Brotes emergentes de leptospirosis del Amazonas colombiano / Emerging leptospirosis outbreaks in the Colombian Department of Amazonas

Se describe por primera vez una serie de nueve casos con clínica indicativa de leptospirosis en el municipio Puerto Nariño en el departamento Amazonas, Colombia. Se muestran evidencias serológicas de exposición con Rickettsia del grupo de las fiebres manchadas. Los casos fueron clínicamente considerados como síndrome febril de origen desconocido. Se descartó infección por dengue y malaria. El diagnóstico de Leptospira se realizó mediante el método de reacción en cadena de la polimerasa en tiempo real. Igualmente, se detectó la presencia de anticuerpos contra rickettsias del grupo de las fiebres manchadas por inmunofluorescencia Indirecta. Finalmente, se realiza revisión del tema(AU)

A description is provided for the first time of a series of nine cases with a clinical examination suggestive of leptospirosis in the municipality of Puerto Nariño, Department of Amazonas, Colombia. Serological evidence is presented of exposure to Rickettsia, spotted fever group. The cases were clinically considered as febrile syndrome of unknown origin. Infection with dengue or malaria was ruled out. Diagnosis of leptospirosis was achieved by real-time polymerase chain reaction. Additionally, indirect immunofluorescence detected the presence of antibodies against rickettsia, spotted fever group. Finally, a review was conducted about the topic(AU)

Phenotypic and Functional Profiles of Antigen-Specific CD4+ and CD8+ T Cells Associated With Infection Control in Patients With Cutaneous Leishmaniasis.

The host immunological response is a key factor determining the pathogenesis of cutaneous leishmaniasis. It is known that a Th1 cellular response is associated with infection control and that antigen-specific memory T cells are necessary for the development of a rapid and strong protective cellular response. The present manuscript reports the analysis of the functional and phenotypic profiles of antigen-specific CD4+ and CD8+ T cells from patients cured of cutaneous leishmaniasis (CL), patients with an active process of cutaneous leishmaniasis, asymptomatic individuals with a positive Montenegro test and healthy donors (HD). Peripheral blood mononuclear cells (PBMCs) from the patients exhibited a lymphoproliferative capacity after stimulation with total soluble protein from either Leishmania panamensis (SLpA) or Leishmania infantum (SLiA) or with a recombinant paraflagellar rod protein-1 (rPFR1). Higher frequencies of antigen-specific TNAIVE cells, mainly following stimulation with rPFR1, were observed in asymptomatic and cured patients than in patients with active cutaneous leishmaniasis, while T cells from patients with active cutaneous leishmaniasis showed a higher percentage of effector memory T cells (TEM for CD4+ T cells and TEMRA for CD8+ T cells). The amount of antigen-specific CD57+/CD8+ TEMRA cells in patients with active cutaneous leishmaniasis was higher than that in cured patients and asymptomatic subjects. Regarding functionality, a more robust multifunctional CD8+ T cell response was detected in cured patients than in those with active cutaneous leishmaniasis. Moreover, cured patients showed a significant increase in the frequency of cells expressing a Th1-type cytotoxic production profile (IFN-γ+/granzyme-B/+perforin+). Patients with an active leishmaniosis process had a significantly higher frequency of CD8+ T cells expressing the inhibitory CD160 and 2B4 receptors than did cured patients. The expression profile observed in cured patients could be indicative of an imbalance toward a CD8+ Th1 response, which could be associated with infection control; consequently, the determination of this profile could be a useful tool for facilitating the clinical follow-up of patients with cutaneous leishmaniasis. The results also suggest a possible exhaustion process of CD8+ T cells associated with the evolution of Leishmania infection.

Morphological and molecular characterization of Paragonimus caliensis Little, 1968 (Trematoda: Paragonimidae) from Medellin and Pichinde, Colombia.

