There is urgent need for having available suitable methods and data regarding the susceptibility levels of antibiotic resistant and sensitive strains of bifidobacteria. Based on a defined standard operation procedure, agar disc diffusion and broth microdilution were compared in order to evaluate the antimicrobial susceptibility profiles of 82 B. pseudolongum and 80 B. thermophilum strains mainly originating from the meat production chain. The methods that were assessed showed interpretable agreement within this study. The disc diffusion zone diameters are highly reproducible making the method a useful alternative to broth microdilution for antimicrobial susceptibility screening of bifidobacteria.