Internal distribution of uranium and associated genotoxic damages in the chronically exposed bivalve Corbicula fluminea.

Uranium (U) internal distribution and involved effects in the bivalve Corbicula fluminea have been studied after direct chronic exposure (90 d, 10 µg.L-1). U distribution was assessed at the subcellular level (Metal Rich Granules -MRG-, pellets and cytosol fractions) in two main organs of the bivalve (gills and visceral mass). Micro-localisation was investigated by TEM-EDX analysis in the gills epithelium. DNA damage in gill and hemolymph samples was measured by the Comet assay. The 90-d exposure period led to a significant increase of U concentration in gills over time (× 5) and a large U quantity in subcellular granules in gills. Finally, a significant increase (× 2) in DNA damage was noted in exposed gills and haemocytes. This study shows that the accumulation levels and consequently the potential toxicity cannot be successfully predicted only on the basis of concentration in water or in tissues and subcellular fractions after chronic exposure.

Genotoxic damages in zebrafish submitted to a polymetallic gradient displayed by the Lot River (France).

Genotoxic effects of a polymetallic pollution gradient displayed by the Lot River and one of its tributary have been assessed on zebrafish Danio rerio. Three methods were compared: RAPD-PCR, the comet assay, and 8-oxo-7, 8-dihydro-2'-deoxyguanosine (8-oxodG) formation. The fishes were exposed for 14 days to waters collected from three stations: Joanis, a site polluted by cadmium (Cd) and zinc (Zn) (mean concentrations: 15 µg Cd/L and 550 µg Zn/L), Bouillac (mean concentrations: 0.55 µg Cd/L and 80 µg Zn/L), and Boisse-Penchot, a reference station (mean concentrations: <0.05 µg Cd/L and 7 µg Zn/L). The quantitative RAPD-PCR methodology proved to be sensitive enough to unmask metal genotoxicity after 3 and 7 days of exposure to Joanis water and after 14 days to Bouillac water, whereas the comet assay only detected DNA damages at the most contaminated station (Joanis). The 8-oxodG quantification was not sensitive enough to be used in zebrafish under these environmental conditions.

A probabilistic assessment of the chemical and radiological risks of chronic exposure to uranium in freshwater ecosystems.

Uranium (U) presents a unique challenge for ecological risk assessments (ERA) because it induces both chemical and radiological toxicity, and the relative importance of these two toxicities differs among the various U source terms (i.e., natural, enriched, depleted). We present a method for the conversion between chemical concentrations microg L(-1)) and radiological dose rates (microGy h(-1)) for a defined set of reference organisms, and apply this conversion method to previously derived chemical and radiological benchmarks to determine the extent to which these benchmarks ensure radiological and chemical protection, respectively, for U in freshwater ecosystems. Results show that the percentage of species radiologically protected by the chemical benchmark decreases with increasing degrees of U enrichment and with increasing periods of radioactive decay. In contrast, the freshwater ecosystem is almost never chemically protected by the radiological benchmark, regardless of the source term or decay period considered, confirming that the risks to the environment from uranium's chemical toxicity generally outweigh those of its radiological toxicity. These results are relevant to developing water quality criteria that protect freshwater ecosystems from the various risks associated with the nuclear applications of U exploitation, and highlight the need for (1) further research on the speciation, bioavailability, and toxicity of U-series radionuclides under different environmental conditions, and (2) the adoption of both chemical and radiological benchmarks for coherent ERAs to be conducted in U-contaminated freshwater ecosystems.

Comparative analysis of gene expression in brain, liver, skeletal muscles, and gills of zebrafish (Danio rerio) exposed to environmentally relevant waterborne uranium concentrations.

