Saúde, ciência e desenvolvimento: a emergência da leishmaniose tegumentar americana como desafio médico-sanitário no Amazonas / Health, science and development: the emergence of American cutaneous leishmaniasis as a medical and public health challenge in Amazonas state, Brazil

Resumo O artigo faz análise histórica da emergência da leishmaniose tegumentar americana como objeto do conhecimento e desafio médico-sanitário no Amazonas desde a década de 1970. Fornece visão geral dessa época, as medidas sanitárias e os estudos científicos realizados no contexto de implantação dos principais projetos de desenvolvimento regionais executados em nome da política de integração nacional do governo federal. Utiliza como metodologia a análise documental de leis, produção científica, relatórios de pesquisa, boletins epidemiológicos e jornais. Os resultados da pesquisa mostram que a doença surgiu no Amazonas associando o grande problema de saúde com mudanças político-econômicas e alterações socioambientais.

Abstract The history of the emergence of American cutaneous leishmaniasis in the Brazilian state of Amazonas since the 1970s is analyzed as an object of knowledge and a medical and public health challenge. An overview of the period is provided, including the public health measures and scientific studies undertaken in the context of the execution of large-scale regional developments pursued in the name of national integration by the federal government. The methodology uses documental analysis of laws, the scientific literature, research reports, epidemiological bulletins, and newspapers. The results show that American cutaneous leishmaniasis emerged as a major health problem in Amazonas in close association with the political, economic, and socioenvironmental changes seen in the period.

Identificación de especies de Leishmania en pacientes derivados al Instituto Nacional de Salud del Perú / Identification of Leishmania species in patients derived to the National Institute of Health, Peru

RESUMEN En el Perú, la leishmaniasis es una enfermedad metaxénica que representa un serio problema de salud pública, debido a su amplia distribución y al número de personas en riesgo de contraer la enfermedad, siendo la población vulnerable principalmente las personas de bajos recursos económicos. El estudio se realizó a partir de pacientes que fueron derivados al Instituto Nacional de Salud entre el 2006 y el 2011 para que se les realizara el diagnóstico especializado. La identificación de la especie de Leishmania infectante se desarrolló mediante el análisis de las curvas de disociación (HRMA) obtenidas a partir del ADN genómico de promastigotes y amastigotes, lo que permitió identificar las especies de Leishmania (Viannia) braziliensis, Leishmania (V.) guyanensis, Leishmania (V.) peruviana como las más prevalentes, además de Leishmania (V.) lainsoni y Leishmania (L.) amazonensis.

ABSTRACT In Peru, leishmaniasis is a metaxenic disease that represents a serious public health problem, due to its wide distribution and the number of people in danger of contracting the disease, being the vulnerable population mainly those with low economic resources. The study was conducted from patients who were derived to Peru's National Institute of Health between 2006 and 2011 so that the specialized diagnosis could be carried out. The identification of the species of infectious Leishmania was developed through the analysis of the High-Resolution Melting Analysis obtained from the genomic DNA of promastigotes and amastigotes, which allows to identify the species of Leishmania (Viannia) braziliensis, Leishmania (V.) guyanensis, Leishmania (V.) peruviana as more prevalent, in addition to Leishmania (V.) lainsoni and Leishmania (L.) amazonensis.

Mucosal leishmaniasis: A forgotten disease, description and identification of species in 50 Colombian cases / Leishmaniasis mucosa: una enfermedad olvidada, descripción e identificación de especies en 50 casos colombianos

Abstract Introduction: Mucosal leishmaniasis has a progressive course and can cause deformity and even mutilation in the affected areas. It is endemic in the American continent and it is mainly caused by Leishmania (Viannia) braziliensis. Objective: To describe a series of mucosal leishmaniasis cases and the infectious Leishmania species. Materials and methods: We included 50 patients with a clinical diagnosis of mucosal leishmaniasis and parasitological confirmation, and we described their clinical and laboratory results. We performed species typing by PCR-RFLP using the miniexon sequence and hsp70 genes; confirmation was done by sequencing. Results: The median time of disease evolution was 2.9 years (range: 1 month to 16 years). The relevant clinical findings included mucosal infiltration (94%), cutaneous leishmaniasis scar (74%), total loss of the nasal septum (24%), nasal deformity (22%), and mucosal ulceration (38%). The symptoms reported included nasal obstruction (90%), epistaxis (72%), rhinorrhea (72%), dysphonia (28%), dysphagia (18%), and nasal pruritus (34%). The histopathological study revealed a pattern compatible with leishmaniasis in 86% of the biopsies, and amastigotes were identified in 14% of them. The Montenegro skin test was positive in 86% of patients, immunofluorescence in 84%, and culture in 8%. Leishmania (V.) braziliensis was identified in 88% of the samples, L. (V) panamensis in 8%, and L. (V.) guyanensis and L. (L.) amazonensis in 2% respectively. Conclusion: In this study, we found a severe nasal disease with destruction and deformity of the nasal septum in 25% of the cases, probably associated with late diagnosis. Leishmania (V.) braziliensis was the predominant species. We described a case of mucosal leishmaniasis in Colombia caused by L. (L.) amazonensis for the first time.

