Microbially derived off-flavor is a major problem in apple juice production as it diminishes the sensory quality of the juice significantly. Fifteen relevant off-flavor compounds that are formed in apple juice, for example, by the strains Alicyclobacillus acidoterrestris and Actinomycetes (Streptomyces ssp.) were investigated with respect to their sensory relevance. The odor threshold values (i.e., detection and recognition values) were determined for all compounds in the matrix apple juice. Odor threshold values for fenchyl alcohol are reported here for the first time. The obtained values were set in relation to the limits of detection and quantification of a previously published GC-MS method. Eight tainted apple juice samples were analyzed for the presence of the 2 strains and the 15 off-flavor compounds. Both strains could be found in the samples; the presence of Streptomyces ssp. as spoilage bacteria of apple juice is reported for the first time. In samples with distinct off-flavor, 2-isopropyl-3-methoxypyrazine, 2-isobutyl-3-methoxypyrazine, 2-methylisoborneol, 1-octen-3-ol, fenchyl alcohol, geosmin, and guaiacol as well as 2,6-dibromophenol were determined in concentrations higher than the detection threshold.