Un caso de Chagas agudo en edad pediátrica / Acute Vector-transmitted Chagas in Pediatric Age. A Case Reportt

Presentación del caso. Se trata una niña de siete años de edad, originaria de una zona rural del departamento de San Miguel, quién consultó por presentar fiebre y edema bipalpebral derecho indoloro, de seis semanas de evolución, sin otros síntomas acompañantes. El estudio para el diagnóstico de enfermedad de Chagas fue realizado en una clínica privada; la Inmunoglobulina M para Chagas tuvo un resultado positivo, luego, se realizó la microscopía directa mediante gota al fresco y técnica de Strout con resultado negativo. En las intervenciones comunitarias se identificó la presencia del vector y la positividad del mismo, así como el diagnóstico de un caso crónico en otro miembro de la familia. Intervención terapéutica. Se indicó tratamiento con nifurtimox 150 mg cada ocho horas por 60 días y se realizó el seguimiento clínico de la evolución y control de efectos secundarios del tratamiento y exámenes de laboratorio. Evolución clínica. Evolucionó con leve disminución del apetito, se manejó con protectores gástricos. El concentrado de Strout y la gota al fresco resultaron negativos y los demás exámenes de laboratorio se mantenían en los rangos normales.

Case presentation. A seven-year-old female patient, with no previous medical history, originally from a rural area of the department of San Miguel, who presented febrile process plus long-term right bipalpebral edema of six weeks of evolution, without accompanying symptoms. Immunoglobulin M for Chagas was positive, direct microscopy by fresh drop and Strout technique was performed with negative results. In community interventions, the presence of the vector and its positivity were identified, as well as the diagnosis of a chronic case in another family member. Treatment. The patient was treated with nifurtimox 150 mg every eight hours for 60 days, subsequent controls were performed to investigate side effects of the treatment, and control tests. Outcome. With the treatment, the patient evolved with a slight decrease in appetite, and was managed with gastric protectors. Strout's concentrate and fresh gout were negative and the other laboratory tests were within normal ranges.

Efectos de la infección del Trypanosoma cruzi en ratones de las cepas Balb/c y NIH / Effects of Trypanosoma cruzi infection in Balb/c and NIH mouse strains

La enfermedad de Chagas es una infección causada por el parásito Trypanosoma cruzi y transmitida por el vector Triatoma dimidiata, conocido en El Salvador como «chinche picuda¼. Esta enfermedad siempre ha sido de interés científico en modelos animales. Objetivo. Identificar el efecto de la infección por Trypanosoma cruzi en ratones de diferentes cepas (BALB/c y NIH) y sexo. Metodología. Se establecieron ocho grupos: cuatro infectados con Trypanosoma cruzi y cuatro grupos no infectados, distribuidos por cepa y sexo, con cinco ratones por grupo. Durante seis semanas se registró el peso corporal de los ratones. Además, se prepararon muestras de sangre de los grupos infectados en láminas para realizar los conteos de parasitemia. Al final del estudio, se extrajeron el bazo y el corazón de ambos grupos para los análisis estadísticos. Resultados. Los grupos infectados mostraron un incremento de peso en comparación a sus grupos controles. En la cepa NIH, las hembras presentaron una mayor parasitemia, mientras que en la cepa BALB/c fueron los machos los de mayor parasitemia. Los órganos de los grupos infectados fueron significativamente más grandes comparados a los de los grupos de control, excepto en el corazón de la cepa BALB/c. Respecto al peso de los órganos, se observaron diferencias significativas únicamente en el corazón de los machos de la cepa BALB/c, mientras que en el bazo ocurrió lo contrario. Conclusión. Los machos de la cepa BALB/c son más susceptibles al Trypanosoma cruzi, presentando niveles de parasitemia más altos entre los grupos estudiados

Chagas disease is an infection caused by the parasite Trypanosoma cruzi and transmitted by the vector Triatoma dimidiata, known in El Salvador as "chinche picuda". This disease has always been of scientific interest in animal models. Objective. Identify the effect of Trypanozoma cruzi infection in mice of different strains (BALB/c and NIH) and sex. Methodology. Eight groups were established: four infected with Trypanosoma cruzi and four uninfected groups, distributed by strain and sex, with five mice per group. The body weight of the mice was recorded for six weeks. In addition, blood samples from the infected groups were prepared on slides for parasitemia counts. At the end of the study, the spleen and heart were extracted from both groups for statistical analyses. Results. The infected groups showed an increase in weight compared to their control groups. In the NIH strain, females had higher parasitemia, whereas in the BALB/c strain, males had higher parasitemia. The organs of the infected groups were significantly larger compared to those of the control groups, except in the heart of the BALB/c strain. Regarding organ weight, significant differences were observed only in the heart of the male BALB/c strain, while the opposite was true for the spleen. Conclusion. Males of the BALB/c strain are more susceptible to Trypanosoma cruzi, presenting higher levels of parasitemia among the groups studied.

