We sought to determine the current level of resistance in Haemophilus influenzae and Streptococcus pneumoniae, the primary pathogens of pediatric conjunctivitis. Between January 1997 and March 1998, we prospectively cultured acute conjunctivitis in 250 ambulatory pediatric patients from rural Kentucky whose average age was 24.3 months. In those 250 cases, 106 H. influenzae (42% of the total) and 75 S. pneumoniae (30% of the total) pathogens were isolated, with no growth or no pathogen resulting in 79 cases (32% of the total). Beta-lactamase was detected in 60 (69%) of 87 tested strains of H. influenzae. Among 65 isolates of S. pneumoniae tested for penicillin susceptibility, 44 (68%) were susceptible, 17 (26%) were resistant, and 4 (6%) were intermediate. Conjunctivitis with acute otitis media was observed in 97 patients (39%), and H. influenzae was recovered in 57% of these 97 cases. As for in vitro activity, ciprofloxacin, ofloxacin, and tetracycline were the most active; and gentamicin, tobramycin, polymyxin B-trimethoprim, and polymyxin B-neomycin were intermediately active. Sulfamethoxazole possessed no activity against either pathogen. Beta-lactamase production was detected in 69% of H. influenzae strains, which still remains the primary causative pathogen of both conjunctivitis and conjunctivitis-otitis syndrome. Penicillin-nonsusceptible S. pneumoniae was observed in 32% of 65 patients with S. pneumoniae conjunctivitis, with most strains being penicillin resistant.Acute conjunctivitis is the most common ocular infection in childhood, usually affecting children younger than 6 years old with a peak incidence between 12 and 36 months (3). Pediatric acute conjunctivitis is diagnosed by clinical signs of ocular purulent discharge or hyperemia of bulbar conjunctiva. The etiology of this infection has been documented as bacterial in 54 to 73% of pediatric cases (3,20). The pathogens predominantly recovered include nontypeable Haemophilus influenzae (44 to 68% of cases) and Streptococcus pneumoniae (7 to 21% of cases). Concomitant infection with acute otitis media (AOM) has been coined the conjunctivitis-otitis syndrome by Bodor (4) and is associated with H. influenzae in 20 to 73% of cases and with S. pneumoniae in 12 to 20% of cases (4, 10). Other rare bacterial pathogens of conjunctivitis include Moraxella catarrhalis, Streptococcus mitis, and Streptococcus pyogenes (10,22).Clinicians nearly always empirically treat acute conjunctivitis with topical antimicrobial therapy. The disease is mild, cultures are rarely obtained because of expense, and culture results are reported days later (13). Compared with placebo, topical therapy with polymyxin-bacitracin ointment has been shown to reduce by half the duration of symptoms and to achieve a 2.5-fold increase in rate of bacteriologic eradication at days 8 to 10 (31 versus 79%, respectively) (6). Furthermore, selecting among the multitude of available topical antimicrobials to treat conjunctivitis has been based on either sparse in vitro data or on earlier limited c...