Contamination of beer arises in 50% of all events at the late stages of production, the filling area. Hereby, biofilms, being consortia of microorganisms embedded in a matrix composed of extracellular polymeric substances, play a critical role. To date, most studies have focused on the presence of (biofilm forming) microorganisms within this filling environment. Our aim was to characterize the microbial status as well as the presence of possible biofilms at a can filling line for beer by determining the presence of microorganisms and their associated matrix components (carbohydrates, proteins and extracellular DNA (eDNA)). Targeted qPCR confirmed the presence of microorganisms at ten sites during operation and three after cleaning (from 23 sites respectively). The evaluation of carbohydrates, eDNA and proteins showed that 16 sites were positive for at least one component during operation and four after cleaning. We identified one potential biofilm hotspot, namely the struts below the filler, harboring high loads of bacteria and yeast, eDNA, carbohydrates and proteins. The protein pattern was different than that of beer. This work deepens our understanding of biofilms and microorganisms found at the filling line of beer beverages at sites critical for production.
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