Influence of sexual hormones on Chagas disease / Influencia de las hormonas sexuales en la enfermedad de Chagas

Abstract Objective: Analyze sex hormone's influence during Chagas disease. Methods: Male and female BALB/c mice were divided into six groups, four experimental (sham, orchiectomized, orchiectomized and supplemented with estradiol, orchiectomized supplemented with testosterone, oophorectomized, oophorectomized and supplemented with estradiol, and oophorectomized and supplemented with testosterone), and two control (healthy and intraperitoneally with T. cruzi strain NINOA infected). Clinical data were recorded daily, parasitemia was evaluated using a Neubauer chamber during the infection, and heart histopathological analysis was performed using the paraffin embedding technique. To analyze parasitemia curves and the area under the parametric curves, two-way ANOVA test was performed to correlate groups' data. P-values < 0.05 were considered statistically significant. Results: Higher mortality rates, cardiomegaly, hepatomegaly, ascites, edema, higher parasitemia levels, more amastigote nests, and more severe inflammatory infiltrate were found in higher testosterone concentration mice, whereas in higher estradiol concentration groups, paresia, prostration, edema, and necrosis were found. Conclusions: Our results showed that testosterone increased infection severity, whereas estradiol had the opposite effect. This research improves the understanding of sex hormones´ infuence upon this infection to contribute with the handling of Chagas´ disease.

Resumen Objetivo: Analizar la influencia de las hormonas durante la enfermedad de Chagas. Métodos: Se separaron grupos de ratones macho y hembras BALB/c, todos infectados con T. cruzi (cepa NINOA), 4 grupos experimentales de machos (Sham, orquidectamizados, orquidectimezados y suplementados con estradiol, orquidectamizaos y suplementados con testosterona). 4 grupos experimentales de hembras (oforectomizadas, oforectomizadas y suplementadas con estradiol, oforectomizadas y suplementadas con testosterona y sham), and y dos grupos control para cada sexo (sin infección e infectados intraperitonealmente con T. cruzi (cepa NINOA). Los datos clínicos fueron registrados diariamente, la parasitemia fue evaluada durante toda la infección utilizando una cámara de Neubauer y el análisis histopatológico del corazón fue realizada con la técnica de inclusión en parafina. Para el análisis de las curvas de parasitemia y el área bajo la curva, se realizó una prueba de ANOVA de dos vías, p < 0.05 fueron considerados estadísticamente diferentes. Resultados: Las mayores tasas de mortalidad, cardiomegalia, hepatomegalia y mayor infiltrado inflamatorio, se encontró en los ratones con una mayor concentración de testosterona. En contraste los ratones con mayor concentración de estradiol presentaron paresia, postración edema y necrosis. Conclusiones: Nuestros resultados ponen en manifiesto que la testosterona incrementa la severidad del curso de la enfermedad de Chagas, mientras que el estradiol tuvo el efecto opuesto. Este trabajo mejora el entendimiento del rol que juegan las hormonas sexuales en esta infección para contribuir en un mejor manejo de la enfermedad de Chagas.

GSK-3 kinase a putative therapeutic target in trypanosomatid parasites

ABSTRACT Trypanosomatids are an important group of parasites that predominate in tropical and subtropical areas of the planet, which cause diseases that are classified as forgotten and neglected by the world health organization. In this group of parasites, we find Trypanosoma cruzi, Trypanosoma brucei, Trypanosoma brucei rhodesiense and Leishmania spp, for which there is no vaccine available, and its control has focused mainly on pharmacological treatment. Due to the poverty situation where these diseases are found and the biological complexity of these parasites, there are multiple variables to control, including the diversity of species, the complexity of their life cycles, drug resistance, cytotoxicity, the limited use in pregnant women, the high costs of treatment and the little-known pharmacological mechanisms of action, among others. It is therefore necessary to find new strategies and approaches for the treatment of these parasitic diseases. Among these new approaches is the rational search for new targets based on the allosteric inhibition of protein kinases, which have been little studied in trypanosomatids. Among these kinases, we find Glycogen Synthase Kinase-3 (GSK-3), a kinase of great pharmacological interest, which is under intense basic and clinical research by pharmaceutical companies for the treatment of cancer. This kinase, highly studied in the PI3K/AKT/mTOR pathway signaling in humans, has an orthologous gene in these parasites (GSK-3 s), which has proven to be essential for them in response to different challenges; Therefore, it is notable to increase research in this kinase in order to achieve a broad structural and functional characterization in the different species of trypanosomatids.

Improving in vitro screening compounds anti-Trypanosoma cruzi by GFP-expressing parasites

BACKGROUND Conventional microscopic counting is a widely utilised method for evaluating the trypanocidal effects of drugs on intracellular amastigotes. This is a low-cost approach, but it is time-consuming and reliant on the expertise of the microscopist. So, there is a pressing need for developing technologies to enhance the efficiency of low-cost anti-Trypanosoma cruzi drug screening. OBJECTIVES In our laboratory, we aimed to expedite the screening of anti-T. cruzi drugs by implementing a fluorescent method that correlates emitted fluorescence from green fluorescent protein (GFP)-expressing T. cruzi (Tc-GFP) with cellular viability. METHODS Epimastigotes (Y strain) were transfected with the pROCKGFPNeo plasmid, resulting in robust and sustained GFP expression across epimastigotes, trypomastigotes, and intracellular amastigotes. Tc-GFP epimastigotes and intracellular amastigotes were exposed to a serial dilution of benznidazole (Bz). Cell viability was assessed through a combination of microscopic counting, MTT, and fluorimetry. FINDINGS The fluorescence data indicated an underestimation of the activity of Bz against epimastigotes (IC50 75 µM x 14 µM). Conversely, for intracellular GFP-amastigotes, both fluorimetry and microscopy yielded identical IC50 values. Factors influencing the fluorimetry approach are discussed. MAIN CONCLUSIONS Our proposed fluorometric assessment is effective and can serve as a viable substitute for the time-consuming microscopic counting of intracellular amastigotes.

