The aim of the study was to evaluate the toxic impact of wastewater from sites 1 and 2 of Tung Dhab drain in the state of Punjab, India, on fish behaviour, morphology and gill histopathological biomarkers in comparison to control group. Static non-renewal tests were conducted for 96 h to determine LC50 of the wastewater for both sites using five concentrations (6.25-100%). Fish were regularly noticed for any deviation in behaviour and external morphology. Physico-chemical analysis of wastewater was done using standard methods recommended by APHA/AWWA/WEF (2005). Chronic toxicity tests were conducted for 15 and 30 days with sublethal concentrations of wastewater (50-90% of LC50) and gill histopathology was assessed. Wastewater near a paper mill was more toxic as observed from LC50 values of 72.45%. There was evident deterioration of water quality as the recorded values of some parameters were higher than the standard discharge limits. The test fish exhibited increased air gulping and surfacing, erratic movements initially and decreased opercular movements as the exposure period increased. Morphological observations include increased body colouration, mucus secretion, scale loss and haemorrhages on the skin and lower lip. Alterations in the gill histology such as complete lamellar fusion, epithelial lifting and intraepithelial oedema, haemorrhages, lamellar necrosis and aneurysm were noted in the test fish. Results demonstrate that the fish exposed to wastewater from both sites showed significantly greater change in gill organ index (IG) as compared to control fish for 15 and 30 days.