Paragonimiasis is a subacute to chronic inflammatory granulomatous lung disease caused by the genus Paragonimus. In Latin America Paragonimus mexicanus Miyazaki & Ishii, 1968 is the only confirmed species to cause human infections. Paragonimus caliensis Little, 1968 is an uncommon species often regarded as a synonym of P. mexicanus. Recently, the study of two types of Paragonimus metacercariae from Costa Rica has provided new molecular and morphological evidence that P. caliensis is a separate species from P. mexicanus. In the present study, molecular, morphological and phylogenetic tools have been used to characterize two populations of Paragonimus located at west of Medellin, Antioquia and at Pichinde, Valle del Cauca (type locality of P. caliensis), Colombia. Adults and metacercariae obtained from Medellin, and metacercariae from Pichinde were analyzed. For morphological observations we used light microscopy and scanning electron microscopy (SEM). Morphology of metacercariae and adults matched with the holotype of P. caliensis. The number and arrangement of sensory papillae in the acetabulum region differs from the morphotypes reported for P. caliensis in Costa Rica. Two morphotypes in branching patterns of ovary and two morphotypes in branching patterns of testes were identified. The main morphological differences between P. caliensis and P. mexicanus corresponded to the size of gonads and their relative positions in the body, and the occasional presence of a cyst wall in P. caliensis metacercariae. The molecular and phylogenetic analyses (using nuclear ribosomal ITS2 and partial cytochrome c oxidase subunit 1 CO1 sequences) confirmed that P. caliensis from the type locality is the same species from Medellin and Costa Rica. Furthermore, these analyses also suggest genetic as well as geographical separation of P. caliensis populations between Colombia and Costa Rica. Currently, P. mexicanus and P. caliensis are sympatric in the Colombian Pacific bioregion, and specific diagnosis based on their egg size is not possible. Therefore, it is necessary to determine the biogeographic distribution ranges of both species and to implement molecular techniques to establish the role of P. caliensis in human paragonimiasis in Colombia.

Phylogenetic signal at the Cytb-SertRNA-IG1-ND1 mitochondrial region in Anopheles (Kerteszia) neivai Howard, Dyar & Knab, 1913 / Señal filogenética de la región Cytb- SertRNA -IG1-ND1 en Anopheles (Kerteszia) neivai Howard, Dyar & Knab, 1913

Abstract Introduction: Mitochondrial DNA has proven its utility for the study of insect evolution. Genes such as cytochrome b (Cytb) and the transfer gene for serine (SertRNA) can be used to compare closely related organisms. Objective: The phylogenetic utility of Cytb-SertRNA-IG1-ND1 was tested for polymorphisms, and secondary structure modeling in SertRNA was done to detect possible cryptic species in Anopheles neivai. Materials and methods: Specimens from Colombia, Guatemala, and the type locality in Panamá were collected and sequenced for specimen comparison based on DNA polymorphisms, and secondary structure modeling for the SertRNA gene. Results: Thirty-six sequences for A. neivai and A. pholidotus were obtained. Conclusions: Polymorphic variants were detected in A. neivai for Cytb-SertRNA-IG1- ND1. Despite this variation in A. neivai, cryptic species could not be detected.

Resumen Introducción. El ADN mitocondrial ha demostrado su utilidad para el estudio de la evolución en los insectos. Existen algunos genes mitocondriales como el citocromo b (Cytb) y el gen de transferencia para el aminoácido serina (SertRNA) que pueden usarse en el diagnóstico de especies estrechamente relacionadas. Objetivo. Explorar la utilidad filogenética de la región Cytb-SertRNA-IG1-ND1 para detectar posibles especies crípticas en Anopheles neivai. Materiales y métodos. Se recolectaron especímenes en Colombia, Guatemala y en la localidad tipo en Panamá, los cuales se secuenciaron y se compararon mediante el polimorfismo de ADN en toda la región y mediante la simulación de estructuras secundarias del gen SertRNA. Resultados. Se obtuvieron las secuencias de especímenes de A. neivai (34) y A. pholidotus (2). Conclusiones. Se detectaron algunos polimorfismos para la regiónCytb-SertRNA-IG1-ND1 en A. neivai, pero no así especies crípticas.

In vitro and in vivo antileishmanial activity of Artemisia annua L. leaf powder and its potential usefulness in the treatment of uncomplicated cutaneous leishmaniasis in humans

ABSTRACT INTRODUCTION: Cutaneous leishmaniasis (CL) is a tropical disease that affects millions of individuals worldwide. The current drugs for CL may be effective but have serious side effects; hence, alternatives are urgently needed. Although plant-derived materials are used for the treatment of various diseases in 80% of the global population, the validation of these products is essential. Gelatin capsules containing dried Artemisia annua leaf powder were recently developed as a new herbal formulation (totum) for the oral treatment of malaria and other parasitic diseases. Here, we aimed to determine the usefulness of A. annua gel capsules in CL. METHODS: The antileishmanial activity and cytotoxicity of A. annua L. capsules was determined via in vitro and in vivo studies. Moreover, a preliminary evaluation of its therapeutic potential as antileishmanial treatment in humans was conducted in 2 patients with uncomplicated CL. RESULTS: Artemisia annua capsules showed moderate in vitro activity in amastigotes of Leishmania (Viannia) panamensis; no cytotoxicity in U-937 macrophages or genotoxicity in human lymphocytes was observed. Five of 6 (83.3%) hamsters treated with A. annua capsules (500mg/kg/day) for 30 days were cured, and the 2 examined patients were cured 45 days after initiation of treatment with 30g of A. annua capsules, without any adverse reactions. Both patients remained disease-free 26 and 24 months after treatment completion. CONCLUSION: Capsules of A. annua L. represent an effective treatment for uncomplicated CL, although further randomized controlled trials are needed to validate its efficacy and safety.

