The effect of the sponge sample collection site on the recovery of multiple indicator organisms from beef carcass surfaces was evaluated to simplify and validate our previous sampling method for ease of implementation as a general protocol. Sponge samples were collected at three beef processing plants using hot water or acidic antimicrobials as interventions. Two 4,000-cm(2) samples were collected from preevisceration carcasses (n = 248), one from the inside and outside round area (top site) and one from the navel-plate-brisket-foreshank area (bottom site). One-half of the samples (n = 124) were collected before a wash cabinet intervention and the other half after the intervention. The numbers of total aerobic bacteria, Enterobacteriaceae, coliforms, and Escherichia coli were determined for one-half of each individual sponge sample. The other halves of the sponges were combined to represent a top plus bottom 8,000-cm(2) SAMPLE: For the preintervention carcasses, 4,000-cm(2) samples collected from the top or bottom sites of the carcasses were not significantly different (P > 0.05) from each other or from the 8,000-cm(2) combined sample in recovery of the indicator organisms. Significant reductions of indicator organisms were observed in all three types of sponge samples after intervention; however, samples collected from the bottom site recovered less organisms (P < 0.05) compared with samples of the other types. These results suggested that samples collected from either the top or the bottom site of the carcasses with this method are suitable for monitoring indicator organisms as long as the same sampling site is consistently used.