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Protected Designation of Origin (PDO) labeling of cheeses has been established by the European Union (EU) as a quality policy that assures the authenticity of a cheese produced in a specific region by applying traditional production methods. However, currently used scientific methods for differentiating and establishing PDO are limited in terms of time, cost, accuracy and their ability to identify through quantifiable methods PDO fraud. Cheese microbiome is a dynamic community that progressively changes throughout ripening, contributing via its metabolism to unique qualitative and sensorial characteristics that differentiate each cheese. High Throughput Sequencing (HTS) methodologies have enabled the more precise identification of the microbial communities developed in fermented cheeses, characterization of their population dynamics during the cheese ripening process, as well as their contribution to the development of specific organoleptic and physio-chemical characteristics. Therefore, their application may provide an additional tool to identify the key microbial species that contribute to PDO cheeses unique sensorial characteristics and to assist to define their typicityin order to distinguish them from various fraudulent products. Additionally, they may assist the cheese-makers to better evaluate the quality, as well as the safety of their products. In this structured literature review indications are provided on the potential for defining PDO enabling differentiating factors based on distinguishable microbial communities shaped throughout the ripening procedures associated to cheese sensorial characteristics, as revealed through metagenomic and metatranscriptomic studies. Conclusively, HTS applications, even though still underexploited, have the potential to demonstrate how the cheese microbiome can affect the ripening process and sensorial characteristics formation via the catabolism of the available nutrients and interplay with other compounds of the matrix and/or production of microbial origin metabolites and thus their further quality enhancement.

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Halloumi is a semi-hard cheese produced in Cyprus for centuries and its popularity has significantly risen over the past years. High-throughput sequencing (HTS) was applied in the present research to characterize traditional Cyprus Halloumi bacterial diversity. Eighteen samples made by different milk mixtures and produced in different areas of the country were analyzed, to reveal that Halloumi's microbiome was mainly comprised by lactic acid bacteria (LAB), including Lactobacillus, Leuconostoc, and Pediococcus, as well as halophilic bacteria, such as Marinilactibacillus and Halomonas. Additionally, spore forming bacteria and spoilage bacteria, were also detected. Halloumi produced with the "traditional" method, had significantly richer bacterial diversity compared to Halloumi produced with the "industrial" method. Variations detected among the bacterial communities highlight the contribution of the initial microbiome that existed in milk and survived pasteurization, as well as factors associated with Halloumi manufacturing conditions, in the final microbiota composition shaping. Identification and characterization of Halloumi microbiome provides an additional, useful tool to characterize its typicity and probably safeguard it from fraud products that may appear in the market. Also, it may assist producers to further improve its quality and guarantee consumers' safety.

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