DETECCIÓN DE ANTICUERPOS ANTI-Leishmania (Trypanosomatidae) EN POBLACIONES CANINAS DEL DEPARTAMENTO DE SUCRE, COLOMBIA / Detection of Anti-Leishmania Antibodies (Trypanosomatidae) in Canine Populations in the Department of Sucre, Colombia

El objetivo de la investigación fue determinar la prevalencia de anticuerpos anti-Leishmania en perros del departamento de Sucre, Colombia. Se analizaron 122 perros del área rural de los municipios de Ovejas, Sampués y Sincelejo, mediante la técnica de inmunofluorescencia indirecta (IFI). Anticuerpos anti-Leishmania fueron detectados en el 69,6 % (IC95 % = 61,1 - 78,2) de la población canina estudiada. En el municipio de Sincelejo la seropositividad fue del 72 % (IC95 % = 61,1 - 82,8), en el municipio de Sampués del 68,4 % (IC95 % = 52,3 - 84,5) y en el municipio de Ovejas del 55,5 % (IC95 % = 21,2 - 86,3). Los animales seropositivos fueron principalmente caninos jóvenes y adultos. El 17,6 % de los perros seropositivos mostró signos clínicos compatibles con leishmaniasis canina, entre los cuales la onicogrifosis, alopecia y caquexia fueron los más comunes. La alta frecuencia de caninos con anticuerpos a Leishmania detectada en el presente estudio, pone de manifiesto la hiperendemicidad de la leishmaniasis canina en esta zona del país, así como el alto riesgo de brotes epidémicos de la enfermedad.

The aim of this survey wasto determine prevalence of antibodies against Leishmania in dogs from Sucre, Colombia. We analyzed 122 dogs in rural areas from municipalities of Sampués, Ovejas and Sincelejo, by indirect immunofluorescence (IFA). Anti-Leishmania antibodies were detected in 69.6 % (95 % CI = 61.1 to 78.2) of the selected canine population. In the municipality of Sincelejo, Sampues and Ovejas, the seropositivities were 72 % (95 % CI = 61.1 to 82.8), 68.4 % (95 % CI = 52.3 to 84.5) and 55.5 % (95 % CI = 21.2 to 86.3) respectively. Seropositive animals were mainly young and adult dogs. The 17.6 % of seropositive dogs showed clinical signs compatible with canine leishmaniasis; onychogryphosis, alopecia and cachexia were the most common ones. The detected high frequency of canines with antibodies against Leishmania in the present study, highlights the hyperendemicity status of canine leishmaniasis in this area of ​​the country and the high risk of disease outbreaks.

Lista actualizada de flebotomíneos (Diptera: Psychodidae: Phlebotominae) de la región cafetera colombiana / An updated checklist of Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) from the Colombian Andean coffee-growing region

Se presenta una lista actualizada de especies de flebotomíneos en zonas cafeteras de la región andina colombiana. Con base en la revisión y verificación taxonómica, se registraron 53 especies presentes en 12 departamentos. Además de los registros obtenidos con base en un muestreo intensivo en cinco departamentos, se recopilaron los datos existentes en trabajos publicados y en la revision taxonómica de los especímenes de la zona pertenecientes a la colección entomológica del Programa de Estudio y Control de Enfermedades Tropicales (PECET). El listado comprende los géneros Brumptomyia (2 especies), Lutzomyia (50 especies) y Warileya (1 especie). Con base en este trabajo se confirmaron 11 nuevos registros de especies en la región cafetera colombiana, entre los cuales es relevante Lutzomyia panamensis , especie de importancia médica no registrada previamente en esta zona. En total, 18 especies de las registradas poseen hábitos antropofílicos o están relacionadas con la transmisión de Leishmania spp.

