Characterization of endocrine disruptors from a complex matrix using estrogen receptor affinity columns and high performance liquid chromatography–high resolution mass spectrometry

Complex mixtures of contaminants with potential adverse effects on human health and wildlife are found in the environment and in the food chain. These mixtures include numerous anthropogenic compounds of various origins and structures, which may behave as endocrine disruptors. Mixture’s complexity is further enhanced by biotic and abiotic transformations. It is therefore necessary to develop new strategies allowing the identification of the structure of known, as well as unknown, nuclear receptor (NR) ligands present in complex matrices. We explored the possibility to use NR-based affinity columns to characterize the presence of bioactive molecules in environmental complex mixtures. Estrogen receptor α (ERα)-based affinity columns were used to trap and purify estrogenic substances present in surface sediment samples collected in a French river under mixed anthropogenic pressure. We combined biological, biochemical and analytical approaches to characterize the structure of ligands retained on columns and demonstrate the presence of known active molecules such as bisphenol A and octylphenol, but also of unexpected ERα ligands (n-butylparaben, hydroxyl-methyl-benzofuranone). High resolution mass spectrometry results demonstrate that ERα affinity columns can be used for the isolation, purification and identification of known as well as unknown estrogenic contaminants present in complex matrices.     

Does radioactive contamination affect the shell morphology of the pond snail <Emphasis Type="Italic">Lymnaea stagnalis</Emphasis> in the exclusion zone of the Chernobyl NPP (Ukraine)?

A distinct external shell abnormality, coarse seam, was found in a high percentage of the pond snail Lymnaea stagnalis (L.) collected near Chernobyl Nuclear Power Plant (NPP), Ukraine. The negative cytogenetical and hematological effects of long-term irradiation on pond snails in the Chernobyl Exclusion Zone have been repeatedly reported; consequently, it has been assumed that radioactivity may also affect shell morphology. However, due to the absence of this shell abnormality in pond snails collected from two radioactive sites in Russia, it can be concluded that the appearance of abnormal shells in snail population near Chernobyl NPP cannot directly be explained by the radioactive contamination.     

Alteration of shell nacre micromorphology in blue mussel Mytilus edulis after exposure to free-ionic silver and silver nanoparticles

This study describes the morphology of inner shell surface (ISS) of the blue mussel Mytilus edulis Linnaeus after short-term exposures to radiolabeled silver in free-ionic (^110mAg⁺) and engineered nanoparticulate (^110mAgNPs, <40 nm) phases. Radiolabeled silver in starting solutions was used in a similar low concentration (∼15 Bq mL⁻¹) for both treatments. After exposure experiments radiolabeled silver was leached from the ISS using HCl. It concentration for shells from both treatments was ∼0.5 Bq mL⁻¹. Whole ISS of young individuals and prismatic layer of adults showed no evidence of any major alteration process after silver uptake. However, the nacre portion of adult mussels exposed to both treatments revealed distinct doughnut shape structures (DSS) formed by calcium carbonate micrograins that covered the surface of aragonite tablets. Scanning electron microscope (SEM) imaging revealed the existence of only minor differences in DSS morphology between mussels exposed to Ag⁺ and AgNPs. From literature survey, DSS were also found in bivalves exposed to Cd²⁺. The DSS occurring in a specimen of a field-collected bivalve is also shown. Formation of distinctive DSS can be explained by a disturbance of the shell calcification mechanism. Although the occurrence of DSS is not exclusively associated with metal bioavailability to the mussels, the morphology of DSS seems to be linked to the speciation of the metal used in the uptake experiments.