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4. ELISA with recombinant antigen Lb6H validated for the diagnosis of American tegumentary leishmaniasis

Author

Valencia-Portillo, Ruth Tamara, primary, Lindoso, José Angelo, additional, Celeste, Beatriz Julieta, additional, Bittencourt, Amanda Azevedo, additional, de Brito, Maria Edileuza Felinto, additional, Duthie, Malcolm Scott, additional, Guderian, Jeffery, additional, Guerra, Jorge, additional, Oliveira, Ana Lúcia Lyrio, additional, Reed, Steven, additional, Rocha, Mussya Cisotto, additional, Santos, Nicolle Tayná, additional, Silveira, Fernando Tobias, additional, Goto, Hiro, additional, and Sanchez, Maria Carmen Arroyo, additional

Published
2024
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5. Nocturnal activity and forestry-urban dispersal of phlebotomine sand flies (Diptera: Psychodidae) from an ecotourism park in Amazonian Brazil

Author

Neves, Danielly Mota, Sánchez Uzcátegui, Yetsenia del Valle, Silveira, Fernando Tobias, and Vasconcelos dos Santos, Thiago

Abstract

Phlebotomine sand flies are insects of notorious importance in public health, mainly due to their involvement in the transmission of Leishmaniaprotozoa. Their flight activity occurs predominantly in the twilight/night period, being stimulated mainly by the need to search for food and reproduction. Despite being naturally wild, some species are able to invade anthropized environments. Present work aimed to assess the nocturnal activity and forestry-urban dispersal of phlebotomine sand flies from an ecotourism park in Belém, Amazonian Brazil. The study area comprised a horizontal transect, extending from a forest park to the neighboring urban environment, in Belém. Sampling was conducted with night-operating light traps. Nocturnal activity was assessed through time-set captures with a collection bottle rotator in the forest environment. Dispersal was assessed through captures carried out along the transect, starting from the forest edge (0 m), extending to the urban environment (50–200 m), phlebotomine sand flies were identified. Abundance, richness, diversity, and sampling sufficiency were estimated. Fourteen species were recorded in the surveyed environments, with Nyssomyia antunesi, Trichophoromyia brachipyga, and Trichophoromyia ubiquitalisbeing the most abundant. Nocturnal activity behavior of Ny. antunesioccurred between 8 PM and 4 AM while that of Th. brachipygaand Th. ubiquitalisoccurred between 2 AM and 6 AM. In the urban environment, the 150 m site presented the highest abundance. Bichromomyia flaviscutellatawas sampled in all sites. Gravid females of Pressatia chotiand Bi. flaviscutellatawere sampled in the urban environment. Putative differential activity between the species herein assessed and their urban dispersal observed are worthy of note, adding data for supporting vector surveillance at a local scale.

Published
2025
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9. Anti-leishmanial activity of Eleutherine plicata Herb. and predictions of isoeleutherin and its analogues

Author

Albuquerque, Kelly Cristina Oliveira de, primary, Veiga, Andreza do Socorro Silva da, additional, Silveira, Fernando Tobias, additional, Campos, Marliane Batista, additional, Costa, Ana Paula Lima da, additional, Brito, Ananda Karolyne Martins, additional, Melo, Paulo Ricardo de Souza, additional, Percario, Sandro, additional, Molfetta, Fábio Alberto de, additional, and Dolabela, Maria Fâni, additional

Published
2024
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11. Ground-dwelling invertebrates from an Urban park in the Brazilian Amazon, with particular reference to Phlebotominae (Diptera: Psychodidae)

Author

Sánchez Uzcátegui, Yetsenia del Valle, Pacheco Colmenares, Rafael Antonio, Neves, Danielly Mota, Silveira, Fernando Tobias, Vasconcelos dos Santos, Thiago, and Póvoa, Marinete Marins

Abstract

The ground-dwelling invertebrate fauna from an urban park in Belém, Amazonian Brazil, with particular reference to the subfamily Phlebotominae (Diptera: Psychodidae), were characterized. Sampling was performed from March 2022 to May 2023, with 10 emergence traps installed in 2 microhabitats, 5 in each 1: “M1,” which included surrounding (up to 1.5 m) trees with tabular roots, and “M2,” which included no trees with tabular roots. Invertebrates trapped in adhesive papers were assessed on 2 occasions/cycles (D21/D42). During 10 sampling cycles (10 traps/cycle = 100 samples), 6,490 invertebrates were captured (M1, n= 4,203; M2, n= 2,287) and classified into 5 classes and 21 orders, with Diptera (n= 2,309; 35.6%) being the most abundant. Twenty-nine specimens of the following phlebotomine species were captured: Nyssomyia antunesi(M1, n= 10; M2, n= 3), Trichophoromyia ubiquitalis (M1, n= 6; M2, n= 1), Th. brachipyga(M1, n= 0; M2, n= 2), Bichromomyia flaviscutellata(M1, n= 2; M2, n= 1) and 4 unidentified specimens (M1, n= 2; M2, n= 2). The male/female ratio was 1.08. Fractional vegetation cover was compared, and the physiochemical characteristics of the soil were compared between the microhabitats. Only temperature showed significant differences. A weak positive correlation was found between phlebotomines and other dipterans and between temperature and the amount of organic matter in the soil. Both sampled microhabitats were shown to be suitable for the development and maintenance of different invertebrates, mainly dipterans. The composition of ground-emerging phlebotomine species was similar to that previously surveyed with light traps, including species associated with the transmission of Leishmaniaspp.

