Agmatine, the decarboxylation product of arginine, is the precursor of putrescine - a harmful biogenic amine (BA) - that can accumulate in dairy products via bacterial metabolism involving the agmatine deiminase (AGDI) pathway. This first requires agmatine be produced via the decarboxylation of arginine and it remains unknown which microorganisms are responsible for this prior decarboxylation step. In addition, agmatine, as other BA, plays different physiological roles including those of co-transmitter and neuromodulator. Preclinical and clinical studies have shown agmatine to have a neuroprotective effect, rendering it of therapeutic interest being agmatine-producing bacteria proposed as psychobiotics. The identification of BA-producing microorganisms is based on the rise in pH due to the consumption of H+ during such decarboxylation reactions. However, in the detection of agmatine-producing microorganisms in cheese, this would lead to false positives since many bacteria possess arginine deiminase activity; this produces ornithine and ammonium from arginine, which also increases the pH. To overcome this problem, a whole-cell biosensor based on a previously developed agmatine-inducible transcription system was designed, and a protocol optimized for the successful identification of agmatine-producing microorganisms in cheese. The application of this protocol in cheese samples allowed for the isolation of agmatine-producing microorganisms identified as Hafnia spp. and unravels, for first time, the capacity of Hafnia paralvei to produce agmatine. This finding evidence the potential role of Hafnia spp. in putrescine accumulation in dairy products.