship between these two species. The Bayesian and distance-based methods clustered the strains of L. aethiopica among African strains of L. tropica, while the factorial correspondence analysis indicated a clear separation between the two species. There was no correlation between

  20. Preparation of live attenuated leishmania parasites by using laser technology

    Science.gov (United States)

    Hussain, Nabiha; Alkhouri, Hassan; Haddad, Shaden

    2018-05-01

    Leishmaniasis is a parasitic disease of humans, affecting the skin, mucosal and/or internal organs, caused by flagellate protozoa Leishmania of the Trypanosomatidae family. Leishmania would be one for which a vaccine could be developed with relative ease. Many studies mount an effective response that resolves the infection and confers solid immunity to reinfection and suggesting that infection may be a prerequisite for immunological memory. Genetically altered live attenuated parasites with controlled infectivity could achieve such immunological memory. Recent concepts include use of genetically modified live-attenuated Leishmania parasites, and proteomics approach for the search of a cross-protective leishmanial vaccine that would ideally protect against both cutaneous and visceral forms of the disease. No licensed vaccine is available till date against any form of leishmaniasis. The present study evaluated role of laser technology in development of a safe live Leishmania vaccine, a vaccine is a biological preparation that improves immunity to a particular disease, and is often made from weakened or killed forms of LPs. The parasite culture was expanded in RPMI 1640 medium with 10% fetal calf serum (FCS) and grown until stationary phase for experiments. 80 samples of leishmania promastigotes (Culture media of LPs) were exposed to Nd:YAG laser (wavelength 1064 nm, single spot or double) with different outputs powers (7w, 100 Hz, 99.03w/cm2, 0.99 J/cm2 and 8 w, 100 Hz, 113.18w/cm2 1.13J/cm2)) for suitable exposer times. The effect of semiconductor laser (wavelength 810 nm, 7w, 2000 Hz, 99.03w/cm2, 0.05 J/cm2) or (7 w, 500 Hz, 99.03 w/cm2, 0.2J/cm2) single spot or double with long exposure times. The viability of Leishmania parasites was measured using XTT method; viable parasites were decreased with long exposure times. XTT test referred both these wavelengths were effective in killing percentage of Leishmania promastigotes, the remaining were devoid flagellum that

  1. Níveis de dano econômico para decisão de controle de capim-arroz (Echinochloa spp. em arroz irrigado (Oryza sativa Economic threshold level for barnyardgrass (Echinochloa spp. control decision in flooded rice (Oryza sativa

    Directory of Open Access Journals (Sweden)

    L. Galon

    2007-12-01

    Full Text Available O nível de dano econômico (NDE para monitoramento das infestações de capim-arroz em lavouras de arroz irrigado permite adotar medidas de manejo na cultura e usar herbicidas de maneira racional. Objetivou-se com o trabalho determinar NDE para capim-arroz, presente em lavouras de arroz irrigado, calculado na base de um único ano, em função de populações de capim-arroz, de cultivares de arroz e de épocas de entrada de água na lavoura. Para isso, foram conduzidos dois experimentos em delineamento experimental completamente casualizado, sem repetição. No primeiro experimento, os tratamentos foram constituídos de seis cultivares de arroz: BRS-Atalanta e IRGA 421 (ciclo muito curto, IRGA 416, IRGA 417 e Avaxi (ciclo curto e BRS-Fronteira (ciclo médio; e dez populações de capim-arroz. No segundo experimento usaram-se como tratamentos épocas de início da irrigação: 1, 10 e 20 dias após aplicação dos tratamentos herbicidas (DAT; e populações de capim-arroz. Os NDEs estimados para o capim-arroz variaram em função das práticas de manejo adotadas na cultura do arroz irrigado. Os NDEs estimados para os cultivares de arroz IRGA 416 e 417 foram superiores aos dos demais cultivares quando em competição com capim-arroz. O atraso da entrada de água diminui os NDEs para o cultivar de arroz BRS-Pelota. Aumentos nas perdas de produtividade por unidade de planta daninha, no potencial de produtividade da cultura, no valor do produto colhido e na eficiência do herbicida e diminuição do custo de controle reduziram os NDEs, tornando econômica a adoção de práticas de manejo sob baixas populações de capim-arroz em competição com arroz irrigado.Economic weed threshold (EWT for monitoring barnyardgrass infestation in flooded rice farms allows the adoption of crop management measurements and a rational use of herbicide. The objective of this study was to determine EWT for barnyardgrass based on a single year in function of

  2. Validación de una metodología multirresiduo para la determinación de residuos de plaguicidas en fresa (Fragraria spp. por cromatografía de gases

    Directory of Open Access Journals (Sweden)

    Mauricio Montaño Garcés

    2011-03-01

    Full Text Available En este estudio se describe la validación de una metodología analítica multirresiduo para la determinación simultáneade 19 compuestos organoclorados, organofosforados y organonitrogenados en fresa. Los plaguicidas se extrajeroncon acetato de etilo, los extractos se limpiaron por cromatografía de permeación por geles (GPC, el análisis se llevó a cabo por cromatografía de gases de alta resolución (CGAR con inyección en modo splitless pulsado y detección simultánea por microcaptura electrónica ( Normal 0 21 false false false ES X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} μ-ECDy nitrógeno-fósforo (NPD acopiadosen paralelo. La metodología es específica, selectiva, precisa y robusta. Las curvas de calibración en extracto de matriz fortificado son lineales en un rango de concentración de 0.04-5.00 mg/kg, con cantidad mínima detectable (CMD entre 0.007-0.50 mg/kg y cantidad mínima cuantificable (CMC entre 0.01-1.00 mg/kg. Los porcentajes de recuperación estuvieron entre el 80-110 %, en el rango de calibración para la mayoría de plaguicidas  evaluados. Con respecto a la distribución del analito en la muestra, se demostró su homogeneidad. Se aplicó la  metodología a muestras reales, encontrándose principalmente residuos de Captan por debajo de los límites m

  3. [Molecular typing of Leishmania (Leishmania) amazonensis and species of the subgenus Viannia associated with cutaneous and mucosal leishmaniasis in Colombia: A concordance study].

    Science.gov (United States)

    Ovalle-Bracho, Clemencia; Camargo, Carolina; Díaz-Toro, Yira; Parra-Muñoz, Marcela

    2018-03-15

    Multilocus enzyme electrophoresis (MLEE) is the reference standard for the characterization of Leishmania species. The test is restricted to specialized laboratories due to its technical complexity, cost, and time required to obtain results. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is used to identify Leishmania species. To establish the concordance between the two tests as identifying methods for circulating species in Colombia. A total of 96 isolates from patients with cutaneous or mucosal leishmaniasis were selected and identified by MLEE and PCR-RFLP with miniexon and hsp70 as the molecular targets, which were used sequentially. Restriction enzymes HaeIII and BccI were similarly applied. Cohen's kappa coefficient and the 95% confidence interval (CI) were calculated. The kappa coefficient and the 95% CI between MLEE and PCR-RFLP displayed "very good" concordance with a coefficient of 0.98 (CI95%: 0.98 to 1.00). The identified species were Leishmania Viannia braziliensis, Leishmania Viannia panamensis, Leishmania Viannia guyanensis and Leishmania Leishmania amazonensis. A total of 80 of the 96 isolates were sequenced and the results obtained by PCR-RFLP were confirmed. Due to the concordance obtained between tests results with the amplification of the genes miniexon and hsp70, PCR-RFLP is proposed as an alternative for identifying circulating Leishmania species in Colombia.

  4. Leishmania chagasi/infantum : further investigations on Leishmania tropisms in atypical cutaneous and visceral leishmaniasis foci in Central America

    NARCIS (Netherlands)

    Campos Ponce, M.; Ponce, C.; Ponce, E; Maingon, R.D.

    2005-01-01

    In Central America, apparently genetically identical Leishmania chagasi/infantum parasites cause cutaneous (CL) and visceral leishmaniasis (VL), the latter being more frequent in young children. The present study investigated if there were pathology-related differences in virulence between Honduran

  5. Leishmania chagasi/infantum: further investigations on Leishmania tropisms in atypical cutaneous and visceral leishmaniasis foci in Central America.

    NARCIS (Netherlands)

    Campos Ponce, M.; Ponce, C.; Ponce, E.; Maingon, R.D.

    2005-01-01

    In Central America, apparently genetically identical Leishmania chagasi/infantum parasites cause cutaneous (CL) and visceral leishmaniasis (VL), the latter being more frequent in young children. The present study investigated if there were pathology-related differences in virulence between Honduran

  6. Differentiation of Leishmania (Viannia) panamensis and Leishmania (V.) guyanensis using BccI for hsp70 PCR-RFLP.

    Science.gov (United States)

    Montalvo Alvarez, Ana Margarita; Nodarse, Jorge Fraga; Goodridge, Ivón Montano; Fidalgo, Lianet Monzote; Marin, Marcel; Van Der Auwera, Gert; Dujardin, Jean-Claude; Bernal, Iván Darío Velez; Muskus, Carlos

    2010-05-01

    Leishmania panamensis and Leishmania guyanensis are two species of the subgenus Viannia that are genetically very similar. Both parasites are usually associated with cutaneous leishmaniasis, but also have the potential to cause the mucocutaneous form of the disease. In addition, the study of foci and consequently the identification of vectors and probable reservoirs involved in transmission require a correct differentiation between both species, which is important at epidemiological level. We explored the possibility of identifying these species by using restriction fragment length polymorphisms (RFLP) in the gene coding for heat-shock protein 70 (hsp70). Previously, an hsp70 PCR-RFLP assay proved to be very effective in differentiating other Leishmania species when HaeIII is used as restriction enzyme. Based on hsp70 sequences analysis, BccI was found to generate species-specific fragments that can easily be recognized by agarose gel electrophoresis. Using the analysis of biopsies, scrapings, and parasite isolates previously grouped in a cluster comprising both L. panamensis and L. guyanensis, we showed that our approach allowed differentiation of both entities. This offers the possibility not only for identification of parasites in biological samples, but also to apply molecular epidemiology in certain countries of the New World, where several Leishmania species could coexist. Copyright 2009 Royal Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. All rights reserved.

  7. Actividad contra Leishmania sp. (Kinetoplastida: Trypanosomatidae de plantas en una Reserva Biológica de Costa Rica

    Directory of Open Access Journals (Sweden)

    Misael Chinchilla-Carmona

    2014-09-01

    Full Text Available La leishmaniosis es una enfermedad muy importante para el ser humano pero su tratamiento es bastante difícil. Por esta razón muchos investigadores han venido buscando plantas que contengan componentes químicos activos contra esta parasitosis. En Costa Rica no se tienen estudios al respecto y por eso el objetivo de este estudio fue la búsqueda de componentes activos contra Leishmania sp. en plantas de Costa Rica; 67 especies de la Reserva Biológica Alberto Manuel Brenes (REBAMB fueron seleccionadas para realizar este trabajo. Para ello se prepararon extractos crudos hidro-alcohólicos de material fresco o desecado de raíz, tallo, hojas maduras o tiernas, flores y frutos inmaduros o maduros. Usando pruebas presuntivas y luego específicas, se analizó el efecto de tales extractos sobre una cepa de Leishmania (OCR. Se consideraron plantas promisorias solamente aquellas en que al menos una de sus partes presentara un CI50<100µg/mL. Las plantas seleccionadas fueron: Bocconia frutescens, Clematis dioica,Cordia megalantha, Eugenia austin-smithii, Guarea bullata, Guateria tonduzii, Mikania holwayana, Nectandra membranacea,Neurolaena lobata, Persea povedae, Piper auritum, Rollinia pittieri, Solanum arboreum, Tetrorchidium euryphyllum, Witheringia solanacea y Zanthoxylum juniperinum. Existió una ligera tendencia de positividad mayor para los extractos frescos y la actividad se presentó en una y hasta más de cuatro partes de la planta. La mayoría de los extractos activos no fueron tóxicos. Se discute la importancia de estos nuevos hallazgos, en relación con el nuevo conocimiento científico y su proyección en el tratamiento de la leishmaniosis.

  8. The use of kDNA minicircle subclass relative abundance to differentiate between Leishmania (L.) infantum and Leishmania (L.) amazonensis.

