The microbiological signature of human cutaneous leishmaniasis lesions exhibits restricted bacterial diversity compared to healthy skin.

Localised cutaneous leishmaniasis (LCL) is the most common form of cutaneous leishmaniasis characterised by single or multiple painless chronic ulcers, which commonly presents with secondary bacterial infection. Previous culture-based studies have found staphylococci, streptococci, and opportunistic pathogenic bacteria in LCL lesions, but there have been no comparisons to normal skin. In addition, this approach has strong bias for determining bacterial composition. The present study tested the hypothesis that bacterial communities in LCL lesions differ from those found on healthy skin (HS). Using a high throughput amplicon sequencing approach, which allows for better populational evaluation due to greater depth coverage and the Quantitative Insights Into Microbial Ecology pipeline, we compared the microbiological signature of LCL lesions with that of contralateral HS from the same individuals.Streptococcus, Staphylococcus,Fusobacterium and other strict or facultative anaerobic bacteria composed the LCL microbiome. Aerobic and facultative anaerobic bacteria found in HS, including environmental bacteria, were significantly decreased in LCL lesions (p < 0.01). This paper presents the first comprehensive microbiome identification from LCL lesions with next generation sequence methodology and shows a marked reduction of bacterial diversity in the lesions.

The Extraordinary Case of a Woman with a 30-Year-Long Diffuse Leishmaniasis Cured with One Single Ampoule of Intranasal Pentavalent Antimoniate.

Infection with Leishmania amazonensis and L. mexicana may lead to diffuse cutaneous leishmaniasis. The cure is exceptional, especially for the strange case of this lady. Case report: The patient acquired the disease in childhood and remained with lesions for over 30 years, albeit several treatments. She worsened after a pregnancy, developing disseminated lesions. Miltefosine with amphotericin B and pentamidine resulted in remission. Lesions reappeared after one year, accompanied by intra-nasal infiltration of the disease. The nasal spraying of a single ampoule of pentavalent antimoniate resulted in the sustained disappearance of the nasal symptoms and all the cutaneous lesions. After over eight years, she remains disease-free, albeit under renal replacement therapy. The high nasal mucosal antimonial concentration may explain the long-lasting cure via new MHC class I epitope-specific CD8+ cell clones against L. amazonensis present in the nasal mucosa.

Identification of Leishmania (Viannia) species and clinical isolates of Leishmania (Leishmania) amazonensis from Brazil using PCR-RFLP of the heat-shock protein 70 gene reveals some unexpected observations.

Hsp70 is a cytoplasmic heat-shock protein, encoded by a multicopy tandemly repeated gene that has recently been gaining popularity as a valuable marker for typing Leishmania species. In this study, we used a previously described hsp70 PCR-RFLP method for identifying Brazilian Leishmania isolates. We identified two distinct L. (L.) amazonensis hsp70 alleles that resulted in two different RFLP patterns. Also, we found RFLP polymorphisms amongst L. (Viannia) naiffi strains. The profiles of both L. (V.) shawi and L. (V.) lindenbergi were very similar to those of other L. (Viannia) species. The observations described herein reflect the polymorphism found within species of Leishmania and indicate that results from this hsp70 PCR-RFLP method should be used with caution when typing isolates from clinical cases of leishmaniasis and Leishmania species from Brazil.

Experimental model of chronic osteomyelitis caused by Leishmania (L) amazonensis.

Experimental animal models have been used for the study of the physiopathogenesis of leishmaniasis, on some occasions with success, while in other situations such as bone alterations that accompany tegumentary leishmaniasis, especially in diffuse cutaneous form (DCL), the mechanisms are still unknown. In the present study, we determined these alterations in an animal model susceptible to Leishmania (L) amazonensis. Amastigotes of L. (L) amazonensis isolated from patients with diffuse cutaneous leishmaniasis (DCL) were inoculated into the hind paws of eight BALB/c mice, macroscopic and histopathological aspects were analyzed. After 90 and 120 days of evolution, histopathological analysis demonstrated a mononuclear cell infiltrate rich in plasma cells and intense parasitism of intra- and extra-medullary macrophages, with areas of bone necrosis and discrete involvement of cartilaginous tissue. The results show that the inflammatory process developed during L. (L) amazonensis infection might cause bone tissue destruction and secondarily affect the joints.

Polymerase chain reaction (PCR) is highly sensitive for diagnosis of mucosal leishmaniasis.

