The bactericidal action of three formulations of a povidone-iodine (PVI) complex in vitro, in vivo, and in the presence of competing organic matter was evaluated. Bacterial organisms included Staphylococcus aureus ATCC 25923 and 25 clinical isolates of Staph. aureus, designated KU 1-25. For the in vitro studies, 1.0 mL of bacterial inoculum containing 10(7) organisms was introduced into 9.0 mL of chemically stable 10% and 1% PVI formulations in sterile culture tubes, and 1.0-mL samples were withdrawn at set intervals. Samples were plated by using standard techniques and incubated for 24 hours, after which colony-forming units were counted. For in vivo studies, 0.1 mL of 10(6) Staph. aureus ATCC 25923 or KU inoculum was deposited on the dorsum of the hand of healthy human subjects. This area was wiped with a cotton swab saturated with 1%, 2.5%, or 10% PVI formulations. Samples were taken at 15 and 30 seconds after application of the iodophor. To test the bactericidal activity of the three formulations in the presence of a competing substrate, a swab soaked with sterile sheep's blood was applied to the skin and allowed to dry. The percentage of 10(4) Staph. aureus inoculum recovered allowed for comparison of the three products. In vitro, the 1% PVI formulation was bactericidal for 10(7) Staph. aureus within two minutes, as compared with the four minutes required by 10% PVI. On the skin contaminated with 10(6) organisms, the rates of killing within 30 seconds were comparable for both solutions.(ABSTRACT TRUNCATED AT 250 WORDS)