Seroprevalencia de Trypanosoma cruzi en niños de Veracruz, México: línea epidemiológica de base para un modelo de control fundamentado de la transmisión activa de la enfermedad de Chagas / Seroprevalence of Trypanosoma cruzi among children from Veracruz, Mexico: Epidemiological baseline for a control model based on Chagas disease active transmission

Resumen Introducción. En el 2021, la Secretaría de Salud de México y la Organización Panamericana de la Salud lanzaron una iniciativa para interrumpir la transmisión vectorial intradomiciliaria de Trypanosoma cruzi, fundamentada en la prevalencia de la enfermedad de Chagas en la población infantil. El estado mexicano de Veracruz fue el pionero de esta iniciativa. Objetivo. Estimar la seroprevalencia de infección por T. cruzi en menores de 15 años de localidades rurales de Veracruz, México. Materiales y métodos. Se identificaron ocho localidades prioritarias para la serología basal del municipio de Tempoal, Veracruz. Entre junio y agosto de 2017, se recolectaron muestras de sangre en papel filtro de 817 individuos para su tamizaje mediante un inmunoensayo enzimático de tercera generación. Los casos reactivos del tamizaje se confirmaron mediante pruebas de hemaglutinación indirecta, ensayo de inmunoabsorción ligado a enzimas e inmunofluorescencia indirecta en muestras de suero. Se calculó la seroprevalencia y su intervalo de confianza (IC) del 95 %. Resultados. En las localidades de Citlaltépetl, Cornizuelo, Cruz de Palma y Rancho Nuevo se confirmaron casos de la enfermedad de Chagas en menores de 15 años con una seroprevalencia de 1,9 % (IC 95 % = 1,12-3,16). Conclusiones. Los resultados indican que estas comunidades presentan transmisión reciente de T. cruzi y permiten establecer una línea epidemiológica de base para el diseño e implementación de un modelo dirigido a aquellas áreas geográficas con transmisión activa. Se espera que dicho modelo contribuya a la eliminación de la transmisión vectorial intradomiciliaria del tripanosomátido en México.

Abstract Introduction. In 2021, the Secretaría de Salud de México and the Pan American Health Organization launched an initiative to interrupt intra-domiciliary vector transmission of Trypanosoma cruzi based on the prevalence of Chagas disease in children. The Mexican State of Veracruz was leading this initiative. Objective. To estimate the seroprevalence of T. cruzi infection among children under 15 years of age from rural areas of Veracruz, México. Materials and methods. We identified eight localities of high priority from the Municipality of Tempoal, Veracruz, for baseline serology. Blood samples were collected on filter paper from 817 individuals between June and August 2017, for screening with a third-generation enzyme immunoassay. Reactive cases were confirmed by indirect hemagglutination, enzyme-linked immunosorbent assay, and indirect immunofluorescence tests on peripheral blood serum samples. We calculated seroprevalence and 95% confidence intervals (CI). Results. We confirmed Chagas disease cases in children under 15 years of age with a seroprevalence of 1,9% (95 % CI = 1,12-3,16) in the localities of Citlaltepetl, Cornizuelo, Cruz de Palma and Rancho Nuevo. Conclusions. These results indicate recent transmission of T. cruzi in these communities and allow to establish an epidemiological baseline for the design and implementation of a model focused on geographical areas with active transmission to advance toward the elimination of intra-domiciliary vector transmission of this parasite in Mexico.

Drug screening and development cascade for Chagas disease: an update of in vitro and in vivo experimental models

Chagas disease is a tropical neglected disease that affects millions of people worldwide, still demanding a more effective and safer therapy, especially in its chronic phase which lacks a treatment that promotes substantial parasitological cure. The technical note of Romanha and collaborators published in 2010 aimed establish a guideline with the set of minimum criteria and decision gates for the development of new agents against Trypanosoma cruzi with the focus on developing new antichagasic drugs. In this sense, the present review aims to update this technical note, bringing the state of the art and new advances on this topic in recent years.

Molecular identification of new Trypanosoma evansi type non-A/B isolates from buffaloes and cattle in Indonesia / Identificação molecular de novos isolados de Trypanosoma evansi tipo não A/B em búfalos e gado na Indonésia

Abstract Trypanosoma evansi is reportedly divided into two genotypes: types A and B. The type B is uncommon and reportedly limited to Africa: Kenya Sudan, and Ethiopia. In contrast, type A has been widely reported in Africa, South America, and Asia. However, Trypanosoma evansi type non-A/B has never been reported. Therefore, this study aims to determine the species and genotype of the Trypanozoon subgenus using a robust identification algorithm. Forty-three trypanosoma isolates from Indonesia were identified as Trypanosoma evansi using a molecular identification algorithm. Further identification showed that 39 isolates were type A and 4 isolates were possibly non-A/B types. The PML, AMN-SB1, and STENT3 isolates were likely non-A/B type Trypanosoma evansi isolated from buffalo, while the PDE isolates were isolated from cattle. Cladistic analysis revealed that Indonesian Trypanosoma evansi was divided into seven clusters based on the gRNA-kDNA minicircle gene. Clusters 6 and 7 are each divided into two sub-clusters. The areas with the highest genetic diversity are the provinces of Banten, Central Java (included Yogyakarta), and East Nusa Tenggara. The Central Java (including Yogyakarta) and East Nusa Tenggara provinces, each have four sub-clusters, while Banten has three.

