Toxicity of lanthanides on various fish cell lines.

The growing use of Lanthanides in new technologies has increased their anthropogenic releases into the aquatic environment over the last decades. However, knowledge on their ecotoxicological impacts is still incomplete, especially with regard to biological effects of Lanthanides mixtures and the possible regular variation in toxicity along the Lanthanides series. The present study evaluated the individual toxicity of all Lanthanides and the toxicity of mixtures of three of them, namely Neodymium (Nd3+), Gadolinium (Gd3+), and Ytterbium (Yb3+) on Danio rerio fibroblast-like cells (ZF4). Individual and mixtures toxicity of Neodymium (Nd3+) and Ytterbium (Yb3+) were also assessed on Danio rerio hepatic cells (ZFL) and Oncorhynchus mykiss epithelial cells (RTgill-W1). The measured Lanthanide concentrations were close to the nominal ones in the culture media of ZF4, ZFL, and RTgill-W1 cells (85-99%). A toxic impact was observed on the three fish cell lines exposed to all Lanthanides tested individually. RTgill-W1 appeared as the less sensitive cells, compared to the two others. Four Lanthanides, Erbium (Er3+), Thulium (Tm3+), Ytterbium (Yb3+) and Lutetium (Lu3+) showed a higher toxicity than the others on ZF4 cells but no correlation could be established between the toxicity of Lanthanides and the order of the elements within the Lanthanides series. Exposures to binary mixtures highlighted the presence of synergistic effects on cell viability for all cell lines.

Variations of anthropogenic gadolinium in rivers close to waste water treatment plant discharges.

Rare earth element (REE) concentrations were determined for 22 sites sampled during two water periods: high flow in winter and low flow in summer. Shale-normalized REE patterns of all samples displayed positive gadolinium (Gd) anomalies. They revealed a widespread contamination of anthropogenic Gd (Gdant) from waste water treatment plant (WWTP) outputs to catchment areas used for drinking water. No significant variations in Gdant were observed between the two flow water periods, but differences in the Gd anomalies were present. However, these differences seem to be associated rather with seasonal variations in the river flow rate than with the release of GdAnt from WWTPs. In proximity to WWTP discharges, strong GdAnt variations ranged from few nanograms per litre to more than 80 µg L-1 and rarely showed a repetitive pattern day after day during 14 days. These concentrations were diluted into the river stream and measured around 10 ng L-1 close to the catchment areas used for drinking water. A principal component analysis (PCA) using the GdAnt concentrations and some classical physicochemical parameters (pH, water temperature, total alkalinity (TA), total organic carbon (TOC), biochemical and chemical oxygen demand (BOD and COD), Cl-, NO3- and SO42-) allowed a site separation according to the level of Gd contamination, highlighting that the highest GdAnt concentrations were found in the north of the region Lorraine (France) where the population density is high and most of the MRI examinations are performed.

Bioaccumulation of gadolinium in freshwater bivalves.

In this study, the presence of anthropogenic gadolinium (Gd) was evaluated in rivers, close to wastewater treatment plant outputs. Then, one site was selected for in situ experiments to assess the bioaccumulation of Gd in the digestive gland and in the gills of two bivalves (Dreissena rostriformis bugensis and Corbicula fluminea). For both organisms, the results suggested that the bioaccumulation of Gd can be observed when organisms are exposed in a geogenic and anthropogenic Gd mixture. In order to observe if Gd can bioaccumulate in tissues of bivalves when the ion is only present as the main anthropogenic speciation of Gd, i.e., Gd-contrast agents (Gd-CAs), the gadoteric acid was used for a laboratory experiment. In this case, the presence of Gd was clearly detected in a significant amount in the digestive glands of D. rostriformis bugensis and C. fluminea while low concentrations are measured in the gills. For the first time, these results clearly showed that Gd can bioaccumulate in bivalve tissues even when it is only present as Gd-CAs. Biochemical activities were measured in the digestive gland and in the gills of the bivalves to assess the effects of Gd-CA bioaccumulation. No significant variations were observed in the gills. Concerning the digestive gland, after 7 days of exposure at 10 µg L-1 of Gd as Gd-CA speciation, GST activity in D. rostriformis bugensis and lipid hydroperoxide and mitochondrial electron transfer system in C. fluminea had increased. The results suggest an acclimation of the organisms to the presence of Gd-CAs in the medium within less than 21 days.

Investigating the emerging role of comparative proteomics in the search for new biomarkers of metal contamination under varying abiotic conditions.