We evaluated the use of polymerase chain reaction (PCR) for diagnosis of mucosal leishmaniasis (ML) in an endemic area in Acre, Brazil, where Leishmania braziliensis is present. Leishmania DNA was detected 34 of 35 cases, yielding a positivity rate of 97.1%, which was higher than the positivity rates for all of the other diagnostic methods studied, namely Montenegro skin test (MST), anti-Leishmania serological testing and microscopic examination of lesion biopsy specimens. These findings have led us to propose guidelines for the diagnosis of ML that use PCR as the principal method of parasitological confirmation of cases.

Imaging exams of bone lesions in patients with diffuse cutaneous leishmaniasis (DCL).

We studied bone lesion alterations in three patients with diffuse cutaneous leishmaniasis (DCL) by imaging exams (radiography and scintigraphy) and histopathology. Two patients had bone lesions of distal extremities of hands and feet, and one infiltrating plaques in the skin. The study was conducted at three specialized centers (Presidente Dutra Hospital/Nucleus of Tropical Pathology, UFMA-MA; Gonçalo Moniz Research Center-FIOCRUZ-BA; Laboratory of Pathology of Infectious Diseases (LIM-50), University of São Paulo, SP). Three-phase bone scintigraphy demonstrated high sensitivity and specificity for bone lesions, showing increased uptake of the radiopharmaceutical at sites of active lesions. In contrast, radiography demonstrated lytic lesions, cortical destruction and local osteopenia of the bone extremeties in two patients. Histopathological analysis showed sequestration with presence of amastigote forms of Leishmania (osteomyelitis), mononuclear cells and macrophages containing amastigote forms of Leishmania in one patient. These preliminary data indicate that imaging exams (radiography and scintigraphy) are important in the evaluation of bone lesions in diffuse cutaneous leishmaniasis and should be included in the routine histopathological diagnosis of the disease and follow-up of bone lesions.

Visceral leishmaniasis in pregnancy: a case report.

Visceral leishmaniasis (VL) is endemic in the island of São Luis, State of Maranhão, Brazil. Despite an increase in the number of VL cases, the frequency of the disease is low among pregnant women. We present here the case of a pregnant woman followed up by our group, who was treated with amphotericin B with excellent outcome.

[Involvement of the renal function in patients with visceral leishmaniasis (kala-azar)]. / Envolvimento da função renal em pacientes com leishmaniose visceral (calazar).

UNLABELLED: Evaluation of the renal involvement of visceral leishmaniasis (kala-azar) aiming at verifying clinical and laboratorial repercussions of the renal implication on VL. Laboratory analysis was performed of blood and urine samples collected immediately after confirming diagnosis through the finding of Leishmania in bone marrow. Two (18.1%) patient presented complications associated with kala-azar. Five patients (45.4%) presented macroscopic hematuria and one case (N 9) clinical manifestations compatible with an acute nephritis syndrome. Laboratorial data of urine exam showed: proteinuria in 10 (90.9%) patients, hematuria in 7 (63.6%) and leukocyturia in 6 (54.5%) cases. Nine (81.8%) patients presented high levels of microalbuminuria, characterizing glomerular lesion. The presence of proximal tubulopathy, measured through retinol binding protein observed in 5 (45.4%) cases. CONCLUSION: renal involvement was manifested in most subjects, contributing to the severity of the disease.

[Fatal case during treatment of american tegumentary leishmaniasis with sodium stibogluconate bp 88 (shandong xinhua)]. / Obito durante tratamento da leishmaniose tegumentar americana com stibogluconato de sódio bp 88 (shandong xinhua).

The authors report the occurrence of a fatal case in patient with cutaneous leishmaniasis in the municipality of Caxias, MA. Male patient, 22 years old, road sweeper, presented with an ulcer in left leg, diagnosed as cutaneous leishmaniasis and treated with sodium stibogluconate BP88 (Sb+5) (Shandong Xinhua) at a dose of 10mg/Sb+5/kg/day/20 days. After dose three he presented arthralgia, myalgia, nausea and weakness. During the therapy there was an aggravation of the symptoms with abdominal pain and irradiation into the thorax. After dose seven he presented a picture of associated dyspnea and thoracic pain of mild intensity. At dose nine there was further worsening of the picture, nevertheless the therapy was continued up to dose 11, when the patient's state deteriorated to such an extent that he was hospitalized in the intensive care unit. Exams: erythrocytes, 4.4 million; hemoglobin, 10.6%; hematocrits, 35%; white blood cells 26,400, basophiles and myelocytes (0); segmented leukocytes, 59%; lymphocytes 30%; monocytes 2%; platelets (normal); glucose, 42mg%; urea, 73mg%; creatinine, 2.4mg%; and ECG (blockade of right branch). The patient died from cardiorespiratory insufficiency. The current report underscores the need to clarify health workers regarding the use of Sb+5 and also to remind the Health Ministry to verify the quality and origin when acquiring new products.

