A dysflagellar mutant of Leishmania (Viannia) braziliensis isolated from a cutaneous leishmaniasis patient

Background: Parasites of the Leishmania genus alternate between the flagellated extracellular promastigote stage and intracellular amastigotes. Here we report the characterization of a Leishmania isolate, obtained from a cutaneous leishmaniasis patient, which presents peculiar morphological features. Methods: The parasite was cultured in vitro and characterized morphologically using optical and electron microscopy. Identification was performed based on monoclonal antibodies and internal ribosomal spacer typing. In vitro macrophage cultures, murine experimental models and sand fly infections were used to evaluate infectivity in vitro and in vivo. Results: The isolate was identified as Leishmania (Viannia) braziliensis. In the atypical promastigotes grown in culture, a short flagellum surrounded or interrupted by a protuberance of disorganized material was observed. A normal axoneme was present close to the basal body but without elongation much further outside the flagellar pocket. A disorganized swelling at the precocious end of the axoneme coincided with the lack of a paraflagellar rod structure. The isolate was able to infect macrophages in vitro, induce lesions in BALB/c mice and infect Lutzomyia longipalpis. Conclusions: Notwithstanding the lack of an extracellular flagellum, this isolate infects macrophages in vitro and produces lesions when inoculated into mice. Moreover, it is able to colonize phlebotomine sand flies. Considering the importance attributed to the flagellum in the successful infection and survival of Leishmania in the insect midgut and in the invasion of macrophages, these findings may bring new light into the infectious mechanisms of L. (V.) braziliensis.

Leishmania Metacyclogenesis Is Promoted in the Absence of Purines

Leishmania parasites, the causative agent of leishmaniasis, are transmitted through the bite of an infected sand fly. Leishmania parasites present two basic forms known as promastigote and amastigote which, respectively, parasitizes the vector and the mammalian hosts. Infection of the vertebrate host is dependent on the development, in the vector, of metacyclic promastigotes, however, little is known about the factors that trigger metacyclogenesis in Leishmania parasites. It has been generally stated that "stressful conditions" will lead to development of metacyclic forms, and with the exception of a few studies no detailed analysis of the molecular nature of the stress factor has been performed. Here we show that presence/absence of nucleosides, especially adenosine, controls metacyclogenesis both in vitro and in vivo. We found that addition of an adenosine-receptor antagonist to in vitro cultures of Leishmania amazonensis significantly increases metacyclogenesis, an effect that can be reversed by the presence of specific purine nucleosides or nucleobases. Furthermore, our results show that proliferation and metacyclogenesis are independently regulated and that addition of adenosine to culture medium is sufficient to recover proliferative characteristics for purified metacyclic promastigotes. More importantly, we show that metacyclogenesis was inhibited in sand flies infected with Leishmania infantum chagasi that were fed a mixture of sucrose and adenosine. Our results fill a gap in the life cycle of Leishmania parasites by demonstrating how metacyclogenesis, a key point in the propagation of the parasite to the mammalian host, can be controlled by the presence of specific purines.

Cutting Edge: Brazilian Pemphigus Foliaceus Anti-Desmoglein 1 Autoantibodies Cross-React with Sand Fly Salivary LJM11 Antigen

The environmental factors that contribute to the development of autoimmune diseases are largely unknown. Endemic pemphigus foliaceus in humans, known as Fogo Selvagem (FS) in Brazil, is mediated by pathogenic IgG4 autoantibodies against desmoglein 1 (Dsg1). Clusters of FS overlap with those of leishmaniasis, a disease transmitted by sand fly (Lutzomyia longipalpis) bites. In this study, we show that salivary Ags from the sand fly, and specifically the LJM11 salivary protein, are recognized by FS Abs. Anti-Dsg1 monoclonal autoantibodies derived from FS patients also cross-react with LJM11. Mice immunized with LJM11 generate anti-Dsg1 Abs. Thus, insect bites may deliver salivary Ags that initiate a cross-reactive IgG4 Ab response in genetically susceptible individuals and lead to subsequent FS. Our findings establish a clear relationship between an environmental, noninfectious Ag and the development of potentially pathogenic autoantibodies in an autoimmune disease. The Journal of Immunology, 2012, 189: 1535-1539.

