Persistence of Bacillus atrophaeus subsp. globigii spores on corroded iron coupons in drinking water was studied using a biofilm annular reactor. Spores were inoculated at 10 6 CFU/ml in the dechlorinated reactor bulk water. The dechlorination allowed for observation of the effects of hydraulic shear and biofilm sloughing on persistence. Approximately 50% of the spores initially adhered to the corroded iron surface were not detected after 1 month. Addition of a stable 10 mg/liter free chlorine residual after 1 month led to a 2-log 10 reduction of adhered B. atrophaeus subsp. globigii, but levels on the coupons quickly stabilized thereafter. Increasing the free chlorine concentration to 25 or 70 mg/liter had no additional effect on inactivation. B. atrophaeus subsp. globigii spores injected in the presence of a typical distribution system chlorine residual (ϳ0.75 mg/liter) resulted in a steady reduction of adhered B. atrophaeus subsp. globigii over 1 month, but levels on the coupons eventually stabilized. Adding elevated chlorine levels (10, 25, and 70 mg/liter) after 1 month had no effect on the rate of inactivation. Decontamination with elevated free chlorine levels immediately after spore injection resulted in a 3-log 10 reduction within 2 weeks, but the rate of inactivation leveled off afterward. This indicates that free chlorine did not reach portions of the corroded iron surface where B. atrophaeus subsp. globigii spores had adhered. B. atrophaeus subsp. globigii spores are capable of persisting for an extended time in the presence of high levels of free chlorine.Contamination of the drinking water distribution system infrastructure with microbiological agents is a current homeland security concern. Should pathogens adhere, persist, detach, and survive in the water column, drinking water safety would be compromised. Vegetative bacterial bioterror agents have been shown to be more susceptible to disinfectants than spores in the planktonic phase (30, 32). However, adhered pathogens detaching from biofilm or corrosion material could be associated with particles, thereby increasing disinfection resistance and the likelihood that they could reach a consumer's tap (4, 16). Sporeforming bacteria could even survive boiling if the procedure were not performed properly (31). Even if pathogen density in the water column were low enough that infection of consumers did not occur, uncertainty surrounding the fate of injected pathogens present in the pipe material and biofilm would assuredly reduce the number of people utilizing municipal potable water. Therefore, understanding how long allochthonous pathogens survive in biofilm and/or pipe material under oligotrophic drinking water conditions, and decontamination of that material, is an important research topic.It is well known that biofilm-associated microorganisms survive longer in the presence of disinfectants than planktonic organisms (8,(20)(21)(22)34). However, information on persistence of microbiological agents in water distribution system pipe material is limite...