P. Gonzalez - Academia.edu (original) (raw)

Papers by P. Gonzalez

Chemosphere, 2014

Since 2007 a remediation process was initiated on a mining site contaminated by metals (Cd and Zn... more Since 2007 a remediation process was initiated on a mining site contaminated by metals (Cd and Zn). Bivalves were exposed for two years to observe early remediation impacts on hydrosystem. Metal concentrations in river and bivalves remained very high in organisms exposed downstream. Bivalves were still sharply genetically impacted by metal contamination. Recovery of aquatic communities after remediation can only be expected in the long term.

Comparative biochemistry and physiology. Toxicology & pharmacology : CBP, Jan 20, 2015

The effect of methylmercury (MeHg) on glass eels' propensity to migrate, mitochondrial activi... more The effect of methylmercury (MeHg) on glass eels' propensity to migrate, mitochondrial activity and antioxidative defence systems was investigated. Marine glass eels were first sorted in an experimental flume according to their response to dusk. Fish responding to the decrease in light intensity by ascending in the water column and moving with or against the flow were considered as having a high propensity to migrate (migrant). Glass eels still sheltering at the end of the 24h catching period were considered as having a low propensity to migrate and were called non-migrant. Migrant and non-migrant glass eels were then individually tagged and exposed to isotopically enriched (201)MeHg (50ngL(-1)) for 11days. The effect of contamination was studied on muscle fibre structure, and the expression level of genes involved in mitochondrial activity and antioxidative defence systems. To investigate the effect of MeHg on glass eel behaviour, migrant and non-migrant glass eels were sorted ...

Environmental pollution (Barking, Essex : 1987), 2014

This study aimed to assess the recovery capacity of the freshwater bivalve Corbicula fluminea sub... more This study aimed to assess the recovery capacity of the freshwater bivalve Corbicula fluminea subjected to industrial metal discharges (Cd, Zn). After a 24-day exposure in a metal-contaminated river, bivalves were transferred and maintained in the laboratory for one year under metal-free conditions. Metal accumulation, metallothionein production and genetic expressions of genes involved in metal stress were studied. Results demonstrated the high persistence of Cd in tissues (only 73% eliminated after 365 days) whereas Zn was rapidly depurated. The Cd half-life was estimated around 240 days. Metallothioneins were strongly induced within the 28 first days of decontamination, then decreased by 45% after 365 days. The metal exposure of bivalves led to a significant gene induction. After 28 days, most of the genes were no longer overexpressed, suggesting that the bivalves may withstand small amounts of non-essential metals in their tissues without showing signs of detrimental effects on ...

International Journal of Molecular Sciences, 2012

Methylmercury (MeHg) is a potent neurotoxin, and human beings are mainly exposed to this pollutan... more Methylmercury (MeHg) is a potent neurotoxin, and human beings are mainly exposed to this pollutant through fish consumption. We addressed the question of whether a diet mimicking the fish consumption of Wayanas Amerindians from French Guiana could result in observable adverse effects in mice. Wayanas adult men are subjected to a mean mercurial dose of 7 g Hg/week/kg of body weight. We decided to supplement a vegetarian-based mice diet with 0.1% of lyophilized Hoplias aimara fish, which Wayanas OPEN ACCESS

Chemosphere, 2015

This study aimed to demonstrate the recovery potential of Danio rerio after Cd and Zn contaminati... more This study aimed to demonstrate the recovery potential of Danio rerio after Cd and Zn contaminations. The 75-day decontamination failed to completely eliminate Cd whereas Zn was quickly depurated. Cd induced over-expressions of genes involved in detoxification and against oxidative stress. Zn binary contamination with Cd provided protective effects on Cd-induced toxicity in Danio rerio. After 14-30 d of depuration, genes were no longer over expressed in response to Cd.

Environmental Toxicology and Chemistry, 2006

Biological indicators can be used to assess polluted sites, but their success depends on their sp... more Biological indicators can be used to assess polluted sites, but their success depends on their specificity. The aim of the present study was to identify a specific cadmium biomarker of the freshwater bivalve Corbicula fluminea. Differential display technique was used to identify new molecular indicators for cadmium exposure. Gene expression of C. fluminea transplanted into a cadmium-and zinc-contaminated river was compared to gene expression of bivalves from a reference site. One differentially expressed band was identified, and the entire cDNA sequence coding for the ribosomal protein S9 (rpS9) was cloned. The rpS9 gene expression was studied by real-time reverse transcription-polymerase chain reaction. After controlled cadmium (15 g/L) and/or zinc (1 mg/L) laboratory contaminations of the bivalves, this gene was shown to be up-regulated by cadmium and downregulated by zinc. A 48-d transplantation of animals into a polymetallic field contamination in the Lot river basin (Midi-Pyrénées region, France) showed that rpS9 gene regulation was correlated with the fluctuating cadmium and zinc water contents. To our knowledge, the existence of a gene that is up-regulated by cadmium and repressed by zinc is demonstrated here for the first time.

