Epidemiological aspects of American and visceral cutaneous leishmaniasis in the city of Cametá, Pará, Amazon / Aspectos epidemiológicos da leishmaniose tegumentar americana e visceral na cidade de Cametá, Pará, Amazônia / Aspectos epidemiológicos de la leishmaniasis cutánea americana y visceral en la ciudad de Cametá, Pará, Amazonas

Background and objectives: leishmaniases are anthropozoonosis considered a major public health problem in tropical regions and endemic in some areas of constant expansion. This study aimed to assess the main epidemiological aspects of American tegumentary leishmaniasis (ATL) and visceral leishmaniasis (VL) in the municipality of Cametá, in the state of Pará, from 2007 to 2017. Methods: this is a descriptive-exploratory analysis, of time series, with data collected in the Department of Epidemiological Surveillance of the Department of Sanitary Surveillance of Cametá. Statistical calculations were performed, and, for the coefficient of incidence of ATL and VL, the standard formula was used to obtain the indicator. Results: a total of 94 and 294 cases of ATL and VL were reported, with the highest incidence rate in 2008. The disease affected all established age groups, with high frequency in children under ten years of age for VL (n=174), and between 20 and 30 years of age, for ATL (n=71). The disease was more prevalent in males (ATL (89.4%) and VL (58.2%)), because men are more related to economic activities. Conclusion: considering the high number of rural cases, it is noteworthy that reporting in urban areas is also worrisome, in addition to the livelihood of local families, because it has made them vulnerable to the disease. Furthermore, there is concern about the possible expansion and change in the pattern of ATL in the municipality. The Municipal Department as well as the epidemiological surveillance must pay attention to promote investments and campaigns to combat and treat this important disease.(AU)

Justificativa e objetivos: as leishmanioses são antropozoonoses consideradas um grande problema para a saúde pública em regiões tropicais e endêmicas em algumas áreas de constante expansão. Este estudo teve como objetivo avaliar os principais aspectos epidemiológicos da leishmaniose tegumentar americana (LTA) e leishmaniose visceral (LV) no município de Cametá, no estado do Pará, no período de 2007 a 2017. Métodos: trata-se de uma análise descritiva-exploratória, de série temporal, com dados coletados no Departamento de Vigilância Epidemiológica da Secretaria de Vigilância Sanitária de Cametá. Realizaram-se os cálculos estatísticos, e, para o coeficiente de incidência de LTA e LV, utilizou-se a fórmula padrão para a obtenção do indicador. Resultados: foram notificados 94 e 294 casos de LTA e LV, com maior taxa de incidência em 2008. A doença atingiu todas as faixas etárias estabelecidas, com alta frequência nos menores de dez anos para LV (n=174), e, entre 20 e 30 anos de idade, para LTA (n=71). A doença foi mais prevalente no sexo masculino (LTA (89,4%) e LV (58,2%)), em virtude dos homens estarem mais relacionados com as atividades econômicas. Conclusão: em vista do alto número de casos rurais, ressalta-se que a notificação em área urbana também é preocupante, além dos meios de subsistência das famílias locais, pois vem tornando-as vulneráveis para o adoecimento. Ademais, há a preocupação com a possível expansão e mudança no padrão da LTA no município. A Secretaria Municipal, bem como de vigilância epidemiológica, deve atentar-se a promover investimentos e campanhas de combate e tratamento deste importante agravo.(AU)

Justificación y objetivos: las leishmaniasis son antropozoonosis consideradas un importante problema de salud pública en las regiones tropicales y endémicas en algunas zonas de constante expansión. Este estudio tuvo como objetivo evaluar los principales aspectos epidemiológicos de la leishmaniasis tegumentaria americana (LTA) y la leishmaniasis visceral (LV) en el municipio de Cametá, en el estado de Pará, de 2007 a 2017. Métodos: se trata de un análisis descriptivo-exploratorio, de serie temporal, con datos recolectados en el Departamento de Vigilancia Epidemiológica de la Secretaría de Vigilancia Sanitaria de Cametá. Se realizaron cálculos estadísticos y, para el coeficiente de incidencia de LCA y LV, se utilizó la fórmula estándar para obtener el indicador. Resultados: se reportaron 94 y 294 casos de LTA y LV, con la mayor tasa de incidencia en 2008. La enfermedad afectó a todos los grupos de edad establecidos, con alta frecuencia en menores de diez años para LV (n=174), y entre 20 y 30 años. años de edad, para LTA (n=71). La enfermedad fue más prevalente en el sexo masculino (LTA (89,4%) y VL (58,2%)), debido a que los hombres están más relacionados con actividades económicas. Conclusión: dado el alto número de casos rurales, cabe señalar que la notificación en las zonas urbanas también es motivo de preocupación, además de los medios de subsistencia de las familias locales, ya que las ha vuelto vulnerables a la enfermedad. Además, existe preocupación por la posible expansión y cambio en el patrón de LTA en el municipio. La Secretaría Municipal, así como la de vigilancia epidemiológica, debe prestar atención a promover inversiones y campañas para combatir y tratar este importante problema.(AU)

Leishmaniasis cutánea en Colombia: una mirada epidemiológica / Cutaneous leishmaniasis in Colombia: an epidemiological overview

La leishmaniasis resulta una enfermedad infectocontagiosa causada por un protozoo del género Leishmania. Esta constituye un problema de salud importante para diversas regiones de América Latina. El aislamiento social impuesta a raíz de la pandemia de COVID-19 favoreció que disminuyera la incidencia de esta entidad, pero las actividades sociales de la normalidad se retomaron paulatinamente, imponiéndose que se retomen las medidas de seguimiento y control correspondientes.

Leishmaniasis is an infectious disease caused by a protozoan of the genus Leishmania. Leishma-niasis constitutes a significant health problem for various regions of Latin America. The social isolation imposed because of the COVID-19 pandemic favored a decrease in the incidence of this entity. However, regular social activities were gradually resumed, imposing the correspon-ding monitoring and control measures.

Chemical composition, in vitro larvicidal and antileishmanial activities of the essential oil from Citrus reticulata Blanco fruit peel / Composição química, atividades larvicida e leishmanicida in vitro do óleo essencial da casca do fruto de Citrus reticulata Blanco

Numerous studies have investigated the chemical composition and biological activities of essential oils from different Citrus species fruit peel, leaves and flowers. This paper aims to investigate the chemical composition, larvicidal and antileishmanial activities of essential oil from Citrus reticulata fruit peel (CR-EO). CR-EO was obtained by hydrodistillation in a Clevenger-type apparatus and its chemical composition was analyzed by GC-MS and GC-FID. Limonene (85.7%), ɣ-terpinene (6.7%) and myrcene (2.1%) were identified as its major components. CR-EO showed high activity against promastigote forms of Leishmania amazonensis (IC50 = 8.23 µg/mL). CR-EO also exhibited high larvicidal activity against third instar Aedes aegypti larvae at a lethal concentration (LC50 = 58.35 µg/mL) and 100% mortality at 150 µg/mL. This study suggests, for the first time, the potential use of CR-EO against this important mosquito-borne viral disease caused by the genus Aedes.

