Background: Chronic dacryocystitis is an inflammation of the lacrimal sac which usually occurs because of the obstruction of the nasolacrimal duct. Objective: The primary objective of the study was to know its clinico-bacteriological profile. Materials and Methods: This prospective observational study was carried out over a period of one year to know the current bacterial flora involved in chronic dacryocystitis. All specimens were processed for isolation and identification of bacterial pathogens according to the standard microbiological techniques. The antibiotic susceptibility pattern was done by Kirby-Bauer disc diffusion method according to the Clinical Laboratory Standards Institute guidelines. Results: Among total 100 cases of chronic dacryocystitis, the highest percentage of infections in relation to age was noted in the age group of 50-60 years and the prevalence rate was higher in female patients compared with male. Out of total chronic dacryocystitis cases, 72% cases were found to be culture positive. Among positive culture, gram positive organisms were higher than that of Gram-negative. Staphylococcus epidermidis was the most frequently isolated and Streptococcus pyogenes was least isolated. The highest percentage of positive samples was found in the mucopurulent discharge followed by extensive purulent and clear fluid, respectively. In Grampositive isolates, chloramphenicol was the most sensitive followed by cefazolin, ciprofloxacin, ofloxacin, cephalexin, vancomycin, gentamycin, and tobramycin, respectively, whereas in Gram-negative isolate, chloramphenicol and nalidixic acid were equally effective followed by ofloxacin, gentamycin, cephalexin, cefazolin and ciprofloxacin, respectively. Conclusion: S. epidermidis was the most frequently isolated bacteria, and chloramphenicol was the most susceptible drug in Gram-positive and Gram-negative bacteria.