Canine visceral leishmaniosis in an area of fishing tourism, Bonito Municipality, Mato Grosso do Sul, Central-West Brazil / Leishmaniose visceral canina em área de turismo pesqueiro, município de Bonito, Mato Grosso do Sul, Centro-Oeste do Brasil

ABSTRACT: The study aimed to investigate Leishmania infection in a population of dogs (92 animals) in a fishing area of Bonito Municipality, and evaluate comparatively the serological methods used, immunoenzymatic assay (ELISA), indirect immunofluorescence antibody test (IFAT), and rapid Dual Path-Platform test (DPP®). Blood and aspirate of bone marrow samples were used and parasitological investigation was also performed, such as parasite isolation in Neal, Novy, Nicolle (NNN) medium culture, Woo technique, Giemsa stained smears and specific identification by polymerase chain reaction (PCR). IFAT revealed 56/92 seropositive, the ELISA 8/92 and the DPP®41/92. Regarding the agreement between the serological tests by the Kappa index, there was a slight agreement between ELISA/IFAT and ELISA/DPP®, and fair agreement in IFAT/DPP®. The NNN culture was positive in three out of 21 dogs, and identified by PCR as Leishmania infantum chagasi. No samples were positive by the Woo technique. Our results showed low agreements between the serological tests recommended by the Brazilian Ministry of Heath, and it is necessary to associate other diagnostic techniques, such as parasitological tests and PCR, to increase the sensitivity for canine visceral leishmaniosis diagnosis, mainly regarding asymptomatic dogs in endemic areas.

RESUMO: O estudo teve como objetivo verificar a infecção por Leishmania em uma população de cães (n=92 animais) de uma área de turismo de pesca no município de Bonito e avaliar comparativamente os métodos sorológicos utilizados, ensaio imunoenzimático (ELISA), reação de imunofluorescência indireta (RIFI) e teste rápido de plataforma dupla (DPP®). Testes parasitológicos também foram realizados, como o isolamento do parasita em meio de cultura Neal, Novy, Nicolle, técnica de Woo, esfregaços em lâminas coradas com Giemsa e identificação específica pela reação em cadeia da polimerase (PCR). A RIFI revelou sorologia positiva de 56/92, ELISA 8/92 e DPP®41/92. Quanto à concordância entre os testes sorológicos pelo índice Kappa, houve um ligeiro acordo entre ELISA/RIFI e ELISA/DPP®, e uma concordância razoável entre RIFI/DPP®. A cultura NNN foi positiva em três cães, e identificada por PCR como Leishmania infantum chagasi. Nenhuma amostra foi positiva pela técnica de Woo. Nossos resultados mostraram baixas concordâncias entre os testes sorológicos recomendados pelo Ministério da Saúde, sendo necessário associar a outras técnicas de diagnóstico, como testes parasitológicos e PCR, para aumentar a sensibilidade ao diagnóstico de leishmaniose visceral canina, principalmente, com relação aos cães assintomáticos em áreas endêmicas.

Phlebotomine fanna (diptera: psychodidae) in anarea of fishing tourism in centtral-western Brazil / Fauna flebotomínea (Diptera: Psychodidae) em área de turismo pesqueiro no Centro-Oeste do Brasil

The aim of this study was to identify behavioral aspects of the sandfly fauna of a fishing tourism area in the municipality of Bonito (MS). Monthly captures were undertaken from December 2009 to November 2010, using automatic CDC type light traps, from 18h00 to 06h00, in a forested area, a savannah area, peridomiciles and animal shelters near peridomiciliary areas. Nyssomyia whitmani was the most frequent out of a total of 6,699 specimens collected, belonging to 16 species, followed by Psathyromyia bigeniculata and Lutzomyia longipalpis, found in all the environments investigated, though in their greatest numbers in the animal shelters. Ny. whitmani exhibited its highest frequencies during the dry months, coincident with the fishing season, when the risk of transmission of cutaneous leishmaniasis for tourists and inhabitants increases. Noteworthy was the finding of two species naturally infected by flagellates: Ny. whitmani and Pa. bigeniculata. The local population and visiting tourists should be warned of the threat posed by leishmaniasis and the health authorities alerted to the need for adopting environmental sanitary measures, especially regarding such animal shelters as they seem to provide favorable conditions to the proliferation, maintenance and breeding opportunities of phlebotomines.

O objetivo deste estudo foi identificar aspectos do comportamento da fauna flebotomínea de área de turismo pesqueiro localizada no município de Bonito (MS). Foram realizadas capturas mensais no período de dezembro de 2009 a novembro de 2010, utilizando armadilhas luminosas automáticas do tipo CDC das 18:00h às 6:00h, em matas, área de cerrado, peridomicílios e abrigos de animais próximos a áreas peridomiciliares. De um total de 6.699 espécimes coletados, pertencentes a 16 espécies, Nyssomyia whitmani foi a mais frequente, seguida de Psathyromyia bigeniculata e Lutzomyia longipalpis, encontradas em todos os tipos de ambientes, porém com maior expressão em abrigos de animais. Ny. whitmani apresentou frequências mais elevadas nos meses mais secos, coincidentes com a estação da pesca, o que eleva o risco de transmissão da leishmaniose tegumentar a turistas e moradores da área. Importante ressaltar o encontro de duas espécies naturalmente infectadas por flagelados: Ny. whitmani e Pa. bigeniculata. A população local e turistas devem ser advertidos da ameaça que representam as leishmanioses e as autoridades de saúde alertadas para adoção de medidas de saneamento ambiental, principalmente com relação aos abrigos de animais, que parecem fornecer condições favoráveis para a proliferação, manutenção e reprodução de flebotomíneos.

