4.4 DISCUSSION
4.4.1 DIETARY VS. WATERBORNE TOXICITY
This study showed that copper accumulated in phytoplankton has a strong toxic effect on E. chloroticus post-settlement, especially given the very low amount of copper present per feed. Interestingly, dietary copper seemed to produce few direct effects on larvae, while carry-over effects were more severe when copper was administered in food rather than in water. Settlement success was on average 30% lower with dietary copper than with waterborne copper. Furthermore, surviving settlers tended to be smaller after 38 d post-settlement when raised in copper-laden food during larval development, while those raised in copper-spiked water had a higher growth and larger final body size than controls. These results show the importance of taking into account dietary intake when evaluating pollutant toxicity.
Other studies evaluating the toxicity of metal-laden diet on zooplankton also found a higher sensitivity to metal when obtained from food rather than from a dissolved source. The copepod Acartia tonsa had a median lethal concentration (LC50) of 43.2μg/L with waterborne silver compared to an LC50 of only 3 μg/L and a median effective concentration (EC50) of 0.1 μg/L with dietary silver (Hook & Fisher 2001a, Bielmyer et al. 2006). Similar results were found for cadmium in copepods, however mercury was more toxic when absorbed from water showing that the relative importance of dietary vs. waterborne exposure varies between metals (Hook & Fisher 2001b).
For carry-over effects, combined exposure to dissolved and dietary copper followed the effect of dietary copper alone; this was as expected, as dietary copper had a stronger effect than dissolved copper on settlement success, and settler size and growth. Surprisingly however, this was also the case for direct effects, where combined dietary and waterborne exposure was less toxic than waterborne exposure alone. It appears that the presence of dietary copper mitigates the toxic effect of waterborne copper, perhaps by triggering acclimation mechanisms such as metal sequestration, as suggested by Hook and Fisher (2002). Acclimation following dietary exposure is also supported by the fact that pulse exposure was worse than chronic exposure. In the field, organisms are likely to be exposed simultaneously to waterborne pollutants and contaminated diet. Therefore, it is crucial to evaluate the toxic effects of both acquisition pathways. However, this is the first study assessing the combined effects of waterborne and dietary metal uptake in invertebrates.
It should be noted that this study was conservative with respect to the algal copper burden. Metal burden per algal cell is higher after 24 – 48 h exposure (Bielmyer et al.2006) and rapid cell division (as in an algal bloom induced by increase nutrients)reduces metal burden per cell (Pickhardt et al. 2002). In this study, algae were cultured for two weeks under high light-nutrient conditions leading to fast algal division. Therefore, metal burden per cell under these conditions is likely to be much lower than encountered in the field for the same water concentration following a rain event, especially in oligotrophic waters.
4.4.2 DIRECT AND CARRY-OVER EFFECTS
Low levels of copper exposure produced both direct and latent effects, in contrast with results from the previous chapter (section 3.3) where no direct effects were detectable. The difference is probably due to the longer exposure in the current study(chronic or four days pulse compared to two days pulse in Chapter 3). The strongest toxic effect observed in this study, however, was a carry-over effect (settlement success).
The main direct effect was a delay in rudiment development at most copper levels, but especially in the Water treatment. The higher in arm growth in presence of copper also reflects a delay in arm development. The difference was predominantly with respect to the pair of arms that develop later in the larval stage (i.e. PD and EO). For these arms, almost no growth was observed in the controls, while increased growth occurred with copper exposure. Nevertheless, final size before settlement was similar across all copper levels, indicating that these arms developed later in the presence of copper. Other direct effects included a slight increase in mortality rate at the High Pulse level and a decrease in normal development at the High Pulse and High Chronic levels, but in the Water treatment only.