Towards a more precise serological diagnosis of human tegumentary leishmaniasis using Leishmania recombinant proteins.

BACKGROUND: Exposure to Leishmania induces a humoral immune response that can be used as a marker of parasite exposure. METHODOLOGY/PRINCIPAL FINDINGS: Herein, ELISA was used to screen sera from patients with Tegumentary Leishmaniasis (TL) against different L. infantum-chagasi-derived recombinant proteins (rHSP70, rH2A, rH2B, rH3, rH4 and rKMP11). Among the recombinant proteins, rHSP70 and rH2A showed the best reactivity against human sera obtained from endemic areas of TL. Receiver-Operator Characteristics (ROC) curve analysis was used to identify the effectiveness of these proteins for serodiagnosis of TL. ROC curves confirmed the superior performance of rHSP70 and rH2A, in comparison to the other tested recombinant proteins. Additionally, we evaluated the specificity of the response to rHSP70 and rH2A by testing sera obtained from patients with Chagas' disease, Tuberculosis, Leprosy or Systemic Lupus Erythematosus. In this case, rHSP70 displayed an increased ability to discriminate diseases, in comparison to SLA. CONCLUSION: Our results raise possibility of using rHSP70 for the serodiagnosis of TL.

The value of the otorhinolaryngologic exam in correct mucocutaneous leishmaniasis diagnosis.

An increase in mucocutaneous leishmaniasis (ML) cases in northern (Brazil) motivated this study. In 44 ML patients with clinical diagnosis, only 13 parasitologically confirmed cases exhibited mucosal lesion suggestive of ML. Other conditions involving nasal manifestations are frequently confounded with ML. Therefore, otorhinolaryngologic examination is important in the clinical management of ML.

Efeito leishmanicida in vitro de Stachytarpheta cayennensis (Rich. ) Vahl (Verbenaceae) / In vitro leishmanicidal effect of Stachytarpheta cayennensis (Rich. ) Vahl (Verbenaceae)

A atividade anti-Leishmania do extrato hidroalcoólico de Stachytarpheta cayennensis, espécie utilizada popularmente no tratamento de lesões cutâneas causadas por Leishmania sp, foi testado em ensaios in vitro utilizando formas promastigotas de Leishmania braziliensis e L. amazonensis. O extrato hidroalcoólico foi preparado a partir das folhas secas e utilizado em culturas de L. amazonensis e L. braziliensis nas concentrações de 500 a 32,5 æg/mL. Após 24 horas as formas promastigotas foram quantificadas para o cálculo da CI50. A citotoxicidade do extrato foi avaliada também em culturas de macrófagos peritoneais. O extrato apresentou efeito leishmanicida dose e espécie-dependente para promastigotas de Leishmania sendo mais eficaz para L. braziliensis. O extrato não apresentou efeito citotóxico quando utilizado nas culturas de macrófagos. Concluiu-se que o extrato hidroalcoólico de S. cayennensis inibe formas promastigotas de Leishmania in vitro o que poderia justificar, pelo menos parcialmente, o uso popular dessa espécie no tratamento de úlceras causadas por Leishmania.

Leishmanicidal activity of the hydroalcoholic extract of Stachytarpheta cayennensis, species that is usually employed in ulcers caused by Leishmania, was evaluated in vitro using Leishmania braziliensis and L. amazonensis promastigotes forms. The hydroalcoholic extract was prepared from dried leaves and used in L. amazonensis and L. braziliensis promastigotes cultures at concentrations of 500 to 32.5 æg/mL. After 24 hours the promastigotes forms were quantified and the IC50 was calculated. The cytotoxicity of the extract was evaluated using peritoneal macrophages. The extract presented a dose and specie-dependent leishmanicidal effect to Leishmania promastigotes, mainly to the L. braziliensis ones. The cytotoxic effect was not observed in macrophage cultures. In conclusion, the hydroalcoholic extract of S. cayennensis inhibits the growing of Leishmania promastigotes forms in vitro accounting for the folk use of this vegetal in skin ulcers caused by Leishmania.

