In vitro and in vivo activity of a hypotoxic copper(I) complex against dermotropic Leishmania species / Atividade in vitro e in vivo de um complexo de cobre(I) hipotóxico contra espécies dermotrópicas de Leishmania

Cutaneous leishmaniasis is a disease caused by protozoa of the genus Leishmania and, currently, the treatment of first choice is meglumine antimoniate. However, due to its limited effectiveness and high toxicity, it is necessary to seek new active principles for leishmaniasis treatment. Metal complexes are gaining importance due to their effectiveness and low toxicity. In this context, the present study aimed to evaluate the in vitro and in vivo antileishmanial activity of the hypotoxic copper(I) complex [HB(pz)3]Cu(PCN). Four dermotropic species of Leishmania were tested with the metal complex and its effectiveness was determined through parasitic viability and infectivity rate, and cytotoxicity was determined using a redox dye (resazurin). For the in vivo tests, hamsters were infected and the lesions treated with a formulated ointment containing the complex, the effectiveness of which was assessed by measuring the diameter of the inoculum/snout location and determining the parasitic load. The results demonstrated moderate toxicity in murine macrophages and human monocytes and better efficacy in Leishmania (V.) braziliensis when compared to the other species tested, with a 50% reduction in the viability of promastigote and amastigote forms (in vitro). General data from daily topical treatment for up to 30 days showed low efficacy for reducing lesions, and no clinical and parasitological cure was observed in the experimental animals. Thus, the [HB(pz)3]Cu(PCN) complex proved to be promising in in vitro studies against L. (V.) braziliensis, and should be further tested in new formulations and new experimental treatment schemes.

A leishmaniose cutânea é uma doença causada por protozoários do gênero Leishmania e, atualmente, o tratamento de primeira escolha é o antimoniato de meglumina. Porém, devido à sua eficácia limitada e alta toxicidade, é necessário buscar novos princípios ativos para o tratamento da leishmaniose. Os complexos metálicos vêm ganhando importância devido à sua eficácia e baixa toxicidade. Nesse contexto, o presente estudo teve como objetivo avaliar a atividade leishmanicida in vitro e in vivo do complexo hipotóxico de cobre(I) [HB(pz)3]Cu(PCN). Quatro espécies dermotrópicas de Leishmania foram testadas com o complexo metálico e sua eficácia foi determinada através da viabilidade parasitária e taxa de infectividade, e a citotoxicidade foi determinada com um corante redox (resazurina). Para os testes in vivo, hamsters foram infectados e as lesões foram tratadas com uma pomada formulada contendo o complexo. A eficácia foi avaliada medindo o diâmetro do inóculo/focinho e determinando a carga parasitária. Os resultados demonstraram toxicidade moderada em macrófagos murinos e monócitos humanos e melhor eficácia em Leishmania (V.) braziliensis quando comparada às demais espécies testadas, com redução de 50% na viabilidade das formas promastigotas e amastigotas (in vitro). Os dados gerais do tratamento tópico diário por até 30 dias mostraram baixa eficácia na redução das lesões, e nenhuma cura clínica e parasitológica foi observada nos animais experimentais. Portanto, o complexo [HB(pz)3]Cu(PCN) mostrou-se promissor em estudos in vitro contra L. (V.) braziliensis, devendo ser empregado em novas formulações e novos esquemas de tratamento experimental.