Published
2024
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13. Assessment of light-emitting diodes for sampling phlebotomines (Diptera: Psychodidae) from an urban park of the Brazilian Amazon

Author

de Souza, Renan Ney Castro, Sánchez Uzcátegui, Yetsenia del Valle, Silveira, Fernando Tobias, and Vasconcelos dos Santos, Thiago

Abstract

The present study aimed to assess different light sources for sampling phlebotomines (Diptera: Psychodidae) from Bosque Rodrigues Alves, a forested park surrounded by the urban area of Belém in the Brazilian Amazon. Centers for Disease Control traps, baited with blue, green, and warm white light-emitting diodes (LEDs), as test group, and incandescent light, as control group, were used. The electromagnetic spectra and luminous intensities of the light sources were characterized. Fractional vegetation cover at each sampling site was also estimated. Abundance, richness, rarefaction curves, Shannon and Simpson diversity indices, phlebotomines/trap/hour, and phlebotomines/trap/night were estimated and compared. The light sources of the test group presented greater luminous intensity than the control, but were similar to each other. There were no differences in vegetation cover at each site. A total of 1,346 phlebotomines comprising 11 species were sampled. The most abundant species were as follows: Nyssomyia antunesi(Coutinho, 1939), Trichophoromyia ubiquitalis(Mangabeira, 1942), Bichromomyia flaviscutellata(Mangabeira, 1942), and Th. brachipyga(Mangabeira, 1942). Light traps with LEDs had richness, abundance, and Shannon diversity indices similar to those obtained with incandescent light. The warm white LED had a higher Simpson’s index than the other light sources. Phlebotomine responses to incandescent light were similar to those to LEDs in most analyses, confirming the applicability of these light sources as alternative devices for entomological surveillance. Low consumption ensures greater autonomy of the traps, providing better operability during fieldwork.

Published
2024
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15. Can domestic dogs be considered a good reservoir of Leishmania (L.) infantum chagasi in an endemic area of nonulcerated cutaneous leishmaniasis in Southern Honduras?

Author

Segura, Gabriela Beatriz Rodriguez, primary, Ochoa, Wilfredo Humberto Sosa, additional, Matta, Vânia Lúcia Ribeiro da, additional, Martínez, Mercedes, additional, Tercero, Carol Rodriguez, additional, Gonzalez, Raquel Romero, additional, Pacheco, Carmen M. Sandoval, additional, Flores, Gabriela V. Araujo, additional, Silveira, Fernando Tobias, additional, Henriquez, Maria Mercedes Rueda, additional, and Laurenti, Márcia Dalastra, additional

Published
2023
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16. Comparative Genomic Analyses of New and Old World Viscerotropic Leishmanine Parasites: Further Insights into the Origins of Visceral Leishmaniasis Agents

Author

Silveira, Fernando Tobias, primary, Sousa Junior, Edivaldo Costa, additional, Silvestre, Rodrigo Vellasco Duarte, additional, Vasconcelos dos Santos, Thiago, additional, Sosa-Ochoa, Wilfredo, additional, Valeriano, Concepción Zúniga, additional, Ramos, Patrícia Karla Santos, additional, Casseb, Samir Mansour Moraes, additional, Lima, Luciana Vieira do Rêgo, additional, Campos, Marliane Batista, additional, da Matta, Vania Lucia, additional, Gomes, Claudia Maria, additional, Flores, Gabriela V. Araujo, additional, Sandoval Pacheco, Carmen M., additional, Corbett, Carlos Eduardo, additional, and Laurenti, Márcia Dalastra, additional

Published
2022
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19. Whole-Genome Sequencing of Leishmania infantum chagasi Isolates from Honduras and Brazil

Author

Silveira, Fernando Tobias, primary, Sousa Junior, Edivaldo Costa, additional, Silvestre, Rodrigo Vellasco, additional, Costa-Martins, André Guilherme, additional, da Costa Pinheiro, Kenny, additional, Sosa Ochoa, Wilfredo, additional, Vasconcelos dos Santos, Thiago, additional, Ramos, Patrícia Karla, additional, Casseb, Samir, additional, da Silva, Sandro Patroca, additional, Valeriano, Concepción Zúniga, additional, Lima, Luciana Vieira, additional, Campos, Marliane Batista, additional, da Matta, Vania Lucia, additional, Gomes, Claudia Maria, additional, Flores, Gabriela Venicia, additional, Pacheco, Carmen Maria, additional, Corbett, Carlos Eduardo, additional, Nakaya, Helder, additional, and Laurenti, Márcia Dalastra, additional

Published
2021
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20. Comparative Genomic Analyses of New and Old World Viscerotropic Leishmanine Parasites: Further Insights into the Origins of Visceral Leishmaniasis Agents.

Author

Silveira, Fernando Tobias, Sousa Junior, Edivaldo Costa, Silvestre, Rodrigo Vellasco Duarte, Vasconcelos dos Santos, Thiago, Sosa-Ochoa, Wilfredo, Valeriano, Concepción Zúniga, Ramos, Patrícia Karla Santos, Casseb, Samir Mansour Moraes, Lima, Luciana Vieira do Rêgo, Campos, Marliane Batista, da Matta, Vania Lucia, Gomes, Claudia Maria, Flores, Gabriela V. Araujo, Sandoval Pacheco, Carmen M., Corbett, Carlos Eduardo, and Laurenti, Márcia Dalastra

Subjects
LEISHMANIASIS, VISCERAL leishmaniasis, GENOMICS, CIRCADIAN rhythms, MOLECULAR clock, DNA polymerases, PARASITES
Abstract

