Effect of sediment turbidity and color on light output measurement for Microtox Basic Solid-Phase Test.

In this work, sediment samples collected from several Spanish harbours were tested with two toxicity procedures, designed for solid samples: the Microtox Basic Solid-Phase Test (BSPT) and a modified procedure of the previous test protocol (mBSPT). According to the BSPT procedure, after initial light readings, pure bacteria were exposed to sediment suspension dilutions and light production was directly measured on suspended sediments without any further manipulation. As measurements are likely to be affected by sediment turbidity and color, a variation in initial light measurement has been here suggested, in order to consider the sample effect at all time readings during the test. Firstly, when sediment suspensions at different concentrations were added to bacteria suspension, immediately the initial light output drastically decayed by more than 50% in signal difference, resulting in a false inhibition, as effect of sample turbidity/color. This effect was more evident at high EC50 values, when slightly or not toxic samples were assessed. Secondly, the comparison of the EC50 obtained with both procedures, demonstrated that the mBSPT produced higher EC50 values (less toxic) than those obtained with the standard procedure. The mBSPT procedure resulted rapid and effective and it could be applied simultaneously with BSPT, in order to better evaluate the toxicity.

Simulating a heavy metal spill under estuarine conditions: effects on the clam Scrobicularia plana.

We describe the effect of heavy metals Zn, Cd, Pb and Cu on the induction of methallothioneins on the clam Scrobicularia plana along a salinity gradient simulated under laboratory conditions. The clams were exposed to constant heavy metal concentrations in a dynamic estuary simulator during a 15-day assay to investigate possible induction of metal-binding proteins in them. The concentration of heavy metals in water was analysed. Clams were analysed for methallothionein concentrations. The speciation of Zn, Cd, Pb and Cu along the salinity gradient was modelled. Zn showed the highest concentrations and its prevalent species was the free ion. Intersite differences have been observed in methallothionein concentration and related to the salinity gradient. It seems that synthesis of methallothioneins is the result of physiological forces acting in concert with the changes in the chemical speciation of metals, owing to the trace metals uptake is controlled by means of an interaction of physiology and physicochemistry.

Toxicokinetics of heavy metals from a mining spill using Carcinus maenas.

The knowledge of the reproduction and growth background related to the shore crab Carcinus maenas promotes the use of this crab as a model crustacean to asses the potential for endocrine disruption in crustaceans. In addition, an enzyme linked inmunosorbent assay (ELISA), sensitive to the shore crab vitellogenin in serial hemolymph samples allows determination of the extent of disruption of the process of vitellogenesis in female crabs and its likely impact on reproductive output. Intermoult females Carcinus maenas were exposed to concentrations of Cd: 3 microgl(-1), Cu: 15 microgl(-1) and Zn: 700 microgl(-1) determined at the Guadalquivir estuary after the Aznalcóllar mining spill, during 21 days. Crab hemolymph samples, were taken every seven days, and analyzed through an ELISA for Carcinus maenas vitellogenin. Vitellogenin concentration along the time was fitted to a first order kinetic approach. Results showed a good correlation among experimental values and estimated ones. Metal exposure resulted in an increase in vitellogenin concentration in hemolymph, especially for cadmium.