Environmental Toxicology and Chemistry, 2006

Quantitative real-time polymerase chain reaction (PCR) was used to compare for the first time the... more Quantitative real-time polymerase chain reaction (PCR) was used to compare for the first time the differential expression of metallothionein (MT) isoform genes, together with biosynthesis of the total MT proteins, in the gills of triploid and diploid juvenile Pacific oyster Crassostrea gigas in response to cadmium (Cd) and zinc (Zn) exposure. Oysters were exposed to Cd (0.133 M), Zn (15.3 M), and CdϩZn for 14 d. Results showed similar response capacities to metal exposures in the two populations. No significant difference was revealed in terms of MT gene expression, MT protein synthesis, and Cd accumulation. However, triploid oysters bioaccumulated Zn 30% less efficiently than diploid oysters. Among the three MT isoform genes, CgMT2 appeared to be more expressed than CgMT1, whereas CgMT3 appeared to be anecdotal (10 6 times lower than CgMT2). CgMT2 and CgMT1 gene expression levels were increased sevenfold in the presence of Cd, whereas Zn appeared to have no effect. A twofold increase in MT protein levels occurred in response to Cd exposure. Discrepancies between mRNA and protein levels suggest that in C. gigas MT are regulated at the transcriptional level, as well as at the translational level.

Environmental Toxicology, 2007

The toxic and genotoxic potential of Cadmium (CdCl 2 ) were evaluated by the micronucleus test (M... more The toxic and genotoxic potential of Cadmium (CdCl 2 ) were evaluated by the micronucleus test (MNT) and comet assay (CA) using amphibian larvae (Xenopus laevis and Pleurodeles waltl). Acute toxicity results showed that Cd is toxic to Xenopus larvae exposed from 2 to 50 mg/L and to Pleurodeles from 5 to 50 mg/L, depending on the nature of the water (reconstituted water containing mineral salts or mineral water MW (Volvic 1 )). The MNT results obtained in MW showed that Cd (2 mg/L) is genotoxic to Xenopus, whereas it was not genotoxic to Pleurodeles at all concentrations tested. The CA established that the genotoxicity of Cd to Xenopus and Pleurodeles larvae depends on the concentration, the exposure times, and the comet parameters (Tail DNA, ETM, OTM, and TL). The CA and MNT results were compared for their ability to detect genotoxic effects, considering the concentrations of Cd applied and the exposure time. The CA showed Cd to be genotoxic from the first day of exposure. In amphibians, the CA appears to be a sensitive and suitable method for detecting genotoxicity such as that caused by Cd. # 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 422-435, 2007.

Environmental Science and Pollution Research, 2010

Ecotoxicology and Environmental Safety, 2012

a b s t r a c t Metallothionein (MT) genes encode crucial metal-binding proteins ubiquitously exp... more a b s t r a c t Metallothionein (MT) genes encode crucial metal-binding proteins ubiquitously expressed in living organisms and which play important roles in homeostasis of essential metals and detoxification processes. Here, the molecular organization of the first metallothionein gene of the edible cockle Cerastoderma edule and its expression after cadmium (Cd) or mercury (Hg) exposures were determined. The resulting sequence (Cemt1) exhibits unusual features. The full length cDNA encodes a protein of 73 amino acids with nine classical Cys-X (1-3) -Cys motifs, but also one Cys-Cys not generally found in molluscan MT. Moreover, characterization of the molecular organization of the Cemt1 gene revealed two different alleles (A1 and A2) with length differences due to large deletion events in their intronic sequences involving direct Short Interspersed repeated Elements (SINE), while their exonic sequences were identical. To our knowledge, such large excision mechanisms have never before been reported in a bivalve gene sequence. After 10 days of Cd exposure at environmentally relevant doses, quantitative real-time PCR revealed a strong induction of Cemt1 in gills of C. edule. Surprisingly, neither induction of the Cemt1 gene nor of MT protein was shown after Hg exposure, despite the fact that this organism is able to bioaccumulate a high amount of this trace metal which is theoretically one of the most powerful inducers of MT biosynthesis.