Visceral leishmaniasis (VL), also known as kala-azar, is an anthropozoonotic disease affecting human populations on five continents. Aetiologic agents belong to the Leishmania (L.) donovani complex. Until the 1990s, three leishmanine parasites comprised this complex: L. (L.) donovani Laveran & Mesnil 1903, L. (L.) infantum Nicolle 1908, and L. (L.) chagasi Lainson & Shaw 1987 (=L. chagasi Cunha & Chagas 1937). The VL causal agent in the New World (NW) was previously identified as L. (L.) chagasi. After the development of molecular characterization, however, comparisons between L. (L.) chagasi and L. (L.) infantum showed high similarity, and L. (L.) chagasi was then regarded as synonymous with L. (L.) infantum. It was, therefore, suggested that L. (L.) chagasi was not native to the NW but had been introduced from the Old World by Iberian colonizers. However, in light of ecological evidence from the NW parasite's enzootic cycle involving a wild phlebotomine vector (Lutzomyia longipalpis) and a wild mammal reservoir (the fox, Cerdocyon thous), we have recently analyzed by molecular clock comparisons of the DNA polymerase alpha subunit gene the whole-genome sequence of L. (L.) infantum chagasi of the most prevalent clinical form, atypical dermal leishmaniasis (ADL), from Honduras (Central America) with that of the same parasite from Brazil (South America), as well as those of L. (L.) donovani (India) and L. (L.) infantum (Europe), which revealed that the Honduran parasite is older ancestry (382,800 ya) than the parasite from Brazil (143,300 ya), L. (L.) donovani (33,776 ya), or L. (L.) infantum (13,000 ya). In the present work, we have now amplified the genomic comparisons among these leishmanine parasites, exploring mainly the variations in the genome for each chromosome, and the number of genomic SNPs for each chromosome. Although the results of this new analysis have confirmed a high genomic similarity (~99%) among these parasites [except L. (L.) donovani], the Honduran parasite revealed a single structural variation on chromosome 17, and the highest frequency of genomic SNPs (more than twice the number seen in the Brazilian one), which together to its extraordinary ancestry (382,800 ya) represent strong evidence that L. (L.) chagasi/L. (L.) infantum chagasi is, in fact, native to the NW, and therefore with valid taxonomic status. Furthermore, the Honduran parasite, the most ancestral viscerotropic leishmanine parasite, showed genomic and clinical taxonomic characteristics compatible with a new Leishmania species causing ADL in Central America. [ABSTRACT FROM AUTHOR]

Published
2023
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21. Evaluation of systemic immunity in atypical cutaneous leishmaniasis caused by Leishmania (L.) infantum chagasi

Author

Laurenti, Márcia Dalastra, primary, Sosa‐Ochoa, Wilfredo, additional, Araujo Flores, Gabriela Venicia, additional, Sandoval Pacheco, Carmen Maria, additional, Tomokane, Thaise Yumie, additional, da Silva Oliveira, Luanda Mara, additional, Zúniga, Concepción, additional, Silveira, Fernando Tobias, additional, and Corbett, Carlos Eduardo Pereira, additional

Published
2021
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25. Natural Leishmania (Viannia) infections of phlebotomines (Diptera: Psychodidae) indicate classical and alternative transmission cycles of American cutaneous leishmaniasis in the Guiana Shield, Brazil

Author

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

Subjects
Phlebotomine, Natural infection, Leishmania, Guiana Shield, Infectious and parasitic diseases, RC109-216
Abstract

From 1996 to 1999 multi-trapping methods (Center of Diseases Control, CDC) light traps, light-baited Shannon traps, and aspiration on tree bases) were used to study the phlebotomine fauna of the “Serra do Navio” region of the Brazilian State of Amapá, which is part of the Guiana Shield. Fifty-three species were identified among 8,685 captured individuals. The following species, associated with the transmission of American cutaneous leishmaniasis in Amazonian Brazil, were captured: Nyssomyia umbratilis (3,388), Psychodopygus squamiventris maripaensis (995), Ny. anduzei (550), Trichophoromyia ubiquitalis (400), Ny. whitmani (291), Ps. paraensis (116), and Bichromomyia flaviscutellata (50). Flagellate infections were detected in 45 flies. Of the 19 parasites isolated in vitro, 15 were Leishmania (Viannia) guyanensis (13 in Ny. umbratilis, 1 in Ny. whitmani, 1 in Ny. anduzei) and three were L. (V.) naiffi (2 in Ps. s. maripaensis, 1 in Ny. anduzei). The results indicate the participation of three phlebotomine species in the transmission of L. (V.) guyanensis and two species in that of L. (V.) naiffi, and show that the same phlebotomine species is involved in the transmission of different Leishmania (Viannia) species in the Guianan/Amazon region. A review of the literature together with the results of the present study, and other published and unpublished results, indicate that eight phlebotomine species potentially participate in the transmission of Leishmania (Viannia) naiffi in Amazonia.

Published
2017
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26. Actividad leishmanicida de Aspidosperma nitidum Benth. Ex Müll. Arg

Author

Veiga, Andreza do Socorro Silva da, Silveira, Fernando Tobias, Marinho, Andrey Moacir do Rosario, Trindade, Rafaela Cabral dos Santos da, Campos, Marliane Batista, and Dolabela, Maria Fani

Subjects
Aspidosperma, Leismania, Extracto, Fraction, Fracción, Fração, Extract, Extrato
Abstract