Resumen Introducción. La leishmaniasis mucosa tiene un curso progresivo y puede causar deformidad e incluso mutilación de las zonas afectadas. Es endémica en el continente americano y es causada principalmente por Leishmania (Viannia) brasiliensis. Objetivo. Describir una serie de casos de leishmaniasis mucosa y las especies de Leishmania infecciosas. Materiales y métodos. Se estudiaron 50 pacientes con diagnóstico clínico de leishmaniasis mucosa y confirmación parasitológica. Se describieron sus características clínicas y los resultados de laboratorio. La tipificación de especies se hizo mediante reacción en cadena de la polimerasa de los polimorfismos de la longitud de los fragmentos de restricción (Restriction Fragment Length Polymorphism Polymerase Chain Reaction, PCR-RFLP) en la secuencia del miniexon y el gen hsp70 y se confirmó por secuenciación. Resultados. La evolución de la enfermedad fue de un mes a dieciséis años (mediana de 2,8 años). Los hallazgos clínicos fueron los siguientes: infiltración mucosa (94 %), cicatriz de leishmaniasis cutánea (74 %), pérdida total del tabique nasal (24 %), deformidad nasal (22 %) y ulceración (38 %). Los síntomas reportados fueron: obstrucción nasal (90 %), epistaxis (72 %), rinorrea (72 %), disfonía (28 %), disfagia (18 %) y prurito nasal (34 %). La histopatología mostró un patrón compatible con leishmaniasis en 86 % de las biopsias y se identificaron amastigotes en 14 % de ellas. La prueba de Montenegro fue positiva en 86 % de los pacientes, la inmunofluorescencia en 84 %, y el cultivo en 8 %. Leishmania (V.) brasiliensis se identificó en 88 % de las muestras, L. (V) panamensis en 8 %, y L. (V.) guyanensis y L. (L.) amazonensis en 2 %, respectivamente. Conclusión. Se encontró enfermedad nasal grave con destrucción y deformidad del tabique nasal en una cuarta parte de los casos, probablemente debido a un diagnóstico tardío. Leishmania (V.) brasiliensis fue la especie predominante. Se describe por primera vez un caso de leishmaniasis mucosa causado por L. (L.) amazonensis en Colombia.

Detección de Leishmania (V) guyanensis en ejemplares de Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) recolectados en pecaríes de collar (Pecari tajacu) / Detection of Leishmania (V) guyanensis in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) collected from Pecari tajacu

Resumen Introducción. En estudios previos se detectó la presencia de Leishmania infantum en Rhipicephalus sanguineus, lo cual planteaba la posibilidad de que R. sanguineus transmitiera la leishmaniasis a una variedad de huéspedes. Objetivo. Identificar Leishmania (Viannia) spp. en garrapatas recolectadas en animales silvestres de una zona endémica para leishmaniasis. Materiales y métodos. Se hicieron 81 extracciones individuales de ADN en las garrapatas recogidas de tres tapires o dantas (Tapirus terrestres) y tres pecaríes de collar (Pecari tajacu) cazados en Madre de Dios, Perú. Las garrapatas recolectadas se identificaron taxonómicamente y se prepararon para la identificación del cinetoblasto (kDNA) de Leishmania (Viannia) spp. mediante reacción en cadena de la polimerasa (PCR), así como de la especie de Leishmania mediante PCR de fusión de alta resolución (High Resolution Melt, HRM). Resultados. Se detectó el kDNA de Leishmania (V) spp. en tres garrapatas silvestres de R. (Boophilus) microplus, Canestrini, 1888, recolectadas en un pecarí de collar cazado en la selva de Madre de Dios. El análisis mediante HRM-PCR evidenció que una de las muestras positivas de kDNA tenía una curva compatible con L. (V) guyanensis. Conclusión. Los resultados evidenciaron la presencia de ADN de L. (V) guyanensis en R. (Boophilus) microplus, probablemente adquirida después de picar al pecarí. Es importante hacer nuevos estudios para aclarar la participación de R. (Boophilus) microplus en la transmisión de la leishmaniasis.