Resumo Trypanosoma evansi é reportado como dividido em dois genótipos: tipos A e B. O tipo B é incomum e reportado como limitado à África: Quênia, Sudão e Etiópia. Em contraste, o tipo A tem sido amplamente relatado na África, América do Sul e Ásia. No entanto, Trypanosoma evansi tipo não-A/B nunca foi relatado. Portanto, este estudo tem como objetivo determinar a espécie e o genótipo do subgênero Trypanozoon, utilizando-se um algoritmo robusto de identificação. Quarenta e três isolados de tripanosoma da Indonésia foram identificados como Trypanosoma evansi, usando-se um algoritmo de identificação molecular. A identificação adicional mostrou que 39 isolados eram do tipo A e 4 isolados eram, possivelmente, do tipo não A/B. Os isolados PML, AMN-SB1 e STENT3 foram, provavelmente, Trypanosoma evansi do tipo não A/B isolado de búfalos, enquanto os isolados de PDE foram isolados de bovinos. A análise cladística revelou que o Trypanosoma evansi indonésio foi dividido em sete grupos baseados no gene do minicírculo gRNA-kDNA. Os clusters 6 e 7 foram divididos cada um em dois subclusters. As áreas com maior diversidade genética são as províncias de Banten, Java Central (incluindo Yogyakarta) e East Nusa Tenggara. As de Java Central (incluindo Yogyakarta) e East Nusa Tenggara têm, cada uma, quatro subgrupos, enquanto Banten tem três.

Diagnóstico de Chagas agudo: experiencia en un hospital pediátrico de tercer nivel de atención / Diagnosis of acute Chagas: experience in a tertiary care children's hospital / Diagnóstico de Chagas agudo: experiência em um hospital pediátrico de terceiro nível de atenção

Resumen Chagas es una infección causada por Trypanosoma cruzi. Actualmente la principal vía de transmisión en la Argentina es la vertical y es prioritario el diagnóstico oportuno. El objetivo de este trabajo fue analizar características epidemiológicas y de seguimiento de pacientes con sospecha de Chagas agudo. Se realizó un estudio observacional, descriptivo y retrospectivo, analizando las historias clínicas de pacientes con pedido de parasitemia entre 2015 y 2022. Se registró resultado, antecedentes epidemiológicos y si se completó el algoritmo diagnóstico. Se analizaron 254 pacientes con una mediana de edad de 6 meses. Veintidós fueron diagnosticados con Chagas; 17 fueron por transmisión vertical, 1 por transmisión vertical/trasplante y 4 por vertical/vectorial. En 126 pacientes se descartó la infección, mientras que en 99 no se completó el algoritmo diagnóstico. Siete pacientes fueron excluidos del estudio. Se diagnosticó Chagas en el 8,7% de los pacientes y se observó pérdida de seguimiento en un 36,2%.

Abstract Chagas is an infection caused by Trypanosoma cruzi. Nowadays, in Argentina vertical transmission is the main source of infection, which makes opportunistic diagnosis mandatory. The aim of this study was to analyse the epidemiological characteristics and follow-up of patients with suspected acute Chagas infection. An observational, descriptive, retrospective study was carried out. Medical records of patients with parasitemia test solicitude in the period between 2015 and 2022 were analysed, and results were registered, along with epidemiological background and if diagnosis algorithm were completed; 254 patients were analysed with a median age of 6 months: 22 patients were diagnosed with Chagas, 17 of whom had a vertical transmission, 1 was vertical/transplant transmission and there were 4 cases of vertical/vectorial transmission. Infection was ruled out in 126 patients, while in 99 children algorithm diagnosis was not completed. Seven patients were excluded from this study. Chagas was diagnosed in 8.7% of the patients studied and there was loss in follow-up in 36.2% of patients.

Resumo Chagas é uma infecção causada pelo Trypanosoma cruzi. Atualmente, na Argentina, a principal via de transmissão é a vertical e é prioritário o diagnóstico oportuno. O objetivo deste trabalho foi analisar as características epidemiológicas e de seguimento de pacientes com suspeita de Chagas agudo. Foi realizado um estudo observacional, descritivo e retrospectivo. Analisaram-se osprontuarios médicos dos pacientes com pedido de parasitemia entre 2015 e 2022. Foram registrados resultado e antecedentes epidemiológicos, e se completou o algoritmo diagnóstico. Analisaram-se 254 pacientes com uma mediana de idade de 6 meses: 22 foram diagnosticados com Chagas; 17 foram por transmissão vertical, 1 transmissão vertical/transplante, e 4 vertical/vetorial. Em 126 pacientes foi descartada a infecção, enquanto que em 99 não se completou o algoritmo diagnóstico. Sete pacientes foram excluídos do estudo. Foi diagnosticado Chagas em 8,7% dos pacientes e se observou perda de seguimento em 36,2%.

Synthesis of (-) - 6, 6'-dinitrohinokinin from hinokinin natural product and in silico and in vitro trypanocidal activity assessment

Chagas disease is a public health problem affecting approximately seven million people worldwide. Thus, there is a need to discover drugs for the adequate treatment of this disease because currently available drugs have serious side effects. Therefore, this study aimed to evaluate the in vitro trypanocidal activity of (-)-6,6'-dinitrohinokinin, obtained from the partial synthesis of (-)-hinokinin, on the trypomastigotes and amastigotes forms. For the trypomastigote assay, blood was collected from mice infected with Trypanosoma cruzi through cardiac puncture at the parasitemic peak. The results show that (-)-6,6'-dinitrohinokinin was effective against the trypomastigote forms, presenting an IC50 of 19.83 µM and lysis percentage values ​​of 78.4% and 69.4% at concentrations of 200 and 100 µM, respectively. Molecular docking calculations indicate that (-)-6,6'-dinitrohinokinin favorably interacts with the amino acids present in the active site of the protein trypanothione reductase, a typical target for anti-trypanosomal drug development. According to the results, the (-)-6,6'-dinitrohinokinin showed more significant trypanocidal activity with IC50 of 1.83 µM than benzonidazole positive control with IC50 of 53.2 µM, showing to be a prototype molecule promising for the development of a new antiparasitic drug.