Numerosos estudos têm investigado a composição química e as atividades biológicas de óleos essenciais extraídos de cascas dos frutos, folhas e flores de diferentes espécies de Citrus. Este trabalho tem como objetivo investigar a composição química e as atividades larvicida e leishmanicida in vitro do óleo essencial das cascas dos frutos de Citrus reticulata (CR-EO). CR-EO foi obtido pela técnica de extração em aparelho Clevenger e sua composição química foi determinada por CG-EM e CG-DIC. Limoneno (85,7%), ɣ-terpineno (6,7%) and mirceno (2,1%) foram identificados como os constituintes majoritários. CR-EO mostrou alta atividade contra as formas promastigota de Leishmania amazonensis (CI50 = 8,23 µg/mL). CR-EO também exibiu alta atividade larvicida contra as larvas do terceiro estágio do Aedes aegypti com concentração letal (CL50 = 58,35 µg/mL) e mortalidade de 100% em 150 µg/mL. Este estudo sugere, pela primeira vez, o uso potencial de CR-EO contra esta importante doença viral transmitida por mosquitos do gênero Aedes.

Molecular survey of Leishmania spp. in skin samples of capybaras (Hydrochoerus hydrochaeris) from different areas of Brazil / Investigação molecular de Leishmania spp. em amostras de pele de capivaras (Hydrochoerus hydrochaeris) de diferentes regiões do Brasil

Leishmaniases comprise a spectrum of diseases caused by protozoan parasites of the genus Leishmania, with some species of rodents being incriminated as reservoirs. The capybara is the largest extant rodent species in the world and is widely distributed in South America. The occurrence of infection by Leishmania spp. was investigated in capybaras captured in Brazil during 2015­2019 from established populations in five highly anthropic areas of the state of São Paulo and two natural areas of the states of Mato Grosso and Mato Grosso do Sul. A total of 186 individuals were captured and subjected to abdominal skin biopsy. All skin samples were Leishmania kDNA-negative, suggesting that capybaras have no role in the transmission cycles of Leishmania species in the studied areas despite the well-known role of other rodents in the life cycle of Leishmania spp.(AU)

As leishmanioses compreendem um espectro de doenças causadas por protozoários do gênero Leishmania e algumas espécies de roedores são incriminadas como reservatórios de Leishmania spp. As capivaras compreendem a maior espécie de roedores existentes e são amplamente distribuídas na América do Sul. Para investigar a ocorrência de infecção por Leishmania spp. em capivaras, durante os anos de 2015-2019 capivaras foram capturadas em cinco áreas antrópicas do estado de São Paulo e em duas áreas naturais dos estados do Mato Grosso e do Mato Grosso do Sul, todos esses ambientes com populações de capivaras estabelecidas. Um total de 186 indivíduos foram capturados e submetidos à biópsia de pele abdominal. Todas as amostras de pele foram negativas para o alvo kDNA, assim, os dados sugerem que nas áreas estudadas as capivaras não têm papel no ciclo de transmissão de espécies de Leishmania spp., apesar do papel bem conhecido de outros roedores no ciclo de vida de Leishmania spp.(AU)

Avaliação do potencial antiparasitário de um Alcaloide Indólico isolado do Coral Marinho Tubastraea tagusensis / Evaluation of the Antiparasitic Potential of an Indole Alkaloid isolated from the Marine Coral Tubastraea tagusensis

Diante da falta de uma terapia eficaz e segura para o tratamento da doença de Chagas (DC) e da leishmaniose visceral, a busca por novos fármacos continua imprescindível. O ambiente marinho é uma rica fonte de produtos naturais bioativos, com aplicações contra parasitas, bactérias e outros patógenos que possam afetar humanos e animais. Entre as diversas classes de produtos naturais, os alcaloides têm um importante papel em aplicações terapêuticas. Neste trabalho, colônias de coral Tubastraea tagusensis foram coletadas, e a extração de seus metabolitos secundários foi realizada utilizando o solvente orgânico. Após fracionamento cromatográfico por diferentes técnicas, um alcaloide indolico foi isolado e elucidado estruturalmente por meio de técnicas espectrométricas e espectroscópicas. O alcaloide 6-bromo-2'-de-N-metillaplisinopsina (MR01), demonstrou atividade contra tripomastigotas e amastigota intracelulares de Trypanosoma cruzi, com valores de Concentração Efetiva 50% de 62 µM e 5 µM, respectivamente...(AU)

Considering the lack of effective and safe therapy for the treatment of Chagas disease (CD) and visceral leishmaniasis, the search for new chemotherapies remains indispensable. The marine environment is a well-known source of bioactive products, with applications against parasites, bacteria and other human / animal pathogenic microorganisms. Among different classes of natural products, alkaloids have a rich history of therapeutic applications. In this work, colonies of the coral Tubastraea tagusensis were collected and their secondary metabolites were extracted with ethyl acetate. Using different chromatographic techniques, an indole alkaloid was isolated and elucidated through spectroscopic and spectrometric techniques, resulting in the 6-bromo-2'-de-Nmethylaplisinopsin (MR01). The compound MR01 demonstrated activity against Trypanosoma cruzi, against both trypomastigote and amastigote forms, resulting in 50% Effective Concentration (EC50) values of 62 µM and 5 µM, respectively. The studies with Leishmania infantum amastigotes showed lack of activity. No cytotoxicity was observed for NCTC cells, to the maximal concentration of 200 µM. The mechanism of action was also explored. After incubation with the trypomastigotes, no alteration was observed in the permeability of the plasma membrane. The compound reduced the ATP and induced mitochondrial depolarization with no alterations in the reactive oxygen species levels. The intracellular calcium levels were reduced after treatment with MR01, including the pH alterations of the acidocalcisomes. Using MALDI-TOF/MS, the protein profile of the parasite was altered to a different manner to that observed for the standard drug benznidazole, suggesting a distinct mechanism of action. In silico studies of the pharmacokinetic / pharmacodynamic properties (PKPD) and physicochemical properties using the SwissADME software, suggested a good drug-like. In conclusion, this study showed for the first time the anti-T. cruzi activity of the alkaloid MR01. In addition, considering the potency and selectiveness, our study can suggest this compound to be explored as a new prototype for CD. (AU)

Chemical study of Adenocalymma axillarum crude leaf extract and isolated compounds