Abstract Introduction: Previous studies identified the presence of Leishmania infantum in Rhipicephalus sanguineus and indicated the possibility that it could transmit leishmaniasis to a variety of hosts. Objective: To identify parasites of Leishmania (Viannia) spp. in ticks collected from wild animals in an endemic area for leishmaniasis. Materials and methods: We performed 81 individual DNA extractions from ticks collected from three Tapirus terrestris and three Pecari tajacu in Madre de Dios, Perú. Ticks were taxonomically identified and they were subsequently prepared to identify Leishmania (Viannia) spp. kDNA by PCR and the species of Leishmania by HRM-PCR. Results: Leishmania (Viannia) kDNA was detected in three wild ticks of the species R. microplus, collected from a collard peccary (P. tajacu) hunted in the forests of Madre de Dios. The HRM-PCR showed that one of the positive samples had a kDNA curve compatible with L. (V) guyanensis. Conclusion: The results showed the presence of L. (V) guyanensis DNA in R. microplus possibly acquired after biting a collarde peccary. Therefore, it is important to design future studies to clarify R. microplus involvement in the transmission of leishmaniasis.

Polymerase chain reaction-based method for the identification of Leishmania (Viannia) braziliensis and Leishmania (Viannia) guyanensis in mucosal tissues conserved in paraffin

ABSTRACTINTRODUCTION: In the Americas, mucosal leishmaniasis is primarily associated with infection by Leishmania (Viannia) braziliensis. However, Leishmania (Viannia) guyanensis is another important cause of this disease in the Brazilian Amazon. In this study, we aimed at detecting Leishmaniadeoxyribonucleic acid (DNA) within paraffin-embedded fragments of mucosal tissues, and characterizing the infecting parasite species.METHODS: We evaluated samples collected from 114 patients treated at a reference center in the Brazilian Amazon by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses.RESULTS: Direct examination of biopsy imprints detected parasites in 10 of the 114 samples, while evaluation of hematoxylin and eosin-stained slides detected amastigotes in an additional 17 samples. Meanwhile, 31/114 samples (27.2%) were positive for Leishmania spp. kinetoplast deoxyribonucleic acid (kDNA) by PCR analysis. Of these, 17 (54.8%) yielded amplification of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identification. Six of the samples were identified as L. (V.) braziliensis, while the remaining 11 were identified as L. (V.) guyanensis.CONCLUSIONS: The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffin-embedded tissues. Moreover, our findings confirm that L. (V.) guyanensisis a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.

Zosteriform cutaneous leishmaniasis due to Leishmania ( Viannia ) panamensis and Leishmania ( Viannia ) braziliensis: Report of three cases / Leishmaniasis cutánea zosteriforme causada por Leishmania ( Viannia) panamensis y Leishmania ( Viannia ) braziliensis: reporte de tres casos

In Colombia, zosteriform leishmaniasis is a little-known and infrequent clinical variant of cutaneous leishmaniasis. Its clinical features include one or more plaques made up of papules and pseudo-vesicles, which conform to a lineal pattern, as well as satellite lesions that affect one or more dermatomes, without crossing the median line. We present three zosteriform cutaneous leishmaniasis cases in which Leishmania panamensis and Leishmania braziliensis were identified as the infective species. In light of the fact that the disease occurs infrequently, diagnosis was reached by taking into account epidemiological and clinical suspicion.

La leishmaniasis zosteriforme es una variante clínica de la leishmaniasis cutánea, infrecuente y poco conocida en Colombia. Clínicamente se caracteriza por una o varias placas conformadas por pápulas y pseudovesículas que siguen un patrón lineal, y por lesiones satelitales que comprometen uno o varios dermatomas sin sobrepasar la línea media. Se presentan tres casos de leishmaniasis cutánea zosteriforme en los que se identificaron Leishmania panamensis y Leishmania braziliensis como especies infectantes. La sospecha epidemiológica derivada de la procedencia de los pacientes, así como la sospecha clínica a partir del reconocimiento de una presentación infrecuente de la enfermedad, permitieron hacer el diagnóstico.

