Evaluation of Methods for Determination of Spore-Formers Responsible for the Late Gas-Blowing Defect in Cheese

Abstract Late gas-blowing is a defect of European specialty cheeses caused by certain spore-forming bacteria of the genus Clostridium . Five most probable number methods for detecting spores were evaluated: a modified Weinzirl, NIZO-Ede, Fryer/Halligan, BB-lactate, and the BB-lactate with modifications using pure cultures of C. butyricum, C. sporogenes , and C. tyrobutyricum inoculated into sterile homogenized milk. Tubes containing media of the different methods were incubated for 14 d at 32°C for C. butyricum and 37°C for C. sporogenes and C. tyrobutyricum . The modified Weinzirl and NIZO-Ede methods took the longest time to indicate gas formation (blowing) and detected the spore-formers less frequently. The Fryer/Halligan and BB-lactate methods, which contain lactate, detected spores quicker. No apparent differences were observed between the Fryer/Halligan and BB-lactate methods. The BB-lactate method was modified by making the medium semisolid by addition of .2% agar, eliminating the use of Durham tubes, and using an agar plug rather than paraffin. These modifications resulted in a simpler, more sensitive method.