An updated list of phlebotomine sand flies species in coffee growing areas in the Colombian Andean region is presented. Fifty three species were reported from 12 departments. In addition, species distribution in the region was derived from specimens obtained during intensive field work in five departments, from previously published studies and from the taxonomic revision of specimens in the entomological collection of the Programa de Estudio y Control de Enfermedades Tropicales (PECET). The list includes the genera Brumptomyia (2 species), Lutzomyia (50 species) and Warileya (1 species). The updated list contains eleven new records in the region under study, including Lutzomyia panamensis , a species of medical importance not recorded previously in this zone. Eighteen of the species are considered to be anthropophilic, and many of them have been implicated in the transmission of leishmaniasis.

Opciones terapéuticas para el manejo de la Leishmaniasis cutánea americana / Treatment options for american cutaneous leishmaniasis

La Leishmaniasis es una enfermedad parasitaria transmitida por insectos vectores. Es un problema de salud preocupante y emergente. Un gran porcentaje de la población en el mundo está expuesta a la enfermedad. La Leishmaniasis cutánea es una forma clínica circunscripta a la piel. Se reportan 1.500.000 nuevos casos por año en el mundo. Se caracteriza por presentar lesiones crónicas y estigmatizantes. Varias opciones de tratamiento se han experimentado con resultados diversos. La Organización Mundial de la Salud (OMS), en el año 2010, actualizó las drogas disponibles para esta enfermedad y elaboró recomendaciones. En Argentina predominan las formas clínicas visceral (LV) y cutánea (LC). El Ministerio de Salud de la Nación propone guías de tratamiento para la LV. El objetivo de esta revisión es analizar las opciones de tratamiento propuestas por la OMS y evaluar su utilización en Argentina para la Leishmaniasis cutánea americana (LCA)

Leishmaniasis is a parasitic disease transmitted by insect vectors. Is an emergent and health concern. A large percentage of the population in the world is exposed to this disease. Cutaneous Leishmaniasis is circumscribed to skin and one million and a half new cases are reported every year in the world. It affects the skin through chronic and stigmatizing lesions. Several treatment options have been used with different results. The World Health Organization (WHO) updated the available drugs to treat Leishmaniasis and made recommendations in the 2010 year. In Argentina, visceral (VL) and cutaneous (CL) Leishmaniasis are the most frequent types of presentation of the disease. Recommended therapeutic options to manage VL, but not for CL, are published by the Ministry of Health of the Nation. The objective of this review is to analyze the treatment options suggested by the WHO and evaluate its use for the treatment of American cutaneous leishmaniasis (ACL) in Argentina

Synthesis of novel analogs of 2-pyrazoline obtained from [(7-chloroquinolin-4-yl)amino]chalcones and hydrazine as potential antitumor and antimalarial agents.

A new series of N-acetyl and N-formyl-pyrazoline derivatives 6 and 7-8 were synthesized by cyclocondensation reaction of [(7-chloroquinolin-4-yl)amino]chalcones with hydrazine hydrate in acetic acid and hydrazine hydrate in formic acid respectively. These compounds were evaluated in vitro as antitumor and as antimalarial agents. Compounds 7b and 8b-e showed remarkable antitumor activity against cancer cell lines, with the most important GI50 values ranging from 0.13 to 0.99 µM. The best antimalarial response was observed for compound 7a with an inhibition percentage of 50.8% for Plasmodium falciparum, a hemolytic capacity of 3.2% and an IC50 of 14.1 µg/mL.

Actividad tripanocida y citotóxica de extractos de plantas colombianas / Trypanocidal and cytotoxic activity of extracts of Colombian plants

Introducción. El tratamiento de la enfermedad de Chagas está basado en sólo dos medicamentos de eficacia limitada y con importantes efectos colaterales. La gran biodiversidad de la flora colombiana hace de la bioprospección una alternativa potencial en la búsqueda de nuevos antiparasitarios. Objetivo. Evaluar in vitro el potencial tripanocida y la citotoxicidad de extractos obtenidos de 23 plantas colombianas. Materiales y métodos. Se obtuvieron extractos de hojas, de tallos o de la planta entera, en solventes de diferente polaridad. La actividad contra epimastigotes y la citotoxicidad se evaluaron por el micrométodo enzimático con MTT. Los extractos activos contra epimastigotes y con baja citotoxicidad se evaluaron también en tripomastigotes y amastigotes intracelulares. Resultados. Se reporta la actividad tripanocida de 13 plantas colombianas y se confirma el efecto biológico de cuatro especies previamente evaluadas. Cuatro extractos activos en epimastigotes también fueron activos en tripomastigotes y, uno de ellos, en amastigotes. Este extracto fue aislado de la planta Hieronyma antioquensis, y presentó CI50 de 3,125, 11,48 y 2,85 µg/ml, e índices de selectividad de 25,7 y 27, para epimastigotes, tripomastigotes y amastigotes, respectivamente. Los resultados sugieren que este extracto es un candidato promisorio para el tratamiento de la enfermedad de Chagas. Conclusión. La flora colombiana es una fuente potencial de nuevas sustancias para la quimioterapia contra la enfermedad de Chagas. El micrométodo enzimático con MTT es una herramienta útil para la tamización de la actividad biológica en epimastigotes y posterior selección para ensayos con otros estadios del parásito.