Drug treatment for leishmaniasis can lead to several adverse reactions, and therapy-resistant isolates of Leishmania have already been found. In this scenario, the search for therapeutic alternatives and alkaloids, isolated from plants belonging to the genus Aspidosperma, has proved to be promising as antiparasitic. The present study evaluated the antipromastigote, amastigote and cytotoxicity activity of the extract and fractions belonging to A. nitidum. The ethanol extract of the shells (EE) was obtained by maceration and submitted to an acid:base partition, with the FN and FA fractions being obtained. In addition, the EE was subjected to fractionation under reflux, obtaining the fractions FrHEX, FrDCL, FrAcOEt and FrMeOH. The antipromastigote activity in L. amazonesis and cytotoxicity to macrophages were evaluated using the viability test (MTT). After the invasion of macrophages by the parasite, the treatment was performed with different sample concentrations during 72 hours, followed by samples staining and reading under the optical microscope. Only FrDCL was moderately active against promastigotes (IC50=105.7 µg/mL). However, EE (IC50 = 23.87 µg/mL) and FA (IC50 = 18.5 µg/mL) were shown to be active against amastigote, whereas in FrDCL (IC50 = 204.5 µg/mL) and FrHEX (IC50 = 143.0 µg/mL) moderate activity was observed. When samples cytoxicity is related to antiamastigote activity, the EE (CC50 = 491.8 µg/mL; IS = 21) seems to be more promising than the FA (CC50 = 209.1 µg/mL; IS = 11). In summary, A. nitidum is promise as a leishmanicide. El tratamiento farmacológico de la leishmaniasis puede provocar varias reacciones adversas y ya se han encontrado cepas de Leishmania resistentes al tratamiento. En este escenario, la búsqueda de alternativas terapéuticas y alcaloides, aislados de plantas pertenecientes al género Aspidosperma, ha demostrado ser prometedora como antiparasitario. El presente estudio evaluó la actividad antipromastigota, amastigota y citotoxicidad del extracto y fracciones pertenecientes a A. nitidum. El extracto etanólico de las cáscaras (EE) se obtuvo por maceración y sometido a un reparto ácido: base, obteniéndose las fracciones FN y FA. Además, el EE se sometió a fraccionamiento a reflujo, obteniendo las fracciones FrHEX, FrDCL, FrAcOEt y FrMeOH. La actividad antipromastigota en Leishmania (Leishmania) amazonensis y la citotoxicidad para los macrófagos se evaluaron mediante la prueba de viabilidad (MTT). Tras la invasión de los macrófagos por el parásito, se realizó el tratamiento con diferentes concentraciones de las muestras durante 72 h, seguido de la fijación y tinción de las muestras y lectura al microscopio óptico. Sólo FrDCL fue moderadamente activo contra promastigote (CI50 = 105,7 ± 1,12 µg / ml). Sin embargo, EE (CI50 = 23,87 ± 0,87 µg / mL) y FA (CI50 = 18,5 ± 0,94 µg / mL) mostraron ser activos contra amastigote, mientras que en FrDCL (CI50 = 204, 5 ± 0,71 µg / mL) y FrHEX (CI50 = 143,0 ± 0,82 µg / ml) se observó una actividad moderada. Cuando la citotoxicidad de las muestras se relaciona con la actividad antiamastigota, se observa que el EE (CC50 = 491,8 + 1,86 µg / mL; IS = 21) parece ser más prometedor que el FA (CC50 = 209,1 + 1, 7 µg / mL; IS = 11). En resumen, A. nitidum se ha mostrado prometedor como leishmanicida. O tratamento medicamentoso da leishmaniose pode acarretar várias reações adversas e já foram encontrados isolados de Leishmania resistentes a terapêutica. Neste cenário, torna-se imprescindível a busca de alternativas terapêuticas e alcaloides, isolados de plantas pertencentes ao gênero Aspidosperma, têm se mostrado promissores como antiparasitário. O presente estudo avaliou a atividade antipromastigota, amastigota e citotoxicidade do extrato e frações pertencentes a A. nitidum. O extrato etanólico das cascas (EE) foi obtido por maceração e submetido a partição ácido:base, sendo obtidas as frações FN e FA. Além disso, o EE foi submetido ao fracionamento sob refluxo, obtendo-se as frações FrHEX, FrDCL, FrAcOEt e FrMeOH. A atividade antipromastigota em L. amazonensis e citotoxicidade para macrófagos foram avaliadas através do ensaio de viabilidade (MTT). Após a invasão dos macrófagos pelo parasito, realizou-se o tratamento com diferentes concentrações das amostras por 72 h, seguida de fixação e coloração das amostras e leitura em microscópio óptico. Somente a FrDCL mostrou-se moderadamente ativa contra promastigota (CI50= 105,7±1,12 µg/mL). No entanto, o EE (CI50= 23,87 ±0,87 µg/mL) e FA (CI50= 18,5 ±0,94 µg/mL) mostraram-se ativos contra amastigota, enquanto que na FrDCL (CI50= 204,5 ± 0,71 µg/mL) e FrHEX (CI50= 143,0 ±0,82 µg/mL) observou-se atividade moderada. Quando se relaciona a citotoxicidade das amostras a atividade antiamastigota, observa-se que o EE (CC50= 491,8 + 1,86 µg/mL; IS= 21) parece ser mais promissor que a FA (CC50= 209,1 + 1,7 µg/mL; IS=11). Em síntese, a A. nitidum mostrou-se promissora como leishmanicida.

Published
2021

28. 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

Author

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, and Silveira Fernando Tobias

Subjects
Leishmania (Viannia) spp., Phlebotomine sand flies, Natural infections, Amazon region, Pará State, Brazil, Infectious and parasitic diseases, RC109-216
Abstract

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.

