We isolated a Bacillus amyloliquefaciens strain present in ultra-high temperature (UHT) pasteurized organic whole milk to ascertain virulence determinants present in this species, as well as the pattern of virulence gene expression over time in a model food (UHT milk) system compared with the type strain Bacillus cereus ATCC14579. The overall goal of this project was to genotypically and phenotypically characterize thermoduric B. amyloliquefaciens virulence potential and the presence of the global regulator effector PlcR. Recovery of bacteria from milk required an enrichment in brain-heart infusion broth, incubated aerobically for >24 h, after which time samples were spread-plated onto tryptic soy agar (TSA) plates to recover Bacillus spp. Resulting colonies were streak plated onto TSA to ensure purity of culture before catalase testing, Gram and spore-staining to presumptively identify to the genus level. Pure cultures were biochemically identified to the species level using the Microgen Bacillus ID system (Hardy Diagnostics), yielding an identification of Bacillus firmus, and validated further using fatty acid profiling and 16S rDNA sequencing, which revealed the isolate to be a strain of B. amyloliquefaciens. To confirm presence of the target genes in each isolate, DNA was extracted from pure cultures during late log phase. Quantified DNA template was used in real-time (SYBR Green-based) PCR with primers specific for each of the target genes: plcR, codY, nheA and hblC, and the16S rRNA gene as a control. PCR results indicated no significant difference existed between average melting temperatures (T_m) of 16s rRNA (P = 0.435) and plcR (P = 0.341) of B. amyloliquefaciens and B. cereus. The applications of this project will be to determine if parameters regarding shipment, storage, and shelf life of UHT organic milk should be revisited, to ensure quality before consumption of product that may harbor thermoduric toxigenic Bacillus spp.