Presença de Toxocara spp. e ancilostomídeos em áreas de peridomicílios de uma comunidade pesqueira no Centro-Oeste do Brasil / Presence of Toxocara spp. and hookworms in peridomiciliary areas of a fishing community in the Mid-west region of Brazil

O objetivo do estudo foi verificar a contaminação do solo de peridomicílios de uma comunidade pesqueira localizadano município de Bonito, Mato Grosso do Sul, Brasil, onde é frequente a ocorrência de casos de Larva migrans cutânea.Utilizando-se das técnicas de centrifugo-flutuação, Baermann-Moraes e centrifugo- sedimentação, amostras de solo efezes de cães foram analisadas e os resultados evidenciaram altos índices de contaminação por ovos de Toxocara spp.,Ancilostomídeos e larvas de nematoides. Estes achados podem estar associados aos casos de Larvas migrans cutânea eàs elevadas taxas de eosinofilia em escolares daquela comunidade.

The aim of this study was to investigate soil contamination in peridomiciliary areas from a fishing community in themunicipality of Bonito, Mato Grosso do Sul state, Brazil, where is often found the occurrence of cases of cutaneous larvamigrans. Using the techniques of zinc sulphate centrifugal flotation, Baermann-Moraes and centrifugal sedimentation,soil samples and feces of dogs were analyzed and the results showed high levels of contamination by Toxocara spp.,hookworm and larvae of nematodes. These findings may be associated with cases of cutaneous larva migrans and highrates of eosinophilia in school children of that community.

Epidemiological factors related to the transmission risk of Trypanosoma cruzi in a Quilombola community, State of Mato Grosso do Sul, Brazil / Aspectos epidemiológicos relacionados ao risco de transmissão de Trypanosoma cruzi em comunidade Quilombola, Estado de Mato Grosso do Sul, Brasil

INTRODUCTION: This work was an epidemiological investigation of the risk of Trypanosoma cruzi transmission in the rural Quilombola community of Furnas do Dionízio, State of Mato Grosso do Sul, Brazil. METHODS: Of the 71 animals examined, seven were captured (two opossums, Didelphis albiventris; four rats, Rattus rattus; and one nine-banded armadillo, Dasypus novemcinctus) and 64 were domestic (one canine, Canis familiaris; five pigs, Sus scrofa; two bovines, Bos taurus; five caprines, Capra sp.; and 51 ovines, Ovis aries). Parasitological tests were performed to detect parasites in the blood and to identify the morphology of flagellates. These methods included fresh examinations, buffy coat tests and blood cultures. Molecular analysis of DNA for identification of trypanosomatids was performed by polymerase chain reaction (PCR) with primers S35 and S36. RESULTS: The parasitological tests showed flagellates in an opossum and two cattle. The molecular tests showed DNA from T. cruzi in an opossum and a pig. Triatoma sordida was the only triatomine species found in the community, and it colonized households (four specimens) and the surrounding areas (124 specimens). Twenty-three specimens tested positive for flagellates, which were subsequently identified as T. cruzi by PCR. CONCLUSIONS: Data analysis demonstrated that T. cruzi has a peridomestic life cycle that involves both domestic and wild mammals.

INTRODUÇÃO: Este trabalho foi uma investigação epidemiológica do risco de transmissão de Trypanosoma cruzi na comunidade rural Quilombola de Furnas do Dionízio, Estado de Mato Grosso do Sul. MÉTODOS: Dos 71 animais examinados, sete foram capturados (dois gambás, Didelphis albiventris; quatro ratos, Rattus rattus; e um tatu, Dasypus novemcinctus) e 64 eram domésticos (um canídeo, Canis familiaris; cinco suínos, Sus scrofa; dois bovinos, Bos taurus; cinco caprinos, Capra sp; e 51 ovinos, Ovis aries). Exames parasitológicos foram realizados para detectar parasitas no sangue e para identificar a morfologia dos flagelados. Estes métodos incluíram exame a fresco, exame do creme leucocitário e hemocultura. A análise molecular de DNA para identificação de tripanossomatídeos encontrados foi feita pela reação em cadeia da polimerase (PCR) com os primers S35 e S36. RESULTADOS: Os exames parasitológicos mostraram flagelados em um gambá e nos dois bovinos. Os testes moleculares mostraram a presença do DNA de T. cruzi em um gambá e um suíno. Triatoma sordida foi a única espécie de triatomíneo encontrada na comunidade colonizando domicílio (quatro espécimes) e peridomicílio (124 espécimes). Vinte e três amostras foram positivas para flagelados e identificados como T. cruzi pela PCR. CONCLUSÕES: A análise dos dados aponta para o ciclo peridoméstico do parasita e envolve tanto animais domésticos como selvagens.