    Science.gov (United States)

    Ceccarelli, Marcello; Galluzzi, Luca; Diotallevi, Aurora; Andreoni, Francesca; Fowler, Hailie; Petersen, Christine; Vitale, Fabrizio; Magnani, Mauro

    2017-05-16

    Leishmaniasis is a neglected disease caused by many Leishmania species, belonging to subgenera Leishmania (Leishmania) and Leishmania (Viannia). Several qPCR-based molecular diagnostic approaches have been reported for detection and quantification of Leishmania species. Many of these approaches use the kinetoplast DNA (kDNA) minicircles as the target sequence. These assays had potential cross-species amplification, due to sequence similarity between Leishmania species. Previous works demonstrated discrimination between L. (Leishmania) and L. (Viannia) by SYBR green-based qPCR assays designed on kDNA, followed by melting or high-resolution melt (HRM) analysis. Importantly, these approaches cannot fully distinguish L. (L.) infantum from L. (L.) amazonensis, which can coexist in the same geographical area. DNA from 18 strains/isolates of L. (L.) infantum, L. (L.) amazonensis, L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, and 62 clinical samples from L. (L.) infantum-infected dogs were amplified by a previously developed qPCR (qPCR-ML) and subjected to HRM analysis; selected PCR products were sequenced using an ABI PRISM 310 Genetic Analyzer. Based on the obtained sequences, a new SYBR-green qPCR assay (qPCR-ama) intended to amplify a minicircle subclass more abundant in L. (L.) amazonensis was designed. The qPCR-ML followed by HRM analysis did not allow discrimination between L. (L.) amazonensis and L. (L.) infantum in 53.4% of cases. Hence, the novel SYBR green-based qPCR (qPCR-ama) has been tested. This assay achieved a detection limit of 0.1 pg of parasite DNA in samples spiked with host DNA and did not show cross amplification with Trypanosoma cruzi or host DNA. Although the qPCR-ama also amplified L. (L.) infantum strains, the C q values were dramatically increased compared to qPCR-ML. Therefore, the combined analysis of C q values from qPCR-ML and qPCR-ama allowed to distinguish L. (L.) infantum and L. (L.) amazonensis in 100% of tested samples

  9. Role of pro-inflammatory cytokine IL-17 in Leishmania pathogenesis and in protective immunity by Leishmania vaccines.

    Science.gov (United States)

    Banerjee, Antara; Bhattacharya, Parna; Joshi, Amritanshu B; Ismail, Nevien; Dey, Ranadhir; Nakhasi, Hira L

    2016-11-01

    The clinical outcome of Leishmania pathogenesis ranges from active skin lesions to fatal visceral dissemination and severely impaired T cell immunity. It is well established that a strong Th1 immune response is protective against cutaneous forms of the disease, however a mixed Th1/Th2 response is most commonly observed against visceral infections as evident from previous studies. Aside from Th1/Th2 cytokines, the pro-inflammatory IL-17 cytokine family plays an important role in the clearance of intracellular pathogens. In Leishmania induced skin lesions, IL-17 produced by Th17 cells is shown to exacerbate the disease, suggesting a role in pathogenesis. However, a protective role for IL-17 is indicated by the expansion of IL-17 producing cells in vaccine-induced immunity. In human visceral leishmaniasis (VL) it has been demonstrated that IL-17 and IL-22 are associated with protection against re-exposure to Leishmania, which further suggests the involvement of IL-17 in vaccine induced protective immunity. Although there is no vaccine against any form of leishmaniasis, the development of genetically modified live attenuated parasites as vaccine candidates prove to be promising, as they successfully induce a robust protective immune response in various animal models. However, the role of IL-17 producing cells and Th17 cells in response to these vaccine candidates remains unexplored. In this article, we review the role of IL-17 in Leishmania pathogenesis and the potential impact on vaccine induced immunity, with a special focus on live attenuated Leishmania parasites. Published by Elsevier Inc.

  10. Unraveling the genetic diversity and phylogeny of Leishmania RNA virus 1 strains of infected Leishmania isolates circulating in French Guiana.

    Science.gov (United States)

    Tirera, Sourakhata; Ginouves, Marine; Donato, Damien; Caballero, Ignacio S; Bouchier, Christiane; Lavergne, Anne; Bourreau, Eliane; Mosnier, Emilie; Vantilcke, Vincent; Couppié, Pierre; Prevot, Ghislaine; Lacoste, Vincent

    2017-07-01

    Leishmania RNA virus type 1 (LRV1) is an endosymbiont of some Leishmania (Vianna) species in South America. Presence of LRV1 in parasites exacerbates disease severity in animal models and humans, related to a disproportioned innate immune response, and is correlated with drug treatment failures in humans. Although the virus was identified decades ago, its genomic diversity has been overlooked until now. We subjected LRV1 strains from 19 L. (V.) guyanensis and one L. (V.) braziliensis isolates obtained from cutaneous leishmaniasis samples identified throughout French Guiana with next-generation sequencing and de novo sequence assembly. We generated and analyzed 24 unique LRV1 sequences over their full-length coding regions. Multiple alignment of these new sequences revealed variability (0.5%-23.5%) across the entire sequence except for highly conserved motifs within the 5' untranslated region. Phylogenetic analyses showed that viral genomes of L. (V.) guyanensis grouped into five distinct clusters. They further showed a species-dependent clustering between viral genomes of L. (V.) guyanensis and L. (V.) braziliensis, confirming a long-term co-evolutionary history. Noteworthy, we identified cases of multiple LRV1 infections in three of the 20 Leishmania isolates. Here, we present the first-ever estimate of LRV1 genomic diversity that exists in Leishmania (V.) guyanensis parasites. Genetic characterization and phylogenetic analyses of these viruses has shed light on their evolutionary relationships. To our knowledge, this study is also the first to report cases of multiple LRV1 infections in some parasites. Finally, this work has made it possible to develop molecular tools for adequate identification and genotyping of LRV1 strains for diagnostic purposes. Given the suspected worsening role of LRV1 infection in the pathogenesis of human leishmaniasis, these data have a major impact from a clinical viewpoint and for the management of Leishmania-infected patients.

  11. Effect of Kelussia odoratissima Mozaff essential oil on promastigot form of Leishmania major (in vitro)

    OpenAIRE

    Pirali Kheirabadi Khodadad; Saei Dehkordi Siavash; Kheibari Parviz

    2015-01-01

    Introduction: Leishmaniasis is a zoonotic disease caused by a protozoan of the genus Leishmania. In this study, the effects of Kelussia odoratissima Mozaff essential oil on the promastigot form of Leishmania major were studied. Methods: In this study, the effects of Kelussia odoratissima Mozaff essential oil on the promastigot form of Leishmania major were assessed by calculating the average number of surviving promastigots after exposure to different concentrations of essential oil, relativ...

  12. DESARROLLO DE UN PROTOCOLO DE PCR CUANTITATIVO EN TIEMPO REAL (QPCR), PARA LA DETECCION Y CUANTIFICACION DE MALEZAS HOLOPARASITAS CUARENTENARIAS (OROBANCHE SPP) QUE CONTAMINAN SEMILLAS Y SUELOS EN CHILE

    OpenAIRE

    Díaz Sánchez, Jorge Eduardo

    2013-01-01

    Las malezas parásitas del género Orobanche constituyen un grave problema para la agricultura a nivel mundial. Chile no está ajeno a esta situación, ya que O. ramosa y O. minor están invadiendo y estableciéndose en importantes zonas agrícolas. Es así como O. ramosa parásita a diversos cultivos (tomate, tabaco y papa) entre la Región Metropolitana y la Araucania (35° a 38° Lat. S) y O. minor a especies forrajeras (trébol, alfalfa y serradella) entre la Región del Maule a la Araucania (35° a 39°...

  13. Acción de distintos coagulantes para la eliminación de Cryptosporidium spp. en el proceso de potabilización del agua The action of different coagulants to remove Cryptosporidium during the process of water treatment

    Directory of Open Access Journals (Sweden)

    B. Abramovich

    2004-06-01

    Full Text Available Cryptosporidium es uno de los microorganismos de mayor interés desde el punto de vista de la Salud Pública y constituye un problema prioritario para las plantas y organismos reguladores de agua. Debido a su pequeño tamaño y a su resistencia a la cloración, la eliminación por el proceso de potabilización es una tarea compleja. En este trabajo se analizó la efectividad de distintos coagulantes utilizados comúnmente en tal proceso para lograr la remoción de los ooquistes. Se trabajó con la prueba de jarras (Jar Test. Se halló que: 1 Los coagulantes con agregado de polímeros coadyuvantes producen remociones de ooquistes superiores a 2 log. 2 Un valor bajo de turbiedad no asegura una remoción óptima de los parásitos. 3 La adición de polielectrolitos al cloruro férrico disminuye la variabilidad tanto en la turbiedad final como en la eliminación de Cryptosporidium.Cryptosporidium is one of the microorganisms of main concern from the point of view of Public Health, being a priority problem for water treatment plants and water regulatory institutions. Due to its small size and resistance to chlorination, Cryptosporidium removal during the process of drinking water treatmentis a hard task. The effectiveness of different coagulants commonly used in the process of removal of oocysts was analyzed. Thetechnique used was the Jar Test. It was found that: 1 coagulants with the addition of polimeric coadjuvants produce over 2 logs of oocyst removal; 2 a low value in turbidity does not necessarily mean optimal parasite removal, and 3 the addition of polyelectrolites to ferric chloride diminishes variability, both in final turbidity and Cryptosporidium removal.

  14. 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

  15. Potensi Trichoderma Spp. Sebagai Agens Pengendali Fusarium Spp. Penyebab Penyakit Layu Pada Tanaman Stroberi

    OpenAIRE

    Dwiastuti, Mutia Erti; Fajri, Melisa N; Yunimar, Yunimar

    2015-01-01

    Layu yang disebabkan oleh Fusarium spp. merupakan salah satu penyakit penting tanaman stroberi (Fragaria x ananassa Dutch.) di daerah subtropika, yang dapat menggagalkan panen. Penelitian bertujuan untuk mempelajari potensi Trichoderma spp. dalam mengendalikan Fusarium spp. Isolat Trichoderma spp. diisolasi dari rizosfer tanaman stroberi dan Fusarium spp. diisolasi dari tanaman stroberi yang mengalami layu fusarium. Isolat cendawan dimurnikan, dikarakterisasi, dan dibandingkan dengan isolat c...

  16. Isolation of an enriched plasma membrame subpellicular microtubule fraction of Leishmania mexicana amazonensis

    Directory of Open Access Journals (Sweden)

    Solange L. Timm

    1980-01-01

    Full Text Available A cell fractionation procedure previously developed for Trypanosoma cruzi was applied to isolated the plasma membrane of promastigotes of Leishania mexicana amazonensis. The cell, swollen in an hypotonic mediun, were disrupted in the presence of a nonionic detergent and the membrane fraction isolated by differencial centrifugation. Electron microscopy showed that the fraction consisted of pieces of the plasma membrane associated with subpellicular microtubules. It was also shown that this fraction is able to induce cell-mediated immune response in mice.Um método de fracionamento subcelular, previamente desenvolvido para Trypanosoma cruzi, foi aplicado para isolar a membrana plasmática de promastigotas de Leishmania mexicana amazonensis. As células, após turgimento em meio hipotônico, foram rompidas na presença de um detergente não iônico e a fração de membrana isolada por centrifugação diferencial. A microscopia eletrônica mostrou consistir a fração de fragmentos de membrana plasmática associados com microtúbulos subpeliculares. Foi também mostrado que esta fração era capaz de induzir resposta celular em camundongos.

  17. Evaluation of a Microculture Method for Isolation of Leishmania Parasites from Cutaneous Lesions of Patients in Peru▿

    Science.gov (United States)

    Boggild, Andrea K.; Miranda-Verastegui, Cesar; Espinosa, Diego; Arevalo, Jorge; Adaui, Vanessa; Tulliano, Gianfranco; Llanos-Cuentas, Alejandro; Low, Donald E.

