Physiological perturbations in juvenile cuttlefish Sepia officinalis induced by subchronic exposure to dissolved zinc.

Although cephalopod early life stage development often occurs in coastal areas where contamination is real and continuous, the physiological perturbations induced by contaminants have been rarely investigated. This study focused on the Zn as it is one of the trace metals the most concentrated in coastal waters, worldwide. As Zn-tolerance limits were unknown in juvenile Sepia officinalis, the aim of this study was to estimate the threshold inducing mortality during the 2-first weeks post-hatching, and to determine its sensitivity using digestive and immune enzymatic assays, as well as growth and behavior follow-up during the first 5weeks post-hatching. Our study highlighted a Zn-mortality threshold lying between 185 and 230µgl(-1), and growth reductions occurring after 5-week at 108µgl(-1) and above, associated with enzymatic perturbations. These results underline a relatively important sensitivity of juvenile cuttlefish to Zn, pointed out by a wide diversity of biomarkers.

Trace metal concentrations in post-hatching cuttlefish Sepia officinalis and consequences of dissolved zinc exposure.

In this study, we investigated the changes of 13 trace metal and metalloid concentrations (i.e. Ag, As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Se, V, Zn) and their subcellular fractionation in juvenile cuttlefish Sepia officinalis reared in controlled conditions between hatching and 2 months post-hatching. In parallel, metallothionein concentrations were determined. Our results highlighted contrasting changes of studied metals. Indeed, As and Fe concentrations measured in hatchlings suggested a maternal transfer of these elements in cuttlefish. The non-essential elements Ag and Cd presented the highest accumulation during our study, correlated with the digestive gland maturation. During the 6 first weeks of study, soluble fractions of most of essential trace metals (i.e. Co, Cr, Cu, Fe, Se, Zn) slowly increased consistently with the progressive needs of cuttlefish metabolism during this period. In order to determine for the first time in a cephalopod how metal concentrations and their subcellular distributions are impacted when the animals are trace metal-exposed, we studied previously described parameters in juveniles exposed to dissolved Zn at environmental (i.e. 50 µg l(-1)) and sublethal (i.e. 200 µg l(-1)) levels. Moreover, oxidative stress (i.e. glutathione S-transferase (GST), superoxide dismutase (SOD) and catalase activities, and lipid peroxidation (LPO)) was assessed in digestive gland and gills after 1 and 2 months exposures. Our results highlighted no or low ability of this stage of life to regulate dissolved Zn accumulation during the studied period, consistently with high sensitivity of this organism. Notably, Zn exposures caused a concentration-dependent Mn depletion in juvenile cuttlefish, and an increase of soluble fraction of Ag, Cd, Cu without accumulation modifications, suggesting substitution of these elements (i.e. Mn, Ag, Cd, Cu) by Zn. In parallel, metallothionein concentrations decreased in individuals most exposed to Zn. Finally, no perturbations in oxidative stress management were detected in gills, whereas modifications of GST, SOD and catalase activity levels were recorded in digestive gland, resulting in an increase of LPO content after a 6-week exposure to low Zn concentration. Altogether, these perturbations are consistent with previously described high sensitivity of juvenile cuttlefish towards Zn. Our results underlined the need to study deeply contamination impact on this animal at this stage of life.