Infección por Trypanosoma spp. en murciélagos capturados en ecótopos urbanos y silvestres de la región Caribe en Colombia / Trypanosoma spp. infection in bats captured in urban and wild ecotopes of the Caribbean region in Colombia

R ESUMEN: Se evaluó la frecuencia de infección por Trypanosoma spp. en murciélagos capturados en ecótopos sil-vestres y urbanos del Departamento del Atlántico, en la región Caribe de Colombia, entre marzo de 2021 y mayo de 2022. Se identificaron taxonómicamente los murciélagos y se determinó sexo, edad relativa y condiciones reproductivas. Se utilizó una muestra de sangre para análisis parasitológico y extracción de ADN para la amplificar una región del ARNr 18S. Se capturaron 125 murciélagos, siendo las familias más abundantes Molossidae (62/125; 49,6%) y Phyllostomidae (43/125; 34,4%). Molossus molossus capturado en ecótopos silvestres mostró una frecuencia de infección del 8,1% (5/61) y 4,1% (3/61) mediante análisis parasitológico y molecular, respectivamente. En comparación, Noctilio albiventris capturado en ecótopos urbanos mostró una frecuencia de infección del 16,6% (2/12) para ambos análisis. Estos hallazgos re-presentan los primeros registros de M. molossus albergando Trypanosoma spp. para el Departamento del Atlántico y de N. albiventris albergando Trypanosoma spp. en Colombia.

Conocimientos, actitudes y prácticas sobre la enfermedad de Chagas en una zona endémica de Boyacá, Colombia

Introducción: Los instrumentos para la obtención de información sobre los conocimientos, actitudes y prácticas de diversas enfermedades son ampliamente utilizados, ya que permiten obtener información clara y detallada de cada uno de los aspectos a indagar. Objetivo: Determinar conocimientos, actitudes y prácticas de la enfermedad de Chagas en una zona endémica de Boyacá, Colombia. Metodología: Estudio transversal que consistió en aplicar un instrumento validado que abordaba datos sociodemográficos, factores epidemiológicos, conocimientos, actitudes y prácticas de la enfermedad de Chagas en un integrante mayor de edad de 341 familias de Miraflores, Boyacá. Se emplearon escalas, óptimo, bueno, regular y malo; para el análisis bivariado se determinó el nivel de conocimientos, actitudes y prácticas con escala favorable y desfavorable. Se determinaron diferencias significativas de factores de riesgo, conocimientos, actitudes y prácticas de la enfermedad entre el grupo de hombres y mujeres encuestados. Resultados: El 68,6 % de la población nació en Miraflores, con un 48 % de escolaridad primaria y pertenecientes a la zona rural en su mayoría (57,2 %); en las mujeres predomina la ocupación de ama de casa, mientras que en los hombres es más frecuente la agricultura. Se identificaron conocimientos (48,1 %) y actitudes (82,1 %) óptimas sobre la enfermedad de Chagas que minimizan el riesgo de contraer la infección por T. cruzi; paradójicamente, se encontraron malas prácticas (61,9 %) en el hogar que no previenen la enfermedad. Se observó relación entre el nivel de escolaridad bajo, sexo femenino, ser menor de 49 años, vivir en zona urbana y actividades del hogar y un conocimiento favorable acerca de la enfermedad de Chagas, aunque sin evidencia estadística. Conclusiones: Se hace necesario incorporar programas que garanticen el aprendizaje y la implementación de actitudes y prácticas favorables contra la enfermedad en habitantes de zonas endémicas.

Introduction: The use of measurement tools to obtain information about knowledge, attitudes and practices of various diseases are widely used, since they allow to collect clear and detailed information of every aspect needing examination. Objective: Determine knowledge, attitudes and practices regarding Chagas disease in an endemic zone in Boyaca, Colombia. Methods: Cross-sectional study that consisted of applying a validated instrument that addressed sociodemographic data, epidemiological factors, knowledge, attitudes and practices of the Chagas disease in an adult member of 341 families from Miraflores, Boyaca. The scale categories used were optimal, good, regular and bad; for the bivariate analysis, the level of knowledge, attitudes and practices was determined with a favorable and unfavorable scale. Significant differences in risk factors, knowledge, attitudes and practices of the disease were determined between the group of men and women surveyed. Results: Sixty-eight percent of the population were born in Miraflores, 48% had primary education and most belonged to the rural area (57.2%); in women the occupation of housewife predominated, in men agriculture was more frequent. Optimal knowledge (48.1%) and attitudes (82.1%) about the Chagas disease that minimize the risk of contracting T. cruzi infection were identified. Paradoxically, bad practices that did not prevent the disease were found at home (61,9%). significant regarding the low level of schooling, female gender, being under 49 years of age, living in an urban area and household activities, all of the above allows a favorable knowledge about Chagas disease, although without statistical evidence. Conclusion: It is necessary to incorporate programs that guarantee learning and implementation of favorable attitudes and practices against the disease in inhabitants of endemic areas.

