To report the microbiological spectrum of acute and chronic dacrocystitis.Retrospective study on 100 patients who presented to the ophthalmic plastic clinic of a tertiary eye care center from May 2011 and April 2013 with acute and chronic dacryocystitis was reviewed for demographic and microbiological profile. The culture results and organisms isolated were recorded.Sixty patients had acute onset and the remaining 40 patients had chronic onset dacryocystitis. The female to male ratio was 1.78. The mean age of patients was 44y. Gram-positive organisms were the most commonly isolated accounting for 54%, and the commonest species isolated was S. aureus in 26%. Percentage of gram positive cultures was higher in chronic dacryocystitis than acute ones (82% vs 48% of positive cultures; P=0.003). Also in culture positive acute dacryocystitis, gram negative species were found in 52% of eyes but only in 18% of chronic dacryocystitis.Gram negative bacteria, culture negative samples, unusual and more virulent organisms are more common in acute dacryocystitis than chronic ones. The results of this study have significant bearing on the treatment of patients with dacrocystitis.