Soiled bedding sentinel programs have long been the cornerstone of rodent health monitoring surveillance. Many recent studies have evaluated methods to replace live animals in these programs; however, the type of ventilated rack being used greatly influences the detection rate of adventitious pathogens. This study evaluated 4 alternative sampling techniques across 5 distinct vivaria and assessed their accuracy in detecting 5 pathogens. Testing was done in an operational (real-world) setting using IVC racks that vent air at the cage level. The 5 agents surveyed were mouse norovirus, spp., spp. , and . Samples were collected for subsequent PCR assays as follows: 1) cages with live sentinels exposed to soiled bedding; 2) filter paper placed on the lid of an unoccupied cage containing soiled bedding; 3) filter paper placed in the bedding of an unoccupied cage that contained soiled bedding; 4) swabs from an unoccupied sentinel cage that contained soiled bedding; and 5) pooled swabs from colony cages admixed with swabs from soiled bedding sentinel mice. Cumulative accuracy for all pathogens of interest was highest with the existing soiled bedding sentinel program, followed by pooled swabs of colony cages mixed with swabs from occupied soiled bedding sentinel cages. Soiled bedding sentinel cages detected mouse norovirus, spp., and S. with the highest accuracy; the pooled swabs were best in detecting Rodentibacter spp. and E. . The findings suggest that with the type of rack and caging used in our facilities, the soiled bedding sentinel method has highest concurrence with the expected health status of an animal room, and the results from this method can be enhanced with the addition of pooled swabs of colony animals.