Clostridium perfringens, Salmonella spp. and Campylobacter spp. are food-borne enteric pathogens that are commonly associated with poultry. The objective of this study was to investigate the effects of supplemental bacitracin and litter management (fresh vs. reused) on the abundance of these pathogens in commercial broiler chickens. Specific quantitative PCR (qPCR) assays were used to quantify C. perfringens, virulent C. perfringens that carried the genes encoding α-toxin (cpa) and NetB-toxin (netB), Salmonella, and Campylobacter in samples of ileal mucosa, cecal content, and litter. Campylobacter was not detected in any of the samples collected. The abundance of Salmonella was not affected by either bacitracin or litter condition. Generic C. perfringens was detected in the ileal mucosa at very low level at 10 days of age but was much higher at 35 days. Chickens reared on reused litter tended to have a lower abundance of generic C. perfringens compared with those reared on fresh litter. In the ileal mucosa, no cpa or netB was detected at day 10 but was detected at day 35 in the chickens that were not fed supplemental bacitracin. Chicks fed supplemental bacitracin had reduced abundance of generic C. perfringens as well as the cpa and netB genes in the ileal mucosa, cecal content, and litters. A strong positive correlation was found between the abundance of all three measurements of C. perfringens. The abundance of Salmonella spp. and C. perfringens was also shown to be correlated. This is the first study that has examined the effect of dietary bacitracin and litter conditions on the prevalence of these three common enteric pathogens. Unless contaminated from previous flocks, reused litter may not necessarily contain significantly greater abundances of C. perfringens or Salmonella.