Published
2016
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29. Anti-leishmania activity of extract and fractions from the stem and leaf of Montrichardia linifera (Arruda) schott (Araceae) against Leishmania amazonensis

Author

Faria, Lara Vinha, primary, Brígido, Heliton Patrick Cordovil, additional, Bentaberry-Rosa, Alexandre Augusto, additional, Correa-Barbosa, Juliana, additional, Silva-Silva, João Victor, additional, Bastos, Mirian Letícia Carmo, additional, Costa, Erica Vanessa Souza, additional, Coelho-Ferreira, Márlia, additional, Silveira, Fernando Tobias, additional, and Dolabela, Maria Fâni, additional

Published
2021
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30. In vitro antileishmanial activity and phytochemical analysis of Casearia javitensis Kunth (Salicaceae)

Author

Silva, Carolina Moraes da, primary, Eiró, Luciana Guimarães, additional, Silva-Silva, João Victor, additional, Corrêa-Barbosa, Juliana, additional, Coelho-Ferreira, Márlia Regina, additional, Silveira, Fernando Tobias, additional, Marinho, Andrey Moacir do Rosário, additional, Almeida, Maria das Graças Carvalho, additional, Brigido, Heliton Patrick Cordovil, additional, and Dolabela, Maria Fâni, additional

Published
2020
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31. Botanical survey and leishmanicidal activity of grown-love

Author

Brandão, Dayse Lúcia do Nascimento, primary, Veiga, Andreza Socorro Silva, additional, Quaresma, Cintia Cardoso, additional, Busman, Dâmaris Vieira, additional, Lins, Alba Lúcia Ferreira de Almeida, additional, Silveira, Fernando Tobias, additional, Campos, Marliane Batista, additional, Percário, Sandro, additional, and Dolabela, Maria Fâni, additional

Published
2020
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32. Atividade antileishmania de Annona glabra L. (Annonaceae)

Author

Br´ígido, Heliton Patrick Cordovil, primary, Silva e Silva, João Victor da, additional, Bastos, Mírian Letícia Carmo, additional, Correa-Barbosa, Juliana, additional, Sarmento, Rosana Moura, additional, Costa, Erica Vanessa Souza, additional, Marinho, Andrey Moacir do Rosário, additional, Coelho-Ferreira, Márlia Regina, additional, Silveira, Fernando Tobias, additional, and Dolabela, Maria Fâni, additional

Published
2020
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35. Blood Feeding Sources of Nyssomyia antunesi(Diptera: Psychodidae): A Suspected Vector of Leishmania(Kinetoplastida: Trypanosomatidae) in the Brazilian Amazon

Author

Pimentel, Amanda Costa, Sánchez Uzcátegui, Yetsenia del Valle, de Lima, Ana Carolina Stocco, Silveira, Fernando Tobias, Vasconcelos dos Santos, Thiago, and Ishikawa, Edna Aoba Yassui

Abstract

Present work aimed to identify blood feeding sources and attempt to detect LeishmaniaDNA in Nyssomyia antunesi, suspected vector of Leishmaniasp., from a park in the urban center of Belém, the capital of Pará State, in the Brazilian Amazon. Entire bodies and gut contents of Ny. antunesiengorged females, previously captured in the urban park with Centers for Disease Control (CDC) light traps and aspiration on tree bases, were subjected to Leishmaniaand vertebrate DNA detection through amplification of the Leishmaniamini-exon and vertebrate cytochrome b (cyt b) gene regions, respectively. The quality of DNA extraction from entire bodies was ensured through amplification of the dipteran cyt b region. The vertebrate cyt b amplicons were sequenced and compared with those available on GenBank. A maximum likelihood phylogenetic tree was constructed to assess the clustering patterns of these sequences. LeishmaniaDNA was not detected. The sequences of 13 vertebrate cyt b amplicons were considered informative, exhibiting similarity and clustering with the following six vertebrate species: Dasyprocta leporina(1), Cuniculus paca(1), Tamandua tetradactyla(4), Choloepus didactylus(4), Pteroglossus aracari aracari(2), Homo sapiens(1). The samples of D. leporinaand C. pacawere obtained from the CDC canopy, whereas the others were by aspiration from tree bases. The present results revealed the eclectic and opportunist blood-feeding behavior of Ny. antunesi, with birds and mammals, these last ones acting as potential reservoirs for Leishmaniaspecies, distributed throughout the vertical forest strata.

Published
2022
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36. Evaluation of systemic immunity in atypical cutaneous leishmaniasis caused by Leishmania (L.) infantum chagasi.

Author

Laurenti, Márcia Dalastra, Sosa‐Ochoa, Wilfredo, Araujo Flores, Gabriela Venicia, Sandoval Pacheco, Carmen Maria, Tomokane, Thaise Yumie, Oliveira, Luanda Mara da Silva, Zúniga, Concepción, Silveira, Fernando Tobias, and Corbett, Carlos Eduardo Pereira

Subjects
LEISHMANIASIS, CUTANEOUS leishmaniasis, IMMUNOGLOBULIN M, HUMORAL immunity, VISCERAL leishmaniasis, CELLULAR immunity, IMMUNITY
Abstract

In some central‐American countries, Leishmania (L.) infantum chagasi infection can cause non‐ulcerated or atypical cutaneous leishmaniasis (NUCL) in addition to the classic clinical form, visceral leishmaniasis (VL). Little is known about the host‐parasite relationship that can contribute to the determination of one or another clinical form. The present study had the objective to evaluate the humoral and cellular immunity in the sera of individuals affected by NUCL to improve the comprehension of this atypical host‐parasite interaction. Based on clinical and laboratory diagnosis, serum of 80 individuals was collected to evaluate the cytokines and immunoglobulins profile of NUCL (n = 47), VL patients (n = 5), and negative controls (n = 28). Cytokines were detected using Cytokine Bead Array (CBA) Human Th1/Th2/Th17 kit according to the manufacturer's instructions; class (IgG and IgM), and subclass of (IgG1 and IgG2) immunoglobulins was evaluated by ELISA using specific antigens. The concentration of TNF‐α, IFN‐γ, IL‐2 and IL‐4 cytokines in NUCL, VL and control was present below the detection threshold of CBA kit. IL‐6, IL‐10 and IL‐17A cytokines was lower in NUCL compared to LV patients. Regarding to immunoglobulins, NUCL patients produced 4.0 times more IgG than the control, while VL patients produced 6.6 times more; and IgM level was 1.6 times higher in NUCL and 2.6 times in VL patients compared to the control. Concerning the immunoglobulins subclass, only VL patients showed positive reaction for IgG1, and IgG2 did not show positive reaction among the groups. The results showed a weak cellular and humoral systemic immune response in NUCL patients. [ABSTRACT FROM AUTHOR]