Antileishmanial activity of extracts from Libidibia ferrea: development of in vitro and in vivo tests / Atividade anti-leishmania de extratos de Libidibia ferrea: desenvolvimento de testes in vitro e in vivo

ABSTRACT Treatment of cutaneous leishmaniasis (CL) is difficult due to the scarce number of drugs able to eliminate completely the intracellular form of the parasite. In the present study, the aim was to evaluate: i) phytochemical properties of extracts from Libidibia ferrea; ii) antileishmanial activity of extracts from L. ferrea against promastigotes and amastigotes of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) guyanensis; iii) the effects of topical treatment using hydrogel containing active extract of L. ferrea on golden hamsters infected with L. (L.) amazonensis. Extracts from leaves, branches and fruits of L. ferrea were obtained with hexane and methanol and were tested by in vitro assays in promastigotes and murine macrophages J774 experimentally infected with amastigotes of Leishmania. Groups of hamsters with CL received topical treatment with a formulation of extract (10%) hydrogels, 50 mg.day-1 for 40 days. In vitro activity of FrMeOH (methanolic extract from fruits without seeds) resulted in significant reduction of viable promastigotes of L. (L.) amazonensis (IC50 of 15.4 µg.mL-1) and demonstrated inhibition potential of amastigote forms of L. (L.) amazonensis and L. (V.) guyanensis and low cytotoxicity in macrophages. The overall data of topical treatment with extract hydrogels (GelFrMeOH) showed that lesion sizes were significantly reduced (42.78%), with low parasite burden by RT-qPCR and culture analysis by microscopy examination, and with histopathological findings such as lower inflammatory cell infiltration 40 days after treatment. Chemical analysis demonstrated FrMeOH contains high levels of phenolic compounds. The results indicate a possible alternative therapy for CL using phytotherapics.

RESUMO O tratamento da leishmaniose cutânea (LC) é de difícil resultado, devido ao escasso número de fármacos capazes de eliminar completamente a forma intracelular do parasita. No presente estudo, objetivou-se avaliar: i) propriedades fitoquímicas dos extratos de Libidibia ferrea; ii) a atividade antileishmania de extratos de L. ferrea contra promastigotas e amastigotas de Leishmania (Leishmania) amazonensis e Leishmania (Viannia) guyanensis; iii) os efeitos do tratamento tópico utilizando hidrogel contendo extrato ativo de L. ferrea em hamsters dourados (Mesocricetus auratus) infectados com L. (L.) amazonensis. Extratos de folhas, galhos e frutos de L. ferrea foram obtidos com hexano e metanol e foram testados por ensaios in vitro contra promastigotas e macrófagos J774 infectados com amastigotas de Leishmania. Grupos de hamsters infectados receberam hidrogel tópico com extrato (10%), 50 mg.dia-1 durante 40 dias. A atividade in vitro de FrMeOH (extrato metanólico dos frutos) mostrou redução significativa de promastigotas de L. (L.) amazonensis (IC50 de 15,4 μg.mL-1), potencial de inibição de formas amastigotas de L. (L.) amazonensis e L. (V.) guyanensis (IC50 303,36 μg.mL-1) e baixa citotoxicidade em células de macrófagos J774. Os resultados destacaram que as lesões cutâneas de animais que receberam tratamento com hidrogel + extrato (GelFrMeOH) apresentaram redução significativa (42,78%), menos úlceração e redução da carga parasitária detectada por RT-qPCR, microscopia e análise em cultura e alterações histopatológicas, como menor infiltrado inflamatório após 40 dias de tratamento. A análise química demonstrou que FrMeOH contém altos níveis de compostos fenólicos. Os resultados apontam para uma possível terapia alternativa para CL utilizando fitoterápicos.

New human case reports of cutaneous leishmaniasis by Leishmania (Viannia) naiffi in the Amazon region, Brazil / Novos relatos de casos de leishmaniose cutânea por Leishmania (Viannia) naiffi na região amazônica, Brasil

Few cases of human cutaneous leishmaniasis (CL) caused by Leishmania naiffi were described in the medical literature. The aim of this study was to report and analyze new cases of L. naiffi in the period between the years 1992 to 2011. The strains were characterized by isoenzyme analysis. All patients assisted had small lesions; ranging from 1.0x1.0 mm and 13.5x11.5 mm. The lesions observed were widely distributed: 55.5% on the lower limb, 5.5% in the abdominal area, 16.6% on upper limb and 22.2% in upper limb and back. Seventy-two percent of patients had ulcerated lesions. Clinical course of the disease varied from 1 to 10 months. According to gender, most infected individuals were men (83.3%). The patients came from Amazonas (10), Pará (01) and Rondônia (01), north States of Brazil. Five patients were infected in experimental stations of the National Institute of Amazonian Research (INPA). Although the results of this study were similar to other reports in the literature, some of the patients had more of one skin lesion. It is also reported the first case of CL caused by L. naiffi in the State of Rondônia and identified an area of disease transmission in the experimental station of INPA.