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

Visceral leishmaniasis (VL) is a zoonotic disease characterized by infection of mononuclear phagocytes by Leishmania chagasi. The primary vector is Lutzomyia longipalpis and the dog is the main domestic reservoir. The control and current treatment of dogs using synthetic drugs have not shown effectiveness in reducing the incidence of disease in man. In attempt to find new compounds with leishmanicidal action, plant secondary metabolites have been studied in search of treatments of VL. This study aimed to evaluate the leishmanicidal activity of Musa paradisiaca (banana tree) and Spondias mombin (cajazeira) chemical constituents on promastigotes and amastigotes of L. chagasi. Phytochemical analysis by column chromatography was performed on ethanol extracts of two plants and fractions were isolated. Thin layer chromatography was used to compare the fractions and for isolation the substances to be used in vitro tests. The in vitro tests on promastigotes of L chagasi used the MTT colorimetric method and the method of ELISA in situ was used against amastigotes besides the cytotoxicity in RAW 264.7 cells. Of the eight fractions tested, Sm1 and Sm2 from S. mombin had no action against promastigotes, but had good activity against amastigotes. The fractions Mp1 e Mp4 of M. paradisiaca were very cytotoxic to RAW 264.7 cells. The best result was obtained with the fraction Sm3 from S. mombin with IC50 of 11.26 mu g/ml against promastigotes and amastigotes of 0.27 mu g/ml. The fraction Sm3 characterized as tannic acid showed the best results against both forms of Leishmania being a good candidate for evaluation in in vivo tests. (C) 2012 Published by Elsevier B.V.

Visceral leishmaniasis in Brazil: trends and challenges

The urbanization of visceral leishmaniasis in Brazil has been related to environmental changes, migration, interaction and spread of sylvatic reservoirs and infected dogs to areas with no transmission, and adaptation of the vector Lutzomyia longipalpis to the peridomiciliary environment. From 1980 to 2005, Brazil recorded 59,129 cases of visceral leishmaniasis, 82.5% of which in the Northeast region. Visceral leishmaniasis gradually spread to other regions of the country: in 1998 these other regions reported 15% of all cases, but by 2005 this proportion had increased to 44%. From 1998 to 2005, indigenous cases were reported in 1,904 different municipalities of the country (34.2%). Reservoir and vector control pose major challenges for disease control, since there is a need for better knowledge of vector behavior in urban areas, and control activities involve high operational costs. In recent years the Brazilian Ministry of Health has supported research on the laboratory diagnosis of infection and disease in humans and dogs, treatment of patients, evaluation of the effectiveness of control strategies, and development of new technologies that could contribute to the surveillance and control of visceral leishmaniasis in the country.

Epidemiological aspects of American cutaneous leishmaniasis and phlebotomine sandfly population, in the municipality of Monte Negro, State of Rondônia, Brazil

INTRODUCTION: This work was carried out on the purpose of identifying the species of phlebotomine sandflies in the municipality of Monte Negro, state of Rondonia, Brazil, that may have been transmitting the American cutaneous leishmaniasis (ACL), and concisely describe epidemiological aspects of disease. METHODS: The epidemiologic and socioeconomical indicators were obtained from government institutions and the local Municipal Secretary of Health. Phlebotomine sandflies were captured using CDC light traps between July 2006 to July 2008. The total of 1,240 of female sandflies were examined by PCR method directed to k-DNA. RESULTS: There has been a significant decrease in the incidence of ACL of about 50% over the last ten years in the municipality. A total of 1,935 specimens of 53 sandfly species were captured, three of the genus Brumptomyia genus and 50 of the genus Lutzomyia. The predominant species was Lutzomyia acanthopharynx, Lutzomyia whitmani, Lutzomyia geniculata and Lutzomyia davisi. None were positive for Leishmania sp. CONCLUSIONS: Four sandflies species were found in the State of Rondonia for the first time: Brumptomyia brumpti, Lutzomyia tarapacaensis, Lutzomyia melloi and Lutzomyia lenti. The presence of Lutzomyia longipalpis, was also captured. Socioeconomical improvement of Brazilian economy and the increase of environmental surveillance in the last 15 years collaborated in the decrease of people exposed to vectors, reducing the incidence of ACL.

Estudo da expansão da Leishmaniose Visceral no estado de São Paulo e de fatores intervenientes