Blood parasites (Kinetoplastida: Trypanosomatidae) infecting birds in the Brazilian Amazon / Parásitos sanguíneos (Kinetoplastida: Trypanosomatidae) que infectan aves en la Amazonía brasileña

Abstract Introduction: Trypanosomes are hemoparasites that can be observed circulating in the peripheral blood of birds. Parasitological studies in birds in their natural environment are neglected, but are important for research relating to transmission, maintenance of the biological cycle, and abundance, among other parasitological aspects. Objective: To describe infections by Trypanosoma sp. in birds in the Brazilian Amazon, as well as the prevalence, morphological and morphometric characteristics of this hemoparasite. Methods: In the Tapajós National Forest, we captured a total of 125 birds, mostly from the order Passeriformes. We obtained blood samples from the ulnar vein using sterile insulin needles, and aliquots of blood using a microhematocrit capillary tube. We made blood smears in triplicate and stained with the Giemsa method. We viewd the morphotypes of the Trypanosoma sp. under the light microscope with objective lenses of 40 X and 100 X. To determine the morphometric characteristics of Trypanosomatidae, we used the Zen Blue Edition 2 software package. Results: We observed the presence of hemoparasites in the trypomastigote form in specimens of Thamnophilidae, Dendrocolaptidae and Conopophagidae, with low prevalence. Only one morphotype of Trypanosoma sp. was detected and measurement. Conclusions: We report the infection by Trypanosoma sp. in species of ecological importance, such as Phlegopsis nigromaculata, endangered in Brazil. The morphology and morphometry of the morphotype found could contribute to more detailed descriptions of these hemoparasites.

Resumen Introducción: Los tripanosomas son hemoparásitos que pueden observarse circulando en la sangre periférica de las aves. Los estudios parasitológicos en aves en el medio natural son escasos, pero son importantes para la investigación relacionada con la transmisión, el mantenimiento del ciclo biológico y la abundancia, entre otros aspectos parasitológicos. Objetivo: Describir infecciones por Trypanosoma sp. en aves de la Amazonia brasileña, así como la prevalencia, características morfológicas y morfométricas de este hemoparásito. Métodos: En la Floresta Nacional de Tapajós, capturamos un total de 125 aves, la mayoría del orden Passeriformes. Obtuvimos muestras de sangre por punción de la vena cubital del ala con agujas estériles de insulina. Con un tubo capilar microhematocrito, obtuvimos alícuotas de sangre. Realizamos frotis de sangre por triplicado y teñimos con el método de Giemsa. Visualizamos los morfotipos de Trypanosoma sp. al microscopio óptico con lentes objetivos de 40 X y 100 X. Para determinar las características morfométricas de Trypanosomatidae, usamos el paquete informático Zen Blue Edition 2. Resultados: Observamos la presencia de hemoparásitos en la forma tripomastigote en ejemplares de la familia de aves Thamnophilidae, Dendrocolaptidae y Conopophagidae, con baja prevalencia. Solo detectamos un morfotipo de Trypanosoma sp. Conclusión: Reportamos la infección por Trypanosoma sp. en especies de importancia ecológica, como Phlegopsis nigromaculata en peligro de extinción en Brasil. La morfología y morfometría del morfotipo encontrado puede contribuir con descripciones más detalladas de estos hemoparásitos.

Contribuições para a mobilização social, comunicação e educação em saúde sobre a doença de Chagas no YouTube ­ percurso e notas sobre o canal Falamos de Chagas / Contributions to social mobilization, communication and health education on Chagas disease on YouTube ­ route and notes on the channel Falamos de Chagas / Aportes a la movilización social, comunicación y educación en salud sobre la enfermedad de Chagas en YouTube ­ trayectoria y notas en el canal Falamos de Chagas

A doença de Chagas crônica afeta seis milhões de pessoas em regiões endêmicas, com 30 mil novos casos anuais ­ logo, espaços de divulgação científica são muito importantes para ofertar informações de qualidade à população. As iniciativas envolvendo o controle da doença de Chagas não podem se limitar às pesquisas com enfoque biológico. Este estudo objetiva apresentar um panorama sobre o processo de construção do canal Falamos de Chagas, no YouTube, sua importância para a comunicação, a informação, a educação em saúde e a mobilização social, bem como refletir sobre a qualidade de uma subamostra de vídeos do canal. Trata-se de um estudo qualitativo, dividido em duas fases: criação do canal e análise qualitativa dos vídeos sobre a doença disponíveis no YouTube. Observamos que existe potencial nas redes sociais, enquanto recurso de comunicação, contudo é preciso cautela, uma vez que se faz necessária a certificação da qualidade do material

Chronic Chagas disease affects six million people in endemic regions, with 30,000 new infected cases an-nually ­ thus, initiatives involving science diffusion are relevant to offer qualified information to the people. Chagas disease control initiatives cannot be limited to the level of biological focused research. This study aims to present an overview of the construction process of the YouTube channel Falando de Chagas, its importance for communication, information, health education and social mobilization, as well as to reflect on the quality of a subsample of videos present in the channel. Qualitative in nature, the study was divided into two stages: construction of the channel and qualitative analysis of videos about the disease available on YouTube. We observed that there is potential for social networks as communication resources, but caution is needed in their use, since the quality of the material needs certification

La enfermedad de Chagas crónica afecta seis millones de personas en regiones endémicas, con 30.000 nuevos casos anuales ­ los espacios de divulgación científica son muy importantes para ofrecer información a la población. Las iniciativas de control de la enfermedad de Chagas no pueden limitarse al nivel de investigación con enfoque biológico. El estudio tiene como objetivo presentar un panorama del proceso de construcción del canal Falando de Chagas, en YouTube, su importancia para la comunicación, información, educación en salud y movilización social, así como reflexionar sobre la calidad de una submuestra de videos presentes en la canal. De naturaleza cualitativa, el estudio se dividió en dos fases: construcción del canal y análisis cualitativo de videos sobre la enfermedad disponibles en YouTube. Observamos que existe potencial para las redes sociales como recurso de comunicación, sin embargo, se requiere cautela en su uso, ya que se requiere certificar la calidad del material