The effects of waterborne uranium (U) exposure on gene expression were examined in four organs (brain, liver, skeletal muscles, and gills) of the zebrafish (Danio rerio). Adult male fish were exposed to three treatments: No added uranium (control), 23 +/- 6 microg U/L, and 130 +/- 34 microg U/L. After 3, 10, 21, and 28 d of exposure and an 8-d depuration period, gene expression and uranium bioaccumulation were analyzed. Bioaccumulation decreased significantly in liver during the depuration phase, and genes involved in detoxification, apoptotic mechanism, and immune response were strongly induced. Among these genes, abcb311, which belongs to the adenosine triphosphate (ATP)-binding cassette transporter family, was induced 4- and 24-fold in organisms previously exposed to 23 +/- 6 and 130 +/- 34 microg U/L, respectively. These results highlight the role of liver in detoxification mechanisms. In gills, at the highest uranium concentration, gpx1a, cat1, sod1, and sod2 genes were up-regulated at day 21, indicating the onset of an oxidative stress. Mitochondrial metabolism and DNA integrity also were affected, because coxI, atp5f1, and rad51 genes were up-regulated at day 21 and during the depuration phase. In skeletal muscles, coxI, atp5f1, and cat were induced at day 3, suggesting an impact on the mitochondrial metabolism and production of reactive oxygen species. In brain, glsI also was induced at day 3, suggesting a need in the glutamate synthesis involved with neuron transmission. No changes in gene expression were observed in brain and skeletal muscles at days 21 and 28, although bioaccumulation increased. During the depuration phase, uranium excretion was inefficient in brain and skeletal muscles, and expression of most of the tissue-specific genes was repressed or unchanged.

Effect of selenate on growth and photosynthesis of Chlamydomonas reinhardtii.

Algal communities play a crucial role in aquatic food webs by facilitating the transfer of dissolved inorganic selenium (both an essential trace element and a toxic compound for a wide variety of organisms) to higher trophic levels. The dominant inorganic chemical species of selenium in freshwaters are selenite (SeO(3)(2-)) and selenate (SeO(4)(2-)). At environmental concentrations, selenite is not likely to have direct toxic effects on phytoplankton growth [Morlon, H., Fortin, C., Floriani, M., Adam, C., Garnier-Laplace, J., Boudou, A., 2005a. Toxicity of selenite in the unicellular green alga Chlamydomonas reinharditii: comparison between effects at the population and sub-cellular level. Aquat. Toxicol. 73(1), 65-78]. The effects of selenate, on the other hand, are poorly documented. We studied the effects of selenate on Chlamydomonas reinhardtii growth (a common parameter in phytotoxicity tests). Growth inhibition (96-h IC(50)) was observed at 4.5+/-0.2 microM selenate (p<0.001), an effective concentration which is low compared to environmental concentrations. Growth inhibition at high selenium concentrations may result from impaired photosynthesis. This is why we also studied the effects of selenate on the photosynthetic process (not previously assessed in this species to our knowledge) as well as selenate's effects on cell ultrastructure. The observed ultrastructural damage (chloroplast alterations, loss of appressed domains) confirmed that chloroplasts are important targets in the mechanism of selenium toxicity. Furthermore, the inhibition of photosynthetic electron transport evaluated by chlorophyll fluorescence induction confirmed this hypothesis and demonstrated that selenate disrupts the photosynthetic electron chain. Compared to the classical 'growth inhibition' parameter used in phytotoxicity tests, cell diameter and operational photosynthetic yield were more sensitive and may be convenient tools for selenate toxicity assessment in non-target plants.

Bioaccumulation, oxidative stress, and neurotoxicity in Danio rerio exposed to different isotopic compositions of uranium.

Experiments were carried out on adult male zebrafish (Danio rerio) to assess early changes induced by waterborne exposure to different isotopic compositions of uranium (depleted uranium associated or not with 233U). Oxidative stress and neurotoxicity were selected as effect endpoints to characterize uranium chemo- and radiotoxicity. Catalase, glutathione peroxidase, and superoxide dismutase activities and total glutathione content of hepatic extracts, as well as brain acetylcholinesterase activity and uranium bioaccumulation, were measured. Oxidative stress induced by uranium exposure led to decreases in superoxide dismutase and catalase activity levels as well as total glutathione content in liver extracts. These perturbations were significantly more marked in 233U-exposed fish. Furthermore, significant increase in acetylcholinesterase activity was observed in brain extracts at the same level, whatever the isotopic composition of uranium.

Selenium bioaccumulation in Chlamydomonas reinhardtii and subsequent transfer to Corbicula fluminea: role of selenium speciation and bivalve ventilation.