Abstract Adenocalymma axillarum (K.Schum.) L.G. Lohmann is a liana belonging to the family Bignoniaceae. In traditional medicine, the genus Adenocalymma is used to treat fever, skin ailments, and body, joint, and facial muscle pains, and it is also applied as cosmetic. Biological assays conducted with the A. axillarum crude leaf ethanol extract have indicated leishmanicidal activity and absence of cytotoxicity. This study aimed to analyze the A. axillarum leaf ethanol crude extract by high-performance liquid chromatography-high-resolution mass spectrometry- diode array detector (HPLC-HRMS-DAD) and to evaluate the leishmanicidal and cytotoxic activities of this crude extract, its fractions, and isolated compounds. HPLC-HRMS-DAD analysis of this extract revealed that it consisted mainly of flavonoids, with nine major compounds. Extract purification yielded 4-hydroxy-N-methylproline, 6-β-hydroxyipolamiide, quercetin-3-O-robinobioside, hyperin, isorhamnetin-3-O-robinobioside, and 3'-O-methylhyperin, which were identified by Nuclear Magnetic Resonance. The isolated compounds were inactive against Leishmania amazonensis promastigotes and human lung fibroblast cells.

Diversity, abundance and ecological importance of plant species for medical use in tropical forest of Tingo Maria, Peru / Diversidad, abundancia e importancia ecológica de especies vegetales de uso medicinal de la selva tropical en Tingo María, Perú

Peru is a megadiverse country due to the large number of animal and plant species. Its diversity derives from the different ecoregions present that developed with geological evolution. Much of its plant diversity is contained in the Peruvian Amazon, which includes a large proportion of plant species, many of them endemic. Of this diversity, many plants have been underestimated, and it is believed that more than 50% of them have not been recorded. These scientific gaps also address medicinal plants, their taxonomic identification, phytochemical bioactives produced, mechanisms of action of phytochemicals, and the metabolic pathways involved. These medicinal plants are active against common diseases such as: protozoa, with emphasis on malaria and leishmania, diabetes, inflammation, hypertension, cancer, infectious diseases (viral, bacterial, and fungal), kidney, liver, diarrhea and other health problems. This work is based on the study of a forest area in the district of Rupa Rupa called Reserve Forest of the Universidad Nacional Agraria de la Selva (BRUNAS) in Tingo María, Peru, which is being highly pressured by the people who live in the surroundings of this forest ecosystem for domestic use, so it is urgent to sensitize the population linked to this natural resource and make known the plants found there with high medicinal potential for the use of the locals, the nation and the world(AU)

El Perú es un país megadiverso debido a la gran cantidad de especies de animales y plantas. Su diversidad se deriva de las diferentes ecorregiones presentes que se fueron desarrollando con evolución geológica. Mucha de su diversidad vegetal está contenida en la Amazonía Peruana que incluye una gran proporción de especies de plantas, muchas de ellas, de carácter endémico. De esta diversidad, muchas plantas han sido subestimada, y se cree que más del 50% de ellas no han sido registrada. Estas lagunas científicas también abordan también a las plantas medicinales, su identificación taxonómica, bioactivos fitoquímicos producidos, mecanismos de acción de los fitoquímicos y las vías metabólicas involucradas. Estas plantas medicinales son activas a enfermedades comunes como: protozoos, con énfasis en la malaria y leishmania, diabetes, inflamación, hipertensión, cáncer, enfermedades infecciosas (virales, bacterianas, y hongos), afecciones renales, hepáticas, diarrea y otros problemas de salud. Este trabajo se basa en el estudio de una zona boscosa del distrito de Rupa Rupa denominada Bosque de Reserva de la Universidad Nacional Agraria de la Selva (BRUNAS) en Tingo María, Perú, la cual esta siendo muy presionada por los pobladores que viven en los alrededores de este ecosistema forestal para uso domésticos, por lo que es urgente sensibilizar a la población vinculada a dicho recurso natural y dar a conocer las plantas allí encontradas con alto potencial medicinal para el uso de los lugareños, de la nación y el mundo(AU)

Planejamento e desenvolvimento de novos compostos com atividade frente a Leishmania spp: Síntese, avaliação da atividade e toxicidade de derivados 5-nitro-2-furfurilidênicos e estudos de relações estrutura-atividade / Design of new compounds against Leishmania spp: synthesis, biological evaluation and toxicity of 5-nitro-2-furfurylidene derivates and structure-activity relationship studies

A leishmaniose é uma zoonose de ampla distribuição mundial, causada pelos parasitas tripanossomatídeos do gênero Leishmania. Infelizmente, o arsenal terapêutico disponível é precário, mas vê-se crescente o interesse científico pela busca do potencial de derivados nitroheterocíclicos como alternativas terapêuticas. Nesse contexto, este trabalho analisou o potencial de derivados 5-nitro-2-furfurilidênicos contra diferentes cepas de Leishmania, assim como investigou um possível modo de ação para esta classe de nitrocompostos. Para tal, a quimioteca foi sintetizada de acordo com publicações prévias do grupo. O potencial de inibição de crescimento das culturas de promastigotas de L. (L.) infantum (Linf) e L. (L.) major (Lmaj) foi determinado, utilizando miltefosina (MILT) (Linf - IC50: 8,28±0,33 µM), anfotericina B (AMB) (Linf - IC50: 0,02±0,002 µM) e nifurtimox (NFX) (Lmaj - IC50: 3,5±0,09 µM) como referência. A maioria dos compostos apresentaram maior potencial que as referênias, destacando o composto 40 (Linf - IC50: 0,2±0,019 µM/ Lmaj - IC50: 0,087 ± 0,001 µM) como mais eficaz. Contra as formas amastigotas intracelulares, para Linf os compostos 40, 13 e 15 foram mais eficazes em reduzir a carga parasitária dos macrófagos infectados que fármacos de referência. Para Lmajor, o composto 40 (IC50: 0,006 ± 0,0003 µM) foi mais ativo que o NFX (IC50: 2,15 ± 0,01 µM). Também foi determinada a atividade da quimioteca frente a enzima nitrorredutase (NTR1), utilizando cepas de T. brucei superexpressantes de NTR1, e os compostos analisados foram até 18 vezes mais eficazes que à cepa wild-type. Ademais, a partir da análise exploratória de dados por análise de componentes principais (PCA) e de grupamentos hierárquicos (HCA), foi reconhecida a influência das propriedades relacionadas com o equilíbrio hidrófilo-lipófilo e da natureza estérica/geométrica das moléculas para atividade anti-Leishmania