    2007-01-01

    Traditional culture of Leishmania spp. is labor intensive and has poor sensitivity. We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-μl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, microculture, 5.2 days in NNN, and 6 days in 10% RPMI (P, 0.009). We have demonstrated that microculture is a more sensitive and time-efficient means of isolating Leishmania parasites from cutaneous lesions than traditional culture. PMID:17881557

  18. Evaluation of a microculture method for isolation of Leishmania parasites from cutaneous lesions of patients in Peru.

    Science.gov (United States)

    Boggild, Andrea K; Miranda-Verastegui, Cesar; Espinosa, Diego; Arevalo, Jorge; Adaui, Vanessa; Tulliano, Gianfranco; Llanos-Cuentas, Alejandro; Low, Donald E

    2007-11-01

    Traditional culture of Leishmania spp. is labor intensive and has poor sensitivity. We evaluated a microculture method for the diagnosis of cutaneous leishmaniasis in consecutive patients presenting to the Leishmaniasis Clinic at the Instituto de Medicina Tropical Alexander von Humboldt, Peru, for evaluation of skin lesions. Lesion aspirates were cultured in duplicate and parallel in traditional culture tubes containing modified Novy-MacNeal-Nicolle (NNN) medium or Roswell Park Memorial Institute medium 1640 with 10% fetal bovine serum (10% RPMI) and in 70-microl capillary tubes containing a mixture of lesion aspirate and 10% RPMI. For sensitivity analysis, the consensus standard was considered to be a positive result in any two of the following four tests: Giemsa-stained lesion smear, culture, kinetoplast DNA PCR, or leishmanin skin test. The outcome measures were sensitivity and time to culture positivity. Forty-five patients with 62 skin lesions were enrolled in the study, of which 53 lesions fulfilled the consensus criteria for a final diagnosis of cutaneous leishmaniasis. Of these 53 lesions, 39 were culture positive: 38 in capillary tubes, 29 in traditional culture tubes with modified NNN medium, and 19 in traditional culture tubes with 10% RPMI medium. The sensitivity of microculture was 71.7%, versus 54.7% for traditional culture with NNN (P, 0.038) and 35.8% with 10% RPMI (P, microculture, 5.2 days in NNN, and 6 days in 10% RPMI (P, 0.009). We have demonstrated that microculture is a more sensitive and time-efficient means of isolating Leishmania parasites from cutaneous lesions than traditional culture.

  19. Amastin Knockdown in Leishmania braziliensis Affects Parasite-Macrophage Interaction and Results in Impaired Viability of Intracellular Amastigotes.

    Directory of Open Access Journals (Sweden)

    Rita Marcia Cardoso de Paiva

    2015-12-01

    Full Text Available Leishmaniasis, a human parasitic disease with manifestations ranging from cutaneous ulcerations to fatal visceral infection, is caused by several Leishmania species. These protozoan parasites replicate as extracellular, flagellated promastigotes in the gut of a sandfly vector and as amastigotes inside the parasitophorous vacuole of vertebrate host macrophages. Amastins are surface glycoproteins encoded by large gene families present in the genomes of several trypanosomatids and highly expressed in the intracellular amastigote stages of Trypanosoma cruzi and Leishmania spp. Here, we showed that the genome of L. braziliensis contains 52 amastin genes belonging to all four previously described amastin subfamilies and that the expression of members of all subfamilies is upregulated in L. braziliensis amastigotes. Although primary sequence alignments showed no homology to any known protein sequence, homology searches based on secondary structure predictions indicate that amastins are related to claudins, a group of proteins that are components of eukaryotic tight junction complexes. By knocking-down the expression of δ-amastins in L. braziliensis, their essential role during infection became evident. δ-amastin knockdown parasites showed impaired growth after in vitro infection of mouse macrophages and completely failed to produce infection when inoculated in BALB/c mice, an attenuated phenotype that was reverted by the re-expression of an RNAi-resistant amastin gene. Further highlighting their essential role in host-parasite interactions, electron microscopy analyses of macrophages infected with amastin knockdown parasites showed significant alterations in the tight contact that is normally observed between the surface of wild type amastigotes and the membrane of the parasitophorous vacuole.

  20. 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].

  1. Post-Genomics and Vaccine Improvement for Leishmania

    Science.gov (United States)

    Seyed, Negar; Taheri, Tahereh; Rafati, Sima

    2016-01-01

    Leishmaniasis is a parasitic disease that primarily affects Asia, Africa, South America, and the Mediterranean basin. Despite extensive efforts to develop an effective prophylactic vaccine, no promising vaccine is available yet. However, recent advancements in computational vaccinology on the one hand and genome sequencing approaches on the other have generated new hopes in vaccine development. Computational genome mining for new vaccine candidates is known as reverse vaccinology and is believed to further extend the current list of Leishmania vaccine candidates. Reverse vaccinology can also reduce the intrinsic risks associated with live attenuated vaccines. Individual epitopes arranged in tandem as polytopes are also a possible outcome of reverse genome mining. Here, we will briefly compare reverse vaccinology with conventional vaccinology in respect to Leishmania vaccine, and we will discuss how it influences the aforementioned topics. We will also introduce new in vivo models that will bridge the gap between human and laboratory animal models in future studies. PMID:27092123

  2. Evolutionary history of Leishmania killicki (synonymous Leishmania tropica) and taxonomic implications.

    Science.gov (United States)

    Chaara, Dhekra; Ravel, Christophe; Bañuls, Anne- Laure; Haouas, Najoua; Lami, Patrick; Talignani, Loïc; El Baidouri, Fouad; Jaouadi, Kaouther; Harrat, Zoubir; Dedet, Jean-Pierre; Babba, Hamouda; Pratlong, Francine

    2015-04-01

    The taxonomic status of Leishmania (L.) killicki, a parasite that causes chronic cutaneous leishmaniasis, is not well defined yet. Indeed, some researchers suggested that this taxon could be included in the L. tropica complex, whereas others considered it as a distinct phylogenetic complex. To try to solve this taxonomic issue we carried out a detailed study on the evolutionary history of L. killicki relative to L. tropica. Thirty-five L. killicki and 25 L. tropica strains isolated from humans and originating from several countries were characterized using the MultiLocus Enzyme Electrophoresis (MLEE) and the MultiLocus Sequence Typing (MLST) approaches. The results of the genetic and phylogenetic analyses strongly support the hypothesis that L. killicki belongs to the L. tropica complex. Our data suggest that L. killicki emerged from a single founder event and that it evolved independently from L. tropica. However, they do not validate the hypothesis that L. killicki is a distinct complex. Therefore, we suggest naming this taxon L. killicki (synonymous L. tropica) until further epidemiological and phylogenetic studies justify the L. killicki denomination. This study provides taxonomic and phylogenetic information on L. killicki and improves our knowledge on the evolutionary history of this taxon.

  3. The current status of phlebotomine sandflies (Diptera: Psychodidae) in Tunisia and their role on Leishmania transmission: A review

    OpenAIRE

    Ahmed Tabbabi; Sajida Sboui; Jabeur Daaboub

    2017-01-01

    In Tunisia, the epidemiological situation of leishmaniasis is characterized by the coexistence in a rather circumscribed territory (165000 km2, including the Sahara) of 4 forms of leishmaniasis caused by 3 species: Leishmania infantum, Leishmania major and Leishmania tropica (L. tropica) (synonymous Leishmania killicki). One of the factors determining the clinical, epidemiological and immunological diversity of leishmanioses is certainly the existence of a vector-parasite specificity of of...

  4. Leishmania major infection in a dog with cutaneous manifestations

    OpenAIRE

    Baneth, Gad; Nachum-Biala, Yaarit; Shabat Simon, Maytal; Brenner, Ori; Gaier, Sarit; Rojas, Alicia; Yasur-Landau, Daniel

    2016-01-01

    Background Leishmania major is a main cause of cutaneous leishmaniasis in humans in an area that stretches from India through Central Asia, the Middle East, to North and West Africa. In Israel, it is a common infection of humans with rodents as the reservoir hosts and Phlebotomus papatasi as its sand fly vector. Findings A 6?months old spayed female mixed breed dog was referred to the Hebrew University Veterinary Teaching Hospital with a large ulcerative dermal lesion on the muzzle, and lesio...

  5. 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

    Roč. 5, č. 6 (2010), s. 1074-1080 ISSN 1754-2189 R&D Projects: GA ČR 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

  6. Microbial stasis of Leishmania enriettii in activated guinea pig macrophages

    International Nuclear Information System (INIS)

    Groocock, C.M.; Soulsby, E.J.L.

    1980-01-01

    Peritoneal exudate cells (PEC) from Leishmania-sensitized guinea pigs were cultured in vitro in the presence (activated) or absence (non-activated) of leishmanial antigen for 24 or 48 hours. These were then labelled with 51 Cr and challenged with 125 I-labelled promastigotes. The changing relationship between the macrophage and the parasite was monitored by observing changes in the ratio of the cell-associated isotopes. Highly significant differences in the ratio change developed during culture. These differences were a result of the activated cultures showing a higher release of 51 Cr and a lower release of 125 I when compared with the non-activated cells, at 12 hours the percentage release of 125 I from the parasite within the activated macrophage was fourfold less than that released by parasites within non-activated cells (9.2% versus 38.3%) and tenfold less than that released from glutaraldehyde-killed organisms phagocytosed by activated macrophages (91.6%). These studies indicate that stasis rather than killing of leishmaniae occurs in the activated macrophage in vitro. Parallel experiments evaluated by the visual counting of leishmaniae within the macrophages support these data. PEC from tuberculin-sensitized guinea pigs activated in vitro by purified protein derivative showed little or no activity against leishmaniae, indicating a specific requirement for this microbial stasis by activated macrophages. As a corollary of this, peritoneal exudate lymphocytes separated from the same preparations of PEC were shown to be specifically reactive to leishmanial antigen by transformation and incorporation of 3 H-thymidine. (author)

  7. Monocyte-Derived Signals Activate Human Natural Killer Cells in Response to Leishmania Parasites

    Science.gov (United States)

    Messlinger, Helena; Sebald, Heidi; Heger, Lukas; Dudziak, Diana; Bogdan, Christian; Schleicher, Ulrike

    2018-01-01

    Activated natural killer (NK) cells release interferon (IFN)-γ, which is crucial for the control of intracellular pathogens such as Leishmania. In contrast to experimental murine leishmaniasis, the human NK cell response to Leishmania is still poorly characterized. Here, we investigated the interaction of human blood NK cells with promastigotes of different Leishmania species (Leishmania major, Leishmania mexicana, Leishmania infantum, and Leishmania donovani). When peripheral blood mononuclear cells or purified NK cells and monocytes (all derived from healthy blood donors from Germany without a history of leishmaniasis) were exposed to promastigotes, NK cells showed increased surface expression of the activation marker CD69. The extent of this effect varied depending on the Leishmania species; differences between dermotropic and viscerotropic L. infantum strains were not observed. Upregulation of CD69 required direct contact between monocytes and Leishmania and was partly inhibitable by anti-interleukin (IL)-18. Unexpectedly, IL-18 was undetectable in most of the supernatants (SNs) of monocyte/parasite cocultures. Confocal fluorescence microscopy of non-permeabilized cells revealed that Leishmania-infected monocytes trans-presented IL-18 to NK cells. Native, but not heat-treated SNs of monocyte/Leishmania cocultures also induced CD69 on NK cells, indicating the involvement of a soluble heat-labile factor other than IL-18. A role for the NK cell-activating cytokines IL-1β, IL-2, IL-12, IL-15, IL-21, and IFN-α/β was excluded. The increase of CD69 was not paralleled by NK cell IFN-γ production or enhanced cytotoxicity. However, prior exposure of NK cells to Leishmania parasites synergistically increased their IFN-γ release in response to IL-12, which was dependent on endogenous IL-18. CD1c+ dendritic cells were identified as possible source of Leishmania-induced IL-12. Finally, we observed that direct contact between Leishmania and NK cells reduced the

  8. Monocyte-Derived Signals Activate Human Natural Killer Cells in Response to Leishmania Parasites

    Directory of Open Access Journals (Sweden)