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

Visceral leishmaniasis (VL) is a severe chronic disease caused by Leishmania (Leishmania) infantum chagasi. Better knowledge on the effects caused by this disease can help develop adequate clinical management and treatment. Parasitological and immunohistochemical studies were performed golden hamsters Mesocricetus auratus infected with bone marrow from individuals with VL in the State of Mato Grosso do Sul, central-west Brazil. The effects of parasitism in the spleen, liver, kidneys, lungs, heart and brain of the animals were examined. Eighteen hamsters were inoculated intraperitoneally, and six healthy animals were used as negative controls. The animals were kept in the animal house and checked for clinical signs. Specimens of each organ were examined for the presence of amastigotes. Immunohistochemical technique was performed in all brain specimens and organs negative on the direct examination of parasites. Direct examination of amastigotes was positive in the spleen and liver of all infected animals; 33.3 percent showed the parasite in the kidneys and lungs, and 16.7 percent in the heart. Parasitic forms were seen in 83.3 percent (15/18) of the brain examined. Immunohistochemistry confirmed the results of the direct examination, except in two specimens of lung tissue and in the brain specimens. Other studies are needed to further clarify the effect of the parasite in the central nervous system.

A leishmaniose visceral (LV) é uma doença crônica grave, causada pelo parasito Leishmania (Leishmania) infantum chagasi. Esclarecer as alterações provocadas pela doença é fundamental para que se adotem condutas clínicas e de tratamento adequadas. Com o objetivo de analisar a infecção experimental em hamsters da linhagem golden, Mesocricetus auratus, infectados com tecido de medula óssea de pacientes com LV no Estado de Mato Grosso do Sul, foram realizados estudos parasitológicos e de imunomarcação. Foi verificada a distribuição do parasitismo no baço, fígado, rim, pulmão, coração e encéfalo desses animais. Foram utilizados 18 hamsters experimentalmente inoculados via intra-peritoneal, e seis animais sadios como controles negativos. Os animais foram mantidos em biotério de experimentação e observados, em busca de alterações clínicas. Com fragmentos de cada órgão, procedeu-se a confecção de lâminas por aposição para pesquisa de amastigotas. Nos órgãos com resultado negativo na pesquisa direta do parasito, e em todas as amostras de encéfalo, foi realizada a técnica de imunohistoquímica. A pesquisa direta de amastigotas foi positiva no baço e fígado de todos os animais infectados; 33,3 por cento apresentaram o parasito em rim e pulmão, e 16,7 por cento no coração. Quando realizada a pesquisa em encéfalo, formas parasitárias foram observadas em 83,3 por cento (15/18) dos animais. A imunomarcação confirmou os resultados da pesquisa direta, exceto em duas amostras de tecido pulmonar e nas amostras de encéfalo. Mais estudos são necessários, para esclarecer o real papel do parasito no sistema nervoso central.

Leishmaniose tegumentar americana: estudo clínico, epidemiológico e laboratorial realizado no Hospital Universitário de Campo Grande, Mato Grosso do Sul, Brasil / American cutaneous leishmaniasis: clinical, epidemiological and laboratory studies conducted at a university teaching hospital in Campo Grande, Mato Grosso do Sul, Brazil

FUNDAMENTOS: Leishmaniose Tegumentar Americana é zoonose de manifestações clínicas variadas, em expansão no Brasil, sendo o estado de Mato Grosso do Sul importante área endêmica. OBJETIVOS: Avaliar clínica, epidemiológica e laboratorialmente pacientes com Leishmaniose Tegumentar Americana , atendidos no Hospital Universitário Maria Aparecida Pedrossian, Universidade Federal de Mato Grosso do Sul, Brasil (HU/UFMS). MÉTODOS: Trata-se de um estudo observacional do tipo transversal com abordagem descritiva e analítica. Foram avaliados, retrospectivamente, dados de pacientes suspeitos de Leishmaniose Tegumentar Americana , atendidos no HU/UFMS de 1998 a 2008, e encaminhados ao Laboratório de Parasitologia/UFMS para complementação diagnóstica. Para a inclusão neste estudo foram considerados critérios clínicos e laboratoriais. RESULTADOS: Quarenta e sete pacientes foram incluídos no estudo. Houve predominância de homens de 45 a 59 anos, com a forma cutânea, lesão única, ulcerada, em áreas expostas do corpo e com duração menor que seis meses. O comprometimento de mucosas foi crescente com o aumento da idade e maior em pacientes que procuraram atendimento tardiamente. Intradermorreação de Montenegro foi o exame de maior sensibilidade e o encontro do parasito mostrou-se mais difícil em lesões antigas. CONCLUSÃO: Suspeição diagnóstica precoce é de extrema importância para diagnóstico preciso. Associação de exame parasitológico e imunológico torna mais seguro o diagnóstico laboratorial.

