Multivariate approach to gill pathology in European sea bass after experimental exposure to cadmium and terbuthylazine

• Guided quantitative expert analysis was adopted to evaluate bass gill pathology.
• Multivariate exploratory data analysis was performed on quantitative data.
• Exposure classes were adequately discriminated by linear discriminant analysis.

The combined use of guided quantitative expert analysis and of multivariate exploratory data analysis is reported as a robust, sensitive and sufficiently specific approach to study European sea bass gill secondary lamellar pathology after exposure to incremental doses of cadmium and terbuthylazine up to 48 h. The following elementary pathological findings were considered: “epithelial lifting”, “epithelial shrinkage”, “epithelial swelling”, “pillar cells coarctation”, “pillar cells detachment”, “channels fusion”, “chloride cells swelling”, and “chloride cells invasion”. The relative spatial extension was determined according to exposure class and data were analyzed by means of canonical correspondence analysis (CCA), linear discriminant analysis (LDA) and canonical variates analysis (CVA). Histologically and ultrastructurally, cellular shrinkage/coarctation prevailed in cadmium exposed lamellae, whereas cellular swelling and epithelial lifting were predominant in terbuthylazine exposed lamellae compared to unexposed fish. Both CCA and CVA permit a good graphical data grouping according to exposure classes by means of the convex hull minimum polygons. This also reveals exposure dose and time gradients in CCA plot. Accordingly, epithelial swelling and epithelial shrinkage were comparatively associated to higher exposure time, whereas epithelial shrinkage and pillar cells coarctation were comparatively associated to higher exposure dose. LDA with only “epithelial shrinkage”, “epithelial swelling” and “pillar cells coarctation” in the model classified correctly 87.5% of the cross-validated cases. A possible pathogenetic relationship between the discriminant elementary lesions and the toxic mode of action at the cellular level of both cadmium and terbuthylazine is also discussed.