    Helena Messlinger

    2018-01-01

    Full Text Available Activated natural killer (NK cells release interferon (IFN-γ, which is crucial for the control of intracellular pathogens such as Leishmania. In contrast to experimental murine leishmaniasis, the human NK cell response to Leishmania is still poorly characterized. Here, we investigated the interaction of human blood NK cells with promastigotes of different Leishmania species (Leishmania major, Leishmania mexicana, Leishmania infantum, and Leishmania donovani. When peripheral blood mononuclear cells or purified NK cells and monocytes (all derived from healthy blood donors from Germany without a history of leishmaniasis were exposed to promastigotes, NK cells showed increased surface expression of the activation marker CD69. The extent of this effect varied depending on the Leishmania species; differences between dermotropic and viscerotropic L. infantum strains were not observed. Upregulation of CD69 required direct contact between monocytes and Leishmania and was partly inhibitable by anti-interleukin (IL-18. Unexpectedly, IL-18 was undetectable in most of the supernatants (SNs of monocyte/parasite cocultures. Confocal fluorescence microscopy of non-permeabilized cells revealed that Leishmania-infected monocytes trans-presented IL-18 to NK cells. Native, but not heat-treated SNs of monocyte/Leishmania cocultures also induced CD69 on NK cells, indicating the involvement of a soluble heat-labile factor other than IL-18. A role for the NK cell-activating cytokines IL-1β, IL-2, IL-12, IL-15, IL-21, and IFN-α/β was excluded. The increase of CD69 was not paralleled by NK cell IFN-γ production or enhanced cytotoxicity. However, prior exposure of NK cells to Leishmania parasites synergistically increased their IFN-γ release in response to IL-12, which was dependent on endogenous IL-18. CD1c+ dendritic cells were identified as possible source of Leishmania-induced IL-12. Finally, we observed that direct contact between Leishmania and NK cells

  9. Eficiência e competitividade de variantes espontâneos isolados de estirpes de Bradyrhizobium spp recomendadas para a cultura da soja (Glycine max Effectiveness and competitiveness of spontaneous mutants isolated from Bradyrhizobium spp strains recommended for soybean crop (Glycine max

    Directory of Open Access Journals (Sweden)

    Fabíola Gomes de Carvalho

    2005-12-01

    Full Text Available O cultivo sucessivo de soja inoculada numa mesma área proporcionou a adaptação de uma população de rizóbios, que podem não ser tão eficientes quanto à capacidade de fixação de N2, mas apresentam alta competitividade, dificultando a introdução de novas estirpes mais eficientes. Com a finalidade de avaliar o desempenho simbiótico (eficiência e competitividade de variantes espontâneos isolados de estirpes de B. japonicum (SEMIA 5079 e SEMIA 5080 e B. elkanii (SEMIA 587 e SEMIA 5019, realizou-se um experimento em casa de vegetação onde os variantes foram inoculados isoladamente e em diferentes combinações entre os variantes e uma estirpe comprovadamente mais competitiva (SEMIA 587 ou SEMIA 5019 a partir da adição de inóculos mistos (1/1; v/v no cultivar de soja BR-16. Por meio da avaliação das variáveis analisadas (nodulação, produção de matéria de seca da parte aérea, N total acumulado na parte aérea e ocupação nodular, foi possível constatar que o determinante da maior eficiência em tratamentos co-inoculados não foi a ocupação nodular de determinada estirpe ou variante presente no inóculo, mas, sim, o tipo de interação (sinérgica ou antagônica predominante no tratamento co-inoculado e que é possível selecionar variantes eficientes e competitivos para a cultura da soja a partir de estirpes parentais que já apresentam características desejáveis para utilização em inoculantes comerciais.The continuous cultivation of inoculated soybean in the same area can determine the soil colonization with a rhizobia population presenting low nitrogen fixation effectiveness. This fact can be a problem for the establishment of a more effective population. A greenhouse experiment was carried out to evaluate the symbiotic effectiveness and competitiveness of spontaneous mutants isolated from B. japonicum (SEMIA 5079 and SEMIA 5080 and B. elkanii (SEMIA 587 and SEMIA 5019 strains. The soybean biovar BR 16 was

  10. A Historical Overview of the Classification, Evolution, and Dispersion of Leishmania Parasites and Sandflies.

    Directory of Open Access Journals (Sweden)

    Mohammad Akhoundi

    2016-03-01

    Full Text Available 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.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?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.

  11. 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

  12. 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

    2016-03-01

    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. 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? 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.

  13. The Dynamics of Lateral Gene Transfer in Genus Leishmania - A Route for Adaptation and Species Diversification.

    Science.gov (United States)

    Vikeved, Elisabet; Backlund, Anders; Alsmark, Cecilia

    2016-01-01

    The genome of Leishmania major harbours a comparably high proportion of genes of prokaryote origin, acquired by lateral gene transfer (LGT). Some of these are present in closely related trypanosomatids, while some are detected in Leishmania only. We have evaluated the impact and destiny of LGT in genus Leishmania. To study the dynamics and fate of LGTs we have performed phylogenetic, as well as nucleotide and amino acid composition analyses within orthologous groups of LGTs detected in Leishmania. A set of universal trypanosomatid LGTs was added as a reference group. Both groups of LGTs have, to some extent, ameliorated to resemble the recipient genomes. However, while virtually all of the universal trypanosomatid LGTs are distributed and conserved in the entire genus Leishmania, the LGTs uniquely present in genus Leishmania are more prone to gene loss and display faster rates of evolution. Furthermore, a PCR based approach has been employed to ascertain the presence of a set of twenty LGTs uniquely present in genus Leishmania, and three universal trypanosomatid LGTs, in ten additional strains of Leishmania. Evolutionary rates and predicted expression levels of these LGTs have also been estimated. Ten of the twenty LGTs are distributed and conserved in all species investigated, while the remainder have been subjected to modifications, or undergone pseudogenization, degradation or loss in one or more species. LGTs unique to the genus Leishmania have been acquired after the divergence of Leishmania from the other trypanosomatids, and are evolving faster than their recipient genomes. This implies that LGT in genus Leishmania is a continuous and dynamic process contributing to species differentiation and speciation. This study also highlights the importance of carefully evaluating these dynamic genes, e.g. as LGTs have been suggested as potential drug targets.

  14. The Dynamics of Lateral Gene Transfer in Genus Leishmania - A Route for Adaptation and Species Diversification.

    Directory of Open Access Journals (Sweden)

    Elisabet Vikeved

    2016-01-01

    Full Text Available The genome of Leishmania major harbours a comparably high proportion of genes of prokaryote origin, acquired by lateral gene transfer (LGT. Some of these are present in closely related trypanosomatids, while some are detected in Leishmania only. We have evaluated the impact and destiny of LGT in genus Leishmania.To study the dynamics and fate of LGTs we have performed phylogenetic, as well as nucleotide and amino acid composition analyses within orthologous groups of LGTs detected in Leishmania. A set of universal trypanosomatid LGTs was added as a reference group. Both groups of LGTs have, to some extent, ameliorated to resemble the recipient genomes. However, while virtually all of the universal trypanosomatid LGTs are distributed and conserved in the entire genus Leishmania, the LGTs uniquely present in genus Leishmania are more prone to gene loss and display faster rates of evolution. Furthermore, a PCR based approach has been employed to ascertain the presence of a set of twenty LGTs uniquely present in genus Leishmania, and three universal trypanosomatid LGTs, in ten additional strains of Leishmania. Evolutionary rates and predicted expression levels of these LGTs have also been estimated. Ten of the twenty LGTs are distributed and conserved in all species investigated, while the remainder have been subjected to modifications, or undergone pseudogenization, degradation or loss in one or more species.LGTs unique to the genus Leishmania have been acquired after the divergence of Leishmania from the other trypanosomatids, and are evolving faster than their recipient genomes. This implies that LGT in genus Leishmania is a continuous and dynamic process contributing to species differentiation and speciation. This study also highlights the importance of carefully evaluating these dynamic genes, e.g. as LGTs have been suggested as potential drug targets.

  15. The Dynamics of Lateral Gene Transfer in Genus Leishmania - A Route for Adaptation and Species Diversification

    Science.gov (United States)

    Vikeved, Elisabet; Backlund, Anders; Alsmark, Cecilia

    2016-01-01

    Background The genome of Leishmania major harbours a comparably high proportion of genes of prokaryote origin, acquired by lateral gene transfer (LGT). Some of these are present in closely related trypanosomatids, while some are detected in Leishmania only. We have evaluated the impact and destiny of LGT in genus Leishmania. Methodology/Principal Findings To study the dynamics and fate of LGTs we have performed phylogenetic, as well as nucleotide and amino acid composition analyses within orthologous groups of LGTs detected in Leishmania. A set of universal trypanosomatid LGTs was added as a reference group. Both groups of LGTs have, to some extent, ameliorated to resemble the recipient genomes. However, while virtually all of the universal trypanosomatid LGTs are distributed and conserved in the entire genus Leishmania, the LGTs uniquely present in genus Leishmania are more prone to gene loss and display faster rates of evolution. Furthermore, a PCR based approach has been employed to ascertain the presence of a set of twenty LGTs uniquely present in genus Leishmania, and three universal trypanosomatid LGTs, in ten additional strains of Leishmania. Evolutionary rates and predicted expression levels of these LGTs have also been estimated. Ten of the twenty LGTs are distributed and conserved in all species investigated, while the remainder have been subjected to modifications, or undergone pseudogenization, degradation or loss in one or more species. Conclusions/Significance LGTs unique to the genus Leishmania have been acquired after the divergence of Leishmania from the other trypanosomatids, and are evolving faster than their recipient genomes. This implies that LGT in genus Leishmania is a continuous and dynamic process contributing to species differentiation and speciation. This study also highlights the importance of carefully evaluating these dynamic genes, e.g. as LGTs have been suggested as potential drug targets. PMID:26730948

  16. Fitness of Leishmania donovani parasites resistant to drug combinations.

    Directory of Open Access Journals (Sweden)

    Raquel García-Hernández

    2015-04-01

    Full Text Available Drug resistance represents one of the main problems for the use of chemotherapy to treat leishmaniasis. Additionally, it could provide some advantages to Leishmania parasites, such as a higher capacity to survive in stress conditions. In this work, in mixed populations of Leishmania donovani parasites, we have analyzed whether experimentally resistant lines to one or two combined anti-leishmanial drugs better support the stress conditions than a susceptible line expressing luciferase (Luc line. In the absence of stress, none of the Leishmania lines showed growth advantage relative to the other when mixed at a 1:1 parasite ratio. However, when promastigotes from resistant lines and the Luc line were mixed and exposed to different stresses, we observed that the resistant lines are more tolerant of different stress conditions: nutrient starvation and heat shock-pH stress. Further to this, we observed that intracellular amastigotes from resistant lines present a higher capacity to survive inside the macrophages than those of the control line. These results suggest that resistant parasites acquire an overall fitness increase and that resistance to drug combinations presents significant differences in their fitness capacity versus single-drug resistant parasites, particularly in intracellular amastigotes. These results contribute to the assessment of the possible impact of drug resistance on leishmaniasis control programs.

  17. Immunopathological Features of Canine Myocarditis Associated with Leishmania infantum Infection

    Directory of Open Access Journals (Sweden)

    Alessandro Costagliola

    2016-01-01

    Full Text Available Myocarditis associated with infectious diseases may occur in dogs, including those caused by the protozoa Neospora caninum, Trypanosoma cruzi, Babesia canis, and Hepatozoon canis. However, although cardiac disease due to Leishmania infection has also been documented, the immunopathological features of myocarditis have not been reported so far. The aim of this study was to examine the types of cellular infiltrates and expression of MHC classes I and II in myocardial samples obtained at necropsy from 15 dogs with an established intravitam diagnosis of visceral leishmaniasis. Pathological features of myocardium were characterized by hyaline degeneration of cardiomyocytes, necrosis, and infiltration of mononuclear inflammatory cells consisting of lymphocytes and macrophages, sometimes with perivascular pattern; fibrosis was also present in various degrees. Immunophenotyping of inflammatory cells was performed by immunohistochemistry on cryostat sections obtained from the heart of the infected dogs. The predominant leukocyte population was CD8+ with a fewer number of CD4+ cells. Many cardiomyocytes expressed MHC classes I and II on the sarcolemma. Leishmania amastigote forms were not detected within macrophages or any other cell of the examined samples. Our study provided evidence that myocarditis in canine visceral leishmaniasis might be related to immunological alterations associated with Leishmania infection.

