Background: The susceptibility to infectious diseases like hepatitis B is high among municipal waste collectors due to improper segregation and during transportation. Objective: The study aimed to estimate the prevalence and factors associated with hepatitis B infection among municipal waste collectors in Puducherry, India. Methods: A cross-sectional study was conducted among waste collectors between July 2021 and December 2021. A semi-structured questionnaire was administered to collect information on socio-demographic, clinical, and behavioral characteristics. Blood sample was collected by fingerstick, and tested for hepatitis B surface antigen (HBsAg) using a rapid testing kit. Multivariable regression analysis was used to identify factors associated with HBsAg positivity. Results: Among the 949 waste collectors interviewed, the mean (standard deviation) age of 49 (8.5) years and 687 (72.4%) were females. The median (IQR) work experience was 6 (5-20) years. Of the 949 waste collectors tested, 31 (3.3%, 95% CI: 2.2%-4.6%) were seropositive for HBsAg and it was 2.1 times higher in males (5.3%) compared to females (2.5%) though not statistically significant (aPR = 0.82, 95% CI: 0.27-2.50; p=0.729). Being married (aPR = 3.06, 95% CI: 1.04-9.02; p=0.042) and history of dental procedures (aPR = 2.51; 95% CI: 1.15-5.46; p=0.020) were significantly associated with HBsAg positivity. Conclusion: Three out of 100 municipal waste collectors had hepatitis B infection. The findings highlight the need for preventive measures and screening programs for hepatitis B among waste collectors.