This study aims at investigating the potential use of comparative proteomics as a multi-marker approach of metal contamination, taking into account the potential confounding effect of water temperature. The major objective was to identify combinations of proteins specifically responding to a given metal, even if included in a metal mixture. The diagnostic approach was performed via the comparative analysis of protein expression on spot mapping provided by adult males of Gammarus pulex (Amphipoda, Crustacea) respectively exposed to arsenate (As), cadmium (Cd) or a binary mixture of these metals (AsCd) at three realistic temperatures (5, 10 and 15°C). Proteomic expression analysis was performed by Differential in-Gel Electrophoresis (2D-DiGE), and completed by an adapted inferential statistical approach. Combinations of under/over-expressed protein spots discriminated the metal identity. However, none of these spots discriminated both the individual metal effect (As or Cd) and its effect in metal mixture (AsCd) whatever the tested temperature. Some limits of the two-dimensional analysis of protein spot maps in G. pulex have been highlighted: (i) the presence of contaminating peptides and/or abundant "déja-vu" proteins which can mask the responses of other proteins of interest or (ii) the presence of post-translational modifications. An optimization of the experimental design (especially during the sample preparation) has been described for future investigations. This study has also highlighted (i) the importance of precisely identifying the protein spots of interest to avoid erroneous interpretations in terms of action mechanisms of chemicals and (ii) the importance of working under controlled laboratory conditions with a temperature close to 10°C. In such conditions, we have demonstrated a higher impact of As than Cd on the energetic metabolism of Gammarus. This As impact is reduced in AsCd mixture confirming the antagonistic interaction of this binary mixture previously observed on G. pulex mortality at 10°C.

Single and combined effects of cadmium and arsenate in Gammarus pulex (Crustacea, Amphipoda): understanding the links between physiological and behavioural responses.

This study aimed at investigating the individual and interactive effects of cadmium (Cd) and arsenate (AsV) in Gammarus pulex (Crustacea, Amphipoda) through the use of several biomarkers. Individuals were exposed for 240 h to two concentrations of AsV or Cd alone, and all the possible binary mixtures of these concentrations of AsV and Cd in a complete factorial design. The pattern of the biomarkers' responses to Cd and AsV alone or in mixture was similar in Gammarus pulex, even if the response intensity varied depending on the tested conditions. G. pulex responded to contamination with increased mobilization of the detoxification systems [i.e. γ-glutamyl-cystein ligase activity (GCL), reduced glutathione content (GSH) and metallothionein concentrations (MT)]. This response seems to imply changes in energy reserve utilization (total lipids and proteins are used prior to glycogen reserves), but also a possible energy reallocation from locomotion to detoxification processes. The observed increase in lipid peroxidation could be relied to the increasing gammarid mortality, despite the higher mobilization of detoxification systems. Even if the outcome of the complex interactions between AsV and Cd remains difficult to unravel, such studies are critically important for better assessing the effects of stressors on organisms, populations and communities in a multi-contamination context of ecosystems.

Antagonistic toxicity of arsenate and cadmium in a freshwater amphipod (Gammarus pulex).

Because toxicants rarely occur alone in the environment, a major challenge in risk assessment is to address the combined effects of chemicals on aquatic organisms. This work is aimed at investigating the joint toxicity action of binary mixtures of cadmium and arsenate on Gammarus pulex. Individuals were exposed during 240 h to four single arsenate or cadmium concentrations and binary mixtures of these metals according to a complete factorial plane. Observed mortality in binary mixtures was compared to observed mortality in single arsenate or cadmium exposures. In addition, interactive effects (antagonistic, additive or synergistic) were evaluated using a predictive model for the theoretically expected interactive effect of chemicals. For all the tested concentration combinations, we observed an antagonist 'between-metals' interaction on G. pulex mortality. This antagonistic effect was more marked for the lowest than for the highest (i.e. 1502.0 µg(AsV) L(-1) and 28.5 µg(Cd) L(-1)) tested concentrations of individual metals in binary mixtures. Metal concentrations in body tissues were evaluated and were significantly lower in binary mixtures than in single metal exposures at similar concentration, especially for combinations corresponding to the highest concentrations of both metals (1502.0 µg(AsV) L(-1) and 28.5 µg(Cd) L(-1)). Results were discussed in terms of (1) mechanisms of uptake and bioconcentration and (2) relationships between metal concentration in gammarid body and observed toxicity.

Comparison of arsenate and cadmium toxicity in a freshwater amphipod (Gammarus pulex).