In Vitro Trypanocidal Effect of the Extract and Isolated Compound of Corymbia torelliana Stem Bark against Trypanosoma brucei brucei (Federe Strain)

African animal trypanosomosis (AAT) is a severe debilitating protozoan disease of livestock and some wild animal species. It is caused by the pathogenic trypanosome species, and transmitted by Tsetse flies. It affects about 60 million cattle in Africa, with about US$1.5 billion in annual losses. Eradication of the disease through chemotherapy and vaccine development has been hindered by parasite antigenic variation and its resistance to drugs. In this study, the crude ethanol stem bark extract of Corymbia torelliana was prepared by cold maceration in 70% Ethanol. It was further purified using the modified Kupchan partition method into n-hexane, Ethyl acetate, N-butanol and Aqueous pure fractions. A pure compound was isolated from the ethyl acetate purified fraction using the column and thin layer chromatographic techniques. The isolated compound was named Makoshi_Mak1. The in vitro antitrypanosomal effects of the crude extract, the purified fractions and the isolated compound were then tested against the blood stream forms of Trypanosoma brucei brucei (Federe strain). 0.2 ml of prepared trypanosome suspension containing about 2x104 parasites was dispensed into each of seven clean test tubes using micropipettes. 0.2ml of prepared Ringer抯 solution was also added to stabilize the parasites. Five varying concentrations of the crude extract, the extract fractions and the isolated compound (Makoshi_Mak1) were added to test tubes 1-5. Test tube 6 and 7 served as the negative and positive controls respectively. Aliquots of the contents of each test tube were examined under the microscope at x400 magnification for parasites motility at different time interval for 2 hours. The crude extract was effective at the concentrations of 12 mg/ml, 1.2 mg/ml,0.12 mg/ml and 0.012 mg/ml. the n-hexane fraction had efficacy at 0.12mg/ml and 0.096mg/ml. The Eethyl acetate fractions were effective at 0.12mg/ml, 0.096 mg/ml, 0.048mg/ml and 0.024mg/ml, the N-butanol fraction had efficacy at 0.12mg/ml and 0.096mg/ml, while the Aqueous fraction was effective at 0.2mg/ml, 0.16mg/ml and 0.08mg/ml. Makoshi_Mak1 was effective in vitro against T. brucei beucei at all the tested concentrations of 0.219mg/ml, 0.109mg/ml, 0.055mg/ml and 0.027mg/ml. Efficacy of the extracts and isolated compound was determined by parasite mortality, and was seen to be concentration dependent. Dead parasites appeared to float freely on the fluid environment, while the lysed parasites were seen as floating debris. The results of the study showed that both the crude extract, the purified extract and the pure isolated compound of Corymbia torelliana had in vitro activity against Trypanosoma brucei brucei in a concentration dependent manner. Corymbia torelliana can then be considered as a potential herbal antitrypanosomal drug candidate.

Diagnóstico de la Enfermedad de Chagas congénito: Aspectos relevantes

Introducción: El presente artículo describe aspectos relevantes entorno de la Enfermedad de Chagas congénita, tales como epidemiología, sintomatología, revisión de casos clínicos y las técnicas diagnósticas. Métodos: Se realizó una revisión de la literatura por medio de bases de datos bibliográficas como PubMed, Science direct, Scopus, Plos One, SciELO, teniendo como criterio de inclusión las publicaciones artículos o comprendidos entre enero de 2013 y enero del año 2022 en idioma español e inglés. Resultados: Se determinó que la prevalencia de la Enfermedad de Chagas congénita aún es un problema de salud pública en áreas endémicas y no endémicas, siendo la serología materna indispensable para dar seguimiento oportuno a los casos. Conclusiones: Los seguimientos diagnósticos actuales difieren en los países endémicos y se están aplicando tamizajes en zonas no endémicas donde migran mujeres procedentes de áreas de trasmisión activa de la Enfermedad Chagásica.

Introduction: This article describes relevant aspects of congenital Chagas disease, such as epidemiology, symptoms, review of clinical cases, and diagnostic techniques. Methods: A review of the literature was carried out through bibliographic databases such as PubMed, Science direct, Scopus, Plos One, SciELO, having as inclusion criteria articles or publications between January 2013 and January 2022 in Spanish and English. Results: It was determined that the prevalence of congenital Chagas disease is still a public health problem in endemic and non-endemic areas, and maternal serology is essential for timely monitoring of cases. Conclusions: Current diagnostic follow-ups differ in endemic countries and screening is being applied in non-endemic areas where women from areas of active transmission of Chagasic disease migrate.

Acute Chagas disease Diagnostic experience through an outbreak / Enfermedad de Chagas aguda Experiencia diagnóstica a través de un brote

Abstract Chagas disease is a public health problem in Latin America, with a tendency to globalize due to migration phenomena. The acute phase of the disease tends to be indolent, but a low percentage of patients have an ominous course, especially when the route of transmission is oral, presenting as a prolonged febrile syndrome in the form of an epidemic outbreak. Early detection of the disease will probably avoid morbid outcomes and, in some cases, death. In this case report we summarize a diagnostic approach by describing the index case in an epidemic outbreak. (Acta Med Colomb 2022; 48. DOI:https://doi.org/10.36104/amc.2023.2727).

