OBJECTIVE To determine the incidence of bacterial contamination of the anterior chamber after phacoemulsification cataract surgery with intraocular lens (IOL) implantation. METHODS Department of Ophthalmology, Royal Prince Alfred Hospital, Sydney, Australia. METHODS Ninety-eight consecutive eyes of 96 patients having phacoemulsification cataract surgery with IOL implantation were included in this prospective study. Two intraoperative anterior chamber aspirates were obtained from each patient, 1 taken at the start and the other at the conclusion of surgery. In addition, preoperative and postoperative conjunctival swabs were acquired. The 4 specimens were cultured using direct culturing techniques under aerobic and anaerobic conditions for 14 days. No preoperative antibiotics were used. RESULTS The incidence of intraoperative anterior chamber contamination was 0% (95% confidence interval, 0%-3.7%) as all intraoperative anterior chamber samples proved culture negative. Sixty-five percent of the preoperative conjunctival swabs were positive for growth, with corynebacteria, coagulase-negative staphylococci, and Propionibacterium acnes being the most frequently cultured organisms. Sixteen percent of the postoperative conjunctival swabs were positive for growth, with corynebacteria and coagulase-negative staphylococci being the most common bacteria. One patient developed culture-positive postoperative endophthalmitis; using pulsed-field gel electrophoresis for further typing, the implicated Staphylococcus epidermidis was indistinguishable from that isolated from the patient's preoperative conjunctival swab. CONCLUSIONS The bacterial contamination rate of the anterior chamber after phacoemulsification and IOL implantation was extremely low. Additional findings support the conjunctiva as being a primary source of bacteria causing postoperative endophthalmitis as well as the ability of povidone-iodine to reduce the conjunctival bacterial load.