Introduction. The treatment of Chagas disease is based on only two drugs with limited efficacy and significant side effects. The rich biodiversity of the Colombian flora makes bio-prospecting a potential alternative in the search for new antiparasitic drugs. Objective. Potential trypanocidal activity and cytotoxicity was assessed in extracts from 23 Colombian plants. Materials and methods. Extracts of leaves, stems, or of the whole plants were obtained in solvents of a range of polarities. The activity against Trypanosoma cruzi epimastigotes and the cytotoxicity were evaluated by the MTT enzymatic micro-method. Extracts active against epimastigotes and with lowcytotoxicity were also tested on trypomastigotes and intracellular amastigotes. Results. Among the extracts, biological activity was confirmed in 4 species. The extracts were active on epimastigotes and trypomastigotes; one was active also against amastigotes. The latter extract was isolated from the plant Hieronyma antioquensis and presented IC50 of 3.1 mg/ml for epimastigotes, 11.5mg/ml for trypomastigotes and 2.9 mg/ml for amastigotes. The selectivity indexes were 25, 7, and 27 respectively. Conclusions. The extract from H. antioquensis proved a promising candidate for Chagas disease treatment. Futhermore, the MTT enzymatic micromethod was a useful tool for screening biological activity on epimastigotes and other stages of the parasite for further extract trials.

A PCR assay for the identification of Leishmania species of the Viannia subgenus / Un ensayo de PCR para identificar especies de Leishmania del subgénero Viannia

We have identified a novel DNA sequence of 500 bp (β500-DNA) on the Leishmania (Viannia) subgenus, located in the intergenic region of one of the loci of the β-tubulin gene family. The sequence analysis showed that this sequence has no homology to any other sequence described so far, including the β-tubulin gene. We improved a specific β500-PCR assay, which generated a PCR product of 375 bp for total genomic DNA from Leishmania strains belonging to the L. (Viannia) subgenus. In contrast, no amplification was found when using genomic DNA from species of L. (Leishmania) subgenus or other organisms. Under our PCR conditions, the lower detection limit was 1 fg when a purified DNA clone (pLgβ4), which contains one copy of the β500-DNA sequence, was used. The β500-DNA PCR assay confirmed the preliminary diagnosis of cutaneous leishmaniasis in clinical samples in which the Montenegro skin test was positive and parasite cultures were negative. The analytical specificity and the sensitivity of the PCR assay provide a tool for epidemiological studies of the disease.

En este trabajo identificamos una nueva secuencia de DNA de 500 pb (β500-DNA) en Leishmania del subgénero Leishmania (Viannia), localizada en la región intergénica de uno de los loci de la familia de los genes de la β tubulina. El análisis de secuencia mostró que β500 no tiene homología con ninguna otra secuencia previamente descrita, incluido el gen de la β tubulina. Nosotros implementamos un ensayo de PCR específico para β500, β500-PCR, que genera un producto de PCR de 375 pb a partir del DNA genómico de cepas de Leishmania pertenecientes al subgénero L. (Viannia). No hubo amplificación alguna cuando se utilizó el DNA genómico de especies del subgénero L. (Leishmania) o el de otros organismos. En las condiciones establecidas, utilizando DNA purificado del clon pLgβ4, que contiene una copia de la secuencia de DNA de β500, el límite de detección más bajo fue de 1 fentogramo. El ensayo β500-PCR confirmó el diagnóstico preliminar de leishmaniasis cutánea en muestras clínicas de pacientes positivas a la prueba de Montenegro y negativas en el cultivo de parásitos. La especificidad y sensibilidad analítica del ensayo de PCR proporciona una herramienta para estudios epidemiológicos de la enfermedad.

Transfusion-transmitted visceral leishmaniasis caused by Leishmania (Leishmania) mexicana in an immunocompromised patient: a case report.