The uptake of Se by the freshwater alga Chlamydomonas reinhardtii and the subsequent transfer to the Asiatic clam Corbicula fluminea was investigated. The objective was to investigate the bioavailability of algal-bound Se for C. fluminea while taking into account Se speciation and bivalve ventilation. First, uptake rates of waterborne Se (selenite, selenate, and selenomethionine) in the algae during a 1-h exposure period were determined for a range of concentrations up to 2,000 microg/L. Fluxes for selenite uptake were constant in the range of concentrations tested, whereas fluxes for selenate and selenomethionine uptake decreased with increasing concentrations, suggesting a saturated transport system at high concentrations (approximately 1,000 microg/L for selenate and 100 microg/L for selenomethionine). These data were used to set the algal contamination for the study of trophic transfer to the clam. Three parameters were studied: The Se form, the algal density, and the Se burden in the algae. The results show that for a fixed algal density, an Se-contaminated algal diet does not modify ventilation. In this case, the driving factor for ventilation is the algal density, with ventilation being enhanced for low algal densities. On the basis of ventilatory flow rate measurements and Se burdens in algae, it was found that bioaccumulation of Se in C. fluminea was proportional to the total quantity of Se passing through the whole organism, but with a lesser extraction coefficient for selenomethionine than for the inorganic forms. These results underline the importance of both physiological factors and speciation in understanding the trophic transfer of Se.

High prevalence of pituitary adenomas: a cross-sectional study in the province of Liege, Belgium.

CONTEXT: Prevalence data are important for assessing the burden of disease on the health care system; data on pituitary adenoma prevalence are very scarce. OBJECTIVE: The objective of the study was to measure the prevalence of clinically relevant pituitary adenomas in a well-defined population. DESIGN: This was a cross-sectional, intensive, case-finding study performed in three regions of the province of Liège, Belgium, to measure pituitary adenoma prevalence as of September 30, 2005. SETTING: The study was conducted in specialist and general medical practitioner patient populations, referral hospitals, and investigational centers. METHODS: Three demographically and geographically distinct districts of the province of Liège were delineated precisely using postal codes. Medical practitioners in these districts were recruited, and patients with pituitary adenomas under their care were identified. Diagnoses were confirmed after retrieval of clinical, hormonal, radiological, and pathological data; full demographic and therapeutic follow-up data were collected in all cases. RESULTS: Sixty-eight patients with clinically relevant pituitary adenomas were identified in a population of 71,972 individuals; the mean (+/- sd) prevalence was 94 +/- 19.3 cases per 100,000 population (95% confidence interval, 72.2 to 115.8). The group was 67.6% female and had a mean age at diagnosis of 40.3 yr; 42.6% had macroadenomas and 55.9% underwent surgery. Prolactinomas comprised 66% of the group, with the rest having nonsecreting tumors (14.7%), somatotropinomas (13.2%), or Cushing's disease (5.9%); 20.6% had hypopituitarism. CONCLUSION: The prevalence of pituitary adenomas in the study population (one case in 1064 individuals) was more than 3.5-5 times that previously reported. This increased prevalence may have important implications when prioritizing funding for research and treatment of pituitary adenomas.

Selenite transport and its inhibition in the unicellular green alga Chlamydomonas reinhardtii.

The influence of time, ambient concentration, and medium composition on selenite (Se(IV)) uptake by the unicellular green alga Chlamydomonas reinhardtii has been investigated. The aims of the performed experiments were to describe the kinetics of accumulation, to characterize transport capacities, to identify key nutrients influencing absorption, and to establish links between speciation and bioavailability. Our results suggested that the adsorbed fraction was negligible compared to the absorbed one. Over the short time scale considered, the absorption was linear with time, with an estimated conductance of approximately 0.2 nmol/m2/h/nM. Uptake was proportional to ambient levels in a broad range of intermediate concentrations (from nM to microM). However, conductances were higher at low concentrations (< nM) and then decreased with increasing concentrations (> microM). These results suggested that a specific but rapidly saturated transport system was involved at low concentrations, coupled with a nonspecific one that was only saturated at high ambient concentrations (approximately mM). The latter could involve transporters used by anionic macronutrients, which is supported by the fact that increasing sulfate and nitrate concentrations induced significant inhibition of Se(IV) uptake. Finally, Se(IV) speciation changes caused by varying pH did not significantly affect bioavailability.

Toxicity of selenite in the unicellular green alga Chlamydomonas reinhardtii: comparison between effects at the population and sub-cellular level.