Poucos casos de leishmaniose cutânea humana (LC) causada por Leishmania naiffi foram descritos na literatura médica. Assim, o objetivo deste estudo foi relatar e analisar novos casos de infecção por L. naiffi no período compreendido entre os anos de 1992 a 2011. As cepas foram caracterizadas por análise de isoenzimas. Todos os pacientes atendidos apresentavam lesões pequenas; variando entre 1,0x1,0 mm e 13,5x11,5 mm. As lesões observadas foram amplamente distribuídas: 55,5% no membro inferior, 5,5% na área abdominal, 16,6% no membro superior e 22,2% no membro superior e para trás. Setenta e dois por cento dos pacientes tiveram lesões ulceradas. O curso clínico da doença variou de 1 a 10 meses. De acordo com o sexo, a maioria dos indivíduos infectados eram homens (83,3%). Os pacientes vieram de Amazonas (10), Pará (01) e Rondônia (01), estados do norte do Brasil. Cinco pacientes foram infectados em estações experimentais do Instituto Nacional de Pesquisas da Amazônia (INPA). Embora os resultados encontrados neste estudo fossem semelhantes às outras descrições na literatura, alguns dos pacientes apresentavam mais de uma lesão cutânea. Também é relatado o primeiro caso de LC causada por L. naiffi no Estado de Rondônia e identificada uma área de transmissão da doença na estação experimental do INPA.

Topical treatment of experimental cutaneous leishmaniasis in golden hamster (Mesocricetus auratus) with formulations containing pentamidine

O tratamento atual para Leishmaniose Tegumentar Americana é realizado com antimoniais pentavalentes como tratamento padrão, e as drogas de segunda escolha incluem pentamidina e anfotericina B, mas essas terapias apresentam efeitos colaterais e requerem administração parenteral. O objetivo deste trabalho foi avaliar formulações tópicas contendo isetionato de pentamidina comercial (IP) no tratamento da leishmaniose cutânea experimental (LC). Hamsters dourados foram infectados no focinho com Leishmania (Leishmania) amazonensis. Grupos de seis animais receberam tratamento tópico com cremes anidro ou emulsões hidratantes, num máximo de 10 dias, com aplicação de 50 mg dia-1. Amostras de tecido de lesões tratadas foram avaliadas por histologia, microscopia eletrônica de transmissão (MET) e cultivo de biópsia. Comparado ao grupo sem tratamento, o tratamento tópico com emulsão hidratante com 10% de IP e ácido úsnico (ACE5AU) mostrou diminuição significativa (P=0,028) nas medidas de lesões, 20 dias após o final do tratamento e 27,37% de redução. O tratamento tópico com emulsão anidra com 10% de IP e ácido úsnico (ACPU) mostrou redução a carga parasitária em Golden hamsters. Este estudo demonstrou a possibilidade de utilizar o tratamento tópico para reduzir o número de parasitas e que este poderia ser associado a outras drogas para tratamento mais rápido e eficaz da leishmaniose cutânea.(AU)

Evaluation of different diagnostic methods of American Cutaneous Leishmaniasis in the Brazilian Amazon.