BACKGROUND: American cutaneous leishmaniasis is a disease with a wide variety of clinical manifestations that is expanding throughout Brazil, the state of Mato Grosso do Sul constituting a significant endemic area. OBJECTIVES: To evaluate the clinical, epidemiological and laboratory characteristics of patients with American cutaneous leishmaniasis. Patients were recruited among those attending the Maria Aparecida Pedrossian Teaching Hospital of the Federal University of Mato Grosso do Sul, Brazil. METHODS: This was a cross-sectional, observational study conducted using a descriptive and analytical approach. Data from patients suspected of having American cutaneous leishmaniasis who were receiving care at this institute between 1998 and 2008 and were referred to the institute's parasitology laboratory for confirmation of diagnosis were evaluated retrospectively. Clinical and laboratory criteria were taken into consideration for the inclusion of patients to the study. RESULTS: Forty-seven patients were included in the study, the majority of whom were male and between 45 and 59 years of age. Most had the cutaneous form of the disease with a single, ulcerated lesion on exposed areas of the body, which had generally been present for periods of less than six months. Mucosal involvement increased with age and was highest in patients who had sought medical care at a later stage. The Montenegro skin test showed the highest sensitivity. Finding the parasite was more difficult in older lesions. CONCLUSION: Suspicion of the disease at an early stage is of extreme importance for a precise diagnosis. A combination of parasitological and immunological tests renders laboratory diagnosis more reliable.

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

INTRODUCTION: The work was conducted to study phlebotomine fauna (Diptera: Psychodidae) and aspects of American cutaneous leishmaniasis transmission in a forested area where Leishmania (Leishmania) amazonensis occurs, situated in the municipality of Bela Vista, State of Mato Grosso do Sul, Brazil. METHODS: The captures were conducted with modified Disney traps, using hamster (Mesocricetus auratus) as bait, from May 2004 to January 2006. RESULTS: Ten species of phlebotomine sandflies were captured: Brumptomyia avellari, Brumptomyia brumpti, Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni and Sciopemyia sordellii. The two predominant species were Ev bourrouli (57.3 percent) and Bi flaviscutellata (41.4 percent), present at all sampling sites. Two of the 36 hamsters used as bait presented natural infection with Leishmania. The parasite was identified as Leishmania (Leishmania) amazonensis. CONCLUSIONS: Analysis of the results revealed the efficiency of Disney traps for capturing Bichromomyia flaviscutellata and the simultaneous presence of both vector and the Leishmania species transmitted by the same can be considered a predictive factor of the occurrence of leishmaniasis outbreaks for the human population that occupies the location.

INTRODUÇÃO: O estudo foi realizado com o objetivo de estudar a fauna de flebotomíneos (Diptera: Psychodidae) e aspectos ligados à transmissão da leishmaniose tegumentar americana em uma área florestal com ocorrência de Leishmania (Leishmania) amazonensis, situada no município de Bela Vista, Estado do Mato Grosso do Sul, Brasil. MÉTODOS: As capturas de flebotomíneos foram realizadas utilizando-se armadilhas tipo Disney modificadas, com isca roedor, Mesocricetus auratus, no período de maio de 2004 a janeiro de 2006. RESULTADOS: As coletas resultaram na identificação de 10 espécies de Phlebotominae: Brumptomyia avellari, Brumptomyia brumpti, Bichromomyia flaviscutellata, Evandromyia bourrouli, Evandromyia lenti, Lutzomyia longipalpis, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni e Sciopemyia sordellii. As duas espécies predominantes foram Ev bourrouli, com 57,3 por cento dos espécimes coletados, e Bi. flaviscutellata, representada por 41,4 por cento e que esteve presente em todos os locais amostrados. Dois hamsters sentinelas adquiriram a infecção natural, sendo os isolados identificados como Leishmania amazonensis. CONCLUSÕES: Os resultados mostram a eficiência das armadilhas Disney para captura de Bichromomyia flaviscutellata, e a presença simultânea de ambos, o vetor e a espécie de Leishmania por ele transmitida pode ser considerada um fator preditor da ocorrência de leishmaniose para a população humana que permanecer nesse local.

HIV/AIDS-associated visceral leishmaniasis in patients from an endemic area in Central-west Brazil

An increase in morbidity associated with visceral leishmaniasis (VL) in human immunodeficiency virus (HIV)/AIDS patients has been described in Africa and the Mediterranean. Despite the high endemicity of VL and HIV-1/AIDS in Brazil, this association has not been thoroughly investigated. Our aim was to evaluate the epidemiologic and clinical characteristics of VL-HIV-1/AIDS cases from Central-west [Mato Grosso do Sul (MS)] Brazil. Medical records of 23 VL-HIV-1/AIDS patients were reviewed. Patients were predominantly adult males (87 percent) and 34.8 percent of the patients were intravenous drug users (IVDU). Leishmaniasis was the first opportunistic infection in 60 percent of the HIV-1 patients. Fever occurred in all patients, although splenomegaly and hepatomegaly were absent in 21.7 percent of the cases. CD4+ T-cell counts were below 200 cells/mm³ in 80 percent of the cases and the counts did not increase after clinical remission despite antiretroviral therapy. The first drug chosen to treat the cases was antimonial, but the therapeutic regimen was altered to amphotericin B in 12 of 17 cases due to side effects. Relapses were reported in 56.5 percent of the patients. IVDU may constitute an important risk factor for the transmission of both diseases in MS. VL-HIV-1/AIDS patients in MS share similar clinical characteristics as those from other endemic regions worldwide. Thus, these findings are critical for improving the surveillance of VL-HIV/AIDS patients.