The toxicity of selenium in aquatic ecosystems is mainly linked to its uptake and biotransformation by micro-organisms, and its subsequent transfer upwards into the food chain. Thus, organisms at low trophic level, such as algae, play a crucial role. The aim of our study was to investigate the biological effects of selenite on Chlamydomonas reinhardtii, both at the sub-cellular level (effect on ultrastructure) and at the population level (effect on growth). The cells were grown under batch culture conditions in well-defined media and exposed to waterborne selenite at concentrations up to 500 microM; i.e. up to lethal conditions. Based on the relationship between Se concentration and cell density achieved after a 96 h exposure period, an EC(50) of 80 microM with a 95% confidence interval ranging between 64 and 98 microM was derived. No adaptation mechanisms were observed: the same toxicity was quantified for algae pre-contaminated with Se. The inhibition of growth was linked to impairments observed at the sub-cellular level. The intensity of the ultrastructural damages caused by selenite exposure depended on the level and duration of exposure. Observations by TEM suggested chloroplasts as the first target of selenite cytotoxicity, with effects on the stroma, thylakoids and pyrenoids. At higher concentrations, we could observe an increase in the number and volume of starch grains. For cells collected at 96 h, electron-dense granules were observed. Energy-dispersive X-ray microanalysis revealed that these granules contained selenium and were also rich in calcium and phosphorus. This study confirms that the direct toxicity of selenite on the phytoplankton biomass is not likely to take place at concentrations found in the environment. At higher concentrations, the link between effects at the sub-cellular and population levels, the over-accumulation of starch, and the formation of dense granules containing selenium are reported for the first time in the literature for a phytoplankton species after exposure to selenite.

Bioaccumulation of waterborne selenium in the Asiatic clam Corbicula fluminea: influence of feeding-induced ventilatory activity and selenium species.

A set of experiments was performed to investigate the bioavailability and the effect of Se on the ventilatory activity of the bivalve Corbicula fluminea, under different conditions of both algal cell densities and dissolved Se chemical forms and concentrations. A first set of experiments was conducted without selenium to investigate the changes in the ventilatory flow rate as a function of the concentration of the unicellular alga Chlamydomonas reinhardtii (10(5)-10(6) cells mL(-1)). For algal concentrations below 2-3x10(5) cells mL(-1), ventilatory activity was highly stimulated whereas it was independent of algal densities for higher values (up to 10(6) cells mL(-1)). To investigate the influence of this first ventilatory drive on selenium contamination process, bivalves were exposed to waterborne selenium at two different algal concentrations, selected to provide contrasting reference ventilatory activities. Three different selenium forms were studied [selenite Se(+IV), selenate Se(+VI) and selenomethionine SeMet] and were added into the water at concentrations of 50 and/or 500 microg L(-1). Each selenium form induced a specific behavioural response, an increase, a decrease or no change of ventilation being observed for Se(+IV), SeMet and Se(+VI), respectively. Selenium accumulation by the organisms was investigated at the organ level for the different exposure conditions. Selenomethionine was the most bioaccumulated form, followed by selenate and selenite, respectively. Despite the bivalves displaying different ventilatory behaviours at low or high algal density, there was no evidence showing reduction or enhancement of Se uptake in the chemical domain investigated.

Influence of metal (Cd and Zn) waterborne exposure on radionuclide (134Cs, 110Ag, and 57Co) bioaccumulation by rainbow trout (Oncorhynchus mykiss): a field and laboratory study.

Field and laboratory experiments were carried out to assess the influence of Cd and Zn on the contamination levels of 110Ag, 57Co, and 134Cs in rainbow trout. During a four-week prior exposure phase, two fish groups were held in tanks in the Lot River (France) at a reference (<0.05 microg Cd/L and 68 microg Zn/L) and at a polluted site (1.5 microg Cd/L and 152 microg Zn/L). During a subsequent phase, organisms were brought back to the laboratory, where the radionuclide accumulation and depuration were studied for 14 and 7 d, respectively. During this second phase, the water used in the experiments was brought back from the two sites on the Lot River in order to work under the same chemical conditions. The potential effect of chronic exposure to stable metals on several biomarkers has been explored: Plasma analysis indicated the disruption of certain variables linked to the energetic metabolism and to the maintenance of the ionic balance. In contrast, no significant disruption of the measured enzyme activities was observed. With regard to the bioaccumulation of radionuclides, concentrations in fish exposed to metals are much lower than those in fish from the control group. Various hypotheses are proposed to link fish metabolic profiles due to metal exposure to the radiocontamination of organisms.