  18. Mitochondrial Proteomics of Antimony and Miltefosine Resistant Leishmania infantum

    Directory of Open Access Journals (Sweden)

    Isabel M. Vincent

    2015-10-01

    Full Text Available Antimony (SbIII and miltefosine (MIL are important drugs for the treatment of Leishmania parasite infections. The mitochondrion is likely to play a central role in SbIII and MIL induced cell death in this parasite. Enriched mitochondrial samples from Leishmania promastigotes selected step by step for in vitro resistance to SbIII and MIL were subjected to differential proteomic analysis. A shared decrease in both mutants in the levels of pyruvate dehydrogenase, dihydrolipoamide dehydrogenase, and isocitrate dehydrogenase was observed, as well as a differential abundance in two calcium-binding proteins and the unique dynamin-1-like protein of the parasite. Both mutants presented a shared increase in the succinyl-CoA:3-ketoacid-coenzyme A transferase and the abundance of numerous hypothetical proteins was also altered in both mutants. In general, the proteomic changes observed in the MIL mutant were less pronounced than in the SbIII mutant, probably due to the early appearance of a mutation in the miltefosine transporter abrogating the need for a strong mitochondrial adaptation. This study is the first analysis of the Leishmania mitochondrial proteome and offers powerful insights into the adaptations to this organelle during SbIII and MIL drug resistance.

  19. 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.

  20. Leishmania and its quest for iron: An update and overview.

    Science.gov (United States)

    Zaidi, Amir; Singh, Krishn Pratap; Ali, Vahab

    2017-01-01

    Parasites of genus Leishmania are the causative agents of complex neglected diseases called leishmaniasis and continue to be a significant health concern globally. Iron is a vital nutritional requirement for virtually all organisms, including pathogenic trypanosomatid parasites, and plays a crucial role in many facets of cellular metabolism as a cofactor of several enzymes. Iron acquisition is essential for the survival of parasites. Yet parasites are also vulnerable to the toxicity of iron and reactive oxygen species. The aim of this review is to provide an update on the current knowledge about iron acquisition and usage by Leishmania species. We have also discussed about host strategy to modulate iron availability and the strategies deployed by Leishmania parasites to overcome iron withholding defences and thus favour parasite growth within host macrophages. Since iron plays central roles in the host's response and parasite metabolism, a comprehensive understanding of the iron metabolism is beneficial to identify potential viable therapeutic opportunities against leishmaniasis. Copyright © 2016 Elsevier B.V. All rights reserved.

  1. Impact of Leishmania metalloprotease GP63 on macrophage signaling

    Science.gov (United States)

    Isnard, Amandine; Shio, Marina T.; Olivier, Martin

    2012-01-01

    The intramacrophage protozoan parasites of Leishmania genus have developed sophisticated ways to subvert the innate immune response permitting their infection and propagation within the macrophages of the mammalian host. Several Leishmania virulence factors have been identified and found to be of importance for the development of leishmaniasis. However, recent findings are now further reinforcing the critical role played by the zinc-metalloprotease GP63 as a virulence factor that greatly influence host cell signaling mechanisms and related functions. GP63 has been found to be involved not only in the cleavage and degradation of various kinases and transcription factors, but also to be the major molecule modulating host negative regulatory mechanisms involving for instance protein tyrosine phosphatases (PTPs). Those latter being well recognized for their pivotal role in the regulation of a great number of signaling pathways. In this review article, we are providing a complete overview about the role of Leishmania GP63 in the mechanisms underlying the subversion of macrophage signaling and functions. PMID:22919663

  2. Passive transfer of leishmania lipopolysaccharide confers parasite survival in macrophages

    International Nuclear Information System (INIS)

    Handman, E.; Schnur, L.F.; Spithill, T.W.; Mitchell, G.F.

    1986-01-01

    Infection of macrophages by the intracellular protozoan parasite Leishmania involves specific attachment to the host membrane, followed by phagocytosis and intracellular survival and growth. Two parasite molecules have been implicated in the attachment event: Leishmania lipopolysaccharide (L-LPS) and a glycoprotein (gp63). This study was designed to clarify the role of L-LPS in infection and the stage in the process of infection at which it operates. The authors have recently identified a Leishmania major strain (LRC-L119) which lacks the L-LPS molecule and is not infective for hamsters or mice. This parasite was isolated from a gerbil in Kenya and was identified phenotypically as L. major by isoenzyme and fatty acid analysis. In this study they have confirmed at the genotype level that LRC-L119 is L. major by analyzing and comparing the organization of cloned DNA sequences in the genome of different strains of L. major. Here they show that LRC-L119 promastigotes are phagocytosed rapidly by macrophages in vitro, but in contrast to virulent strains of L. major, they are then killed over a period of 18 hr. In addition, they show that transfer of purified L-LPS from a virulent clone of L. major (V121) into LRC-L119 promastigotes confers on them the ability to survive in macrophages in vitro

  3. Evaluating the Role of Host AMPK in Leishmania Burden.

    Science.gov (United States)

    Moreira, Diana; Estaquier, Jérôme; Cordeiro-da-Silva, Anabela; Silvestre, Ricardo

    2018-01-01

    The study of host AMP-activated protein kinase (AMPK) activation during Leishmania infection imposes distinct types of techniques to measure protein expression and activation, as well as to quantify, at transcription and translational levels, its downstream targets. The investigation of host AMPK protein modulation during Leishmania infection should primarily be assessed during in vitro infections using as a host murine bone marrow-derived macrophages (BMMos). The infection outcome is assessed measuring the percentage of infected cells in the context of BMMos. To evaluate AMPK activity during infection, the expression of AMPK phosphorylated at Thr172 as well as the transcription and translational levels of its downstream targets are evaluated by quantitative PCR and immunoblotting. The modulation of AMPK activity in vivo is determined specifically in sorted splenic macrophages harboring Leishmania parasites recovered from infected mice using fluorescent-labeled parasites in the infectious inocolum. The modulation of AMPK activity was assessed by AMPK activators and inhibitors and also using AMPK, SIRT1, or LKB1 KO mice models. The infection outcome in BMMos and in vivo was further determined using these two different approaches. To finally understand the metabolic impact of AMPK during infection, in vitro metabolic assays in infected BMMos were measured in the bioenergetic profile using an extracellular flux analyzer.

  4. 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.

  5. Leishmania Surveillance and Diagnostic Capability in Support of the Joint Biological Agent Identification and Diagnostic System (JBAIDS) and Leishmania Vector Surveillance

    Science.gov (United States)

    2013-02-07

    01-10-09 to 07-02-13 ’+. I II L~ J.\\NU :::OU~ Ill L~ :la. l-UI’I I 11J.\\l- I NUIVI~~I1 LEISHMANIA SURVEILLANCE AND DIAGNOSTIC CAPABILITY IN None...SUPPORT OF THE JOINT BIOLOGICAL AGENT IDENTIFICATION AND :lD. l:JI1J.\\NI NUIVI~~I1 DIAGNOSTIC SYSTEM (JBAIDS) None . ./ LEISHMANIA VECTOR...Field Station at Kisumu completed project activities through a resource sharing arrangement with the 59th MDW. Testing of the Leishmania epidemiology

  6. First occurrence of an autochthonous canine case of Leishmania (Leishmania infantum chagasi in the municipality of Campinas, State of São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Elisa San Martin Mouriz Savani

    2011-08-01

    Full Text Available An autochthonous case of visceral leishmaniasis is reported in a dog (Canis familiaris as an apparently natural infection in a non-endemic area. DNA obtained from spleen and liver samples produced the expected fragment in a Leishmania-specific rDNA-based nested-PCR assay. The PCR product, a 490 bp fragment, was sequenced and the nucleotide sequence was identical to that of Leishmania (Leishmania infantum chagasi. These results are surprising since no autochthonous human or canine cases of visceral leishmaniasis have ever been reported in this municipality. This case suggests that natural transmission of this disease is occurring in this area.

  7. Antileishmanial activity of licochalcone A in mice infected with Leishmania major and in hamsters infected with Leishmania donovani

    DEFF Research Database (Denmark)

    Chen, M; Christensen, S B; Theander, T G

    1994-01-01

    This study was designed to examine the antileishmanial activity of the oxygenated chalcone licochalcone A in mice and hamsters infected with Leishmania parasites. Intraperitoneal administration of licochalcone A at doses of 2.5 and 5 mg/kg of body weight per day completely prevented lesion...... development in BALB/c mice infected with Leishmania major. Treatment of hamsters infected with L. donovani with intraperitoneal administration of licochalcone A at a dose of 20 mg/kg of body weight per day for 6 consecutive days resulted in a > 96% reduction of parasite load in the liver and the spleen...... consecutive days resulted in > 65 and 85% reductions of L. donovani parasite loads in the liver and the spleen, respectively, compared with those of untreated control hamsters. These data clearly demonstrate that licochalcone A is a promising lead for the development of a new drug against leishmaniases....

  8. Perfil fitoquímico, actividad anti-Leishmania, hemolítica y toxicológica de Cordia dentata Poir. y Heliotropium indicum L.

    OpenAIRE

    Espitia-Baena, Jorge Eliecer; Robledo-Restrepo, Sara María; Soraya Cuadrado-Cano, Bernarda; Duran-Sandoval, Harriette del Rosario; Gómez-Estrada, Harold Alberto

    2014-01-01

    Introducción: la leishmaniasis constituye un grave problema de salud pública a nivel mundial. En Colombia, la forma clínica cutánea tiene la mayor incidencia y es causada principalmente por especies de Leishmania (Viannia) panamensis. Dado que los tratamientos disponibles para esta enfermedad resultan ineficaces, es necesaria la búsqueda de nuevos agentes terapéuticos en especies vegetales utilizadas por la medicina popular. Objetivo: evaluar el perfil fitoquímico, la actividad hemolítica, ci...

  9. Serum Removal from Culture Induces Growth Arrest, Ploidy Alteration, Decrease in Infectivity and Differential Expression of Crucial Genes in Leishmania infantum Promastigotes.

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    Pedro J Alcolea

    Full Text Available Leishmania infantum is one of the species responsible for visceral leishmaniasis. This species is distributed basically in the Mediterranean basin. A recent outbreak in humans has been reported in Spain. Axenic cultures are performed for most procedures with Leishmania spp. promastigotes. This model is stable and reproducible and mimics the conditions of the gut of the sand fly host, which is the natural environment of promastigote development. Culture media are undefined because they contain mammalian serum, which is a rich source of complex lipids and proteins. Serum deprivation slows down the growth kinetics and therefore, yield in biomass. In fact, we have confirmed that the growth rate decreases, as well as infectivity. Ploidy is also affected. Regarding the transcriptome, a high-throughput approach has revealed a low differential expression rate but important differentially regulated genes. The most remarkable profiles are: up-regulation of the GINS Psf3, the fatty acyl-CoA synthase (FAS1, the glyoxylase I (GLO1, the hydrophilic surface protein B (HASPB, the methylmalonyl-CoA epimerase (MMCE and an amastin gene; and down-regulation of the gPEPCK and the arginase. Implications for metabolic adaptations, differentiation and infectivity are discussed herein.

  10. Serum Removal from Culture Induces Growth Arrest, Ploidy Alteration, Decrease in Infectivity and Differential Expression of Crucial Genes in Leishmania infantum Promastigotes.

    Science.gov (United States)

    Alcolea, Pedro J; Alonso, Ana; Moreno-Izquierdo, Miguel A; Degayón, María A; Moreno, Inmaculada; Larraga, Vicente

    2016-01-01

    Leishmania infantum is one of the species responsible for visceral leishmaniasis. This species is distributed basically in the Mediterranean basin. A recent outbreak in humans has been reported in Spain. Axenic cultures are performed for most procedures with Leishmania spp. promastigotes. This model is stable and reproducible and mimics the conditions of the gut of the sand fly host, which is the natural environment of promastigote development. Culture media are undefined because they contain mammalian serum, which is a rich source of complex lipids and proteins. Serum deprivation slows down the growth kinetics and therefore, yield in biomass. In fact, we have confirmed that the growth rate decreases, as well as infectivity. Ploidy is also affected. Regarding the transcriptome, a high-throughput approach has revealed a low differential expression rate but important differentially regulated genes. The most remarkable profiles are: up-regulation of the GINS Psf3, the fatty acyl-CoA synthase (FAS1), the glyoxylase I (GLO1), the hydrophilic surface protein B (HASPB), the methylmalonyl-CoA epimerase (MMCE) and an amastin gene; and down-regulation of the gPEPCK and the arginase. Implications for metabolic adaptations, differentiation and infectivity are discussed herein.