Leishmaniasis is a worldwide zoonosis caused by trypanosomatid parasites of the genus Leishmania. Unfortunately, the available therapeutic arsenal is precarious, but there is growing scientific interest in searching the potential of nitroheterocyclic derivatives as therapeutic alternatives. In this context, this work analyzed the potential of 5-nitro-2-furfurylidene derivatives against different Leishmania strains, as well as investigated the potential mode of action for this nitro compounds class. To this end, the chemolibrary was synthesized according to our group's previous publications. The growth inhibitory potential potential for promastigote cultures of L. (L.) infantum (Linf) and L. (L.) major (Lmaj) was determined using miltefosine (MILT) (Linf - IC50: 8.28±0.33 µM), amphotericin B (AMB) (Linf - IC50: 0.02±0.002 µM) and nifurtimox (NFX) (Lmaj - IC50: 3.5±0.09 µM) as reference. Most of the compounds were more potent than the references, highlighting compound 40 (Linf - IC50: 0.2±0.019 µM/ Lmaj - IC50: 0.087 ± 0.001 µM) as the most effective. Against intracellular amastigote, for Linf, compounds 40, 13 and 15 were more effective in reducing the parasite load of infected macrophages than reference drugs. For Lmajor, compound 40 (IC50: 0.006 ± 0.0003 µM) was more active than NFX (IC50: 2.15 ± 0.01 µM). The activity against nitroreductase (NTR1) enzyme was determined using overexpressing NTR1 mutant T. brucei strains, and the analyzed compounds were up to 18 times more effective than wild-type. Furthermore, exploratory data analysis using principal component analysis (PCA) and hierarchical clustering (HCA) methods were used. The influence of properties related to the hydrophiliclipophilic balance and the steric/geometric nature of the molecules was associated with the anti-Leishmanial activity

Leishmaniasis cutánea americana: Índices epidemiológicos en Valle Hermoso de la provincia Santo Domingo de Los Tsachilas / American cutaneous leishmaniasis: Epidemiological indices in Valle Hermoso, Santo Domingo de Los Tsachilas province

Las leishmaniasis son causadas por aproximadamente 15 especies de protozoos del género Leishmania. Prevalecen en áreas tropicales y subtropicales del Viejo y Nuevo Mundo, la movilidad humana también los convierte en un problema médico en áreas no endémicas. Las manifestaciones clínicas pueden comprender formas cutáneas que pueden ser localizadas. El diagnóstico y el tratamiento varían según las manifestaciones clínicas, el área geográfica y la especie involucrada. Este artículo destaca aspectos epidemiológicos y clínicos en la provincia de Santo Domingo de los Tsachilas, Ecuador. Se utilizaron fichas clínicas y epidemiológicas de pacientes que presentaron lesiones ulceradas o no, con un tiempo de evolución igual o mayor a 2 semanas. El mayor número de casos están entre los grupos sucesivos de adolescentes de 11 a 19 años y los adultos jóvenes entre 20 y 39 años. El 81,79% de los positivos procedían de áreas rurales o periurbanas. Dos terceras partes de los pacientes eran masculinos. Hubo predominio de lesiones únicas (87,79%); ubicadas en cuello y región cervical, y extremidades inferiores (24,84 y 24,20%, respectivamente). Cerca del 88% de los pacientes no presentaron ningún tipo de sintomatología asociada a la infección parasitaria. El 97,86% de los diagnósticos de laboratorio fueron por observación microscópica directa. 81,80% obtuvieron cura total de las lesiones y el 22% de los pacientes abandonaron la terapia antiparasitaria. Se requieren pruebas rápidas y accesibles que puedan definir las especies de Leishmania, ya que la discriminación tiene una importancia significativa para el pronóstico y tratamientos específicos de la especie(AU)

Leishmaniases are caused by approximately 15 species of protozoa of the genus Leishmania. They prevail in tropical and subtropical areas of the Old and New World human mobility also makes them a medical problem in nonendemic areas. Clinical manifestations may comprise cutaneous forms that may be localized. Diagnosis and treatment vary according to the clinical manifestations, geographic area, and species involved. This article highlights epidemiological and clinical aspects in the province of Santo Domingo de los Tsachilas, Ecuador. Clinical and epidemiological records of patients who presented ulcerated lesions or not, with an evolution time equal to or greater than 2 weeks were used. The largest numbers of cases are between the successive groups of adolescents from 11 to 19 years old and young adults between 20 and 39 years old. 81.79% of the positives came from rural or peri-urban areas. Two thirds of the patients were male. There was a predominance of single lesions (87.79%); located in the neck and cervical region, and lower extremities (24.84 and 24.20%, respectively). About 88% of the patients did not present any type of symptomatology associated with the parasitic infection. 97.86% of laboratory diagnoses were by direct microscopic observation. 81.80% obtained complete cure of the lesions and 22% of the patients abandoned antiparasitic therapy. Rapid and accessible tests that can define Leishmania species are required, as discrimination is of significant importance for species-specific prognosis and treatment(AU)

Anti-leishmanial activity of 29 daily sessions of intralesional-pentavalent antimony administration on Leishmania (Viannia)-infected BALB/c mice / Actividad antileishmania de 29 sesiones diarias de antimonio pentavalente intralesional en ratones BALB/c infectados con Leishmania (Viannia)

Abstract Introduction: Intralesional-pentavalent antimonials (IL-SbV) are recommended for simple cutaneous leishmaniasis (CL). Few treatment sessions (1-5) and drug volumes (1-5 ml each), relative to lesion size (LS), are recommended. There is not a validated IL-SbV protocol using doses calculated as mg/kg body weight and administered over a large number of IL-sessions, with small injection volumes. Objective: The study aim was to determine the efficacy of different concentrations of IL-SbV administered in 29 daily sessions of 100 μL each, on CL infected mice. Methods: Leishmania (Viannia) panamensis and L. (V.) braziliensis-infected mice (N = 6) were treated with 150, 50, and 16.6 mgSbV/kg/day x 29 days. Percentage of lesion area reduction, aesthetic and final (no lesions, no parasites) efficacy and effective dose (ED)50 were determined. In vitro-SbV activity against parasites was evaluated for both species. Results: The ED50 values were 72.2 and 66.3 (at the end of treatment), 54.3 and 37.7 (15-days pt.), and 145.3 and 148.6 (60-days pt.) for each species, respectively. Differences were observed between Leishmania species at 15-days pt., but not later. At 60-day pt., IL-SbV-150 mg showed final cure rates of 66.6 % for L. (V.) panamensis and 33.3 % for L. (V.) braziliensis-infected mice. After 15 days pt., lesion reactivation was observed in some "aesthetically cured" mice. Glucantime was not active in in vitro assays. Conclusions: The IL-SbV use with a dose calculated as mg/kg body weight and administered over a large number of IL-sessions, with small injection volumes each day could be effective against L. (V.) panamensis and L. (V.) braziliensis-CL infection. An appropriate SbV-dose (higher than 150 mg/kg/day x less than 29 days) must be evaluated.

