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We assessed how multi- and univariate models reflect marine environmental health based on macrobenthic community responses to three environmental stressor categories: hydrodynamics, organic enrichment and metal contamination. We then compared the models with the benthic index AMBI (AZTI Marine Biotic Index). Macrobenthic community and physicochemical variables were sampled at 35 sites along Babitonga Bay, a subtropical estuary in Southern Brazil. Distance-based linear modelling identified depth, grain size and organic matter as well as Cu and Zn as key stressors affecting the macrobenthos. Using canonical analysis of principal coordinates (CAP), we developed three multivariate models based on the variability in community composition, creating stress gradients. The metal gradient showed better correlation with the benthic community. Sediment quality indices (Geoaccumulation Index and Contamination Factor) showed a low to moderate contamination status, with higher concentrations for Cr, Ni and Zn at the inner areas of the bay. According to AMBI, Babitonga Bay has a “good” environmental health status, and the AMBI values show stronger correlations with the hydrodynamic and organic enrichment gradients (r = 0.50 and r = 0.47) rather than the metal gradient (r = 0.29). Lumbrineridae polychaetes (not included in the AMBI list) and Scoloplos sp. were negatively related to the metal contamination gradient and were considered sensitive, while Sigambra sp., Magelona papillicornis, the gastropod Heleobia australis and species of the crustacean order Mysida were positively related to the gradient and considered tolerant to higher concentrations of metals in the sediment. Despite the inconsistency in the ecological classification provided by AMBI and its relationship with the metal gradient, our results suggest that the environmental quality was satisfactory for the studied gradients. The metal gradient showed the weakest correlation to AMBI. In such cases, the ecological classification of taxa by the index should be evaluated under the perspective of the action of inorganic genotoxic contaminants represented by metals.