  11. Molecular detection of Leishmania parasites and host blood meal identification in wild sand flies from a new endemic rural region, south of Iran.

    Science.gov (United States)

    Azizi, Kourosh; Askari, Mohammad Bagher; Kalantari, Mohsen; Moemenbellah-Fard, Mohammad Djaefar

    Zoonotic Cutaneous Leishmaniosis (ZCL) remains the most crucial vector-borne public health disease particularly in endemic rural parts of Iran. The main aim of this study is to identify wild sand flies (Diptera: Psychodidae), determine their infection rate, and differentiate their host blood meal sources using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Sand fly populations were caught with sticky paper traps from 10 different villages in the county of Darab, Fars province, southern Iran. Following their species identification, they were used in one step PCR to determine their infection with Leishmania spp. parasites. They were then subjected to PCR-RFLP protocol to identify and differentiate their blood meal sources. Two genera of Phlebotomus and Sergentomyia comprising 13 species of sand flies were identified in this region. From a total of 150 parous female sand flies, encompassing 4 different medically important species, 7 specimens (4.7%) including 6 Phlebotomus papatasi and 1 Phlebotomus bergeroti were infected with Leishmania major. Molecular data indicated that about 32% of female sand flies fed on man, while nearly 43% fed on rodent and canine hosts. Molecular detection is an efficient way of differentiating the source of blood meals in female sand flies feeding on different vertebrate hosts. It is suggested that P. papatasi is not highly anthropophagic and appears to be an opportunistic feeder on man. This species is, however, the primary vector of ZCL in this region.

  12. T-cell responses associated with resistance to Leishmania infection in individuals from endemic areas for Leishmania (Viannia braziliensis

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    Rita C Bittar

    2007-08-01

    Full Text Available Subclinical or asymptomatic infection is documented in individuals living in endemic areas for leishmaniasis suggesting that the development of an appropriate immune response can control parasite replication and maintain tissue integrity. A low morbidity indicates that intrinsic factors could favor resistance to Leishmania infection. Herein, leishmanial T-cell responses induced in subjects with low susceptibility to leishmaniasis as asymptomatic subjects were compared to those observed in cured cutaneous leishmaniasis (CCL patients, who controlled the disease after antimonial therapy. All of them have shown maintenance of specific long-term immune responses characterized by expansion of higher proportions of CD4+ as compared to CD8+ Leishmania reactive T-lymphocytes. Asymptomatic subjects had lower indexes of in vitro Leishmania induced lymphoproliferative responses and interferon-gamma (IFN-gamma production in comparison to CCL patients. On the other hand, interleukin (IL-10 production was much higher in asymptomatics than in CCL, while no differences in IL-5 levels were found. In conclusion, long lived T-cell responses achieved by asymptomatic individuals differed from those who had developed symptomatic leishmaniasis in terms of intensity of lymphocyte activation (proliferation or IFN-gamma and regulatory mechanisms (IL-10. The absence of the disease in asymptomatics could be explained by their intrinsic ability to create a balance between immunoregulatory (IL-10 and effector cytokines (IFN-gamma, leading to parasite destruction without producing skin tissue damage. The establishment of profiles of cell-mediated immune responses associated with resistance against Leishmania infection is likely to make new inroads into understanding the long-lived immune protection against the disease.

  13. Dietas contendo silagem de milho (Zea maiz L. e feno de capim-tifton 85 (Cynodon spp. em diferentes proporções para bovinos Corn silage and tifton 85 bermudagrass hay-based diets for steers

    Directory of Open Access Journals (Sweden)

    Ana Clara Rodrigues Cavalcante

    2004-12-01

    Full Text Available Avaliaram-se o consumo, a digestibilidade, o pH e concentração de amônia ruminais e a taxa de passagem em bovinos alimentados com dietas contendo feno de capim-tifton 85 (TIF e silagem de milho (SM em diferentes proporções. Utilizaram-se quatro animais castrados, com peso médio de 523kg, fistulados no rúmen, distribuídos em um quadrado latino 4 x 4, recebendo 60% de volumoso e 40% de concentrado, na base da matéria seca. O volumoso consistiu das seguintes proporções (%: 100 TIF:0 SM; 67 TIF:33 SM; 33 TIF:67 SM e 0 TIF:100 SM. Os consumos de matéria seca (MS, de matéria orgânica (MO e de matéria orgânica digestível (MOD, de proteína bruta (PB, de fibra em detergente neutro (FDN e de carboidratos totais (CHOT e digeríveis (CHOD não foram influenciados pelas diferentes proporções de feno de capim-tifton 85: silagem de milho nas dietas, registrando-se valores médios de 9,2; 8,7; 6,4; 1,1; 3,7; 7,3 e 5,4 kg/dia, respectivamente. O consumo de extrato etéreo (EE elevou linearmente com o aumento da silagem de milho nas dietas. As digestibilidades aparentes da MS, MO, PB, EE e CHO também não foram influenciadas pelas diferentes proporções de silagem de milho, obtendo-se, respectivamente, valores médios de 71,8; 73,0; 69,5; 69,2 e 73,9%. Para a digestibilidade da FDN, observou-se efeito quadrático, estimando-se valor máximo de 65,2% para dietas contendo 30,98% de silagem de milho. Estimou-se concentração máxima de amônia de 12,0 mg/100 ml e valor mínimo de pH de 5,98 às 2,44 e 6,82 horas após a alimentação, respectivamente. A taxa de passagem não foi influenciada pelas diferentes proporções de feno:silagem, no volumoso, apresentando valor médio de 4,2%/hora.The intake, digestibility, ruminal pH and ammonia concentrations and passage rate were evaluated in steers fed corn silage (CS and Tifton 85 bermudagrass hay (T85H-based diets, at different forage proportions. Four rumen fistulated steers with 523 kg of live

  14. In vivo and in vitro phagocytosis of Leishmania (Leishmania) amazonensis promastigotes by B-1 cells.

    Science.gov (United States)

    Geraldo, M M; Costa, C R; Barbosa, F M C; Vivanco, B C; Gonzaga, W F K M; Novaes E Brito, R R; Popi, A F; Lopes, J D; Xander, P

    2016-06-01

    Leishmaniasis is caused by Leishmania parasites that infect several cell types. The promastigote stage of Leishmania is internalized by phagocytic cells and transformed into the obligate intracellular amastigote form. B-1 cells are a subpopulation of B cells that are able to differentiate in vitro and in vivo into mononuclear phagocyte-like cells with phagocytic properties. B-1 cells use several receptors for phagocytosis, such as the mannose receptor and third complement receptor. Leishmania binds to the same receptors on macrophages. In this study, we demonstrated that phagocytes derived from B-1 cells (B-1 CDP) were able to internalize promastigotes of L. (L.) amazonensis in vitro. The internalized promastigotes differentiated into amastigotes. Our results showed that the phagocytic index was higher in B-1 CDP compared to peritoneal macrophages and bone marrow-derived macrophages. The in vivo phagocytic ability of B-1 cells was also demonstrated. Parasites were detected inside purified B-1 cells after intraperitoneal infection with L. (L.) amazonensis promastigotes. Intraperitoneal stimulation with the parasites led to an increase in both IL-10 and TNF-α. These results highlight the importance of studying B-1 CDP cells as phagocytic cells that can participate and contribute to immunity to parasites. © 2016 John Wiley & Sons Ltd.

  15. Leishmania (Leishmania mexicana en el corregimiento de San Matías, municipio de Gómez Plata, Antioquia, Colombia

    Directory of Open Access Journals (Sweden)

    Diana Sierra

    2006-10-01

    Full Text Available Introducción. La leishmaniaisis es una enfermedad encontrada en focos naturales de infección donde están presentes insectos vectores y mamíferos reservorios deLeishmania. Objetivo. Registrar por primera vez la presencia de Leishmania (Leishmania mexicana Biagi, 1953, en el corregimiento de San Matías, municipio de Gómez Plata, departamento de Antioquia. Materiales y métodos. La especie fue aislada de un paciente con leishmaniasis cutánea localizada e identificada por la técnica de Inmunofluorescencia utilizando anticuerpos monoclonales específicos de especie y electroforesis de enzimas . Resultados y conclusión. Su perfil isoenzimático similar al de las cepas de referencia L. (L. mexicana (MNCY/BZ/62/M379 y L. (L. mexicana (MHOM/BE/82/BEL21, permitió concluír que la especie aislada del paciente es L. (L. mexicana.

  16. Natural infection of phlebotomines (Diptera: Psychodidae) by Leishmania (Leishmania) amazonensis in an area of ecotourism in Central-Western Brazil.

    Science.gov (United States)

    Brilhante, Andreia Fernandes; Nunes, Vânia Lúcia Brandão; Kohatsu, Kleber Augusto; Galati, Eunice Aparecida Bianchi; Rocca, Maria Elizabeth Ghizzi; Ishikawa, Edna Aoba Yassui

    2015-01-01

    Bonito municipality, known as an area of ecoturism, in Mato Grosso do Sul state, Brazil, is also a focus of visceral and cutaneous leishmaniases, with cases registered in both human and canine populations. This study sought to investigate natural infection by flagellate forms of Leishmania in phlebotomines of the urban area of Bonito. Sand flies were collected fortnightly from October 2005 to July 2006 with modified automatic light traps installed in peridomiciles and animal shelters in the center and on the outskirts of the city. The females were dissected and their guts observed under an optical microscope. A total of 1977 specimens were captured, Lutzomyia longipalpis (88.4 %) and Bichromomyia flaviscutelata (3.0 %) being the most frequent species. Bi. flaviscutellata was found infected by flagellates that were identified as Leishmania (Leishmania) amazonensis by indirect immunofluorescence reaction, employing monoclonal antibodies and the biotin-avidin system. This is the first report of natural infection by L. amazonensis in Bi. flaviscutellata in a Brazilian urban area. As Bi. flaviscutellata is only slightly attracted by humans, the transmission of L. amazonensis in the study area may have a zoonotic character; however, the sympatric occurrence of this parasite and Lu. longipalpis should be taken into consideration by the local health authorities since this sand fly has already been found with L. amazonensis DNA in a focus of canine visceral leishmaniasis in Bonito municipality.