Envolvimento da função renal em pacientes com leishmaniose visceral (calazar) / Involvement of the renal function in patients with visceral leishmaniasis (kala-azar)

Envolvimento da funçäo renal em pacientes com leishmaniose visceral (calazar). Apresenta-se estudo prospectivo de 11 pacientes com LV, com o intuito de verificar as repercussöes clínico-laboratoriais da funçäo renal na doença. Realizou-se análises laboratoriais das amostras de sangue e urina, colhidas logo após confirmaçäo diagnóstica, através do encontro de leishmanias no aspirado de médula óssea. Dois (18 por cento) pacientes apresentaram complicaçöes associadas a LV. Cinco (45,4 por cento) apresentaram hematúria macroscópica e em um caso (No.9) manifestaçöes clínicas compatíveis com síndrome nefrítica aguda. Os resultados dos exames de urina mostraram: proteinúria em 10 (90,9 por cento) pacientes, hematúria 7 (63,6 por cento) e leucocitúria em 6 (54,5 por cento) casos. Nove (81,8 por cento) pacientes apresentaram níveis elevados de microalbuminúria caracterizando lesäo glomerular. A presença de tubulopatia proximal medida através da proteína ligadora de retinol, foi observada em 5 (45,4 por cento) casos. Concluiu-se que o envolvimento renal se fez presente na maioria dos pacientes, contribuindo para a gravidade da doença

Obito durante tratamento da leishmaniose tegumentar americana com stibogluconato de sódio bp 88® (shandong xinhua) / Fatal case during treatment of american tegumentary leishmaniasis with sodium stibogluconate bp 88® (shandong xinhua)

Os autores relatam a ocorrência de óbito em paciente portador da forma cutânea da LTA no município de Caxias-MA. Trata-se de paciente do sexo masculino, 22 anos, gari, portador de lesäo ulcerada no membro inferior (perna esquerda), diagnosticado, após encontro do parasita (Leishmania) na lesäo, tratado com stibogluconato de sódio BP88® (Shandong Xinhua) na dose de 10mg/Sb+5/kg/dia/20 dias. Após a 3ª dose apresentou dores articulares, naúseas, mal estar geral. Com a continuaçäo da medicaçäo houve agravamento do quadro com dor epigástrica e no hipocôndrio direito irradiando-se para o hemitórax homolateral. Após a 7ª dose apresentou dispnéia associado à dor torácica de leve intensidade. Na 9ª dose houve piora do quadro, mesmo assim continuou a usar o medicamento até a 11ª dose quando seu estado agravou-se. Foi internado, necessitando de tratamento intensivo. Nos exames realizados apresentou: 4,4 milhöes de eritócitos, 10,6 por cento de hemoglobina, 35 por cento de hematócrito, 26.400 de leucócitos, basófilos e mielócitos (0), 59 por cento de segmentados, 30 por cento de linfócitos, 2 por cento de monócitos, plaquetas (normais), glicose 42mg por cento, uréia 73mg por cento, creatinina (2,4mg por cento), eletrocardiograma (bloqueio de ramo direito). Veio a falecer tendo como causa do óbito, insuficiência cárdio respiratória. O relato atual mostra a necessidade de esclarecimento das equipes de saúde quanto ao uso dos Sb+5 e também lembrar o Ministério da Saúde quando da aquisiçäo de novos produtos, preocupar-se com a qualidade e procedência do mesmo

Infecçäo por Leishmania (Leishmania) chagasi em crianças de uma área endêmica de leishmaniose visceral americana na Ilha de Säo Luís-MA, Brasil / Leishmania (Leishmania) chagasi infection on childhood from endemic area of visceral leishmaniasis in the Säo Luis-MA, island

Realizou-se estudo prospectivo com 648 crianças de zero a cinco anos no município da Raposa-MA, de julho/97 a junho/98, com o objetivo de avaliar as características da infecção por L.(L.)chagasi e verificar se existe associação entre desnutrição e infecção assintomática. Utilizou-se questionário com dados socioeconômicos, ambientais e hábitos de vida; realizou-se Intradermorreação de Montenegro(IDRM) com antígeno de L. amazonensis e Enzyme Linked Immunosorbant Assay(ELISA) para detectar infecção, e exame antropométrico. A prevalência inicial, final e incidência da infecção foram 18,6 por cento, 20,6 por cento e 10,8 por cento pelo IDRM, e 13,5 por cento, 34,4 por cento e 28 por cento pelo ELISA, respectivamente. A prevalência da desnutrição crônica (altura/idade) foi 26 por cento. Não houve associação estatisticamente significante entre desnutrição e infecção assintomática por L. (L.) chagasi. A forma assintomática da doença está presente nas áreas estudadas, necessitando de medidas de controle mais efetivas