Polymerase chain reaction of peripheral blood as a tool for the diagnosis of visceral leishmaniasis in children

The diagnosis of visceral leishmaniasis (VL) generally requires the use of invasive tests for the collection of infected tissue (aspirates of bone marrow, spleen, liver or lymph nodes). This difficulty has led to the search for safer and less painful techniques to confirm the occurrence of the disease in children. Polymerase chain reaction (PCR) is a method that is advantageous in that it allows the use of peripheral blood samples for diagnosis. This paper reports the utilisation of PCR on peripheral blood samples to diagnose VL in 45 children in Mato Grosso do Sul, Brazil. This technique is compared with methods carried out using tissue collected by invasive procedures, including direct microscopy, culture and detection of Leishmania DNA by PCR in bone marrow aspirates. The results show that PCR of peripheral blood provides great sensitivity (95.6 percent) that is similar to that from the PCR of bone marrow aspirates (91.1 percent) and higher than that achieved with microscopy (80 percent) or culture (26.7 percent) methods. PCR of peripheral blood proved to be a suitable tool for the diagnosis of VL in children because it is highly sensitive and safe, with tissue collection being less invasive than in traditional tests.

Mortalidade por leishmaniose visceral: aspectos clínicos e laboratoriais / Mortality due to visceral leishmaniasis: clinical and laboratory characteristics

INTRODUÇÃO: A leishmaniose visceral é uma doença infecciosa sistêmica de ampla distribuição geográfica, caracterizada pelo alto potencial de letalidade. Visando contribuir com a redução da mortalidade, bem como auxiliar profissionais da saúde no manejo clínico dos pacientes portadores desse agravo, este trabalho teve como objetivo investigar as características clínicas e laboratoriais dos casos que evoluíram para o êxito letal em hospitais de Campo Grande, MS, nos anos de 2003 a 2008. MÉTODOS: Foram analisados 55 prontuários de pacientes que tiveram a leishmaniose visceral como causa de óbito. RESULTADOS: Dos 55 pacientes estudados, 37 eram procedentes do município de Campo Grande, sendo 41 (74,5 por cento) do sexo masculino, com predominância da faixa etária acima dos 40 anos. Quanto ao quadro clínico, a febre esteve presente em 89,1 por cento dos casos. A duração da doença desde o início dos sintomas até a hospitalização variou em média 78,2 dias. A leucopenia ocorreu em 85,5 por cento dos pacientes. Comorbidades estiveram presentes em 39 (70,9 por cento) pacientes, sendo a desnutrição e o etilismo as mais frequentes. A confirmação do diagnóstico ocorreu em média 6,7 dias após a internação. O antimoniato pentavalente foi a droga mais utilizada, com 87,5 por cento dos pacientes apresentando algum tipo de reação adversa. Infecções bacterianas ocorreram em 36 pacientes e, em 27 (49 por cento), foram uma das causas do óbito. CONCLUSÕES: Os dados indicam que a identificação precoce dessas características clínicas e laboratoriais no primeiro atendimento ao paciente é de fundamental importância para se reduzir a mortalidade por meio da instituição de medidas terapêuticas e profiláticas eficazes.

INTRODUCTION: Visceral leishmaniasis is a systemic infectious disease of broad geographical distribution, characterized by high potential for lethality. With the purpose of contributing towards reducing mortality and helping healthcare professionals in clinical management of patients with this disease, this paper aimed to investigate the clinical and laboratory characteristics of cases with a fatal outcome in hospitals in Campo Grande, Mato Grosso do Sul, between 2003 and 2008. METHODS: Fifty-five medical files on patients who died due to visceral leishmaniasis were analyzed. RESULTS: Among the 55 patients studied, 37 were from the municipality of Campo Grande; 41 (74.5 percent) were males; and age over 40 years predominated. The patients presented with fever in 89.1 percent of the cases. The duration of the illness from the onset of symptoms to hospitalization was 78.2 days on average. Leukopenia was seen in 85.5 percent of the patients. Comorbidities were present in 39 (70.9 percent) patients; malnutrition and alcoholism were the most frequent of these. Confirmation of the diagnosis occurred on average 6.7 days after admission. Pentavalent antimoniate was the drug most used, and 87.5 percent of the patients presented some type of adverse reaction. Bacterial infections occurred in 36 patients and were one of the causes of death in 27 (49 percent). CONCLUSIONS: The data showed that early identification of these clinical and laboratory characteristics, at the time when patients are first attended, is extremely important for reducing mortality through instituting efficient therapeutic and prophylactic measures.