  17. Leishmaniasis in Turkey: Visceral and cutaneous leishmaniasis caused by Leishmania donovani in Turkey

    NARCIS (Netherlands)

    Özbilgin, Ahmet; Harman, Mehmet; Karakuş, Mehmet; Bart, Aldert; Töz, Seray; Kurt, Özgür; Çavuş, İbrahim; Polat, Erdal; Gündüz, Cumhur; van Gool, Tom; Özbel, Yusuf

    2017-01-01

    In Turkey, the main causative agents are Leishmania tropica (L. tropica) and Leishmania infantum (L. infantum) for cutaneous leishmaniasis (CL) and L. infantum for visceral leishmaniasis (VL). In this study, we investigated leishmaniasis cases caused by L. donovani and established animal models for

  18. Serological and molecular survey of Leishmania infection in dogs from Luanda, Angola

    NARCIS (Netherlands)

    Vilhena, Hugo; Granada, Sara; Oliveira, Ana Cristina; Schallig, Henk D. F. H.; Nachum-Biala, Yaarit; Cardoso, Luís; Baneth, Gad

    2014-01-01

    Canine leishmaniosis (CanL) due to Leishmania infantum is a global zoonosis endemic in more than 70 countries in Europe, North Africa, Asia and America; however, data on this infection is scarce from southern Africa. The aim of this study was to survey dogs in Luanda, Angola, for Leishmania

  19. Development of a dipstick assay for detection of Leishmania-specific canine antibodies

    NARCIS (Netherlands)

    Schallig, Henk D. F. H.; Cardoso, Luís; Hommers, Marieke; Kroon, Nel; Belling, Guus; Rodrigues, Manuela; Semião-Santos, Saul J.; Vetter, Hans

    2004-01-01

    A dipstick assay, based on Leishmania infantum antigen, for the rapid detection of Leishmania-specific antibodies in canine serum samples was developed and evaluated. After determination of optimal dipstick test conditions, test performance was compared with two existing serological tests, i.e., the

  20. Susceptibilidad de genotipos de Solanum spp. al nematodo causante del nudo radical Meloidogyne spp. (chitwood

    Directory of Open Access Journals (Sweden)

    Gelpud Chaves Cristian

    2011-03-01

    Full Text Available

    El cultivo del lulo (Solanum quitoense L. presenta una disminución en su productividad, debido al ataque de patógenos como el nematodo del nudo radical Meloidogyne  spp., en el Departamento  de Nariño (Colombia, se han reportado incidencias cercanas al 79%, y pérdidas del 50%.   En la presente investigación, se colectaron 45 genotipos de (Solanum quitoense  L. en los Departamentos  de Nariño  y Putumayo  y 4 genotipos  silvestres  (S. mammosum, S. hirtum,       S. marginatum  y S. umbellatum buscando fuentes de resistencia al nematodo. Se inocularon 9 plantas de cada genotipo de dos meses de edad con 10000 huevos de Meloidogyne spp., dejando tres testigos por cada material. Las variables evaluadas fueron: altura de planta, severidad, incidencia, peso fresco (tallo y raíz y especies prevalentes de Meloidogyne spp. Se hizo una clasificación de genotipos mediante escala de resistencia y regresión entre la severidad y las demás variables para establecer el efecto de Meloidogyne spp. sobre los genotipos de planta. Los resultados mostraron 100% de incidencia del nematodo en  todos  los  genotipos,  2.04%  genotipos  resistentes,  34.7%  moderadamente  resistentes, 42.8% moderadamente susceptibles, 18.3% susceptibles, y 2.04% altamente susceptibles. El genotipo SQbr05 resistente, no se vio afectado por la severidad, al contrario SQbc04 genotipo susceptible, mostró reducciones significativas en peso fresco de tallo y raIz, (R2 = 0.71 y 0.98,el genotipo silvestre (S. mammosum es altamente susceptible, Meloidogyne incognita presentó 55.31% de presencia. El genotipo SQbr05 es promisorio para ser evaluado en campo.

  1. Differential Recruitment of Dendritic Cells Subsets to Lymph Nodes Correlates with a Protective or Permissive T-Cell Response during Leishmania (Viannia) Braziliensis or Leishmania (Leishmania) Amazonensis Infection.

    Science.gov (United States)

    Carvalho, A K; Carvalho, K; Passero, L F D; Sousa, M G T; da Matta, V L R; Gomes, C M C; Corbett, C E P; Kallas, G E; Silveira, F T; Laurenti, M D

    2016-01-01

    Leishmania (L.) amazonensis (La) and L. (V.) braziliensis (Lb) are responsible for a large clinical and immunopathological spectrum in human disease; while La may be responsible for anergic disease, Lb infection leads to cellular hypersensitivity. To better understand the dichotomy in the immune response caused by these Leishmania species, we evaluated subsets of dendritic cells (DCs) and T lymphocyte in draining lymph nodes during the course of La and Lb infection in BALB/c mice. Our results demonstrated a high involvement of DCs in La infection, which was characterized by the greater accumulation of Langerhans cells (LCs); conversely, Lb infection led to an increase in dermal DCs (dDCs) throughout the infection. Considering the T lymphocyte response, an increase of effector, activated, and memory CD4(+) T-cells was observed in Lb infection. Interleukin- (IL-) 4- and IL-10-producing CD4(+)and CD8(+) T-cells were present in both La and Lb infection; however, interferon- (IFN-) γ-producing CD4(+)and CD8(+) T-cells were detected only in Lb infection. The results suggest that during Lb infection, the dDCs were the predominant subset of DCs that in turn was associated with the development of Th1 immune response; in contrast La infection was associated with a preferential accumulation of LCs and total blockage of the development of Th1 immune response.

  2. Severity of tegumentary leishmaniasis is not exclusively associated with Leishmania RNA virus 1 infection in Brazil

    Directory of Open Access Journals (Sweden)

    Luiza de Oliveira Ramos Pereira

    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 infection 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.

  3. Anti-Leishmania activity of new ruthenium(II) complexes: Effect on parasite-host interaction.

    Science.gov (United States)

    Costa, Mônica S; Gonçalves, Yasmim G; Nunes, Débora C O; Napolitano, Danielle R; Maia, Pedro I S; Rodrigues, Renata S; Rodrigues, Veridiana M; Von Poelhsitz, Gustavo; Yoneyama, Kelly A G

    2017-10-01

    Leishmaniasis is a parasitic disease caused by protozoa of the genus Leishmania. The many complications presented by the current treatment - including high toxicity, high cost and parasite resistance - make the development of new therapeutic agents indispensable. The present study aims to evaluate the anti-Leishmania potential of new ruthenium(II) complexes, cis‑[Ru II (η 2 -O 2 CR)(dppm) 2 ]PF 6 , with dppm=bis(diphenylphosphino)methane and R=4-butylbenzoate (bbato) 1, 4-(methylthio)benzoate (mtbato) 2 and 3-hydroxy-4-methoxybenzoate (hmxbato) 3, in promastigote cytotoxicity and their effect on parasite-host interaction. The cytotoxicity of complexes was analyzed by MTT assay against Leishmania (Leishmania) amazonensis, Leishmania (Viannia) braziliensis, Leishmania (Leishmania) infantum promastigotes and the murine macrophage (RAW 264.7). The effect of complexes on parasite-host interaction was evaluated by in vitro infectivity assay performed in the presence of two different concentrations of each complex: the promastigote IC 50 value and the concentration nontoxic to 90% of RAW 264.7 macrophages. Complexes 1-3 exhibited potent cytotoxic activity against all Leishmania species assayed. The IC 50 values ranged from 7.52-12.59μM (complex 1); 0.70-3.28μM (complex 2) and 0.52-1.75μM (complex 3). All complexes significantly inhibited the infectivity index at both tested concentrations. The infectivity inhibitions ranged from 37 to 85%. Interestingly, the infectivity inhibitions due to complex action did not differ significantly at either of the tested concentrations, except for the complex 1 against Leishmania (Leishmania) infantum. The infectivity inhibitions resulted from reductions in both percentage of infected macrophages and number of parasites per macrophage. Taken together the results suggest remarkable leishmanicidal activity in vitro by these new ruthenium(II) complexes. Copyright © 2017 Elsevier Inc. All rights reserved.

  4. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil

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    Tereza Emmanuelle de Farias Rotondano

    Full Text Available This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100 of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA and polymerase chain reaction (PCR, respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100 of the animals. Babesia vogeli was detected by PCR in ten animals (10% and was correlated with young age (p = 0.007 and thrombocytopenia (p = 0.01. None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.

  5. Survey of Ehrlichia canis, Babesia spp. and Hepatozoon spp. in dogs from a semiarid region of Brazil.

    Science.gov (United States)

    Rotondano, Tereza Emmanuelle de Farias; Almeida, Herta Karyanne Araújo; Krawczak, Felipe da Silva; Santana, Vanessa Lira; Vidal, Ivana Fernandes; Labruna, Marcelo Bahia; de Azevedo, Sérgio Santos; Ade lmeida, Alzira Maria Paiva; de Melo, Marcia Almeida

    2015-01-01

    This study assessed the occurrence of Ehrlichia spp., Babesia spp. and Hepatozoon spp. infections in 100 tick-harboring dogs from a semiarid region of the State of Paraíba, Northeastern Brazil. Blood samples and ticks were collected from the animals, and a questionnaire was submitted to dog owners to obtain general data. Blood samples were used to perform hemogram, direct blood smear and immunological and molecular hemoparasite detection. The 1,151 ticks collected were identified as Rhipicephalus sanguineus; direct smears revealed E. canis-like morulae in the monocytes of 4% (4/100) of the non-vaccinated female dogs, and 34% and 25% of the dogs tested positive for Ehrlichia canis by indirect immunofluorescence assay (IFA) and polymerase chain reaction (PCR), respectively. Blood smear examination revealed Babesia-suggestive merozoites in the erythrocytes of 2% (2/100) of the animals. Babesia vogeli was detected by PCR in ten animals (10%) and was correlated with young age (p = 0.007) and thrombocytopenia (p = 0.01). None of the animals showed Hepatozoon spp. positivity. These results indicate that E. canis is the main tick-borne canine pathogen in the study area and provide the first report of B. vogeli infection in dogs from Paraiba State.

  6. Occurrence of Campylobacter spp. and Cryptosporidium spp. in seagulls (Larus spp.).

    Science.gov (United States)

    Moore, John E; Gilpin, Deidre; Crothers, Elizabeth; Canney, Anne; Kaneko, Aki; Matsuda, Motoo

    2002-01-01

    An investigation was carried out into the prevalence of thermophilic Campylobacter subspecies (spp.) and Cryptosporidium spp. in fresh fecal specimens collected from members of the gull family (Larus spp.) from three coastal locations of Northern Ireland. A total of 205 fresh fecal specimens were collected from gulls, of which 28 of 205 (13.7%) were positive for Campylobacter spp. and none of 205 for Cryptosporidium spp. Of these campylobacters, 21 of 28 (75%) isolates obtained belonged to the urease-positive thermophilic Campylobacter (UPTC) taxon, followed by five of 28 (17.9%) Campylobacter lari and 2/28 (7.1%) Campylobacter jejuni. It is significant that seagulls are the sole warm-blooded animal host of this bacterial taxon in Northern Ireland. It is proposed that physiological adaptation to starvation by gulls may lead to increased concentrations of urea through energy production from protein, yielding increased levels of urea for metabolism by UPTC organisms. In general, the possibility exists that environmental contamination of surface waters with campylobacters might be mediated by wild birds (such as gulls), where such waters are used for recreational purposes or where such waters are consumed untreated, might represent a risk to public health.

  7. Aeromonas spp.: an emerging pathogen?

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    Andrea Bartolini

    2015-12-01

    Full Text Available The aim of this study is to identify and monitor the presence of Aeromonas spp. strains in stool cultures. We analyzed 5564 stool cultures from September 2012 to August 2013. Sixty-three patients were positive for Aeromonas spp. The most frequent symptoms were: diarrhea (46.0% and abdominal pain (12.7%. Pediatric subjects were 28. Samples’ microscopic examination showed leukocytes in 38.1% of cases. It is still controversial whether Aeromonas are responsible for human gastroenteritis, but their presence in faecies of symptomatic patients supports their etiologic role. We propose search for toxins by polymerase chain reaction to identify strains that require an antibiotic therapy.

  8. 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.

    Science.gov (United States)

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

    2010-01-01

    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. The captures were conducted with modified Disney traps, using hamster (Mesocricetus auratus) as bait, from May 2004 to January 2006. 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. 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.

  9. 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, f

  10. 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.

  11. Gentamicin-attenuated Leishmania infantum: a clinicopathological study in dogs.

    Science.gov (United States)

    Daneshvar, Hamid; Molaei, Mohammad M; Afshar, Reza Malekpour; Kamiabi, Hosein; Burchmore, Richard; Hagan, Paul; Phillips, R Stephen

    2009-05-15

    The clinicopathological changes following infection with an attenuated line of Leishmania infantum (L. infantum H-line) were evaluated in mixed breed dogs. Two groups of dogs were infected intravenously (i.v.) or intradermally (i.d.) with L. infantum H-line and two control groups were infected i.v. or i.d. with L. infantum wild-type (L. infantum WT). None of the dogs, which were infected i.v. or i.d. with L. infantum H-line, showed any abnormalities during the observation period. In contrast, two out of three dogs, which were infected i.v. with L. infantum WT, developed clinical signs of disease. In addition, no histopathological changes were seen in the liver and spleen of the dogs infected with the attenuated line of parasite, whereas the histopathological changes in the two dogs infected i.v. with L. infantum WT were severe in form and manifested by infiltration of high numbers of inflammatory cells. No promastigotes were found in cultures set up from spleens and livers of dogs infected with L. infantum H-line at 12 months post-infection, whereas promastigotes were seen in the spleen and liver cultures from 2 dogs infected i.v. with L. infantum WT. Serum levels of total IgG anti-Leishmania antibody were raised in all dogs. The antibody level in the serum of dogs infected i.v. with L. infantum WT was higher than that in dogs infected with L. infantum H-line. These results show no clinicopathological abnormalities in the dogs infected with gentamicin-attenuated L. infantum H-line. Moreover, L. infantum H-line induced IgG anti-Leishmania antibody in the dogs.

