Suchergebnisse

Milk is one of the most important nutritious foods, widely consumed worldwide, either in its natural form or via dairy products. Currently, several economic, health and ethical issues emphasize the need for a more frequent and rigorous quality control of dairy products and the importance of detecting adulterations in these products. For this reason, several conventional and advanced techniques have been proposed, aiming at detecting and quantifying eventual adulterations, preferentially in a rapid, cost-effective, easy to implement, sensitive and specific way. They have relied mostly on electrophoretic, chromatographic and immunoenzymatic techniques. More recently, mass spectrometry, spectroscopic methods (near infrared (NIR), mid infrared (MIR), nuclear magnetic resonance (NMR) and front face fluorescence coupled to chemometrics), DNA analysis (real-time PCR, high-resolution melting analysis, next generation sequencing and droplet digital PCR) and biosensors have been advanced as innovative tools for dairy product authentication. Milk substitution from high-valued species with lower-cost bovine milk is one of the most frequent adulteration practices. Therefore, this review intends to describe the most relevant developments regarding the current and advanced analytical methodologies applied to species authentication of milk and dairy products. ; This work was funded by national funds (FCT, Fundação para a Ciência e Tecnologia) through the strategic funding of CIMO (UIDB/00690/2020) and LAQV-REQUIMTE (UIDB/50006/2020, UIDP/50006/2020). This study was also supported by the European Union through European Regional Development Fund (NORTE-01-0145-FEDER-000052) and SYSTEMIC (Knowledge Hub on Food and Nutrition Security, ERA-Net Cofund ERA-HDHL no. 696295). I.Mafra thanks FCT for funding through the Individual Call to Scientific Employment Stimulus (2021.03670.CEECIND).M. Honrado is grateful to FCT grant 2021.08119.BD, financed by POPH-QREN (subsidized by FSE andMCTES). ; info:eu-repo/semantics/publishedVersion

This work has received funding from the Programa Nacional Apícola 2020-2022 under the project "AUTENT+ Desenvolvimento de abordagens inovadoras com vista à valorização e exploração do potencial de mercado do mel Português". The authors are also grateful to the Foundation for Science and Technology (FCT, Portugal) for financial support by national funds FCT/MCTES to CIMO (UIDB/00690/2020). D. Henriques is supported by the project BeeHappy (POCI-01-0145-FEDER-029871) funded by FEDER (Fundo Europeu de Desenvolvimento Regional) through the program COMPETE 2020—POCI (Programa Operacional para a Competividade e Internacionalização), and by Portuguese funds through FCT and A.R. Lopes by the PhD scholarship funded by the FCT (SFRH/BD/143627/2019). ; Honey is a natural product widely consumed around the globe, not only for its taste and nutritional value, but also for its health benefits. Being a product of high dietary relevance and increasing demand, it has also become a target of economically motivated adulteration. According to the 2014 European Parliament report on the food crisis, fraud in the food chain and the control thereof, honey is among the 10 food products most prone of being adulterated [1]. Up until now, honey authenticity was mainly focused on the issues of sugars addition and botanical and geographical origin. However, recently an increased attention has been paid to the entomological origin of honey. To this aim, different approaches have been proposed to differentiate honey produced by different Apis mellifera subspecies, including those from distinct mitochondrial (mt) DNA lineages [2]. This work aimed to develop a novel real-time PCR method coupled with HRM analysis that allows for the simultaneous differentiation of honeybee from maternal lineages A, M and C, for further application in honey authentication. In this sense, data previously obtained from the mitogenomes of a total of 112 honeybees of different lineages were considered for the development of new DNA markers. Considering the aim of further application in honey, new primer sets were designed to amplify short fragments that included different single nucleotide polymorphisms (SNPs) allowing for HRM application. Three primer sets were proposed, amsCOI-F/amsCOI-R targeting the Cytochrome oxidase I (COI) gene, amsND1-F-amsND1-R targeting the NADH-ubiquinone oxidoreductase chain 1 (ND1) gene and amsCox3-F/amsCox3-R targeting the Cytochrome oxidase subunit III (Cox3) gene. Each primer set was first tested using qualitative PCR using DNA extracted from honeybees of A, M and C mtDNA lineages. After optimizing the real-time PCR conditions, each primer set was tested using a series of mtDNA extracted from honeybees. While amsCOI-F/amsCOI-R allowed only for the separation of the honeybees in two clusters, with lineage C and M clustering together, both the amsCox3-F/amsCox3-R and amsND1-F/amsND1-R set of primers allowed to differentiate the three lineages in separate clusters, with high level of confidence. As future work, the methodology will be assayed in commercial honey samples. ; info:eu-repo/semantics/publishedVersion