Phlebotomine fauna (Diptera: Psychodidae) of an American cutaneous leishmaniasis endemic area in the state of Mato Grosso do Sul, Brazil

The occurrence of an outbreak of cutaneous leishmaniasis associated with Leishmania (Leishmania) amazonensis in the municipality of Bela Vista, state of Mato Grosso do Sul, Brazil, and the absence of information on its vectors in this area led the authors to undertake captures of phlebotomine sand flies, using Shannon traps and automatic CDC light traps, in domiciles, forested areas and animal shelters from February 2004-January 2006. A total of 808 specimens belonging to 18 sandfly species have been identified: Bichromomyia flaviscutellata,Brumptomyia avellari, Brumptomyia brumpti, Brumptomyia sp, Evandromyia aldafalcaoae, Evandromyia cortelezzii, Evandromyia evandroi, Evandromyia lenti, Evandromyia teratodes, Evandromyia termitophila, Lutzomyia longipalpis, Nyssomyia whitmani, Pintomyia christenseni, Psathyromyia aragaoi, Psathyromyia campograndensis, Psathyromyia punctigeniculata, Psathyromyia shannoni and Sciopemyia sordellii. The presence of Lu. longipalpis, Ny. whitmani and Bi. flaviscutellata, vectors of Leishmania chagasi, Leishmania braziliensis and L. amazonensis, respectively, has increased.

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

As leishmanioses são zoonoses endêmicas em Mato Grosso do Sul e têm por agentes etiológicos nessa região Leishmania (Leishmania) chagasi, Leishmania (Leishmania) amazonensis e Leishmania (Viannia) braziliensis. Como método para identificação de espécies de Leishmania, a reação em cadeia da polimerase é uma ferramenta com elevada especificidade e sensibilidade. Analisaram-se 39 isolados de Leishmania criopreservados, obtidos por meio de aspirado medular e/ou biópsia de lesão, conforme a suspeita clínica. Os isolados foram submetidos à extração de DNA e à reação em cadeia da polimerase com os iniciadores: RV1/RV2 para Leishmania (Leishmania) chagasi, a1/a2 para a identificação de Leishmania (Leishmania) amazonensis e b1/b2 para Leishmania (Viannia) braziliensis. Leishmania (Leishmania) chagasi foi a única espécie identificada em 37 casos de leishmaniose visceral. Leishmania (Leishmania) amazonensis foi identificada em dois isolados de pacientes com diagnóstico de leishmaniose tegumentar. Os resultados obtidos confirmam a possibilidade do uso dos três pares de iniciadores como uma ferramenta na caracterização de isolados de Leishmania.

Leishmaniases are endemic zoonoses in the State of Mato Grosso do Sul. Their etiological agents in this region of Brazil are Leishmania (Leishmania) chagasi, Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis. The polymerase chain reaction (PCR) is a tool with high specificity and sensitivity for identifying Leishmania species. This study examined 39 cryopreserved isolates of Leishmania that had been collected by bone marrow aspiration and/or lesion biopsy, depending on the clinical suspicion. The isolates were subjected to DNA extraction and PCR using the following primers: RV1/RV2 for identifying Leishmania (Leishmania) chagasi, a1/a2 for Leishmania (Leishmania) amazonensis and b1/b2 for Leishmania (Viannia) braziliensis.Leishmania (Leishmania) chagasi was the only species identified in the 37 cases of visceral leishmaniasis.Leishmania (Leishmania) amazonensis was identified in two isolates from patients with a diagnosis of cutaneous leishmaniasis. The results obtained confirm that it is possible to use these three pairs of primers as a tool for characterizing Leishmania isolates.

Asymptomatic infection in family contacts of patients with human visceral leishmaniasis in Três Lagoas, Mato Grosso do Sul State, Brazil / Infecção assintomática em contactantes de pacientes com leishmaniose visceral humana em Três Lagoas, Mato Grosso do Sul, Brasil

The Brazilian city of Três Lagoas, Mato Grosso do Sul State, has experienced an urban outbreak of visceral leishmaniasis since 2000. In 2002, due to the increase in the number of cases, 46 families with cases of visceral leishmaniasis were studied to verify the prevalence of asymptomatic infection in household contacts. Indirect immunofluorescence and ELISA showed a 36.4 percent positive infection rate. There were no cases of symptomatic disease among these contacts. There was no statistically significant difference in gender or age. Median age was 21 years, and the 10-19-year age bracket was the most heavily affected (23 percent). As for family characteristics, no differences were observed in schooling or family income; most families (58.7 percent) owned their homes, which were built of masonry (97.8 percent) and had adequate infrastructure. All the families reported what were probably phlebotomine sand flies in the peridomicile. In conclusion, asymptomatic visceral leishmaniasis infection is frequent and occurs in both males and females, regardless of age.