Uso de Cetoconazol® no tratamento da entomoftoromicose cutâneo-mucosa: relato de caso / Use of Ketoconazole for treating mucocutaneous Entomophthoromycosis: a case report

As entomoftoromicoses constituem entidade clínica pertencente ao grupo das zigomicoses, cujos agentes etiológicos são o Conidiobolus coronatus, Conidiobolos incongruus e o Basidiobolos ranarum. Um caso de entomoftoromicose cutâneo-mucosa é descrito em homem de 51 anos de idade, lavrador, procedente da região amazônica do Estado do Maranhão, Brasil. Teve diagnóstico esclarecido por exame anatomopatológico, um ano após as manifestações clínicas iniciais. Como tratamento utilizou-se um dos derivados imidazólicos (cetoconazol®) 400mg/dia divididos em duas tomadas, por 12 meses), mostrando boa tolerância, com resposta favorável. Na última avaliação, 24 meses após início do tratamento, encontrava-se clinicamente curado.

Leishmaniose visceral no estado do Maranhäo, Brasil: a evoluçäo de uma epidemia / Visceral leishmaniasis in the state of Maranhäo, Brazil: evolution of an epidemic

Breve relato da evoluçäo histórica da leishmaniose visceral no estado do Maranhäo, Brasil, avaliando as possíveis causas da expansäo da referida doença no estado, assim como as medidas de controle adotadas pelo Ministério da Saúde, objetivando a diminuiçäo da incidência da mesma.

Mudança do componente cognitivo da atitude de uma populaçäo de regiäo endêmica do sul da Bahia diante da leishmaniose tegumentar / Information-related changes in attitude to mucocutaneous leishmaniasis of a population from an endemic region of the south of Bahia

O caráter mutilante da forma mucosa da leishmaniose tegumentar pode provocar alteraçöes no relacionamento interpessoal. Para identificar o nivel de informaçäo dos portadores da doença e a reaçäo psicológica dos moradores aos doentes de leishmaniose e seu nivel de informaçäo quanto à doença, foi aplicado um questionário na regiäo endêmica do sul da Bahia. Foram identificadas diversas crenças falsas e um nível de rejeiçäo da populaçäo bastante forte que admitiu a existência de relaçäo entre o medo do contágio e a atitude de rejeiçäo. Presume-se que a Comunicaçäo da Informaçäo Persuasiva à populaçäo provocara uma mudança da atitude através da alteraçäo do seu componente cognitivo

Procedência de pacientes portadores de leishmaniose tegumentar americana nas áreas endêmicas de três braços e corte de pedra, Estado da Bahia, Brasil / Origin of patients with american tegumentar leishmaniasis recorded in the endemic areas of Tres Braços and Corte de Pedra, Bahia State, Brazil

De acordo com o cadastro de 1416 pacientes portadores de leishmaniose tegumentar americana (L.T.A) referentes ao período de janeiro de 1976 a maio de 1987, os autores realizaram um levantamento da procedência dos casos dessa parasitose atendidos nas áreas endêmicas de Três Braços e Corte de Pedra no estado da Bahia. Os pacientes procediam de 24 dos 89 municípios que compöem a regiäo cacaueira da Bahia, destacando-se os municípios de Valença, Wenceslau Guimaräes e Teolândia como os de maior prevalência, somando juntos 923 casos (65,1%, com 520 destes residindo em umas poucas localidades contínuas entre si, formando uma área endêmica, com transmissäo ocorrendo provavelmente no peri, intradomicílio e nas lavouras de cacau e cravo localizadas nas encostas da floresta atlântica. Nota-se que a partir de 1983, houve um aumento do número de casos em toda a regiäo, havendo evidências da leishmaniose tegumentar americana comportar-se como uma doença ocupacional em toda a regiäo estudada

Spontaneous regional healing of extensive skin lesions in diffuse cutaneous leishmaniais (DCL)

The authors report a case of diffuse cutaneous leishimaniasis, with longstanding evolution and presenting with diffuse infiltrated lesions rich in amastigotes in the absence of mucosal involvement. In situ characterization with inonoclonal antibodies revealed Leishmania amazonensis. Large regional lesions have presented spontaneous healing without specific therapy. Considering that DCI presents with a defect in the cellular immune response, this fact demonstrate that this patient may develop a regional cellular immune response enough to destroy the parasites and to produce clearing of some lesions.