Pigmented macrophages and related aggregates in the spleen of european sea bass dosed with heavy metals: Ultrastructure and explorative morphometric analysis

The ultrastructure and morphometrics of pigmented macrophages (PMs) were assessed in the spleen of European sea bass experimentally dosed with Cd and Hg. PMs occurred either as solitary cells or as variably structured aggregations, defined as macrophage aggregates (MAs). Light microscopy revealed a high degree of morphological heterogeneity amongst MAs of all experimental groups. At the ultrastructural level, MAs showed a heterogeneous pigment content that was not influenced by the treatment. Cytoplasm rarefaction/vacuolation and euchromatic nuclei, were observed in PMs of dosed fish. Undosed and Cd-dosed samples differ significantly with regard to the following morphometric features: the Minor axis of the best fitting ellipse, Aspect Ratio, and Roundness. In Cd-dosed fish, MAs showed reduced size and complexity. Lacunarity showed significant differences between undosed and both Cd and Hg-dosed samples. These results suggest that heavy metals, and especially Cd, may influence the dynamics of PM aggregation/disaggregation. Variability in splenic MAs was observed both by light and electron microscopy. However, only the morphometric techniques adequately and objectively described the phenomenon, allowing a quantitative/statistical comparison of morphology among experimental groups. These morphometric analyses could be usefully applied in toxicological and ecotoxicological, as well as morpho-functional studies.