Published
2022
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40. Flavopereirine—An Alkaloid Derived from Geissospermum vellosii—Presents Leishmanicidal Activity In Vitro

Author

da Silva e Silva, João Victor, primary, Cordovil Brigido, Heliton Patrick, additional, Oliveira de Albuquerque, Kelly Cristina, additional, Miranda Carvalho, Josiwander, additional, Ferreira Reis, Jordano, additional, Vinhal Faria, Lara, additional, Coelho-Ferreira, Márlia, additional, Silveira, Fernando Tobias, additional, da Silva Carneiro, Agnaldo, additional, Percário, Sandro, additional, do Rosário Marinho, Andrey Moacir, additional, and Dolabela, Maria Fâni, additional

Published
2019
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42. Toll-like receptors 2, 4, and 9 expressions over the entire clinical and immunopathological spectrum of American cutaneous leishmaniasis due to Leishmania (V.) braziliensis and Leishmania (L.) amazonensis

Author

Campos, Marliane Batista, primary, Lima, Luciana Vieira do Rêgo, additional, de Lima, Ana Carolina Stocco, additional, Vasconcelos dos Santos, Thiago, additional, Ramos, Patrícia Karla Santos, additional, Gomes, Claudia Maria de Castro, additional, and Silveira, Fernando Tobias, additional

Published
2018
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43. Serum cytokine responses over the entire clinical-immunological spectrum of human Leishmania (L.) infantum chagasi infection

Author

Ramos, Patr?cia Karla Santos, Carvalho, Karina In?cio, Rosa, Daniela Santoro, Rodrigues, Ana Paula, Lima, Luciana Vieira, Campos, Marliane Batista, Gomes, Claudia Maria C, Laurenti, M?rcia Dalastra, Corbett, Carlos Eduardo, and Silveira, Fernando Tobias

Subjects
Interleucina-6 / imunologia, parasitic diseases, Brasil (BR), Leishmania infantum / imunologia, virus diseases, Leishmaniose Visceral, Interleucina-10 / imunologia, Ecossistema Amaz?nico, Estudos Transversais / m?todos, geographic locations, Citocinas / an?lise
Abstract

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Albert Einstein Israelite Hospital. S?o Paulo, SP, Brazil. Federal University of S?o Paulo. Division of Immunology. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Medical School of Sao Paulo University. Pathology Department. S?o Paulo, SP, Brazil. Medical School of Sao Paulo University. Pathology Department. S?o Paulo, SP, Brazil. Medical School of Sao Paulo University. Pathology Department. S?o Paulo, SP, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Federal University of Para. Tropical Medicine Nucleus. Bel?m, PA, Brazil. The clinical-immunological spectrum of human Leishmania (L.) infantum chagasi infection in Amazonian Brazil was recently reviewed based on clinical, DTH, and IFAT (IgG) evaluations that identified five profiles: three asymptomatic (asymptomatic infection, AI; subclinical resistant infection, SRI; and indeterminate initial infection, III) and two symptomatic (symptomatic infection, SI; American visceral leishmaniasis, AVL; and subclinical oligosymptomatic infection, SOI). TNF-?, IL-4, IL-6, and IL-10 serum cytokines were analyzed using multiplexed Cytometric Bead Array in 161 samples from endemic areas in the Brazilian Amazon: SI [AVL] (21 cases), III (49), SRI (19), SOI (12), AI (36), and a control group [CG] (24). The highest IL-6 serum levels were observed in the SI profile (AVL); higher IL-10 serum levels were observed in SI than in SOI or CG and in AI and III than in SOI; higher TNF-? serum levels were seen in SI than in CG. Positive correlations were found between IL-6 and IL-10 serum levels in the SI and III profiles and between IL-6 and TNF-? and between IL-4 and TNF-? in the III profile. These results provide strong evidence for associating IL-6 and IL-10 with the immunopathogenesis of AVL and help clarify the role of these cytokines in the infection spectrum.

Published
2017

44. Use of recombinant antigens for sensitive serodiagnosis of american tegumentary leishmaniasis caused by different leishmania species

Author

Sato, Camila Massae, Sanchez, Maria Carmen Arroyo, Celeste, Beatriz Julieta, Duthie, Malcolm S, Guderian, Jeffrey, Reed, Steven G, Brito, Maria Edileuza Felinto de, Campos, Marliane Batista, Encarna??o, Helia Valeria de Souza, Guerra, Jorge, Mesquita, Tirza Gabrielle Ramos de, Pinheiro, Suzana Kanawati, Ramasawmy, Rajendranath, Silveira, Fernando Tobias, Souza, Marina de Assis, and Goto, Hiro

Subjects
Prote?nas Recombinantes / gen?tica, Leishmaniose Cut?nea / diagn?stico, Leishmania / imunologia, Anticorpos Antiprotozo?rios / gen?tica, Sensibilidade e Especificidade, Testes Sorol?gicos / m?todos, Ensaio de Imunoadsor??o Enzim?tica / m?todos, Prote?nas Recombinantes / imunologia, Anticorpos Antiprotozo?rios / sangue, Anticorpos Antiprotozo?rios / imunologia
Abstract