Epidemiological studies have been conducted to better understand the dynamics of American Cutaneous Leishmaniasis (ACL) in the Amazon region where distinct species of Leishmania circulate. In endemic areas, the optimal diagnosis must be made in the earlier clinical presentation to avoid the complications of chronic disease. The scarcity of financial support, laboratory infrastructure and trained persons are the major obstacles in this reality. This paper describes the result of performing different diagnostic methods for ACL in Amazonas State between the years 2010 and 2011. The tests used were the intradermal skin test (Montenegro's skin test), ELISA (Enzyme-Linked Immunosorbent Assay), direct examination, culture isolation and identification of Leishmania species. A total of 38 suspected human cases of ACL were diagnosed by different methods, of which 71.0% (n = 27) were positive by direct examination, 75.6% (n = 28) had positivity in the culture isolates and, of these, 54.0% (n = 19) had infection with Leishmania (Viannia) guyanensis. The positivity of the intradermal skin test with the leishmanin solution was observed in 77.0% of cases analyzed and the serology with detection of IgG and IgM showed the presence of antibodies in 100% of exams realized results, showing variation in the titles of antibodies. The success of Leishmaniasis treatment depends on an effective and early diagnosis. Parasitological diagnosis is highly specific, but sensitivity is subject to variation because the tissue distribution of parasites generally is not homogeneous and depends on the specie of parasite. Moreover, parasitological tests require invasive procedures and depend on restrictive conditions for the collection of biological sample, which limit their use in large-scale for epidemiological studies. ELISA has been the most widely used serological method for the diagnosis of Visceral Leishmaniasis (VL) as it is easy to perform and has a low cost. However, flaws in specificity are observed in the diagnosis of cutaneous leishmaniasis. Actually the diagnosis needs to be done as an associated methods depending on the question to be solved.

The role of inflammatory, anti-inflammatory, and regulatory cytokines in patients infected with cutaneous leishmaniasis in Amazonas State, Brazil.

The authors discuss in this paper the role of inflammatory, anti-inflammatory, and regulatory cytokines in patients infected with different species of Leishmania in Amazonas State, Brazil. A comparative analysis was made of serum concentrations of these cytokines in the peripheral blood of 33 patients infected with cutaneous leishmaniasis. The isolates were identified as Leishmania guyanensis, L. naiffi, and L. amazonensis. Most (64%) of the patients were male ranging in age from 18 to 58 years. Protein expression profiles of IL-2, IL-4, IL-6, IL-10, IFN-γ, TNF-α, and IL-17 cytokines were shown to vary significantly between infected and noninfected (control group) individuals and according to the Leishmania species. Infection caused by L. guyanensis accounted for 73% of the cases and patients with this parasite also showed higher concentrations of IL-2, IFN-γ, IL-4, and IL-17 when compared to infection by L. amazonensis. Patients with infection caused by L. naiffi showed higher concentration of the cytokines analyzed when compared to uninfected patients; however, there was no statistically significant difference with the other species analyzed.

Distribuição de casos de leishmaniose tegumentar no município de Rio Preto da Eva, Amazonas, Brasil / Distribution of cases of cutaneous leishmaniasis in the city of Rio Preto da Eva, Amazonas, Brazil

Foram analisados os aspectos clínicos e epidemiológicos de 90 pacientes com leishmaniose tegumentar americana (LTA) no município de Rio Preto da Eva, estado do Amazonas, Brasil, entre os anos de 2005 e 2012. Todos os pacientes apresentavam lesões cutâneas cujas cepas isoladas foram caracterizadas por eletroforese de isoenzimas e identificadas como Leishmania (Viannia) guyanensis (n=80), Leishmania (V.) naiffi (n=3) e Leishmania (Leishmania) amazonensis (n=7). A maior parte dos pacientes era do sexo masculino (70 porcento) e a faixa etária com maior número de casos foi de 11 a 20 anos (31,7 porcento). Em relação à ocupação, a maioria era constituída por agricultores ou caseiros (37,7 porcento). Um percentual de 95 porcento dos casos de LTA era proveniente de focos endêmicos de zonas rurais. O Assentamento Iporá, localizado no km 127 da AM 010, apresentou o maior número de casos (33,3 porcento). Concluímos que a LTA é uma parasitose endêmica de importância no município de Rio Preto da Eva, ocorre principalmente ao longo da rodovia AM 010 e, nos pacientes analisados, a aquisição da infecção ocorreu, majoritariamente, por meio de contato com o ciclo silvestre de transmissão, ressaltando-se, porém, a possibilidade de transmissão da doença no intra e peridomicílio...