Combining diagnostic procedures for the management of leishmaniasis in areas with high prevalence of Leishmania guyanensis / Procedimentos diagnósticos combinados no manejo da leishmaniose em áreas com alta prevalência de Leishmania guyanensis

BACKGROUND: The Amazon region corresponds to approximately 40 percent of the cases of leishmaniasis in Brazil. We report a prospective study with 180 patients conducted in a health care unit that diagnoses 10 percent of the cases of leishmaniasis in the Brazilian Amazon. The study addresses how a combination of procedures improves diagnosis in areas with high prevalence of Leishmania guyanensis. OBJECTIVES: to evaluate diagnostic methods in areas with high prevalence of Leishmania guyanensis. METHODS: All subjects were amastigote-positive by direct microscopic examination of lesion scarifications. We conducted skin biopsy and histopathology, polymerase chain reaction and parasite cultivation. RESULTS: Polymerase chain reaction detected almost ninety percent of infections when two amplification protocols were used (mini-exon and HSP-70). HSP-70 specific polymerase chain reaction matched the sensitivity of parasite cultivation plus histopathology. CONCLUSION: The best combination was polymerase chain reaction plus histopathology, which increased diagnostic sensitivity to 94 percent. Species discrimination by polymerase chain reaction disclosed prevalence of human infections with Leishmania guyanensis of 94 percent and with Leishmania braziliensis of 6 percent for this region.

FUNDAMENTOS: O Amazonas corresponde a aproximadamente 40 por cento dos casos de leishmaniose do país. Nós reportamos um estudo prospectivo com 180 pacientes de uma unidade de saúde que diagnostica 10 por cento dos casos de leishmaniose da amazônia brasileira, com combinação de métodos diagnóstico em área de alta prevalência de Leishmania guyanensis. OBJETIVOS: avaliar métodos diagnóstico da Leishmaniose em área endêmica para Leishmania Amazonensis. MÉTODOS: Todos os pacientes tiveram exame direto positivo com presença de amastigotas. Foi feita também biópsia cutânea, com realização de exame histológico, reação em cadeia da polimerase e cultura. RESULTADO: A reação em cadeia da polimerase detectou aproximadamente 90 por cento de infecção quando foram usados duas técnicas de amplificação (mini-exon and HSP-70). A reação em cadeia da polimerase com HSP-70 foi mais sensível que a cultura associada à histopatologia. CONCLUSÃO: A melhor combinação foi a reação em cadeia da polimerase com histopatologia, com sensibilidade de 94 por cento. A discrimanação das espécies causadoras de infecção humana nessa região mostrou Leishmania guyanensis em 94 por cento dos casos e Leishmania brasiliensis em 6 por cento.

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

Foram caracterizados/identificados por eletroforese de isoenzimas 23 isolados de Leishmania sp de pacientes dos municípios de Rio Preto da Eva e Manaus, analisando-se o grau de similaridade entre os organismos. Os resultados indicaram ocorrência de Leishmania guyanensis e Leishmania naiffi nestes dois ambientes e a heterogeneidade das amostras de Leishmania naiffi.

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.

Single and concomitant experimental infectionsby Endotrypanum spp. and Leishmania (Viannia) guyanensis (Kinetoplastida: Trypanosomatidae) in the Neotropical sand fly Lutzomyia longipalpis (Diptera: Psychodidae)

Lutzomyia longipalpis females received single and mixed infections with Endotrypanum and Leishmania. Two biological parameters were analyzed: the percentage of infected females and the distribution of flagellates in the gut of the females. The principal comparisons were performed between (1) two strains of Endotrypanum, (2) cloned versus primary sample of one strain of Endotrypanum, (3) Endotrypanum versus Leishmania guyanensis, and (4) the pattern of flagellates behaviour by optical microscopy in females with single or mixed infection versus the identification of parasites isolated from digestive tracts by isoenzyme electrophoresis. Flagellates of Endotrypanum showed distinct patterns of infection suggesting that there is variation between and within strains. The distribution of Endotrypanum and L. guyanensis differed significantly in relation to the colonization of the stomodeal valve. In co-infection with L. guyanensis, a large number of flagellates were seen to be plentifully infecting the stomodeal valve in significantly more specimens than in females infected by Endotrypanum only. However, the electrophoretic profiles of isoenzymes of parasites recovered from all co-infected specimens corresponded to Endotrypanum. This suggests that the mere correlation sand fly infection-biochemical analysis of isolates may induce parasitological incorrect consideration.