This work was supported by Funda??o de Amparo ? Pesquisa do Estado de S?o Paulo (grant 2015/22075-8 and fellowship 2016/02622-7 to C.M.S.), by CAPES (fellowship to C.M.S.), by Conselho Nacional de Pesquisa (research fellowship to H.G.), and by Laborat?rios de Investiga??o M?dica (LIM-38-HC, FMUSP). This research was also supported by funding from the Bill and Melinda Gates Foundation under grants OPP49932 and OPP1084251 and by grant R01AI025038 from NIAID, National Institutes of Health. Universidade de S?o Paulo. Instituto de Medicina Tropical de S?o Paulo. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Instituto de Medicina Tropical de S?o Paulo. S?o Paulo, SP, Brazil. Universidade de S?o Paulo. Instituto de Medicina Tropical de S?o Paulo. S?o Paulo, SP, Brazil. Infectious Disease Research Institute. Seattle, WA, USA. Infectious Disease Research Institute. Seattle, WA, USA. Infectious Disease Research Institute. Seattle, WA, USA. Oswaldo Cruz Foundation. Aggeu Magalh?es Research Center. Department of Immunology. Recife, PE, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Funda??o de Medicina Tropical Dr. Heitor Vieira Dourado. Manaus, AM, Brazil. Funda??o de Medicina Tropical Dr. Heitor Vieira Dourado. Manaus, AM, Brazil / Universidade do Estado do Amazonas. Manaus, AM, Brazil. Funda??o de Medicina Tropical Dr. Heitor Vieira Dourado. Manaus, AM, Brazil Funda??o de Medicina Tropical Dr. Heitor Vieira Dourado. Manaus, AM, Brazil Funda??o de Medicina Tropical Dr. Heitor Vieira Dourado. Manaus, AM, Brazil / Universidade Nilton Lins. Manaus, AM, Brazil; Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / Universidade Federal do Par?. N?cleo de Medicina Tropical. Bel?m, PA, Brazil. Universidade de S?o Paulo. Instituto de Medicina Tropical de S?o Paulo. S?o Paulo, SP, Brazil / Oswaldo Cruz Foundation. Aggeu Magalh?es Research Center. Department of Immunology. Recife, PE, Brazil. Universidade de S?o Paulo. Instituto de Medicina Tropical de S?o Paulo. S?o Paulo, SP, Brazil / Universidade de S?o Paulo. Faculdade de Medicina. S?o Paulo, SP, Brazil. American tegumentary leishmaniasis (ATL) (also known as cutaneous leishmaniasis [CL]) is caused by various species of protozoa of the genus Leishmania. The diagnosis is achieved on a clinical, epidemiological, and pathological basis, supported by positive parasitological exams and demonstration of leishmanin delayedtype hypersensitivity. Serological assays are not routinely used in the diagnosis because many are considered to have low sensitivity and the particular Leishmania species causing the disease can lead to variable performance. In the present study, we generated recombinant versions of two highly conserved Leishmania proteins, Leishmania (Viannia) braziliensis-derived Lb8E and Lb6H, and evaluated both in enzyme-linked immunosorbent assays (ELISA). Recombinant Lb6H (rLb6H) had better performance and reacted with 100.0% of the ATL and 89.4% of the VL samples. These reactions with rLb6H were highly specific (98.5%) when compared against those for samples from healthy control individuals. We then assessed rLb6H against sera from ATL patients infected with different species of Leishmania prevalent in Brazil [Leishmania (Leishmania) amazonensis, L. (Viannia) braziliensis, and L. (V.) guyanensis] and samples from patients with other infectious diseases. In analyses of 500 sera, ELISA using rLb6H detected all 219 ATL samples (sensitivity of 100.0%) with an overall specificity of 93.9% (considering healthy individuals and other infectious diseases patients). Only a minority of samples from Chagas disease patients possessed antibodies against rLb6H, and all of these responses were low (with a highest reactivity index of 2.2). Taken together, our data support further evaluation of rLb6H and the potential for its routine use in the serological diagnosis of ATL.

Published
2017

45. Use of Recombinant Antigens for Sensitive Serodiagnosis of American Tegumentary Leishmaniasis Caused by Different Leishmania Species

Author

Sato, Camila Massae, primary, Sanchez, Maria Carmen Arroyo, additional, Celeste, Beatriz Julieta, additional, Duthie, Malcolm S., additional, Guderian, Jeffrey, additional, Reed, Steven G., additional, de Brito, Maria Edileuza Felinto, additional, Campos, Marliane Batista, additional, de Souza Encarnação, Helia Valeria, additional, Guerra, Jorge, additional, de Mesquita, Tirza Gabrielle Ramos, additional, Pinheiro, Suzana Kanawati, additional, Ramasawmy, Rajendranath, additional, Silveira, Fernando Tobias, additional, de Assis Souza, Marina, additional, and Goto, Hiro, additional

Published
2017
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47. NaturalLeishmania(Viannia) infections of phlebotomines (Diptera: Psychodidae) indicate classical and alternative transmission cycles of American cutaneous leishmaniasis in the Guiana Shield, Brazil

Author

de Souza, Adelson Alcimar Almeida, primary, da Rocha Barata, Iorlando, additional, das Graças Soares Silva, Maria, additional, Lima, José Aprígio Nunes, additional, Jennings, Yara Lúcia Lins, additional, Ishikawa, Edna Aoba Yassui, additional, Prévot, Ghislaine, additional, Ginouves, Marine, additional, Silveira, Fernando Tobias, additional, Shaw, Jeffrey, additional, and dos Santos, Thiago Vasconcelos, additional

Published
2017
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50. Molecular diagnosis and frequency of anti-Leishmania infantum chagasi antibodies in dogs in Bel?m/Par?, Brazil

Author

Schwanke, Katiane, Silva, Aryane M. M. da, Pacheco, Adlilton, Bahia, Michele, Silveira, Fernando Tobias, Scofield, Alessandra, and G?es-Cavalcante, Gustavo