INTRODUÇÃO: Leishmaniose visceral (LV) é uma doença negligenciada que afeta milhões de pessoas no mundo e que constitui um grave problema de saúde pública. OBJETIVOS: Descrever no tempo e no espaço, a dispersão de Lutzomyia longipalpis e a expansão da LV no estado de São Paulo (SP); identificar fatores associados a estes processos. MÉTODOS: Foram realizados estudos descritivos, ecológicos e de análise de sobrevida. Informações sobre o vetor e os casos foram obtidas na Superintendência de Controle de Endemias e no Sistema de Informações de Agravos de Notificação para o período de 1997 a 2014. A área de estudo foi composta pelos 645 municípios de SP. Foram produzidos mapas temáticos e de fluxo e calcularam-se incidência, mortalidade e letalidade por LV em humanos (LVH). Utilizou-se a técnica de análise de sobrevida (Curvas de Kaplan-Meier e Regressão de Cox) para a identificação de fatores associados à dispersão do vetor e expansão da LV. RESULTADOS: A partir da detecção de Lu. longipalpis em Araçatuba em 1997, deram-se a ocorrência do primeiro caso canino (LVC) (1998) e o primeiro caso humano (LVH) autóctones (1999) em SP. Até 2014, foi detectada a presença do vetor em 173 (26,8 por cento ) municípios, LVC em 108 (16,7 por cento ) e LVH em 84 (13 por cento ). A expansão dos três fenômenos ocorreu no sentido noroeste para sudeste e se deram a velocidades constantes. Na região de São José do Rio Preto, a dispersão do vetor deu-se por vizinhança com municípios anteriormente infestados, a expansão da LV relacionou-se com os municípios sede das microrregiões e a doença ocorreu com maior intensidade nas áreas periféricas dos municípios. A presença da Rodovia Marechal Rondon e a divisa com o Mato Grosso do Sul foram fatores associados à ocorrência dos três eventos, assim como a presença da Rodovia Euclides da Cunha para presença do vetor e casos caninos, e, presença de presídios para casos humanos. CONCLUSÕES: A dispersão do vetor e da LV em SP iniciou-se, a partir de 1997, próximo à divisa com o estado do Mato Grosso do Sul, avançou no sentido noroeste para sudeste, na trajetória da rodovia Marechal Rondon, e ocorreu em progressão aritmética, com as sedes das microrregiões de SP tendo papel preponderante neste processo. A ocorrência autóctone de LVC e LVH iniciou-se na sequência da detecção de Lu. longipalpis em Araçatuba e de seu espalhamento por SP e não a partir dos locais onde anteriormente ele já estava presente. O uso da análise de sobrevida permitiu identificar fatores associados à dispersão do vetor e a expansão da LV. Os resultados deste estudo podem ser úteis para aprimorar as atividades de vigilância e controle da LV, no sentido de retardar sua expansão e/ou de mitigar seus efeitos, quando de sua ocorrência.

Influência de fatores ambientais na Leishmaniose visceral no Rio Grande do Norte

The Visceral Leishmaniose (LV) disease is endemic in some places in Brazil. It is caused by the protozoa Leishmania chagasi, being transmitted for vector, the phlebotomies, Lutzomyia longipalpis. In virtue of the expansion of the illness in Rio Grande do Norte, it is necessary to evaluate the determinative ambient factors in the proliferation of the vector for better control of the illness. The variable rainfall and the social variables had been analyzed using space regression with two models and the ambient variable of ZANE and the variables analyzed in 205 houses in the cities of Natal, Extremoz, Nísia Floresta, São Gonçalo do Amarante, São Jose do Mipibu, Parnamirim and Macaíba the Person and ML Chi-square were used . The analyses had shown that high rainfall, plain relief, the forest, the humid tropical climate the activities of production culture of sugar cane and fruit culture and the presence of bovines increase the risk of the LV. The work showed that it has space aggregation and that ambient factors influence in the LV in the State

Characteristics of the biological cycle of lutzomyia evandroi Costa Lima & Antunes, 1936 (diptera: psychodidae) under experimental conditions

XIMENES, Maria de Fátima Freire de Melo; MACIEL, Janaína Cunha; JERONIMO, Selma Maria Bezerra. Characteristics of the Biological Cycle of Lutzomyia evandroi Costa Lima & Antunes, 1936 (diptera: psychodidae) under experimental conditions. Memorias do Instituto Oswaldo Cruz, Rio de Janeiro, v.96, n.6, p.883-886, ago. 2001. Disponivel em: Acesso em: 4 out. 2010.

Characteristics of the biological cycle of lutzomyia evandroi Costa Lima & Antunes, 1936 (diptera: psychodidae) under experimental conditions

XIMENES, Maria de Fátima Freire de Melo; MACIEL, Janaína Cunha; JERONIMO, Selma Maria Bezerra. Characteristics of the Biological Cycle of Lutzomyia evandroi Costa Lima & Antunes, 1936 (diptera: psychodidae) under experimental conditions. Memorias do Instituto Oswaldo Cruz, Rio de Janeiro, v.96, n.6, p.883-886, ago. 2001. Disponivel em: Acesso em: 4 out. 2010.

Characteristics of the biological cycle of lutzomyia evandroi Costa Lima & Antunes, 1936 (diptera: psychodidae) under experimental conditions

XIMENES, Maria de Fátima Freire de Melo; MACIEL, Janaína Cunha; JERONIMO, Selma Maria Bezerra. Characteristics of the Biological Cycle of Lutzomyia evandroi Costa Lima & Antunes, 1936 (diptera: psychodidae) under experimental conditions. Memorias do Instituto Oswaldo Cruz, Rio de Janeiro, v.96, n.6, p.883-886, ago. 2001. Disponivel em: Acesso em: 4 out. 2010.