Cadmium is largely documented on freshwater organisms while arsenic, especially arsenate, is rarely studied. The kinetic of the LC50s values for both metals was realized on Gammarus pulex. Physiological [i.e. metal concentration in body tissues, bioconcentration factor (BCF)] effects and behavioural responses (via pleopods beats) were investigated after 240-h exposure. Arsenate LC50 value was 100 fold higher than Cd-LC50 value after 240-h exposure, while concentrations in gammarids were similar for both metals at their respective LC50s. BCF decreased with increasing cadmium concentration while BCF remained stable with increasing arsenate concentration. Moreover, BCF was between 148 and 344 times lower for arsenate than cadmium. A significant hypoventilation was observed for cadmium concentrations exceeding or close to the 240h-LC50(Cd), while gammarids hyperventilated for the lowest arsenate concentrations and hypoventilated for the highest arsenate concentrations. We discussed the relationships between potential action mechanisms of these two metals and observed results.

Cardiovascular diseases and electrocardiogram teletransmission aboard ships: the French TMAS experience.

BACKGROUND: The aim of this study was to analyse emergency calls for teleconsultation received at French TMAS relating to cardiovascular pathologies, to assess the relevance of electrocardiogram teletransmission. MATERIAL AND METHODS: A two-year descriptive and retrospective study from the TMAS medical files database. We selected patients whose telemedical request was related to a possible cardiovascular pathology. The French TMAS receives calls from all kinds of ship (passenger or merchant vessel), and our analysis was conducted by comparing passengers and professional seamen. On board, the caregiver performs the ECG after medical prescription and sends it by satellite to the TMAS. RESULTS: A total of 179 cases of cardiovascular disease were selected, including 79 passengers (PG) (44.1%) and 89 professional seafarers (PS) (49.7%). In 11 cases (6.1%) patient status was not specified. The most frequent diagnoses were chest pain (58 cases) and STEMI (23 cases), followed by NON STEMI (21 cases). An ECG was performed in 70% of cases of chest pain. ECG diagnosed 23 STEMI, and all these patients had been evacuated by a medical team. This diagnosis led to the establishment of antithrombotic therapy. The mortality rate due to cardiovascular diseases was 9.5%. Ten occurred on board and 7 during the evacuation of the patient. Eleven were passengers and 6 were professional seamen. CONCLUSIONS: Calls regarding cardiovascular disease are infrequent but require an effective response. Recording and transmitting an ECG to the TMAS is technically feasible and enables treatment to be started with specific drugs, mainly in the management of STEMI.

Biomarker versus environmental factors: seasonal variations and modelling multixenobiotic defence (MXD) transport activity in transplanted zebra mussels.

The occurrence of biomarker temporal variations linked to environmental factors makes it difficult to distinguish the specific effect of pollution. The present work aims to investigate the seasonal variations of the transport activity of the multixenobiotic defence (MXD), which is used as a biological tool for the monitoring of pollution in aquatic ecosystems. The MXD transport activity was monitored monthly from August 2001 to October 2002 in zebra mussels (Dreissena polymorpha) transplanted to three sites in the Moselle River. The 'efflux method' was used to evaluate functional activity of MXD by assessing rhodamine B efflux with or without an inhibitor (verapamil). Environmental parameters were provided by a French regulatory agency (Water Agency) that monitors river water quality. The results of a principal components analysis describe the seasonal cycle of water characteristics and demonstrate that MXD activity is subjected to significant temporal variations. These data were described with a generalised linear model that enables it to link MXD variability to the seasonal variations of environmental parameters such as temperature or levels of organic contamination. This work proposes a modelling approach and highlights that the occurrence of seasonal variations in MXD response has to be taken into account in the interpretation of in situ monitoring studies.

Identification of multixenobiotic defence mechanism (MXR) background activities in the freshwater bivalve Dreissena polymorpha as reference values for its use as biomarker in contaminated ecosystems.