  12. Recent Advances in Vaccines Against Leishmania Based on Patent Applications.

    Science.gov (United States)

    Thomaz-Soccol, Vanete; Ferreira da Costa, Eduardo Scopel; Karp, Susan Grace; Junior Letti, Luiz Alberto; Soccol, Flavia Thomaz; Soccol, Carlos Ricardo

    2018-01-01

    Leishmaniasis is caused by parasites of the genus Leishmania, and represents a group of chronic diseases with an epidemiological and clinical diversity. The disease is endemic in tropical regions, being found in 98 countries, affecting around 12 million people, with an estimated increase of 1.5 million per year. The present review aims to analyze recent and most important patents regarding development of vaccines to improve immunization against leishmaniasis. For this purpose, the Web of Science - Derwent Innovations Index was consulted. There is also a short description of the licensed vaccines already on the market for commercialization, and a critical opinion on future developments. The data herein presented comprises national and international filings, thus considering the patent's country of origin, and can be used an indicator of a country's technological development regarding a specific field. Several types of vaccines against Leishmania were studied. The main classes comprise: vaccines using live cells (virulent or attenuated); dead cells; containing recombinant protein; using DNA of the parasite. United States (74 patents) leads the ranking of patent applications for vaccines against Leishmania, followed by Brazil (36 patents), which is an endemic region of leishmaniasis with 20,000 human cases of cutaneous leishmaniasis and over 3,000 cases of visceral form. This review showed that there is still a lot of space for development regarding the creation of a feasible, effective vaccine against leishmaniasis. The scientific community appears to be taking steps in the right direction, though. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  13. Disseminated feline leishmaniosis due to Leishmania infantum in Southern France.

    Science.gov (United States)

    Ozon, C; Marty, P; Pratlong, F; Breton, C; Blein, M; Lelièvre, A; Haas, P

    1998-02-28

    A fortuitously discovered case of feline leishmaniosis is reported. The parasites were found in the skin and the bone marrow of a domestic female cat that spontaneously died after a few weeks of evolution. Serological tests for FeLV, FIV and PIF virus detection gave negative results. By using Western blot serology, a characteristic pattern of leishmaniosis was obtained and by performing an isoenzyme electrophoresis, a Leishmania infantum MON-1 strain was identified. The same zymodeme is implicated in most of the canine and human leishmaniosis in Southern Europe. A study on the prevalence of asymptomatic feline leismaniosis is foreseen.

  14. Analysis of kinetoplast cytochrome b gene of 16 Leishmania isolates from different foci of China: different species of Leishmania in China and their phylogenetic inference

    Science.gov (United States)

    2013-01-01

    Background Leishmania species belong to the family Trypanosomatidae and cause leishmaniasis, a geographically widespread disease that infects humans and other vertebrates. This disease remains endemic in China. Due to the large geographic area and complex ecological environment, the taxonomic position and phylogenetic relationship of Chinese Leishmania isolates remain uncertain. A recent internal transcribed spacer 1 and cytochrome oxidase II phylogeny of Chinese Leishmania isolates has challenged some aspects of their traditional taxonomy as well as cladistics hypotheses of their phylogeny. The current study was designed to provide further disease background and sequence analysis. Methods We systematically analyzed 50 cytochrome b (cyt b) gene sequences of 19 isolates (16 from China, 3 from other countries) sequenced after polymerase chain reaction (PCR) using a special primer for cyt b as well as 31 sequences downloaded from GenBank. After alignment, the data were analyzed using the maximum parsimony, Bayesian and netwok methods. Results Sequences of six haplotypes representing 10 Chinese isolates formed a monophyletic group and clustered with Leishmania tarentolae. The isolates GS1, GS7, XJ771 of this study from China clustered with other isolates of Leishmania donovani complex. The isolate JS1 was a sister to Leishmania tropica, which represented an L. tropica complex instead of clustering with L. donovani complex or with the other 10 Chinese isolates. The isolates KXG-2 and GS-GER20 formed a monophyletic group with Leishmania turanica from central Asia. In the different phylogenetic trees, all of the Chinese isolates occurred in at least four groups regardless of geographic distribution. Conclusions The undescribed Leishmania species of China, which are clearly causative agents of canine leishmaniasis and human visceral leishmaniasis and are related to Sauroleishmania, may have evolved from a common ancestral parasite that came from the Americas and may have

  15. Methylobacterium spp. as an indicator for the presence or absence of Mycobacterium spp.

    OpenAIRE

    Falkinham III, Joseph O.; Williams, Myra D.; Kwait, Rebecca; Lande, Leah

    2016-01-01

    Objective/Background: A published survey of bacteria in showerhead biofilm samples revealed that Methylobacterium spp. and Mycobacterium spp. seldom coexisted in biofilms. Method: To confirm that information, biofilm samples were collected from household plumbing of Mycobacterium avium patients and Methylobacterium spp. and M. avium numbers were measured by direct colony counts. Results: The results demonstrated that if Methylobacterium spp. were present, Mycobacterium spp. were absent,...

  16. NKT cell activation by Leishmania mexicana LPG: Description of a novel pathway.

    Science.gov (United States)

    Zamora-Chimal, Jaime; Fernández-Figueroa, Edith A; Ruiz-Remigio, Adriana; Wilkins-Rodríguez, Arturo A; Delgado-Domínguez, José; Salaiza-Suazo, Norma; Gutiérrez-Kobeh, Laila; Becker, Ingeborg

    2017-02-01

    NKT cells have been associated with protection against Leishmania donovani, yet their role in infections with Leishmania mexicana has not been addressed, nor has the activation pathway been defined after stimulation with Leishmania mexicana lipophosphoglycan (LPG). We analyzed the activation of NKT cells and their cytokine production in response to Leishmania mexicana LPG. Additionally we compared NKT-cell numbers and cytokine profile in lymph nodes of skin lesions induced by Leishmania mexicana in BALB/c and C57BL/6 mice. We show that LPG activates NKT cells primarily through the indirect pathway, initiating with TLR2 stimulation of dendritic cells (DC), thereby enhancing TLR2, MHC II, and CD86 expressions and IL-12p70 production. This leads to IFN-γ production by NKT cells. C57BL/6 mice showed enhanced DC activation, which correlated with augmented IFN-γ production by NKT cells. Additionally, infected C57BL/6 mice showed elevated percentages of NKT cells with higher IFN-γ and IL-4 production in lymph nodes. We conclude that the response of NKT cells towards Leishmania mexicana LPG initiates with the indirect activation, after binding of LPG to TLR2 in DC. This indirect activation pathway enables NKT cells to produce IFN-γ during the innate phase of Leishmania infection, the magnitude of which differs between mouse strains. Copyright © 2016 The Authors. Published by Elsevier GmbH.. All rights reserved.

  17. Structurally optimized analogs of the retrograde trafficking inhibitor Retro-2cycl limit Leishmania infections.

    Science.gov (United States)

    Craig, Evan; Huyghues-Despointes, Charles-Eugene; Yu, Chun; Handy, Emma L; Sello, Jason K; Kima, Peter E

    2017-05-01

    In infected mammalian cells, Leishmania parasites reside within specialized compartments called parasitophorous vacuoles (LPVs). We have previously shown that Retro-2, a member of a novel class of small retrograde pathway inhibitors caused reduced LPV sizes and lower parasite numbers during experimental L. mexicana sp. infections. The purpose of this study was to determine if structural analogs of Retro-2cycl reported to have superior potency in the inhibition of retrograde pathway-dependent phenomena (i.e., polyomavirus cellular infection by polyomavrius and Shiga toxin trafficking in cells) are also more effective than the parent compound at controlling Leishmania infections. In addition to their effects on LPV development, we show that two optimized analogs of Retro-2cycl, DHQZ 36 and DHQZ 36.1 limit Leishmania amazonensis infection in macrophages at EC50 of 13.63+/-2.58μM and10.57+/-2.66μM, respectively, which is significantly lower than 40.15μM the EC50 of Retro-2cycl. In addition, these analogs caused a reversal in Leishmania induced suppression of IL-6 release by infected cells after LPS activation. Moreover, we show that in contrast to Retro-2cycl that is Leishmania static, the analogs can kill Leishmania parasites in axenic cultures, which is a desirable attribute for any drug to treat Leishmania infections. Together, these studies validate and extend the published structure-activity relationship analyses of Retro-2cycl.

  18. Genome-wide mapping reveals single-origin chromosome replication in Leishmania, a eukaryotic microbe.

    Science.gov (United States)

    Marques, Catarina A; Dickens, Nicholas J; Paape, Daniel; Campbell, Samantha J; McCulloch, Richard

    2015-10-19

    DNA replication initiates on defined genome sites, termed origins. Origin usage appears to follow common rules in the eukaryotic organisms examined to date: all chromosomes are replicated from multiple origins, which display variations in firing efficiency and are selected from a larger pool of potential origins. To ask if these features of DNA replication are true of all eukaryotes, we describe genome-wide origin mapping in the parasite Leishmania. Origin mapping in Leishmania suggests a striking divergence in origin usage relative to characterized eukaryotes, since each chromosome appears to be replicated from a single origin. By comparing two species of Leishmania, we find evidence that such origin singularity is maintained in the face of chromosome fusion or fission events during evolution. Mapping Leishmania origins suggests that all origins fire with equal efficiency, and that the genomic sites occupied by origins differ from related non-origins sites. Finally, we provide evidence that origin location in Leishmania displays striking conservation with Trypanosoma brucei, despite the latter parasite replicating its chromosomes from multiple, variable strength origins. The demonstration of chromosome replication for a single origin in Leishmania, a microbial eukaryote, has implications for the evolution of origin multiplicity and associated controls, and may explain the pervasive aneuploidy that characterizes Leishmania chromosome architecture.

  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.

  20. The flagellar protein FLAG1/SMP1 is a candidate for Leishmania-sand fly interaction.

    Science.gov (United States)

    Di-Blasi, Tatiana; Lobo, Amanda R; Nascimento, Luanda M; Córdova-Rojas, Jose L; Pestana, Karen; Marín-Villa, Marcel; Tempone, Antonio J; Telleria, Erich L; Ramalho-Ortigão, Marcelo; McMahon-Pratt, Diane; Traub-Csekö, Yara M

    2015-03-01

    Leishmaniasis is a serious problem that affects mostly poor countries. Various species of Leishmania are the agents of the disease, which take different clinical manifestations. The parasite is transmitted by sandflies, predominantly from the Phlebotomus genus in the Old World and Lutzomyia in the New World. During development in the gut, Leishmania must survive various challenges, which include avoiding being expelled with blood remnants after digestion. It is believed that attachment to the gut epithelium is a necessary step for vector infection, and molecules from parasites and sand flies have been implicated in this attachment. In previous work, monoclonal antibodies were produced against Leishmania. Among these an antibody was obtained against Leishmania braziliensis flagella, which blocked the attachment of Leishmania panamensis flagella to Phlebotomus papatasi guts. The protein recognized by this antibody was identified and named FLAG1, and the complete FLAG1 gene sequence was obtained. This protein was later independently identified as a small, myristoylated protein and called SMP1, so from now on it will be denominated FLAG1/SMP1. The FLAG1/SMP1 gene is expressed in all developmental stages of the parasite, but has higher expression in promastigotes. The anti-FLAG1/SMP1 antibody recognized the flagellum of all Leishmania species tested and generated the expected band by western blots. This antibody was used in attachment and infection blocking experiments. Using the New World vector Lutzomyia longipalpis and Leishmania infantum chagasi, no inhibition of attachment ex vivo or infection in vivo was seen. On the other hand, when the Old World vectors P. papatasi and Leishmania major were used, a significant decrease of both attachment and infection were seen in the presence of the antibody. We propose that FLAG1/SMP1 is involved in the attachment/infection of Leishmania in the strict vector P. papatasi and not the permissive vector L. longipalpis.