Establecimiento, mantenimiento y productividad de una colonia de laboratorio de Lutzomyia spinicrassa Morales, Osorno-Mesa, Osorno y Hoyos, 1969 (Diptera: Psychodidae) en Colombia

El flebótomo Lutzomyia spinicrassa es vector de Leishmania braziliensis y tiene amplia distribución en plantaciones de café en Colombia y Venezuela. Metodología: Se estableció una colonia en condiciones de laboratorio a partir de 600 hembras de L. spinicrassa capturadas en el campo y mantenidas a temperatura de 23º C y humedad relativa de 70%. El tiempo de desarrollo desde huevo hasta adulto osciló entre 58 y 78 días, en promedio 11 semanas. Se compararon parámetros poblacionales de la especie obtenidos a partir de cinco generaciones sucesivas mantenidas en grupos, con una generación criada individualmente. Resultados: Se obtuvieron los siguientes parámetros en cada condición experimental: tasa neta de reproducción (6,92 y 7 hembras por hembra por generación), tasa intrínseca de incremento poblacional (0,17 y 0,18 hembras por hembra por semana) y tasa finita de incremento poblacional (1,06 y 1,19 individuos por hembra por semana). Conclusión: Estos datos sugieren que la colonia de L. spinicrassa tuvo un incremento constante durante las seis generaciones analizadas.

Spatial and temporal patterns of occurrence of Lutzomyia sand fly species in an endemic area for cutaneous leishmaniasis in the Atlantic Forest region of northeast Brazil

Sand fly populations of different ecological niches in the Amaraji endemic American Cutaneous Leishmaniasis (ACL) focus of the Pernambuco Atlantic Forest region of northeastern Brazil were monitored spatiotemporally. Lutzomyia whitmani was dominant in all niches but occurred in smaller numbers in forested locations. L. whitmani was significantly less seasonal than the other species, being present throughout the year while other species were more abundant between February and April. These results suggest that L. whitmani may potentially be the principal vector of ACL in the region, even though the sand fly fauna was diverse: 88% were L.whitmani and 12% belonged to 11 other species. Two other species, L. complexa (1.3%) and L. migonei (0.8%), considered to be ACL vectors in other regions, were also present. This detailed picture of the sand fly population`s abundance and spatiotemporal distribution provides a basis for future modeling studies of forecasting sand fly activity patterns and ACL occurence.

Phlebotominae (Diptera: Psychodidae) em área de transmissão de leishmaniose tegumentar americana no litoral norte do Estado de São Paulo, Brasil

A região do litoral norte do Estado de São Paulo registrou 14, 30 e 104 casos de leishmaniose tegumentar americana nos anos de 1993, 1994 e 1995 respectivamente. Com objetivo de caracterizar a fauna e a distribuição sazonal e horária das espécies de flebotomíneos foram realizadas coletas quinzenais de formas adultas durante o período de dezembro de 1995 a novembro de 1996, utilizando-se armadilhas luminosas do tipo CDC, no interior da residência, no peridomicílio e na mata durante 12 horas a partir do crepúsculo vespertino. No peridomicílio foi utilizada, durante 6 horas também a partir do crepúsculo vespertino, armadilha de Shannon instalada a 100 metros da casa. A cada 3 meses esta armadilha foi utilizada durante 12 horas. Foram observadas flutuações das densidades populacionais, bem como as ocorrências intra e extradomiciliar das espécies predominantes. Lutzomyia intermedia foi a espécie mais abundante nas diferentes armadilhas utilizadas e nos ambientes investigados.

Description of Two New Sand Fly Species Related to Nyssomyia antunesi (Diptera, Psychodidae, Phlebotominae)

Both sexes of two new Brazilian phlebotomine sand flies of the genus Nyssomyia Barretto 1962 [= Lutzomyia (Nyssomyia) sensu Barretto], Nyssomyia delsionatali n. sp. and Nyssomyia urbinattii n. sp., presenting close affinity with Nyssomyia antunesi (Coutinho), are described and illustrated. N. delsionatali n. sp was captured on the edge of a riparian Amazonian forest on the Juruena river in the northwest of Mato Grosso state and N. urbinattii n. sp in a riparian Amazonian forest on the Teles Pires river between Mato Grosso and Para states. Some measurements of both sexes of N. antunesi and illustration of the male genitalia and the female spermathecae as well as an identi_cation key for males and females of the genus Nyssomyia are provided.