O Município de Três Lagoas, Mato Grosso do Sul, Brasil, foi alvo de uma epidemia de leishmaniose visceral a partir de 2000. Em 2002, devido ao incremento de casos, estudou-se 46 famílias que apresentavam um caso de doença para verificar-se o percentual de positividade de infecção assintomática por leishmaniose visceral em contactantes. Encontrou-se 36,4 por cento de positividade pelos testes sorológicos Reação de Imunofluorescência Indireta e/ou imunoenzimático ELISA, sem diferença estatisticamente significativa quanto ao sexo e faixa etária. A mediana de idade foi de 21 anos, sendo a faixa etária mais acometida de 10 a 19 anos (23 por cento). Quanto às características familiares não observaram-se diferenças quanto ao nível de instrução e renda familiar; a moradia, em sua maioria, era própria (58,7 por cento), em alvenaria (97,8 por cento), com infra-estrutura adequada. Todas as famílias relataram a presença de provável flebotomíneo no peridomicílio. Conclui-se que a infecção assintomática por leishmaniose visceral é freqüente, ocorrendo em homens e mulheres, independente de faixa etária.

Modification of Disney trap for capture of sand flies (Diptera: Psychodidae: Phlebotominae)

This paper describes the modifications made to the original model of the Disney trap, with a view to easier handling of the same, greater practicability in the collection of sand flies, protection of the animal bait and durability of the trap in the field.

Sensitivity and specificity of polymerase chain reaction in Giemsa-stained slides for diagnosis of visceral leishmaniasis in children

The aim of this study was to evaluate the sensitivity and specificity of polymerase chain reaction (PCR) in the detection of Leishmania DNA in archived Giemsa-stained bone marrow slides for diagnosis of visceral leishmaniasis (VL), and to compare PCR with conventional diagnostic techniques, like direct microscopy and parasite culture. Specimens of archived Giemsa-stained bone marrow slides from 91 patients with VL and from 79 controls with other diseases or conditions were studied. PCR showed the highest sensitivity (92.3 percent) and had good specificity (97.5 percent). Direct examination detected 79.1 percent and culture 59 percent of positive samples. In addition, PCR was able to detect VL in 16 of 19 patients (84.2 percent) with negative microscopy. PCR in Giemsa-stained bone marrow slides is a suitable tool for confirming diagnosis in patients with VL and may be useful in the diagnosis of difficult cases. Slide smears are easily stored, do not require special storage conditions such as low temperatures, and can be easily mailed to centers where PCR is available, making it an excellent option for diagnosis in the field.

Comparison of conventional methods for diagnosis of visceral leishmaniasis in children of the Center-West Region of Brazil

In Brazil, sophisticated techniques currently employed for diagnosis of visceral leishmaniasis, such as polymerase chain reaction-based assays, are only available in major research centers, whereas conventional methods are still used in many areas where the disease occurs. In the state of Mato Grosso do Sul, in the country's Center-West Region, visceral leishmaniasis has recently emerged in many cities, and duration of the disease, from the onset of symptoms to diagnosis, has been short. Considering that results of diagnostic tests may depend on the phase of the disease, we compared direct examination of bone marrow aspirates (BMAs), BMA culture, and serology by Indirect Immunofluorescence Antibody Test (IFAT) for diagnosis in children, according to time of evolution (< 30 days or >30 days) and to spleen size (< 5 cm or > 5 cm) at admission. Duration of the illness did not interfere with test positivity: direct smear examination and IFAT were positive in more than 80 percent of patients, as was culture in around 60 percent. Results of positive microscopy, however, where predominant in patients with larger spleens. Thanks to the association of traditional techniques, only a few patients had to begin a treatment trial without confirming the diagnosis. Conventional methods for diagnosis of visceral leishmaniasis are still indispensable in our region, and training professionals in basic techniques should be incremented. The highest sensitivity in laboratory diagnosis among the cases investigated was that obtained with a combination of BMA direct examination and IFAT, nearing 100 percent.

Abundance of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) and urban transmission of visceral leishmaniasis in Campo Grande, state of Mato Grosso do Sul, Brazil

The outspread and urbanization of visceral leishmaniasis (VL) in Campo Grande, state of Mato Grosso do Sul, lead us to undertake the present study over diversity and abundance of sand flies in the urban area to compare with previous search carried out during 1999/2000, before the identification of the disease in the human population.The captures were carried out with automatic light traps, weekly, from February 2004 to February 2005 on three sites including a forested area (Zé Pereira), two peridomicilies (shelters of domestic animals and cultivation areas), and intradomicilie. In the present study 110 collections were obtained during 13 months for 1320 h of collections, resulting in 5004 specimens, 3649 males and 1355 females belonging to the 20 following species: Brumptomyia avellari, Brumptomyia sp., Bichromomyia flaviscutellata, Evandromyia lenti, E. termitophila, E. cortelezzii, E. borrouli, Lutzomyia sp., L. longipalpis, Micropygomyia quinquefer, N. antunesi, N. whitmani, Pintomyia christenseni, Pi. damascenoi, Psathyromyia aragaoi, Ps. campograndensis, Ps. hermanlenti, Ps. shannoni, Pychodopygus claustrei, and Sciopemyia sordellii. L. longipalpis was the most abundant species in the anthropic environment with 92.22 percent of the captures. This shows an increase of sixty times in the density of L. longipalpis compared to the last sand fly evaluation in 1999/2000. The high density of L. longipalpis in Campo Grande is the main factor of risk in transmission of the disease to human in the urban area. The capture of N. antunesi, typical specie from Amazonian region, in Mato Grosso do Sul is reported for the first time.