Resumen Introducción: Los antimoniales pentavalentes aplicados intralesionalmente (IL-SbV) se recomiendan para el tratamiento de la leishmaniasis cutánea (LC) simple. Se recomiendan pocas sesiones (1-5) y volúmenes (1-5 ml cada uno) en relación con el tamaño de la lesión (LS). No existe un protocolo de IL-SbV validado que utilice dosis calculadas según el peso corporal (en mg/kg) y administradas durante varias sesiones en pocos volúmenes de inyección. Objetivo: El objetivo del estudio fue determinar la eficacia de diferentes concentraciones de IL-SbV administradas en 29 sesiones diarias de 100 μL cada una, en ratones con LC. Métodos: Ratones infectados con L. (V.) panamensis y L. (V.) braziliensis (N = 6) fueron tratados intralesionalmente con 150, 50 y 16,6 mg SbV/kg/día x 29 días. Se determinó el porcentaje de reducción del área de la lesión, la eficacia estética y final (sin lesiones, sin parásitos) y la dosis efectiva (DE)50. Adicionalmente de evaluó la actividad in vitro del SbV. Resultados: Los valores de DE50 fueron 72.2 y 66.3 (al final del tratamiento), 54.3 y 37.7 (15 días pt) y 145.3 y 148.6 (60 días pt) para cada especie. Se encontraron diferencias entre las especies sólo a los 15 días pt. La eficacia del tratamiento IL-SbV-150 mg, 60 días pt., fue de 66.6 y 33.3 % en ratones infectados con L. (V.) panamensis L. (V.) braziliensis respectivamente. Después de 15 días pt., se observó reactivación de la lesión en algunos ratones "estéticamente curados". Glucantime no fue activo in vitro. Conclusiones: El uso intralesional de SbV con una dosis calculada en mg/kg de peso corporal y administrada durante varias sesiones, con pequeños volúmenes de inyección cada día, podría ser eficaz en LC por L. (V.) panamensis y L. (V.) braziliensis. Dosis adecuadas de SbV (superiores a 150 mg/kg/día x 20) deben evaluarse.

Leishmania (L.) amazonensis infection impairs reproductive and fetal parameters in female mice / La infección por Leishmania (L.) amazonensis afecta parámetros reproductivos y fetales en ratones hembras

ABSTRACT Leishmaniasis is a group of parasitic zoonotic diseases caused by intracellular protozoans belonging to the genusLeishmania. Little is known about the effects that this parasitosis may have on the reproductive parameters and pregnancy of infected humans and pets. This study aimed to evaluate the influence of chronic cutaneous leishmaniasis caused byLeishmania (Leishmania) amazonensison reproductive and fetal parameters using a female murine model. A control group of female BALB/c mice and a group infected withL. (L.) amazonensiswere mated with healthy males. Clinical parameters were monitored during the pre-mating and gestational periods. Female mice were euthanized on day 19 of gestation, when the fetuses were weighed and their length measured and embryonic resorptions and fetal death were recorded. We observed five fetal deaths and three embryonic resorptions in the infected group. Furthermore, there was a decrease in fertility in the infected group (26.32%). The weight of the offspring from infected mothers was lower than that in the control group (1.019±0.035g and 1.163±0.032g,p<0.01). Fetal length was reduced in the infected group (3.71±0.05cm in the control group and 3.40±0.06cm in the infected groupp<0.001). This study shows that cutaneous leishmaniasis caused byL. (L.) amazonensisimpairs reproductive and fetal parameters in mice.

RESUMEN La leishmaniasis comprende un grupo de enfermedades zoonóticas parasitarias causadas por protozoos intracelulares pertenecientes al géneroLeishmania. Poco se conoce sobre los efectos que esta parasitosis puede tener sobre los parámetros reproductivos y la gestación en humanos y en otras especies infectadas. El objetivo de este estudio fue determinar la influencia de la leishmaniasis cutánea crónica, causada porLeishmania (Leishmania) amazonensis, en parámetros reproductivos y fetales. Se apareó un grupo control y un grupo infectado de ratones hembra BALB/c (previamente inoculado conL. (L.) amazonensis) con machos sanos. Se analizaron parámetros clínicos durante los períodos pregestacional y gestacional. Las hembras fueron eutanasiadas en el día 19 de gestación, momento en el cual se pesaron y midieron los fetos y se registraron las reabsorciones embrionarias y las muertes fetales. Se observaron 5 muertes fetales y 3 reabsorciones embrionarias en el grupo infectado. Además, hubo una disminución en la fertilidad de este último grupo (26,32%). Por otra parte, el peso de la descendencia de madres infectadas fue menor que el del grupo control (1,019±0,035g y 1,163±0,032g, respectivamente,p<0,01). Por último, la longitud fetal se redujo en el grupo infectado (3,71±0,05cm en el grupo control y 3,40±0,06cm en el grupo infectado,p<0,001). Este estudio muestra que la leishmaniasis cutánea causada porL. (L.) amazonensisafecta los parámetros reproductivos y fetales en ratones.

Evaluación de la toxicidad de chalconas sintéticas con potencial anti-Leishmania en ratones BALB/c / Toxicity assessment of synthetic chalcones with antileishmanial potential in BALB/c mice

RESUMEN Objetivo: Evaluar la toxicidad de tres chalconas sintéticas administradas por vía intraperitoneal en ratones BALB/c. Materiales y métodos: La dosis letal media (DL50) se estimó por el método Up-and-Down de Dixon. La toxicidad subcrónica de las chalconas se evaluó a 20 y 40 mg/kg por 21 días. Se evaluó el efecto tóxico a nivel de comportamiento, fisiológico, bioquímico e histológico. Resultados: La chalcona 43 generó moco en las heces, daño visceral (hígado) y alteración en el coeficiente de órganos (riñón, p = 0,037 y cerebro, p = 0,008) en comparación con el grupo control. Además, en el análisis histológico se observó que esta chalcona produjo edema, inflamación y necrosis en los órganos evaluados, aunque no hubo diferencia significativa con el control. Todos los parámetros bioquímicos no difirieron significativamente entre los grupos de tratamiento a dosis de 40 mg/kg y el control. Conclusiones: La DL50 para las tres chalconas fue superior a 550 mg/kg de peso corporal. Las chalconas 40 y 42 son relativamente no tóxicas. Ambas pueden considerarse seguras para la aplicación vía intraperitoneal en ratones BALB/c y, en consecuencia, son posibles candidatas para ser usadas en el tratamiento contra las leishmaniosis.

