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Chagas disease, caused by the parasite Trypanosoma cruzi, affects over 6 million people, mainly in Latin America. Two different clinical phases, acute and chronic, are recognised. Currently, 2 anti-parasitic drugs are available to treat the disease (nifurtimox and benznidazole), but diagnostic methods require of a relatively complex infrastructure and trained personnel, limiting its widespread use in endemic areas, and the access of patients to treatment. New diagnostic methods, such as rapid tests (RDTs) to diagnose chronic Chagas disease, or loop-mediated isothermal amplification (LAMP), to detect acute infections, represent valuable alternatives, but the parasite's remarkable genetic diversity might make its implementation difficult. Furthermore, determining the efficacy of Chagas disease treatment is complicated, given the slow reversion of serological anti-T. cruzi antibody reactivity, which may even take decades to occur. New biomarkers to evaluate early therapeutic efficacy, as well as diagnostic tests able to detect the wide variety of circulating genotypes, are therefore, urgently required. To carry out studies that address these needs, high-quality and traceable samples from T. cruzi-infected individuals with different geographical backgrounds, along with associated clinical and epidemiological data, are necessary. This work describes the framework for the creation of such repositories, following standardised and uniform protocols, and considering the ethical, technical, and logistic aspects of the process. The manual can be adapted according to the resources of each laboratory, to guarantee that samples are obtained in a reproducible way, favouring the exchange of data among different work groups, and their generalizable evaluation and analysis. The main objective of this is to accelerate the development of new diagnostic methods and the identification of biomarkers for Chagas disease., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Martínez-Peinado et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

We aimed to characterize the genetic constitution of natural T. cruzi populations involved in an Oral Chagas Disease (OCD) outbreak at a rural school of the community of Chichiriviche de la Costa, Venezuela, which affected patients did not respond to the etiological treatment. Peripheral blood samples and/or hemocultures were obtained from twenty-nine OCD patients at time of diagnosis or along nine years of Post-treatment (Tx) follow-up. The IgG serology, T. cruzi discrete typing units (DTU), satellite DNA-qPCR parasitic loads, and minicircle signatures were determined at Pre-Tx and after Tx. The serological titles and parasitic loads changed after treatment, with a significant decrease of IgG titers (Spearman's r value= -0.961) and median parasite loads from 2.869 [IQR = 2.113 to 3.720] to 0.105 [IQR = -1.147 to 1.761] log10 par eq. /mL at Pre-Tx and Post-Tx, respectively, suggesting infection evolution from acute to chronic phase, without seroconversion or parasitological eradication, which was indicative of treatment failure. All patients were infected with T. cruzi DTU I populations. At Pre-Tx their median Jaccard genetic distances were 0.775 [IQR = 0.708 to 0.882], decreasing in genetic variability towards the end of follow-up (Mann-Whitney U test p= 0.0031). Interestingly, no Post-Tx minicircle signature was identical to its Pre-Tx counterpart population in a same patient, revealing selection of parasite subpopulations between the primary infection and Post-Tx. The parasitic populations isolated from hemocultures showed a lower number of bands in the minicircle signatures with respect to the signatures obtained directly from the patients' blood samples, demonstrating a process of parasitic selection and reduction of the population variability that initially infected the patients. Decrease of parasitic loads after treatment as well as Pre- and Post-Tx intra-TcI diversity might be a consequence of both, natural evolution of the acute infection to the chronic phase and persistence of refractory populations due to Tx selection., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Muñoz-Calderón, Díaz-Bello, Alarcón de Noya, Noya-González and Schijman.)

American trypanosomiasis is one of the few native parasites of this continent. As a zoonosis, Trypanosoma cruzi infects about 180 species out of 25 families of mammals. Its regular transmission is through triatomines, which can easily transmit parasites either by the skin route (contamination of mammals skin with their feces) or by oral route (ingestion of food contaminated with complete triatomines or their feces) and additionally through haematogenous via (congenital and transfusional) and by tissues (transplants). The oral route, which seems to be the ancestral form of transmission to wild and domestic mammals, has recently become more important after the success achieved in the control of domicile vectors using residual pesticides. From its initial diagnosis in 1967, tens of oral outbreaks have been diagnosed mostly in the Brazilian Amazon and subsequently in other four countries in South America. Environmental imbalance caused by man through the invasion and deforestation of woodlands, results in reduction of biodiversity of mammals as food source for triatomines, affecting the "dilution effect" of T. cruzi in the nature increasing the risk of human infection. On the other hand, triatomines invade houses looking for new blood sources. One of the consequences of domiciliated triatomines is the food contamination spread, especially in home-made juices, which has been the source of infection of most oral outbreaks. Other biotic and abiotic factors help to explain the recent increase of oral transmission outbreaks of Chagas disease, distributed in nine eco-regions of America., (Copyright © 2015 Elsevier B.V. All rights reserved.)

The NHEPACHA Iberoamerican Network, founded on the initiative of a group of researchers from Latin American countries and Spain, aims to establish a research framework for Chagas disease that encompasses diagnosis and treatment. For this purpose, the network has created a questionnaire to gather relevant data on epidemiological, clinical, diagnostic, and therapeutic aspects of the disease. This questionnaire was developed based on a consensus of expert members of the network, with the intention of collecting high-quality standardized data, which can be used interchangeably by the different research centers that make up the NHEPACHA network. Furthermore, the network intends to offer a clinical protocol that can be embraced by other researchers, facilitating comparability among published studies, as well as the development of therapeutic response and progression markers. Author summary: In this work, we present a new clinical questionnaire for the standardized evaluation and collection of clinical data from patients with acute or chronic Trypanosoma cruzi infection, as well as a uniformed database to collect it. The questionnaire has been produced after consensus from clinical experts belonging to the NHEPACHA network, a coalition of clinical research institutions in Latin America and Spain. This tool will facilitate the collection and sharing of high-quality clinical and paraclinical data useful to annotate sample collections, and allow clinically meaningful comparisons between different cohorts. [ABSTRACT FROM AUTHOR]