Introduction: Blood and body fluid spillage continues to be a public health problem, despite, advances in our understanding and control of infections. So, this study was done to compare the knowledge in Spill Management among trained and untrained health care workers based on which measures can be taken to reduce the hazards in hospital. Methods: A Cross-Sectional Study in Teaching Hospital, Kakinada was conducted from September to October 2019 using Semi-structured questionnaire & data was collected from a convenient sampling of  202 Health Care Workers consisting  of Interns & PGs, Staff Nurses, Student Nurses & Lab Technicians, MNOs, Sanitation Staff, & Others divided in to three  groups. Data was obtained from those present in duty at that time and who gave consent. Those who were not willing to participate in the study were excluded. Ethical approval was taken from the Institutional Ethics Committee. Data obtained was entered in Excel 2016, analysed using SPSS20 software and presented as Percentage Distribution, Mean & Standard Deviations and Significance applied wherever necessary. Results:   86.14% (174) had knowledge about Hand hygiene. Knowledge on discarding used materials in the yellow bin was observed in 77.23% (156)  overall. Only 27.23% (55) were aware of proper use of PPEs.  The right sequence of steps of Spill Management was mentioned by 16.34% subjects overall. 38.61% (78) received training on Spill Management. Hypochlorite was the most used disinfectant for cleaning the spill, as mentioned by 55.94% (113 out of 202). Only 30.2% (61) told they were aware of “Spill Management kit”, of whom, 62.29% (38 of 61) received training (p value obtained is 0.0002). Conclusion: Spill management is seen highest among Group I compared to Group II. So, trainings must be conducted on Quarterly basis to update and sustain knowledge level among all groups of health care workers as this is a dynamic group.