ABSTRACT Objective: To evaluate the toxicity of three synthetic chalcones administered intraperitoneally to BALB/c mice. Materials and methods: The median lethal dose (LD50) was estimated by Dixon's Up-and-Down method. Subchronic toxicity of chalcones was evaluated at 20 and 40 mg/kg for 21 days. Behavioral, physiological, biochemical, and histological toxic effects were evaluated. Results: Chalcone 43 produced mucus in feces, visceral damage (liver) and alterations in organ coefficient (kidney, p = 0.037 and brain, p = 0.008) when compared to the control group. In addition, histological analysis showed that this chalcone produced edema, inflammation and necrosis in the evaluated organs, although there was no significant difference with the control. None of the biochemical parameters differed significantly between the treatment groups at 40 mg/kg dose and the control. Conclusions: The LD50 for all three chalcones was greater than 550 mg/kg of body weight. Chalcones 40 and 42 were found to be relatively non-toxic. Both can be considered safe for intraperitoneal application in BALB/c mice and, consequently, are potential candidates for use in the treatment of leishmaniasis.

Plantar ulcer as an atypical manifestation of cutaneous leishmaniasis

Abstract Cutaneous leishmaniasis is characterized by ulcers with raised edges and a granular bottom, mainly on the lower limbs. This is a case report of a male patient with an ulcer on the left plantar region. The diagnosis was confirmed by positive PCR for L. braziliensis and the presence of amastigotes of Leishmania sp. in the histopathological examination. After treatment with Glucantime, the patient showed full healing of the ulcer. The unusual location of the ulceration calls attention to atypical presentations of leishmaniasis, and the importance of histopathological examination and PCR, leading to the appropriate diagnosis and treatment.

Avaliação da tendência temporal e da prevalência de leishmaniose visceral canina e humana dentre os anos de 2010 a 2019 em municípios de Minas Gerais ­ Brasil / Evaluation of the temporal trend and prevalence of canine and human visceral leishmaniosis between 2010 and 2019 in municipalities of Minas Gerais - Brazil / Evaluación de la tendencia temporal y de la prevalencia de leishmaniosis visceral canina y humana entre los años 2010 a 2019 en los municipios de Minas Gerais ­ Brasil

Leishmanioses são causadas por protozoários do gênero Leishmania, parasitos que infectam grande número de mamíferos, incluindo o homem. A Leishmaniose Visceral (LV) é a forma mais severa da doença e invariavelmente leva ao óbito, se não diagnosticada e tratada precocemente. O objetivo deste estudo foi realizar uma análise de natureza documental, descritiva e analítica, de abordagem quantitativa das informações contidas no banco de dados Fundação Ezequiel Dias/Gerenciador de Ambientes Laboratoriais (FUNED/GAL), da Secretaria Estadual de Saúde de Minas Gerais. Os dados epidemiológicos que compõe este estudo são registros de casos positivos e negativos da cidade de Patos de Minas-MG e 18 municípios das mesorregiões do Triângulo Mineiro e Alto Paranaíba, Noroeste de Minas e Norte de Minas. No período compreendido entre janeiro de 2010 e junho de 2019 foram notificados um total de 1170 indivíduos suspeitos de Leishmaniose Visceral Canina e/ou Humana. 304 (25,98%) indivíduos receberam o resultado positivo, enquanto 866 foram negativos, e em alguns casos, inconclusivos. Os dados obtidos no estudo revelaram a tendência temporal crescente e alta prevalência da doença, mostrando que a doença está em expansão na região estudada onde o cão é o principal reservatório doméstico da doença, permanecendo como principal elo de ligação entre o protozoário e o hospedeiro humano.(AU)

Leishmaniasis are caused by protozoa of the genus Leishmania, parasites that infect a large number of mammals, including humans. Visceral Leishmaniasis (VL) is the most severe form of the disease and invariably leads to death if not diagnosed and treated early. The objective of this study was to carry out a documentary, descriptive and analytical analysis, with a quantitative approach to the information contained in the Fundação Ezequiel Dias/Manager of Laboratory Environments (FUNED/GAL) database from the Minas Gerais State Health Secretariat. The epidemiological data that make up this study are records of positive and negative cases in the city of Patos de Minas - MG and of 18 municipalities in the mesoregions of the Triângulo Mineiro and Alto Paranaíba, Northwest of Minas and North of Minas. In the period between January 2010 and June 2019, a total of 1170 individuals suspected of Canine and/or Human Visceral Leishmaniasis were notified. A total of 304 (25.98%) individuals received a positive result, while 866 were considered negative, and in some cases, inconclusive. The data obtained in the study revealed the growing temporal trend and high prevalence of the disease, showing that the disease is expanding in the studied region where the dog is presented as the main domestic reservoir of the disease, remaining as the main link between the protozoan and the human host.(AU)

Leishmaniosis son causadas por protozoos del género Leishmania, parásitos que infectan a un gran número de mamíferos, incluyendo el hombre. La Leishmaniosis Visceral (LV) es la forma más grave de la enfermedad e invariablemente conduce a la muerte, si no se la diagnostica y la trata a tiempo. El objetivo de ese estudio fue realizar un análisis de naturaleza documental, descriptivo y analítico, de enfoque cuantitativo de las informaciones contenidas en la base de datos Fundação Ezequiel Dias/Gerente de Ambientes de Laboratorio (FUNED /GAL), de la Secretaría de Salud del Estado de Minas Gerais. Los datos epidemiológicos que conforman este estudio son registros de casos positivos y negativos en la ciudad de Patos de Minas-MG y 18 municipios de las mesorregiones del Triângulo Mineiro y Alto Paranaíba, Noroeste de Minas y Norte de Minas. En el período comprendido entre enero de 2010 y junio de 2019, se notificó a un total de 1170 personas sospechosas de Leishmaniosis Visceral Canina y/o Humana. 304 (25,98%) individuos recibieron un resultado positivo, mientras que 866 fueron negativos y, en algunos casos, no concluyentes. Los datos obtenidos en el estudio revelaron la tendencia creciente temporal y la alta prevalencia de la enfermedad, mostrando que la enfermedad se está expandiendo en la región estudiada donde el perro es el principal reservorio doméstico de la enfermedad, permaneciendo como principal conexión entre el protozoo y el hospedero humano.(AU)

Leishmania Game: tecnologia educativa para prevenção/ensino de leishmaniose visceral / Leishmania Game: educational technology for prevention/teaching visceral leishmaniasis

