Heavy metals are polluting the freshwater ecosystems with hazardous impacts on aquatic animals especially fish. Considerable amounts of Cadmium (Cd) in aquatic ecosystems are posing a serious threat to sustainable growth of aquaculture industry. Objective: To investigate acute toxicity of Cd (96-hour LC50 and lethal concentrations) was determined by utilizing probit analysis method for the freshwater fish Labeo rohita. Methods: Different organs of fish (liver, gills and muscles) were also analyzed for accumulation of Cd by inductively coupled plasma mass spectrometry (ICP-MS). Results: Acute toxicity of Cd (96-hour LC50) for Labeo rohita was calculated as 159.59 mg/L. Bioaccumulation of Cd in different tissues of fish was examined in the following order liver>gills> muscles. Comet assay was performed for the evaluation of DNA damage by different sub-lethal doses (1/2, 1/3, 1/4 of LC50) of Cd. Conclusions: This research will aid in control of heavy metals contamination and conservation of fish species Labeo rohita in natural aquatic habitats of Pakistan.