In Brazil, orally acquired T. cruzi infection has become the most relevant transmission mechanisms from public health perspective. Around 70% of new Chagas disease cases have been associated with consumption of contaminated food or beverages. Açai (Euterpe oleracea and Euterpe precatoria) is currently one of the most commercialized Amazonian fruits in the Brazilian and international markets. Therefore, it has become important to incorporate in the production process some procedures to measure out effective hygiene and product quality control required by global market. Molecular methods have been developed for rapid detection and quantification of T. cruzi DNA in several biological samples, including food matrices, for epidemiological investigation of Chagas disease and food quality control. However, a high-performance molecular methodology since DNA extraction until detection and quantification of T. cruzi DNA in açai berry pulp is still needed. Herein, a simple DNA extraction methodology was standardized from the supernatant of açai berry pulp stabilized in a 6M Guanidine-HCl/0.2M EDTA buffer. In addition, a multiplex real time qPCR assay, targeting T. cruzi DNA and an Exogenous Internal Positive Control was developed and validated, using reference from all T. cruzi DTUs and commercial samples of açai pulp, from an endemic municipality with previous history of oral Chagas disease outbreak. Thus, a high-sensitivity qPCR assay, that could detect up to 0.01 parasite equivalents/mL in açai, was reached. As of the 45 commercial samples analyzed, 9 (20%) were positive for T. cruzi. This high-sensitive, fast, and easy-to-use molecular assay is compatible with most of the laboratories involved in the investigations of oral Chagas disease outbreaks, representing an important tool to the epidemiology, control, and surveillance of Chagas disease.

O presente artigo descreve a análise de um indicador natural de pH obtido a partir do tubérculo do cará-roxo (Dioscorea trifida), espécie muito conhecida na região Norte e nordeste do Brasil. O objetivo foi testar o potencial do extrato como indicador natural de substâncias ácidas e básicas e otimizar seu método de extração por meio de maceração utilizando diferentes solventes de extração. A amostra utilizada no presente estudo foi obtida no município de Coari, Amazonas. Diferentes parâmetros de extração foram utilizados: o material seco e in natura, macerados em temperatura ambiente, e o solvente etanol e hidroalcoólico. A capacidade indicadora dos extratos foi testada com soluções com pH de 1 a 14 e analisada pela absorbância na região UV (300 à 700 nm), por meio de espectrofotômetro. Os resultados evidenciaram uma significativa eficiência dos extratos como indicador natural, com coloração característica conforme a acidez do meio (ácido – vermelho violeta, básico – azul violeta e neutro - violeta). O extrato hidroalcoólico a partir da amostra seca e triturada apresentou melhor desempenho, com uma maior absorção no comprimento de onda característico. Conclui-se que o cará-roxo é uma alternativa para ser utilizada nas aulas de Ciências, em razão do processo de extração simples, com materiais de baixo custo, sendo uma técnica reprodutível e ambientalmente adequada, somado a um bom desempenho de mudança de coloração em soluções com diferentes valores pH. Assim, esse extrato se apresenta uma possibilidade de substituição dos indicadores tradicionais disponíveis no mercado.

Penicillium meliponae, a recently described and rare species, was isolated as an endophytic fungus from the Amazonian plant Duguetia sthelechantha, and has been proven to be a pigment producer. Considering the high productivity of this species and the lack of data on its chemical composition, the present study aimed to characterize the chemical profile of P. meliponae and evaluate the influence of agitation and the use of different culture media. For this purpose, liquid chromatography coupled with mass spectrometry (LC-MS/MS) and molecular networking were used, allowing the identification of 17 azaphilone molecules with sclerotiorin-like skeletons, becoming the first chemical report of this species. In addition, the different production patterns in the tested culture media were indicative that this species is sensitive to changes in the composition of the carbon source and to the presence of agitation. Furthermore, this work contributes to the fragmentation mechanisms of the different possible structural arrangements for azaphilones of the sclerotiorin type and serves as a repository of information on the gas-phase behavior of this type of metabolite in mass spectrometry experiments and will assist future studies aimed at the discovery of azaphilones.

Currently, approximately 70% of new cases of Chagas disease (CD) in Brazil are attributed to oral transmission, particularly through foods such as açaí, bacaba, and sugarcane juice, primarily in the northern and northeastern regions of the country. This underscores the imperative need to control the spread of the disease. The methods utilized to conduct quality control for food associated with outbreaks and to assess the potential for the oral transmission of CD through consuming açaí primarily rely on isolating the parasite or inoculating food into experimental animals, restricting the analyses to major research centers. While there are existing studies in the literature on the detection and quantification of T. cruzi DNA in açaí, the evaluation of parasites’ viability using molecular methods in this type of sample and differentiating between live and dead parasites in açaí pulp remain challenging. Consequently, we developed a molecular methodology based on RT-qPCR for detecting and quantifying viable T. cruzi in açaí pulp samples. This protocol enables the stabilization and preservation of nucleic acids in açaí, along with incorporating an exogenous internal amplification control. The standardization of the RNA extraction method involved a simple and reproducible approach, coupled with a one-step RT-qPCR assay. The assay underwent validation with various T. cruzi DTUs and demonstrated sensitivity in detecting up to 0.1 viable parasite equivalents/mL in açaí samples. Furthermore, we investigated the effectiveness of a bleaching method in eliminating viable parasites in açaí samples contaminated with T. cruzi by comparing the detection of DNA versus RNA. Finally, we validated this methodology using açaí pulp samples positive for T. cruzi DNA, which were collected in a municipality with a history of oral CD outbreaks (Coari-AM). This validation involved comparing the detection and quantification of total versus viable T. cruzi. Collectively, our findings demonstrate the feasibility of this methodology in detecting viable forms of T. cruzi in açaí pulp samples, emerging as a crucial tool for monitoring oral outbreaks of Chagas disease resulting from açaí consumption.

The Amazon hosts countless plant species, many still unknown to science. Among catalogued species, hundreds are edible, yet most are rarely consumed or locally restricted. Known in Brazil as PANCs (“Plantas Alimentícias Não Convencionais”), these unconventional food plants, these species can enhance food security, especially for vulnerable populations. Beyond nutrition, studies reveal their nutraceutical potential due to specialized metabolites such as polyphenols, terpenes, and alkaloids, which enable applications in supplements, cosmetics, and herbal formulations. In this context, Theobroma mariae (Mart.) K. Schum., or “cacau-jacaré”, is a little-known fruit-bearing plant with promising applicability. This study annotated its volatile and non-volatile compounds in pulp and seeds using headspace solid-phase microextraction gas chromatography coupled with mass spectrometry (HS-SPME/GC-MS) and ultra-high-performance liquid chromatography-high resolution tandem mass spectrometry (UHPLC-HRMS/MS), respectively. A total of 72 volatile and 32 non-volatile compounds were annotated. Fatty acids predominated in the pulp volatilome, whereas theacrine was the main volatile compound in the seeds. Phenolic compounds dominated the non-volatile fraction, notably flavonoid-C-glycosides, O-glycosides, and O-glucuronides. The findings highlight T. mariae as a valuable yet underutilized Amazonian fruit, whose chemical profile resembles that of common cacao and offers potential for novel applications in food, nutraceutical, and cosmetic formulations.