BACKGROUND: Transfusion-transmitted leishmaniasis is an increasing problem in areas where visceral and cutaneous leishmaniases are endemic. CASE REPORT: This article reports a case of transfusion-transmitted fatal visceral leishmaniasis (VL) caused by Leishmania (Leishmania) mexicana in a 42-year-old male resident of northwestern Colombia who after developing a terminal renal failure due to lupus nephritis received a renal transplant and multiple transfusions. RESULTS: Multiple intracellular Leishmania amastigotes were demonstrated in both renal biopsy and marrow aspirates. Cultures of the parasite were obtained in NNN medium and the identification of the species was done both by direct immunofluorescence and polymerase chain reaction-restriction fragment length polymorphism. CONCLUSIONS: This is the first report of a VL case produced by L. (L.) mexicana in Colombia, which usually is a dermotropic species. Our report suggests that although leishmaniasis is transmitted by the bite of phlebotomine sand flies, Leishmania parasite may be transmitted by blood transfusion, complicating the clinical course of organ transplant and being fatal.

Trece registros nuevos de Lutzomyia (Diptera: Psychodidae) para el departamento de Vichada, Orinoquia Colombiana / Thirteen new records of Lutzomyia (Diptera: Psychodidae) for the department of Vichada, Colombian Orinoquia

Se informa el primer hallazgo de Lutzomyia trinidadensis (Newstead, 1922), L. bettinii Feliciangeli, Ramírez-Pérez & Ramírez, 1988, L. flaviscutellata (Mangabeira, 1942), L. yuilli Young & Porter, 1972, L. saulensis (Floch & Abonnenc, 1944), L. runoides (Fairchild & Hertig, 1953), L. ayrozai (Barretto & Coutinho, 1940), L. claustrei Abonnenc, Leger & Fauran, 1979, L. walkeri (Newstead, 1914), L. preclara Young & Arias, 1984, L. (geniculata) sp., L. (squamiventris) sp., y L. (Evandromyia) sp., en el departamento de Vichada, Colombia. También se confirma la presencia de L. antunesi (Coutinho, 1939) y L. aragaoi (Costa Lima, 1932) en este departamento. Los flebotomíneos fueron recolectados en los municipios de Puerto Carreño, Cumaribo, y La Primavera, con trampas de luz tipo CDC, trampa Shannon y atrayente humano, y mediante búsqueda activa en lugares de reposo. Se discute la importancia epidemiológica de estas especies de flebotomíneos en relación con la leishmaniosis cutánea.

Lutzomyia trinidadensis (Newstead, 1922), L. bettinii Feliciangeli, Ramírez-Pérez & Ramírez, 1988, L. flaviscutellata (Mangabeira, 1942), L. yuilli Young & Porter, 1972, L. saulensis (Floch & Abonnenc, 1944), L. runoides (Fairchild & Hertig, 1953), L. ayrozai (Barretto & Coutinho, 1940), L. claustrei Abonnenc, Leger & Fauran, 1979, L. walkeri (Newstead, 1914), L. preclara Young & Arias, 1984, L. (geniculata) sp., L. (squamiventris) sp., and L. (Evandromyia) sp., are recorded for the first time in the department of Vichada, Colombia. In addition, the presence of L. antunesi (Coutinho, 1939) and L. aragaoi (Costa Lima, 1932) in this department is confirmed. Sand flies were collected in the municipalities of Puerto Carreño, Cumaribo, and La Primavera, using CDC light traps, Shannon trap, and human bait, and by active search in resting places. The epidemiological importance of these sand fly species is discussed in relation with the cutaneous leishmaniasis.

Two cases of visceral leishmaniasis in Colombia resistant to meglumine antimonial treatment / Dois casos de leishmaniose visceral da Colômbia, resistentes ao tratamento com antimoniato de meglumina

Visceral leishmaniasis (VL) affects over 500 000 people worldwide each year. The disease occurs in the Mediterranean basin, Central and South America and is caused by Leishmania infantum (syn L. chagasi). VL is an endemic disease in Colombia, particularly along the Caribbean coast and the Magdalena River Valley and 90% of VL cases occur in children under the age of five. The first line of treatment is chemotherapy with pentavalent antimonial compounds, including sodium stibogluconate (Pentostam®) and meglumine antimoniate (Glucantime®). These compounds are the ones most used in Colombia, at a dose of 20 mg/kg/day for 28 days. Nevertheless resistance of L. infantum to pentavalent antimonials is becoming an important problem. No cases of VL resistant to pentavalent antimonial compounds have previously been reported from Colombia. This report describes the two cases of VL resistance to antimonial compounds in a girl and a boy who did not respond to previous treatment with Pentacarinat® and Glucantime® regimens but were treated successfully with liposomal amphotericin B. Based on our findings, we recommend liposomal amphotericin B as the first line of treatment for VL due to its low toxicity, shorter administration period and the low price obtained by WHO.