Foco emergente de leishmaniose visceral em Mato Grosso do Sul / Emergent outbreak of visceral leishmaniasis in Mato Grosso do Sul State

Este trabalho, realizado a partir de outubro de 2000 até janeiro de 2003, descreve as características clínico-epidemiológicas da leishmaniose visceral em Três Lagoas, Mato Grosso do Sul. Foram confirmados 149 casos da doença, com predominância do sexo masculino (71,1 por cento). A faixa etária mais acometida foi de 0 a 4 anos (42 por cento). Quanto ao quadro clínico, estiveram presente febre (97,3 por cento), esplenomegalia (85,9 por cento) e anemia (75,8 por cento). Em 32 (21,5 por cento) pacientes ocorreu infecção associada, predominando a pneumonia. Sobre as alterações laboratoriais observou-se mediana de hemoglobina de 8mg/dl e de leucócitos de 3.100 cel/mm³. O esfregaço de medula foi positivo em 90,6 por cento. Dos 97,9 por cento de pacientes tratados, 78,2 por cento utilizaram antimoniato pentavalente. Ocorreram 8 por cento de óbitos, metade deles portadores de infecção associada. O conjunto destes dados sugere mudanças no padrão fisiográfico de ocorrência da leishmaniose visceral na localidade, com expansão e urbanização da doença, necessitando atenção para o diagnóstico e tratamento precoces.

This study, realized from October 2000 to January 2003 describes the clinical epidemiological characteristics of visceral leishmaniasis in Três Lagoas, Mato Grosso do Sul State, Brazil. A total of 149 cases were confirmed, with a predominance of the male gender (71.1 percent). The principal age group was aged 0 to 4 years old (42 percent). The clinical picture included fever (97.3 percent), esplenomegaly (85.9 percent) and anemia (75.8 percent). Associated infections were seen in 32 patients (21.5 percent), pneumonia being most common. Changes registered in the laboratory included a median hemoglobin level of 8mg/dl and 3,100 leucocytes/mm³. Bone marrow smears were positive in 90.6 percent of patients. Of the 97.9 percent patients treated, 78.2 percent used pentavalent antimony. Mortalities occurred in 8 percent of cases, half of these with associated infection. Taken together, these data suggest changes in the physiographical occurrence of visceral leishmaniasis in this locality, with expansion and urbanization of the disease, requiring greater attention for early diagnosis and treatment.

Phlebotomines (Diptera: Psychodidae) in forested areas of the Serra da Bodoquena, state of Mato Grosso do Sul, Brazil

Investigation was undertaken on the behaviour of the phlebotomine fauna in caves, forests, and anthropic environments of the Serra da Bodoquena, between January 1998 and January 2000. This paper reports on the phlebotomines captured in forested areas with automatic light traps (ALT), Shannon traps (ST), aspiration (AN), at natural resting sites and by human attractiveness (HA) during 24 h. The diversity and abundance of the species were investigated with ALT installed at 16 points (ground level) and 6 in the canopy. Natural infection by flagellates was investigated in females captured with ST, AN, and HA. The sand fly fauna was represented by 23 species. Twenty-two of these were captured with ALT, 15 of them on the western side, and 20 on the eastern. Lutzomyia longipalpis and Nyssomyia whitmani were the most abundant on the former and this species together with Lutzomyia almerioi on the latter side. On the eastern side the ecotopes located close to caves rendered a significantly greater number (P < 0.01) of specimens than did more distant sites. On this side Lu. almerioi contributed with 56 percent of the total number of specimens. Lu. almerioi females were predominantly attracted by humans (96.4 percent) and by ST (93.2 percent) and three of the 2173 dissected (0.138 percent) presented natural infection by flagellates. The attraction of Lu. almerioi to humans occurred during all seasons, predominantly in the summer, and in nocturnal and diurnal periods. Thus it is bothersome to inhabitants of and visitors to the Bodoquena ridge and a potential vector of flagellates.

Ocorrência de leishmaniose tegumentar americana no Estado do Mato Grosso do Sul associada à infeccão por Leishmania (Leishmania) amazonensis / Occurrence of American tegumentary leishmaniasis in the Mato Grosso do Sul State associated to the infection for Leishmania (Leishmania) amazonensis

São relatados nove casos de leishmaniose tegumentar americana ocorridos no ano de 2001 em uma unidade de treinamento militar localizada no município de Bela Vista, Estado de Mato Grosso do Sul. Parasitas obtidos de lesões de seis pacientes foram isolados em cultura e posteriormente identificados através da análise de isoenzimas como sendo Leishmania (Leishmania) amazonensis. Esta é a primeira evidência da presenca desta espécie de parasita em Mato Grosso do Sul.