In this study, biological responses and contaminant levels in biological tissues were investigated in grass goby fish specimens (Zosterisessor ophiocephalus) collected from five stations in a moderately polluted ecosystem, namely the Bizerte lagoon on the north coast of Tunisia. The following biomarkers were measured: muscular acetylcholinesterase (AChE), hepatic ethoxyresorufin-O-deethylase (EROD), glutathione-S-transferase (GST), catalase (CAT), lipoperoxidation (TBARS), condition factor (CF), and hepatosomatic index (HSI). These measurements were taken in parallel with the content of Organochlorine pesticides (OCPs), Polychlorinated biphenyls (PCBs), Polycyclic aromatic hydrocarbons (PAHs) and trace metals (As, Cr, Cu, Mn, Pb, V, Zn, Ag, Cd, Co and Ni) in muscle tissue. Total PAH concentrations ranged from 20.09 ± 0.68 to 105.77 ± 42.58 ng g(-1) dw, PCB from 33.19 ± 6.25 to 126.28 ± 7.37 ng g(-1) dw, OCP from 11.26 ± 1.62 to 19.17 ± 2.06 ng g(-1) dw, and metals from 107.83 ± 1.83 to 187.21 ± 2.00 mg/kg dw. The highest levels of pollutants and biomarkers were observed at station S1, located in the Bizerte channel. Elevated EROD, GST and CAT activities, as well as TBARS levels in liver were positively correlated with tissue contaminant levels at station S1. Significant negative correlations were also found between hexachlorobenzene (HCB) and dichlorodiphenyltrichloroethane (DDTs) body burden with AChE activity in muscle at station S2. The integration of biological responses and contaminant tissue content indicated that certain areas of the Bizerte lagoon, notably station S1, are significantly impacted by various human activities, which likely represent a threat for aquatic wildlife. On the basis of these results, and due to its ecological characteristics, the grass goby appears a suitable indicator species for pollution biomonitoring in coastal marine areas along the Mediterranean Sea.