We analyzed the clinical and epidemiological characteristics of 90 patients with Cutaneous Leishmaniasis (CL) in Rio Preto da Eva, Amazonas state, Brazil, between 2005 and 2012. All patients had skin lesions, whose isolates were characterized by isoenzyme electrophoresis and identified as Leishmania (Viannia) guyanensis (n=80), Leishmania (V.) naiffi (n=3) and Leishmania(Leishmania) amazonensis (n=7). Most patients were male (70 percent) and the age group with the highest number of cases was 11 to 20 years old (31.7 percent). In terms of occupation, most were farmers or home caretakers (37.7 percent). Around 95 percent of cases of CL were from rural areas of endemic foci and the settlement known as Iporá, located at KM 127 of the AM 010 road, had the highest number of cases (33.3 percent). We conclude that American egumentary Leishmaniasis is a parasitic disease endemic in Rio Preto da Eva (Amazonas State), occurring mainly along the 010 AM Highway, and that infection was mainly acquired through contact with the sylvatic transmission cycle, although the possibility of disease transmission peridomiciliary was also analysed...

Diversity and distribution of sandflies (Diptera: Psychodidae: Phlebotominae) in a military area in the state of Amazonas, Brazil.

This study reports the distribution, ecotopes and fauna diversity of sandflies captured in five training bases on a military reserve in Manaus, state of Amazonas (AM). A total of 10,762 specimens were collected, which were distributed among 58 species, with the highest number recorded at Base Instruction 1 (BI1). A higher rate of species richness was found at the Base Instruction Boina Rajada and low levels of diversity associated with a high abundance index with the clear dominance of Lutzomyia umbratilis, Lutzomyia ruii and Lutzomyia anduzei were found at BI1. The abundance of Lu. umbratilis raises the possibility of outbreaks of American cutaneous leishmaniasis by the main vector of the disease in AM.

Diversity and distribution of sandflies (Diptera: Psychodidae: Phlebotominae) in a military area in the state of Amazonas, Brazil

This study reports the distribution, ecotopes and fauna diversity of sandflies captured in five training bases on a military reserve in Manaus, state of Amazonas (AM). A total of 10,762 specimens were collected, which were distributed among 58 species, with the highest number recorded at Base Instruction 1 (BI1). A higher rate of species richness was found at the Base Instruction Boina Rajada and low levels of diversity associated with a high abundance index with the clear dominance of Lutzomyia umbratilis, Lutzomyia ruii and Lutzomyia anduzei were found at BI1. The abundance of Lu. umbratilis raises the possibility of outbreaks of American cutaneous leishmaniasis by the main vector of the disease in AM.

Ocorrência de flebotomíneos (Diptera: Psychodidae: Phlebotominae) no ambiente peridomiciliar em área de foco de transmissão de leishmaniose tegumentar no município de Manaus, Amazonas / Occurrence of sandflies (Diptera: Psychodidae: Phlebotominae) in the peridomestic environment in an area of transmission focus for cutaneous Leishmaniasis in Manaus, Amazon

Leishmaniasis is an antropozoonosis with natural foci, in which infection is maintained in wild ecotopes among the population of vectors and their vertebrate hosts. This study identified sand fly species involved in transmission of Leishmaniasis in the east area of Manaus, in households in the neighborhood of Cidade de Deus, where there are outbreaks of human and animal disease. The vicinity is near to Adolpho Ducke forest reserve. In the study, 122 specimens were captured at different times, using the CDC light traps. The most commonly encountered species of phlebotomine sandflies were Lutzomyia umbratilis, L. rorotaensis and L. olmeca nociva. Near one of the residences where the collections were made, there was disposal of organic waste, a fact that favors the attraction of animal reservoirs in the forest reserve, thus enabling the occurrence of the disease transmission cycle in the peridomicile.