A leishmaniose visceral (VL) afeta aproximadamente 500000 pessoas anualmente no mundo. A doença ocorre no mediterrâneo, na América Central e na América do Sul, sendo causada por Leishmania infantum (syn. L. chagasi). Na Colômbia VL é uma doença endêmica, presente no litoral do Caribe e no Vale do rio Magdalena sendo que 90% de casos de VL ocorrem em crianças menores de cinco anos. O principal tratamento é a quimioterapia com compostos de antimoniais pentavalentes, incluindo stibogluconato de sódio (Pentostam®) e antimoniato de meglumina (Glucantime®). Estes compostos são os mais usados na Colômbia em dosagem de 20 mg/kg/dia durante 28 dias. Entretanto, a resistência de L. infantum aos antimoniais pentavalentes está se tornando problema importante. Na Colômbia não existiam relatos de casos de VL resistentes aos antimoniais pentavalentes. Este trabalho descreve os dois primeiros casos colombianos de VL resistentes aos compostos antimoniais em uma menina e um menino, que foram tratados com regime de Pentamidina e Glucantime®, e demonstra o sucesso obtido no tratamento com anfotericina B liposomal. Em conclusão, sugerimos como primeira opção de tratamento a anfotericina B liposomal porque é altamente efetiva no tratamento da VL, dada sua baixa toxicidade, curtos períodos de administração e o baixo preço obtido pela organização Médicos Sem Fronteiras.

Presencia de infestación por Fasciola hepatica en habitantes del valle de San Nicolás, oriente antioqueño / Presence of infection by Fasciola hepatica in inhabitants from the valle de San Nicolás, eastern Antioquia

Introducción. En estudios previos se han encontrado prevalencias hasta de 90% de fasciolosis hepática en el ganado bovino del valle de San Nicolás en el oriente antioqueño. Objetivo. En este estudio se determinó la presencia de infestación en la población humana que está en contacto con el ganado de la región. Materiales y métodos. La presencia de anticuerpos en muestras de suero de 61 personas residentes de la zona y cuyos trabajos exigían contacto constante con ganado bovino, se determinó mediante una prueba tipo ELISA específica para Fasciola. hepatica. También se determinaron las conductas de riesgo para la adquisición del parásito, al igual que los antecedentes clínicos para la población estudiada. Resultados. Tres de los 61 sueros fueron positivos para la presencia de anticuerpos específicos para F. hepatica que corresponde a una prevalencia de infestación de 4,9%. Los tres individuos con muestras positivas relataron manipular pastos para ganadería y beber agua de las quebradas aledañas a las zonas de pastoreo. Sólo una de las personas con suero positivo refirió haber presentado ictericia en los últimos seis meses. Conclusiones Los resultados documentan la presencia de F. hepatica en la población humana que se encuentra en contacto con ganado infectado y permiten identificar las conductas de riesgo para la adquisición de la enfermedad en los habitantes de la zona. Es importante conocer la incidencia y prevalencia de la infestación, tanto en la población humana como animal, para adoptar medidas de prevención de las personas que están en contacto con el ganado enfermo, así como para tratar las personas enfermas.

Introduction: The Programa de Estudio y Control de Enfermedades Tropicales (PECET) at the Universidad de Antioquia has found a prevalence of 90% of hepatic fasciolasis in cattle from valle de San Nicolás, eastern Antioquia. Objective: In this study the presence of infection in the human population that is in contact with livestock was determined. Materials and methods: The presence of antibodies in serum samples from 61 residents of the area and whose jobs require constant contact with cattle was determined by an ELISA test. Risk behaviors for acquisition of the parasite as well as the clinical and symptoms for the study population were also determined.Results: Three of the 61 sera tested were positive for the presence of specific antibodies to F. hepatica, corresponding to a prevalence of infection of 4.9%. These three subjects reported manipulating pasture for livestock and drinking water from streams near grazing fields. Only one person referred jaundice in the last six months. Conclusions: The findings of this study document the presence of F. hepatica in the human population that is in contact with infected livestock and identify the risk behaviors for acquisition of the disease among the inhabitants of the area. It is important to know the incidence and prevalence of infection in both human and animal population, to adopt preventive measures for those that are in contact with sick cattle, and treatment of infected people.