Introdução: a leishmaniose visceral, mais conhecida como calazar, é uma doença crônica causada por protozoários de Leishmania infantum e é uma doença caracterizada como endêmica no município de Pau dos Ferros, estado do Rio Grande do Norte. Dessa forma, a construção de uma tecnologia educativa pode ser uma ferramenta auxiliadora no processo de ensino e aprendizagem dos alunos. Objetivo: construir e validar conteúdo e aparência de jogo didático no ensino de leishmaniose visceral para alunos do ensino fundamental. Metodologia: trata-se de um estudo metodológico de abordagem quantitativa. O jogo foi confeccionado a partir de um esboço feito à mão pela autora e posteriormente entregue a um profissional responsável pelo design. Para validar o jogo, foi aplicado um protocolo de validação para 10 juízes que validaram o jogo de acordo com as expectativas curriculares e cognitivas dos alunos dos anos finais do ensino fundamental. Os dados coletados foram organizados no Microsoft® Excel® e posteriormente apresentados em gráficos e tabelas para análise final. Resultados: no que diz respeito à média geral da tecnologia educacional, obteve-se média 4,6, que corresponde a 92% de concordância dos juízes. Dessa maneira, comprova-se que todos os itens da tecnologia educacional "Leishmania" foram validados pelos juízes obtendo média superior ao estipulado. Conclusão: todos os itens apresentados no protocolo de validação foram considerados adequados na avaliação dos juízes, o que garante a confiabilidade do instrumento para ser utilizado. Dessa forma, o jogo pode ser utilizado com segurança por professores para avaliar o conhecimento dos alunos acerca da leishmaniose visceral.(AU)

Introduction: visceral leishmaniasis, better known as calazar, is a chronic disease caused by protozoa of Leishmania infantum and is a disease characterized as endemic in the municipality of Pau dos Ferros, state of Rio Grande do Norte. Thus, the construction of an educational technology can be a helping tool in the teaching and learning process of students. Objective: to construct and validate content and appearance of didactic game in the teaching of visceral leishmaniasis for elementary school students. Methodology: this was a methodological study with a quantitative approach. The game was handmade from a sketch by the author and delivered to a professional responsible for the design. To validate the game, an authentication protocol was applied to 10 judges who validated the game according to the curricular and cognitive expectations of students in the final years of elementary school. The collected data were organized in MicrosoftTM ExcelTM and later presented in graphs and tables for final analysis. Results: regarding the overall average of educational technology, obtained a score of 4.6, corresponding to 92% of the judges' agreement. Thus, it is proven that all items of educational technology "Leishmania" were validated by the judges obtaining an average higher than stipulated. Conclusion: all items presented in the validation protocol were considered adequate in the evaluation of the judges, which guarantees the reliability of the instrument to be used. Thus, the game can be safely used by teachers to assess students' knowledge about visceral leishmaniasis.(AU)

Characterization of proteinuria in treated and untreated dogs naturally infected with Leishmania sp / Caracterização da proteinúria de cães tratados e não tratados, naturalmente infectados com Leishmania sp

In the search for an early biomarker of renal injury, this study aimed to determine the urinary protein profile of dogs with leishmaniasis without treatment and treated as determined by Brazilian legislation. The identification of proteinuria, its classification and the circumstances in which it takes place instigated this study. For this, 30 dogs from an outpatient clinic at a Veterinary Hospital in Belo Horizonte were evaluated. All animals underwent clinical and laboratory tests, which included renal biomarkers. The proteins were characterized using the SDS-page electrophoresis technique, and thus, a urinary protein profile was developed comparing patients considered clinically healthy with dogs infected with leishmaniasis that were under treatment and with untreated infected dogs. The results showed that the hematological and biochemical parameters showed similar behavior between the groups of healthy dogs and dogs with leishmaniasis treated, however a very heterogeneous pattern of urinary proteins can be observed and differed between healthy animals and animals with leishmaniasis, as well as between treated and untreated animals. The results suggest that the classification of proteinuria can be a tool that helps in the staging of animals infected with L. infantum and can differentiate them as to the severity of existing kidney injuries.(AU)

Na busca por um biomarcador precoce de injúria renal, este trabalho teve como objetivo determinar o perfil proteico urinário de cães infectados com leishmaniose sem tratamento e tratados conforme determina a legislação brasileira. A identificação da proteinúria, sua classificação e as circunstâncias em que ocorrem instigaram este estudo. Para tanto, foram avaliados 30 cães oriundos do atendimento clínico ambulatorial de um Hospital Veterinário em Belo Horizonte. Todos os animais passaram por exame clínico e laboratorial, que incluíram biomarcadores renais. As proteínas foram caracterizadas através da técnica de eletroforese por SDS-PAGE, e assim, foi elaborado um perfil proteico urinário comparando pacientes considerados clinicamente hígidos, com cães infectados por Leishmania (L.) infantum e que estavam sob tratamento e cães infectados não tratados. Os resultados demonstraram que os parâmetros hematológicos e bioquímicos apresentaram comportamento semelhante entre os grupos de cães hígidos e de cães infectados com L. infantum tratados, entretanto um padrão muito heterogêneo de proteínas urinárias pode ser observado e diferiu entre animais hígidos e animais com leishmaniose, assim como entre os animais tratados e não tratados. Os resultados sugerem que a classificação da proteinúria pode ser uma ferramenta que auxilia no estadiamento de animais infectados por L. infantum podendo diferenciá-los quanto à gravidade de lesões renais existentes.(AU)

In situ cellular immune response in non-ulcerated skin lesions due to Leishmania (L) infantum chagasi infection

Skin lesions of patients affected by non-ulcerated cutaneous leishmaniasis (NUCL) caused by L. (L.) infantum chagasi are characterized by lymphohistiocytic inflammatory infiltrate associated with epithelioid granuloma and scarce parasitism. However, the in situ cellular immune response of these patients is unclear. Therefore, the aim of the present study was to characterize the cellular immune response in the skin lesions of patients affected by NUCL. Methods Twenty biopsies were processed by immunohistochemistry using primary antibodies to T lymphocytes (CD4, CD8), NK cells, B lymphocytes, macrophages, nitric oxide synthase and interferon-gamma. Results Immunohistochemistry revealed higher expression of all cellular types and molecules (IFN-γ, iNOS) in the dermis of diseased skin compared to the skin of healthy individuals (p < 0.05). Morphometric analysis performed in the skin lesions sections showed the predominance of CD8+ T lymphocytes in the mononuclear infiltrate, followed by macrophages, mostly iNOS+, a response that could be mediated by IFN-γ. Conclusion Our study improves knowledge of the cellular immune response in non-ulcerated or atypical cutaneous leishmaniasis caused by L. (L.) infantum chagasi in Central America and pointed to the pivotal participation of CD8+ T lymphocytes in the host defense mechanisms against the parasite in patients with NUCL.(AU)

Comparison of serological and molecular tests to investigate Leishmania spp. infections in stray dogs from an area of intense visceral leishmaniasis transmission in Brazil / Comparação de testes sorológicos e moleculares para investigar infecções por Leishmania spp. em cães errantes de uma área de intensa transmissão de leishmaniose visceral no Brasil

