Background: To investigate the microbiologic spectrum of dacryocystitis in adult and pediatric groups, specifically the microbiologic differences between chronic dacryocystitis with nasolacrimal duct obstruction (NLDO) and acute dacryocystitis in pediatric group. Methods: This retrospective study was reviewed for demographic and microbiologic profile of dacryocystitis. The culture results were reported. Results: Sixty-four adults and one hundred and five pediatrics with dacryocystitis were included in this study. Of all adults, only chronic dacryocystitis with NLDO was observed. Of all pediatric patients, 89 had chronic dacryocystitis with NLDO and 16 had acute dacryocystitis. Gram positive and negative isolates were numerically equal in adult group (both 36(48.65%)), while gram positive isolates were the major organism in pediatric group (71(58.68%)). Streptococcus pneumonia was the most common isolate in both adult (11(14.86%)) and pediatric (30(24.79%)) dacryocystitis. For both pediatric subgroups, gram positive isolates were the major organism (59(57.84%) for chronic dacryocystitis with NLDO and 12 (63.16%) for acute dacryocystitis). However, the leading isolates in those two subgroups were distinct, with Streptococcus pneumonia (29(28.43%)) being most common in chronic dacryocystitis with NLDO and Staphylococcus aureus (8(42.11%)) being most common in acute dacryocystitis. Conclusions: In adult group, gram negative isolates were more common in dacryocystitis than before. In pediatric group, gram positive isolates were still the major infection pathogen. Moreover, the more virulent organisms were more common in acute dacryocystitis than chronic dacryocystitis with NLDO.