Background:Postoperative endophthalmitis is one the most serious complications of cataract surgery. The majority of causative organisms in this destructive infection come from the patient’s own periocular flora. Efforts have been made to reduce the virulence of organisms in the eyelid and conjunctiva with perioperative topical antibiotics, preparation of surgical field, covering eyelids and conjunctival surface with 5% povidone–iodine solution and intracameral antibiotics at the time of surgery to minimize the risk of endophthalmitis.Objectives:We assessed the effect of subconjunctival injection of cefazolin and pouring povidone-iodine on the conjunctiva bacterial colony forming units (CFU) in phacoemulsification cataract surgery.Patients and Methods:In this prospective, randomized, double-blind clinical trial, 122 patients having phacoemulsification cataract surgery with clear corneal incision and topical anesthesia were randomized into two groups including group 1 (subconjunctival injection of cefazolin) and group 2 (recipients of a drop of povidone-iodine). Cultures were collected from the bulbar conjunctiva at the injection site and from the corresponding location in the patient’s eye, three different times.Results:The mean of eyelid samples on blood and chocolate agars, on the day after compared to the day before the surgery in group 1 showed a 52% and 56% reduction. These values were 58% and 50% in group 2 (P < 0.05). The mean CFU of conjunctiva before and at the end of surgery on blood and chocolate agars showed 57% and 56% reduction in group one and 51% and 52% reduction in group 2 (P < 0.05). While comparing mean CFU of conjunctiva at the end and one day post-surgery (interval of 14 ± 2 hours) showed 27% and 27% increase in group 1 and 20% and 21% increase in group 2 (P < 0.05), which reflects conjunctival flora proliferation during the early postoperative period.Conclusions:Due to the good tolerance of patients towards topical anesthesia, pouring a drop of povidone-iodine 10% seems to be a simple and acceptable method to reduce the growth of microorganisms of the conjunctiva.