Background: Conjunctivitis is an inflammation of the outermost layer of the white part of the eye and the inner surface of the eyelid. Conjunctivitis is a global economic burden due to its common occurrence, contagiousness and potentially debilitating morbidities. Some reports indicate that 50-75% of acute conjunctivitis is caused due to the bacteria. In adults Staphylococcus, S. pneumoniae and H. influenza are the common pathogens causing bacterial conjunctivitis. Aim: The aim of the present study to determine the bacteriological profile of acute conjunctivitis. Materials and Methods: This prospective observational study was done the Department of Ophthalmology, Nalanda Medical College and Hospital, Patna, Bihar, India, for 10 months. This is prospective observational study with evaluation of demographic factors, associated comorbid conditions and finding causative organism i.e. bacteria gram staining culture methods and biochemical reaction. Results: Out of 100 cases high number of cases of acute conjunctivitis was seen in the age group of 25–35. A male preponderance is noted with 65 males and 35 females. According to our study acute conjunctivitis was observed 70 patients were from low socioeconomic status and 30 cases were from middle socioeconomic status. All 100 patients underwent Gram stain and bacterial culture out of 100, 71 patients was culture positive and other was negative. The most common organism isolated Coagulase positive staphylococci 43% followed Klebsiella pneumoniae with 13%, Pseudomonas 6%, Diptheroids 3% and least Alkaligenes fecalis was 2%. Out of 100 patients 45 patients had a involvement of both eyes which is 45% and 55 patients had involvement of only one eye which is 55%. Conclusion: we conclude, bacteriological evaluation of conjunctivitis provides to the ophthalmologist a working knowledge of the causal microbes, their common presentations, clinical course and antibiotic sensitivity patterns along with confirming the clinical diagnosis. It also helps to avert the use of inappropriate medications and reduce the risk of drug resistant strains. To achieve this end, public awareness particularly of conjunctivitis, its cause, routes of spread and medical management should be sought.