The aim of this study was to establish knowledge about the most suitable substrate for cultivating different kinds of anaerobic beer spoilage bacteria in the brewing industry. During the study, two other aspects were investigated; if the sample volume affects the results and which anaerobic beer spoilage bacteria are present in the micro flora at the brewery. The method used was standard cultivation with four different substrates; Universal Beer Agar, Nachweismedium Bierschädliche Bakterien, Raka Ray and VLB S7-S. During the identification of bacteria in the micro flora, a method using PCR was used. None of the substrates showed a significant advantage over the others.