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.
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 Lysis 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.Author SummaryOral transmission of Chagas disease has acquired an increasingly importance on the disease epidemiology. Most of the orally acquired Chagas Disease cases are related to the consumption of fresh foods or drinks, as sugar cane juice, açai berry pulp and bacaba wine, contaminated with triatomines or its feces. In Brazil, it has recently caused numerous outbreaks and has been linked to unusually severe acute infections. So far, the evaluation of the potential for oral transmission of Chagas disease through the consumption of açai-based products is mostly determined by clinical or parasitological methods. Despite the recent advances, a highly sensitive, reproductible and properly validated real time PCR assay for the molecular diagnostic of T. cruzi in açai pulp samples is still missing. Herein, a simple and reproducible multiplex real-time PCR assay was developed to the detection and quantification of T. cruzi DNA in açai pulp samples. This methodology, that includes a simple step for sample stabilization and DNA extraction based on silica-membrane spin columns, can be useful for analyzing orally transmitted acute Chagas disease outbreaks.
Nos últimos anos, os estudos no ensino de Ciências têm expressado uma preocupação em possibilitar a construção de conhecimentos científicos com intuito de permitir ao indivíduo agir criticamente e consciente na sociedade, exercendo plenamente sua cidadania. Verifica-se, no entanto, que inúmeros problemas são enfrentados pelos docentes na metodologia a seguir na abordagem dos conteúdos ministrados em sala de aula. Desta forma, este trabalho teve como objetivo uma abordagem diferenciada no ensino de Ciências, agregando conhecimento científico ao conhecimento tradicional dos alunos. A metodologia empregada apresentou caráter qualitativo e quantitativo, com aplicação de questionário inicial e final contendo questões abertas e fechadas. O trabalho foi desenvolvido em uma escola Municipal no interior do Amazonas (Alvarães), no 9º ano do Ensino Fundamental, com aulas expositivas, atividade lúdica e aula de campo. Por meio deste trabalho pode proporcionar um ensino dialogado e dinâmico com os alunos, relacionando conhecimento científico e conhecimento empírico dos alunos, como forma de valorização das experiências cotidianas dos alunos e conservação da cultura local. Pode-se conciliar e relacionar ciência-tecnologia-sociedade e ambiente (CTSA), através da produção de farinha de mandioca, buscando promover um aprendizado significativo
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.
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.
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