Resumen La enfermedad de Chagas constituye un problema de salud pública en Latinoamérica con tendencia a la globalización, debido a los fenómenos de migración. La fase aguda de la enfermedad suele ser indolente, pero un bajo porcentaje de pacientes tienen un curso ominoso, especialmente cuando la ruta de transmisión es oral, presentándose como un síndrome febril prolongado en forma de brote epidémico. La detección temprana de la enfermedad probablemente evite desenlaces de morbilidad y en algunos casos la muerte. En este reporte de caso resumimos un enfoque diagnóstico a través de la descripción del caso índice de un brote epidémico. (Acta Med Colomb 2022; 48. DOI:https://doi.org/10.36104/amc.2023.2727).

Biochemical blood profile in water buffaloes: alterations related to natural infection by Trypanosoma spp / Perfil bioquímico do sangue em búfalos: alterações relacionadas à infecção natural por Trypanosoma spp

ABSTRACT Blood and biochemical profiles of animals can be used to evaluate the physiological state of an individual and relate it to that individual's health. In many countries, water buffaloes are currently evaluated using blood parameters from other bovine species. To accurately interpret the biochemical results from individual animals, species-specific reference ranges should be established. One hundred and twenty-eight water buffaloes, without clinical signs of disease and of different ages, were analyzed, and reference intervals were calculated (95% CI; p < 0.05). The data were analyzed according to three age groups (calves, young animals, and adults) and the infection status (infected or uninfected with Trypanosoma spp.). All the biochemical values of total serum proteins, aspartate aminotransferase, gamma globulin, urea nitrogen, calcium, and phosphorus were included; these are parameters that are generally affected by parasites or by infection with hemotropic agents. Natural infection with Trypanosoma spp. gave rise to differences (p < 0.05) in terms of increased calcium and decreased total protein, alkaline phosphatase, and aspartate aminotransferase. Thus, it can be inferred that the infection follows a chronic course in the affected animals, in this case, young animals or young animals.

RESUMO Introdução: Os perfis sanguíneos e bioquímicos de animais podem ser usados para avaliar o estado fisiológico de um indivíduo e relacioná-lo com a saúde desse indivíduo. Em muitos países, os búfalos são atualmente avaliados usando parâmetros sanguíneos de outras espécies bovinas. Para interpretar com precisão os resultados bioquímicos de animais individuais, os intervalos de referência específicos da espécie devem ser estabelecidos. Método: Foram analisados 128 búfalos, sem sinais clínicos da doença e de diferentes idades, e calculados os intervalos de referência (IC 95%; p < 0,05). Os dados foram analisados de acordo com três faixas etárias (bezerros, novilhos e adultos) e o estado de infecção (infectado ou não por Trypanosoma spp.). Todos os valores bioquímicos de proteínas séricas totais, aspartato aminotransferase, gamaglobulina, nitrogênio ureico, cálcio e fósforo foram incluídos; estes são parâmetros que geralmente são afetados por parasitas ou por infecção com agentes hemotrópicos. Resultados: A infecção natural por Trypanosoma spp. originaram diferenças (p < 0,05) em termos de aumento de cálcio e diminuição de proteína total, fosfatase alcalina e aspartato aminotransferase. Com isso, pode-se inferir que a infecção segue um curso crônico nos animais acometidos, neste caso, animais jovens ou novilhos.

An atypical Trypanosoma lewisi infection in a 22-day-old neonate from India: An emergent zoonosis

Reports on atypical human trypanosomiasis, caused by Trypanosoma lewisi, are rare and so far a total of 19 reports on human infection with animal trypanosomes, which includes nine cases from Trypanosoma lewisi exist. Trypanosoma lewisi, a Stercorarian trypanosoma of rats, is transmitted by the fecal contamination of the wound or the bite caused by rat flea Ceratophyllus fasciatus. We report here an atypical neonatal infection of T. lewisi in a 22-day-old infant from Agra. The infant presented with a history of high fever, poor appetite, and lethargy for 3 days. The hematological parameters were normal except for a low platelet count. A high C-reactive protein (CRP) concentration of 70.49 mg/L indicated marked inflammation. The Leishman-stained thin blood smears were microscopically positive for the hemoflagellate. Based on the morphological features and further confirmed by polymerase chain reaction (PCR) assay, the hemoflagellate was identified as T. lewisi. Symptomatic treatment and antibiotic therapy helped in an uneventful recovery of the patient.

Exploring the performance of Escherichia coli outer membrane vesicles as a tool for vaccine development against Chagas disease

BACKGROUND Vaccine development is a laborious craftwork in which at least two main components must be defined: a highly immunogenic antigen and a suitable delivery method. Hence, the interplay of these elements could elicit the required immune response to cope with the targeted pathogen with a long-lasting protective capacity. OBJECTIVES Here we evaluate the properties of Escherichia coli spherical proteoliposomes - known as outer membrane vesicles (OMVs) - as particles with natural adjuvant capacities and as antigen-carrier structures to assemble an innovative prophylactic vaccine for Chagas disease. METHODS To achieve this, genetic manipulation was carried out on E. coli using an engineered plasmid containing the Tc24 Trypanosoma cruzi antigen. The goal was to induce the release of OMVs displaying the parasite protein on their surface. FINDINGS As a proof of principle, we observed that native OMVs - as well as those carrying the T. cruzi antigen - were able to trigger a slight, but functional humoral response at low immunization doses. Of note, compared to the non-immunized group, native OMVs-vaccinated animals survived the lethal challenge and showed minor parasitemia values, suggesting a possible involvement of innate trained immunity mechanism. MAIN CONCLUSION These results open the range for further research on the design of new carrier strategies focused on innate immunity activation as an additional immunization target and venture to seek for alternative forms in which OMVs could be used for optimizing vaccine development.