A leishmaniose é uma antropozoonose com focos naturais, nos quais a infecção é mantida em ecótopos silvestres entre a população de vetores e seus hospedeiros vertebrados. Neste estudo foram identificadas espécies de flebotomíneos envolvidas na transmissão de leishmaniose tegumentar no Bairro Cidade de Deus, onde há focos da doença humana e animal. O bairro está situado à margem da reserva florestal Adolpho Ducke. Foram capturados 122 espécimens em épocas variadas, utilizando-se armadilhas luminosas do tipo CDC. As espécies de flebotomíneos mais encontradas foram Lutzomyia umbratilis, L. rorotaensis e L. olmeca nociva. Nas proximidades de uma das residências onde foram realizadas as coletas de flebotomíneos, verificou-se descarte de resíduos orgânicos (lixo), o que possibilita a atração de reservatórios silvestres presentes na reserva florestal favorecendo a ocorrência do ciclo de transmissão da doença no peridomicílio.

Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) in urban rainforest fragments, Manaus -- Amazonas State, Brazil.

The non-flooded upland rainforest fragment in the Federal University of Amazonas Campus is considered one of the world's largest urban tropical woodland areas and Brazil's second largest one in an urban setting. It is located in the city of Manaus, State of Amazonas at 03° 04' 34″ S, 59° 57' 30″ W, in an area covering nearly 800 hectares. Forty-one (41) sand fly species belonging to genus Lutzomyia were found attaining a total of 4662 specimens collected. Lutzomyia umbratilis was the dominant species at all heights, followed by Lutzomyia anduzei and Lutzomyia claustrei. The fauna alpha diversity index showed to be 6.4, which is not much lower than that reported for areas of continuous forest in this Amazonian region. This data provides additional evidence on Phlebotomine sand flies found to transmit Leishmania and other trypanosomatids to humans and other animals circulating in this area. This is the first study being reported on sand flies collected in an urban rainforest fragment in Amazonia.

Estudo de uma série de casos de leishmaniose tegumentar americana no município de Rio Preto da Eva, Amazonas, Brasil / Case series study of american cutaneous leishmaniasis in Rio Preto da Eva municipality, Amazonas State, Brazil

A leishmaniose tegumentar americana (LTA) é uma zoonose causada por flagelados do gênero Leishmania. Em áreas de desmatamento e extrativismo, a LTA atinge o homem quando este entra em contato com focos zoonóticos. No Brasil, a LTA tem caráter endêmico e está distribuída em todos os estados da Federação. No estado do Amazonas, segundo dados da Secretaria Estadual de Saúde (2005), os municípios de Manaus e Rio Preto da Eva estão em primeiro e segundo lugar, respectivamente, em número de casos registrados. Em Rio Preto da Eva, cerca de 80por cento dos casos de LTA são provenientes de assentamentos rurais ao longo da Rodovia AM-010. Foram diagnosticados 113 casos de LTA em 141 pacientes examinados ao longo de um ano, em todas as faixas etárias e de ambos os sexos, na Unidade Mista de Saúde de Rio Preto da Eva. Houve predomínio do sexo masculino (68,1por cento), adultos jovens com idades variando entre 20 e 39 (40,4por cento) anos, agricultores (49,1por cento), todos com a forma cutânea da doença principalmente em membros inferiores (35por cento). A realização de estudos de mapeamento das áreas de risco, assim como a identificação dos agentes causadores e transmissores da leishmaniose podem indicar propostas mais efetivas para o controle da doença.