Subjects
C?es, Leishmaniose Visceral / diagn?stico, Imunofluoresc?ncia / /utiliza??o, Leishmania infantum / parasitologia, Rea??o em Cadeia da Polimerase / m?todos, Testes Sorol?gicos / m?todos, Bel?m (PA)
Abstract

Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Laborat?rio de Parasitologia Animal. Castanhal, PA, Brasil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Laborat?rio de Parasitologia Animal. Castanhal, PA, Brasil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Laborat?rio de Parasitologia Animal. Castanhal, PA, Brasil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Laborat?rio de Parasitologia Animal. Castanhal, PA, Brasil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Laborat?rio de Parasitologia Animal. Castanhal, PA, Brasil. Universidade Federal do Par?. Instituto de Medicina Veterin?ria. Laborat?rio de Parasitologia Animal. Castanhal, PA, Brasil. A leishmaniose visceral ? uma enfermidade cujo agente etiol?gico no Brasil ? o protozo?rio Leishmania infantum chagasi. Os c?es s?o considerados reservat?rios urbanos da doen?a, sendo indicadores da ocorr?ncia de casos humanos. O presente trabalho teve como objetivo diagnosticar a infec??o por L. infantum chagasi em c?es domiciliados e errantes do munic?pio de Bel?m, estado do Par?, atrav?s da rea??o em cadeia da polimerase (PCR) e da rea??o de imunofluoresc?ncia indireta (RIFI), empregando dois ant?genos distintos. Amostras de sangue venoso de c?es adultos, sem distin??o de sexo ou ra?a, de diferentes bairros e ?pocas do ano da cidade de Bel?m-PA, foram colhidas em tubos sem e com anticoagulante para obten??o do soro e do DNA, respectivamente. Esses animais foram divididos em dois grupos: c?es errantes capturados pelo Centro de Controle de Zoonoses (Grupo A) e c?es domiciliados (Grupo B). Os soros foram analisados atrav?s do teste de RIFI para pesquisa de IgG utilizando-se dois ant?genos distintos: 1) ant?geno do kit Bio-Manguinhos/FIOCRUZ (Ag-PRO) contendo formas promastigotas de Leishmania sp. (complexo major-like); 2) Ant?geno do Instituto Evandro Chagas (Ag-AMA) constitu?do por formas amastigotas de L. infantum chagasi. A avalia??o dos dois ant?genos foi realizada com as amostras reagentes a partir da titula??o 1:80. J? a PCR foi realizada a partir do DNA extra?do do sangue total dos animais e amplificado utilizando-se os iniciadores RV1e RV2. Das 335 amostras analisadas, 10,4 por cento (35/335) foram reagentes na RIFI (Ag-PRO) e 0,9 por cento (3/335) reagiram com o Ag-AMA. A distribui??o das amostras positivas se deu da seguinte forma: Grupo A 14,8 por cento (25/169) com Ag-PRO e 1,2 por cento (2/169) com Ag-AMA; Grupo B 6 por cento (10/166) com Ag-PRO e 0,6 por cento (1/166) com Ag-AMA; sendo que todas as amostras positivas pelo teste de RIFI com o Ag-AMA tamb?m reagiram com o Ag-PRO e em nenhuma das amostras foi detectado o DNA de L. infantum chagasi. Os achados do presente estudo indicam que Bel?m ainda pode ser considerada ?rea n?o end?mica para leishmaniose visceral canina e que a natureza do ant?geno influencia no resultado da RIFI para a pesquisa de anticorpos anti-L. infantum chagasi em c?es, sendo que a RIFI que utiliza formas promastigotas de Leishmania major-like como ant?geno deve ser utilizada com cautela como m?todo diagn?stico confirmat?rio em estudos epidemiol?gicos em ?reas n?o end?micas para LVC. Visceral leishmaniasis is a disease whose etiological agent in Brazil is Leishmania infantum chagasi. Dogs are considered urban reservoirs of the disease, being an indicator of the human cases occurrence. The present study aimed to diagnose L. infantum chagasi infection in stray and owned dogs in Bel?m, Par? State, by polymerase chain reaction (PCR) and indirect immunofluorescence assay (IFA) using two different antigens. Venous blood samples from adult dogs, regardless of gender or breed, of different neighborhoods in Bel?m-PA, were collected in tubes with and without anticoagulant to obtain DNA and serum, respectively. These animals were divided into two groups: stray dogs captured by the Center for Zoonosis Control (Group A) and owned dogs (Group B). Sera were analyzed by IFA testing for IgG using two different antigens: 1) Bio-Manguinhos/Fiocruz antigen kit (Ag-PRO) containing promastigotes of Leishmania sp. (Complex Major-Like), 2) Instituto Evandro Chagas Antigen (Ag-AMA) consisting of amastigotes of L. infantum chagasi. The evaluation of the two antigens was performed considering positive the reactions above the 1:80 dilution. Already PCR was performed with DNA isolated from whole blood of animals and amplified with the primers RV1 and RV2. Of the 335 samples analyzed, 10.4 per cent (35/335) were positive by IFA (Ag-PRO) and 0.9 per cent (3/335) with the Ag-AMA. The distribution of positive samples is given as follows: Group A 14.8 per cent (25/169) with Ag-PRO and 1.2 per cent (2/169) with Ag-AMA; Group B 6 per cent (10/166) with Ag-PRO and 0.6 per cent (1/166) with Ag-AMA, being that all samples positive by IFA with Ag-AMA also reacted with Ag-PRO, and none of the samples detected DNA of L. infantum chagasi. The findings of this study indicate that Bel?m can still be considered non-endemic area for canine visceral leishmaniasis and the nature of the antigen influences the result of the IFA for the detection of anti-L. infantum chagasi antibodies in dogs, and the IFA using promastigotes of Leishmania major- -like antigen should be used with caution as a confirmatory diagnostic on epidemiological studies in non-endemic areas.

Published
2014

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