PCR-RFLP y RAPD para la tipificación de Leishmania neotropical / PCR-RFLP and RAPD for typing neotropical Leishmania

Introducción. El análisis de la longitud de los fragmentos de restricción del producto amplificado y el estudio del ADN polimórfico amplificado al azar han demostrado ser herramientas útiles para la tipificación de Leishmania. Objetivos. Estudiar la utilidad de las técnicas moleculares para la identificación y tipificación de cepas de referencia de Leishmania spp. del Nuevo Mundo y valorar su aplicabilidad a muestras clínicas. Materiales y métodos. Se aplicó PCR para amplificar el gen que codifica la cisteíno-proteinasa B, y el análisis de la longitud de los fragmentos de restricción del producto amplificado utilizando ácido desoxirribonucleico de 16 cepas de referencia de Latinoamérica y de muestras clínicas de pacientes colombianos con leishmaniasis, y la técnica del ácido desoxirribonucleico polimórfico amplificado al azar utilizando ocho cepas de referencia. Se establecieron los patrones de bandas en cada caso. Resultados. Se obtuvo producto de amplificación en la PCR para Leishmania braziliensis, L. peruviana, L. panamensis y L. guyanensis. Para el resto, no fue posible amplificar el gen con los cebadores utilizados. La restricción mostró un patrón de bandas común para L. peruviana, L. guyanensis y L. panamensis, mientras L. braziliensis, presentaba un perfil individual único. El análisis de restricción del producto amplificado generó un patrón de bandas similar en los cinco pacientes estudiados, que se correspondía con el patrón generado por L. peruviana, L. guyanensis o L. panamensis. Mediante la amplificación al azar se obtuvieron patrones de bandas reproducibles con todas las cepas estudiadas, que posibilitaron la diferenciación. Se discuten las ventajas y limitaciones de ambos procederes. Conclusiones. El combinar ambas metodologías resultaría útil para identificar especies de importancia médica, tomando en cuenta sus ventajas y desventajas.

Primer informe de Lutzomyia yuilli Young & Porter, 1972 y Lutzomyia triramula (Fairchild & Hertig 1952) (Diptera: Psychodidae) en el departamento de Caldas, Colombia / First record of Lutzomyia yuilli Young & Porter, 1972 and Lutzomyia triramula (Fairchild & Hertig 1952) (Diptera: Psychodidae) in Caldas departament, Colombia

Durante estudios de foco en varios municipios del departamento de Caldas, Colombia, donde se han presentado casos de leishmaniosis cutánea, se recolectaron nueve especies de Lutzomyia: Lutzomyia gomezi (Nitzulescu 1931), Lu. ayrozai (Barreto & Coutinho 1940), Lu. panamensis (Shannon 1926), Lu. bifoliata Osorno, Morales, Osorno & Hoyos, 1970, Lu. trapidoi (Farchild & Hertig 1952), Lu. yuilli Young & Porter, 1952, Lu. triramula (Fairchild & Hertig 1952), Lu. atroclavata (Knab 1913) y Lu. trinidadensis (Newstead 1922). Dos de estos taxones constituyen nuevos registros para el departamento, Lu. yuilli del subgénero Nyssomyia, y Lu. triramula del subgénero Trichopygomyia. Los individuos de Lu. yuilli se obtuvieron mediante el uso de trampas de luz tipo Shannon y CDC en el municipio de Norcasia, mientras que los de Lu. triramula se recolectaron usando trampas de luz y adhesivas en los municipios de Norcasia y Victoria. Se discute la importancia de este hallazgo teniendo en cuenta la limitada distribución de Lu. triramula en el territorio colombiano y la capacidad vectorial de Lu. yuilli.

Nine species of Lutzomyia were collected during the research of various cutaneous leishmaniasis foci in the department of Caldas, Colombia: Lutzomyia gomezi (Nitzulescu, 1931), Lu. ayrozai (Barretto & Coutinho 1940), Lu. panamensis (Shannon 1926), Lu. bifoliata Osorno, Morales, Osorno & Hoyos, 1970, Lu. trapidoi (Fairchild & Hertig 1952), Lu. yuilli Young & Porter, 1972, Lu. triramula (Fairchild & Hertig 1952), Lu. atroclavata (Knab 1913), and Lu. trinidadensis (Newstead 1922). Two of these species constitute a new record for the department, Lu. yuilli of the subgenus Nyssomyia, and Lu. triramula of the subgenus Trichopygomyia. The specimens of Lu. yuilli were collected using Shannon and CDC light traps in the municipality of Norcasia, while the specimens of Lu. triramula were collected using light and sticky traps in both Norcasia and Victoria municipalities. The importance of this finding, taking into account the limited distribution of Lu. triramula in the Colombian territory and the vector capacity of Lu. yuilli is considered.