Leishmania (Viannia) guyanensis induces low immunologic responsiveness in leishmaniasis patients from an endemic area of the Brazilian Amazon Highland.

Cutaneous leishmaniasis caused by Leishmania (Viannia) guyanensis (CL-Lguy) is endemic in the Brazilian Amazon, differing from L. braziliensis infection in clinical, diagnostic, and therapeutic aspects. T-cell reactivity to leishmanial antigens possibly involved in the pathogenesis of CL-Lguy was studied herein. Variable lymphoproliferative responses (LPRs) to Leishmania antigens were found among the 23 studied patients, and 50% of them showed low or no response to these antigens. Active disease was associated with an enrichment of leishmanial-reactive T lymphocytes, mainly TCD4(+). High and low interferon (IFN)-gamma producers were observed. TNF-alpha, interleukin (IL)-10, and IL-5 were consistently detected. CL-Lguy displayed low antibody response in comparison to L. braziliensis patients. CL caused by L. braziliensis presented positive LPRs and higher IFN-gamma production but undetectable IL-5. L. guyanensis seems to induce a down-regulation of the immune system compared with L. braziliensis. This finding could explain some aspects of clinical presentation of CL-Lguy, such as high tissue parasite burden and frequent resistance to therapy.

Fauna de flebotomíneos (Diptera: Psychodidae: Phlebotominae) em terra firme e planície fluvial na área de influência do gasoduto Coari-Manaus, Amazonas, Brasil / Fauna of Sandflies (Diptera: Psychodidae: Phlebotominae) on terra firme and fluvial lowland in areas under the influence of the Coari-Manaus gas pipeline, Amazonas, Brazil

Este trabalho teve como objetivo realizar um levantamento preliminar da entomofauna de flebotomíneos nas áreas de influência do gasoduto Coari-Manaus (AM). As coletas foram realizadas no período de 4 a 8 de agosto (2003), em cinco dos sete municípios da área de influência do gasoduto, utilizando-se de armadilhas de luz do tipo CDC. Foram capturados 205 indivíduos, distribuídos em 25 espécies, pertencentes a oito subgêneros (151 – 73,7%) e quatro grupos (54 – 26,3%). A espécie Lutzomyia umbratilis foi a mais numerosa encontrando-se 21,0% (43) do total coletado. Nestas coletas foi registrada pela primeira vez no Amazonas e segunda no Brasil a espécie Lutzomyia preclara, coletada nos municípios de Caapiranga e Manacapuru.

The object of this work was to make a preliminary survey of the entomofauna sandflies in the areas under the influence of the Coari-Manaus (AM) gas pipeline. The collections were made from 4 to 8 August 2003, in five of the seven municipal districts under the influence of the gas pipeline, using light traps of the type CDC. We captured 205 individuals, distributed in 25 species, belonging to eight subgenera (151 – 73.7%) and four groups (54 – 26.3%). The Lutzomyia umbratilis was the most numerous species found (43 – 21.0%). The species Lutzomyia preclara, collected in the municipal districts of Caapiranga and Manacupuru, was registered for the first time in the state of Amazonas, and the second time in Brasil.

[Isoenzymatic characterization of human isolates of Leishmania sp (Kinetoplastida: Trypanosomatidae) from the municipalities of Rio Preto da Eva and Manaus, State of Amazonas]. / Caracterização isoenzimática de isolados humanos de Leishmania sp (Kinetoplastida: Trypanosomatidae) dos municípios de Rio Preto da Eva e Manaus, Estado do Amazonas.

Twenty-three isolates of Leishmania sp from patients in the municipalities of Rio Preto da Eva and Manaus were characterized and identified by means of isoenzyme electrophoresis and the degree of similarity between the organisms was analyzed. The results indicated that Leishmania guyanensis and Leishmania naiffi were present in these two environments and that the Leishmania naiffi samples were heterogenous.