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Craft beer is more susceptible to microbial spoilage because it does not have a pasteurization or filtration process, with lactic acid bacteria (LAB) being the most common beer spoilage microorganism. The aim of this study was to isolate LAB in a craft brewery and their characterization from a food safety and microbiological quality perspective, with a special focus on their abilities to produce biogenic amines (BA) and spoil the beer. The results of 60 monitored points inside the craft brewery showed that LAB associated with the craft brewing processes belonged to Lactobacillus, Pediococcus, and Leuconostoc genera, and most of them were detected in the filling area, which can lead to secondary contamination. Two isolates of L. brevis showed the most significant beer spoilage ability because they could grow in more acidic conditions, at a higher hop and alcohol content, and they displayed horA, horC, and hitA genes, which spoiled the vast majority of the tested beers. In addition, the aforementioned L. brevis isolates showed the highest BA production., This study was supported by grants AGL2015-64522-C2-1-R and PID2019-108851RB-C21 (Spanish Ministry of Science and Innovation), and ALIBIRD-CM 2020 P2018/BAA-4343 (Comunidad de Madrid). M. R-S thanks to the “President of the Republic Fellowship” from the Ministry of Education of the Republic of Peru.