The biological defence mechanism called MXR or MXD for multixenobiotic resistance or defence protects cells against the entry and the accumulation of xenobiotics. As the defence is modulated by man made chemicals, MXR is used as a biomarker of organisms' exposure to environmental contamination. However, to reliably assess and evidence MXR induction, the use of a reference level is required. In this context, we focused on MXR background level in a freshwater bivalve, the zebra mussel Dreissena polymorpha, in order to propose its use as a reference during MXR evaluation. We monitored the MXR transport activity in mussels collected either in a natural population or in a caged population and then transplanted to clean water in the laboratory. The results showed that MXR activity was decreased to its baseline level after an eight to nine day depuration period (13.1+/-3.1; 7+/-2.6; 13.7+/-3.9 pmol RB min(-1)org(-1) after three experiments of laboratory depurations). Moreover, significant MXR induction was measured in depurated zebra mussels exposed to contaminated sites (39.6+/-3.7; 59.2+/-20.3 pmol RB min(-1)org(-1) after two experiments of field exposure), showing that the laboratory depuration did not affect the induction potential. The MXR responses (decrease as well as increase) occurred in few days and were highly significant, highlighting its reactivity in zebra mussels. Finally, this paper confirms the usefulness of MXR as a tool in biomonitoring studies and provides a protocol for field experiments that enables to establish and use the background level of MXR activity as a reference.

[Multixenobiotic defense mechanism (MDMX) in bivalves]. / Le mécanisme de défense multixénobiotique (MDMX) chez les bivalves.

Multixenobiotic defence mechanism (MXDM) consists in a cellular system that functions as membrane extrusion pumps effluxing organic compounds out of the cells. In bivalves, it represents a primordial protection against toxic effects of organic xenobiotics in preventing their cellular accumulation. It has raised attention during the last decade for its potential to be used as a biomarker of pollution. This article reviews the fundamental knowledge on the MXDM system in bivalves and the methods proposed to assess its activity. Finally, it reviews the major results of laboratory and field studies that enabled to hypothesise that MXDM could be used as a biomarker of environmental stress rather than of pollutant exposure.

Response of multixenobiotic defence mechanism in Dreissena polymorpha exposed to environmental stress.

Multixenobiotic defence mechanism (MXDM) has been recently described as a first line defence system in many aquatic organisms and it has raised attention for its potential as an early biomarker of exposure to environmental stress. In order to investigate the relevance of this biomarker in the freshwater zebra mussel Dreissena polymorpha, we examined its response to exposure and to cessation of exposure. For this purpose, depurated zebra mussels were transplanted to natural rivers or exposed to medium experimentally contaminated with fuel oil 2 in the laboratory. After the exposure period, mussels were submitted to depuration in clean water in the laboratory. The level of MXDM activity was assessed by the efflux method that allows a simple and fast measure of the rate of rhodamine B efflux. MXDM activity was induced following exposure and induction appears earlier after laboratory exposure (3 days) than after in the field exposure (3-14 days). The level of MXDM response could not be directly correlated to exposure dose in the laboratory and to isolated physicochemical parameters or AOX levels in the field. The behaviour of MXDM activity seems to reflect the occurrence of exposure to a global stress rather than to specific compounds. Our results highlighted the possibility to monitor MXDM response in the freshwater zebra mussel. In spite of the difficulty to interpret responses resulting from the lack of specificity of response, multixenobiotic defence mechanism displays qualities to represent a biomarker of general stress in freshwater ecosystems.

Changes in kinetics of cerebral auto-regulation with head-down bed rest.

Thoraco-cephalic fluid shift induced by weightlessness may influence cerebral autoregulation. Our objective was to assess effects of simulated weightlessness by a 7-day head-down bed rest (HDBR) on the kinetics of cerebral blood flow (CBF) autoregulation in eight healthy women (27.9 +/- 0.9 years). This was studied by transcranial Doppler (TCD) of the middle cerebral artery (MCA) during the sudden decrease in blood pressure (BP) induced by quickly deflating thigh cuff aftera 4-min arterial occlusion before (D - 3), during (D2, D5) and after the HDBR (D + 1). BP (Finapres) and MCA maximal blood flow velocity were continuously recorded. Cerebrovascular resistance (CR) was expressed as the ratio of mean BP to mean MCA velocity. The CR slope was defined as changes in CR per second during the BP decrease. The magnitude of the relative decrease in mean BP and MCA velocity as well as the CR slope did not differ significantly before, during and after the HDBR, showing no major impairment of cerebral autoregulation during short-term HDBR. The time to maximum decrease in CR (T1 in s), corresponding to the maximum vasodilation was reduced on D2 (7-2 +/- 0.6) versus D - 3 (9.9 +/- 1-3), D5 (9-6 +/- 0.8) and R + 1 (11.7 +/- 11) probably as a result of the fluid shift. We also looked if the responses during the thigh cuff release differed in women according to their tolerance to the 10 min stand test performed after the HDBR: T1 was larger in the five women who presented orthostatic intolerance suggesting that some differences in cerebral autoregulatory responses may be related to orthostatic intolerance.