Abstract The aim of this study was to investigate the level of exposure to Leishmania infection in stray dogs in an area of intense visceral leishmaniasis transmission in the state of Pernambuco, Brazil. Blood samples from 178 dogs were analyzed using serological and molecular assays: rapid immunochromatographic test (ICT), enzyme-linked immunosorbent assay (ELISA), immunofluorescence antibody test (IFAT), and conventional and quantitative polymerase chain reaction (cPCR and qPCR). Positivity values obtained with serological tests were 71.4% (127/178), 70.2% (125/178), and 50.6% (90/178) using ICT, ELISA, and IFAT, respectively, with 38.8% (69/178) of the dogs were simultaneously positive for all three tests. The positivity values obtained with cPCR and qPCR were 20.2% (36/178) and 38.8% (69/178), respectively, with 11.8% (21/178) testing positive in both molecular assays. Overall, 87.1% (155/178) were positive for anti-Leishmania spp. antibodies and/or Leishmania spp. DNA. Positivity to one or more tests was statistically associated with lymphadenomegaly, skin lesions, lymphocytosis, anemia and hyperproteinemia. The results of this study revealed a high level of exposure to Leishmania in stray dogs in an area of intense human visceral leishmaniasis transmission, suggesting that dogs play a role as reservoirs in the transmission cycle of this zoonosis.

Resumo O objetivo deste estudo foi investigar o nível de exposição à infecção por Leishmania em cães errantes de uma área de transmissão intensa de leishmaniose visceral no estado de Pernambuco. Amostras de sangue de 178 cães foram avaliadas por testes sorológicos e moleculares: teste imunocromatográfico (IC), ensaio imunoenzimático (ELISA), teste de imunofluorescência de anticorpos (RIFI), reação em cadeia da polimerase convencional e quantitativa (cPCR e qPCR). Os valores de positividade obtidos com os testes sorológicos foram 71,4% (127/178), 70,2% (125/178) e 50,6% (90/178) com IC, ELISA e RIFI, respectivamente. E com 38,8% (69/178) dos cães mostraram-se simultaneamente positivos aos três testes. Os valores de positividade obtidos por cPCR e qPCR foram 20,2% (36/178) e 38,8% (69/178), respectivamente. E com 11,8% (21/178) dos cães foram positivos em ambos os testes moleculares. No geral, 87,1% (155/178) foram positivos para anticorpos anti-Leishmania spp. e/ou material genético de Leishmania spp. A positividade a um ou mais testes foi estatisticamente associada à linfonodomegalia, lesões cutâneas, linfocitose, anemia e hiperproteinemia. Os resultados do estudo revelam um alto nível de exposição à Leishmania spp. em cães errantes em uma área de transmissão intensa de leishmaniose visceral humana, sugerindo que eles podem desempenhar um papel como reservatórios no ciclo de transmissão dessa zoonose.

Polymerase chain reaction using conjunctival swab samples for detecting Leishmania DNA in dogs / Reação em cadeia da polimerase usando amostras de Swab de conjuntiva para detecção de DNA de Leishmania em cães

Abstract The dog is the main domestic reservoir of Leishmania and font of infection for the vector, constituting an important host for the transmission of the parasite to humans. Non-invasive collection of swab samples for leishmaniasis diagnosis has been a promising alternative. This study analyzed the positivity of polymerase chain reaction (PCR) for the diagnosis of canine leishmaniasis in conjunctiva samples. DNA extraction was performed using SDS 20% and PCR was performed using 13A/13B primers that amplify 120-bp of Leishmania kDNA. Of the 77 dogs analyzed, 50 (64.93%) had ocular changes: 25 (32.47%) dogs had periocular lesion, 41 (53.25%) dogs had purulent eye discharge, and 17 (22.08%) dogs had both signals. PCR was positive in 35 dogs (45.45%), and there was no significant difference between dogs with and without ocular signals (p=0.4074). PCR positivity was significant higher in dogs without periocular injury (p=0.0018). Conjunctive PCR, a less invasive, fast, and painless collection technique, is indicated to complement the diagnosis, especially in dogs without periocular injury, independent of the presence of purulent eye discharge.

Resumo O cão é o principal reservatório doméstico de Leishmania e também fonte de infecção para o vetor, constituindo um importante hospedeiro para a transmissão do parasita ao homem. A coleta não invasiva de amostras em swab para diagnóstico das leishmanioses tem sido uma alternativa promissora. Este estudo analisou a positividade da reação em cadeia da polimerase (PCR) para o diagnóstico de leishmaniose canina em amostras de conjuntiva. A extração do DNA foi realizada com SDS 20%. A PCR foi realizada com primers 13A/13B que amplificam 120-pb do kDNA de Leishmania. Dos 77 cães analisados, 50 (64,93%) tiveram alterações oculares; 25 (32,47%) cães tiveram uma lesão periocular; 41 (53,25%) tiveram secreção ocular purulenta e 17 (22,08%) cães tiveram ambos os sinais. A PCR foi positiva em 35 cães (45,45%) e não houve diferença significativa em cães com e sem sinais oculares (p = 0,4074). A positividade da PCR foi significativamente maior em cães sem lesão periocular (p = 0,0018). PCR em conjuntiva, uma técnica de coleta menos invasiva, rápida e indolor, é indicada para complementar o diagnóstico, principalmente em cães sem lesão periocular, independentemente da presença de secreção ocular purulenta.

Evaluation of cytochrome b sequence to identify Leishmania species and variants: the case of Panama

BACKGROUND The genetic heterogeneity of Leishmania parasites is a major factor responsible for the wide variety of Leishmania-associated manifestations. Consequently, understanding the genetic make-up of Leishmania species using suitable molecular markers is an important component of realising local and regional scale disease risk. The cytochrome b (cytb) is frequently used to type New World Leishmania species. However, its potential to discriminate Leishmania species and variants requires further evaluation. OBJECTIVES To explore the capacity of cytb gene to identify New World Leishmania species and variants and to develop an approach able to type local Leishmania species and variants. METHODS We retrieved 360 partial and complete Leishmania cytb gene sequences publicly available in GenBank database to study all single nucleotide polymorphisms (SNPs) across the cytb gene that differentiate New World Leishmania species. This information was used to develop an approach based upon the polymorphisms found in a DNA segment of 948bp. We also compared the typing results found with this technique with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) profiling obtained using HSP70 gene as target. One hundred Panamanian isolates were used to both typed Leishmania species and assess local genetic variability. FINDINGS We found complete agreement between our cytb approach and the PCR-RFLP profiling method based on HSP70 for Leishmania species identification. Ninety-two isolates were identified as L. panamensis, although other Viannia species were found circulating at a lower frequency. Three L. panamensis haplotypes were identified in Panamanian provinces. We also provide an initial report of L. guyanensis haplotypes circulating in Panama. MAIN CONCLUSIONS Cytb gene sequence encompasses key main SNPs that aid to identify Leishmania species. The cytb approach developed with this information was able to identify and assess genetic variability of local Leishmania species found in this study.