Ecotoxicology and Environmental Safety, 2010

This study aimed to assess the effect of two dietary cadmium (Cd) levels (C1: 1 mg kg À 1 ; C10: ... more This study aimed to assess the effect of two dietary cadmium (Cd) levels (C1: 1 mg kg À 1 ; C10: 10 mg kg À 1 ) on bird Anas platyrhynchos exposed for 10, 20 and 40 days (5 animals per experimental condition). Ducks were able to accumulate high amounts of Cd, especially in kidneys (after 40 days: C1 8.1 7 1 mg kg À 1 , C10 37.7 7 4.3 mg kg À 1 ). After 40 days, the lowest Cd level triggered oxidative stress and stimulated mitochondrial metabolism. At the same time, highest amounts of Cd (C10 group) only triggered repression of genes encoding for catalase and acetyl-CoA carboxylase, with repression factors of 1/50 and 1/5, respectively. High dose exposures were then associated with the repression of genes encoding for antioxidant, whereas low dose exposure triggered their induction. In contrast, the onset of MT gene expression appeared quickly for the C10 group even if a time delay was observed between gene expression and protein accumulation. Through the comparison of A. platyrhynchos and Cairina moschata, the response to Cd toxicity appeared species-dependent. Discrepancies between species could be explained by differential utilization of MT. This pathway of detoxification seemed sufficient to counter Cd toxicity.

Ecotoxicology and Environmental Safety, 2012

Because of their high capacity to accumulate contaminants such as persistent organic pollutants a... more Because of their high capacity to accumulate contaminants such as persistent organic pollutants and heavy metals, aquatic sediments are considered as a long-term source of contamination for aquatic organisms. In compliance with the increasing interest both for sediment quality evaluation and the use of fish early life stage (ELS) toxicity assays, we proposed an embryo-larval test to evaluate embryotoxicity and genotoxicity of sediment-bound contaminants. Pre-blastula stage medaka (Oryzias latipes) embryos were exposed by static sediment contact to two model heavy metals (cadmium and copper) at environmental concentrations during the whole 10-day embryonic development. Lethal and sub-lethal effects were recorded in both embryos and larvae for 20 days post fertilisation (dpf) using several global toxicity and phenotypic endpoints. The comet assay was also performed on medaka prolarvae to evaluate genotoxic effects of the tested chemicals. Environmental concentrations of cadmium (Cd) and copper (Cu) did not affect embryo and larval survival. However, both heavy metals significantly induced morphological abnormalities, particularly spinal and cardiovascular deformities. Cd but not Cu induced tachycardia. Both heavy metals induced a significant increase in DNA damage at all tested concentrations. Resulting LOEC values for Cd and Cu corresponded to 1.9 and 8.5 mg/g d.w. sediment, respectively. Although metal bioavailability is probably lower for naturally contaminated sediments, the relatively low toxicity thresholds for both Cd and Cu raise the question of possible risk for fish embryos developing in direct contact to sediments. This study demonstrates the applicability, sensitivity and relevance of the Japanese medaka embryo-larval assay (MELA) to evaluate sediment hazardous potency at environmental concentrations of heavy metals.

Ecotoxicology and Environmental Safety, 2011

Ecotoxicological studies aim to assess the potential environmental risks of various products. Thi... more Ecotoxicological studies aim to assess the potential environmental risks of various products. This implies the use of various biological models and tests on live animals. In case of handling fish and mammals, ethical rules have to be respected. The use of anaesthesia is considered to be the best way to ensure animal welfare. Eugenol and electro-narcosis are among the most popular chemical and physical anaesthetics used in fisheries and by field biologists. In this study, the genetic and endocrine impacts of these anaesthetics were assessed in order to establish whether the use of such methods could skew the results of ecotoxicological studies. Twenty yellow European eels (Anguilla anguilla) were submitted to Eugenol (50 mg/L) and electro-narcosis until they reached a level of deep anaesthesia, while 20 other eels were kept aware. Five anaesthetized and five unanaesthetized eels were sacrificed and analysed directly after treatment and after 1, 7 and 21 days of recovery. At the brain level, Eugenol triggered an increase in the transcription level of genes encoding proteins involved in oxidative stress responses (catalase expression 2.5-fold, mitochondrial superoxide dismutase expression 3-fold), probably due to a hypoxic event during anaesthesia. Later impacts were detected in muscles 21 days after anaesthesia (ATP synthase subunit 6 3-fold, NADH deshydrogenase subunit 5 4-fold and mitochondrial superoxide dismutase 3-fold increased) revealing oxidative stress from an accrued mitochondrial respiratory metabolism. Hormone dosages showed that the use of Eugenol reduced the release of plasma cortisol during anaesthesia. However, this impact seemed to be reversible within one day. In case of electro-narcosis, no significant variation in transcriptional levels could be detected between anaesthetized and unanaesthetized eels. Our results suggest that the use of Eugenol as an aesthetic in ecotoxicological studies measuring gene expression or plasma cortisol concentration is not appropriate, while electro-narcosis does not seem to have any impact, at least on the parameters taken into consideration in this study.

Ecotoxicology, 2010

In environment, birds often fast in connection with breeding, migration or drastic climatic condi... more In environment, birds often fast in connection with breeding, migration or drastic climatic conditions and need to mobilize lipid reserves during these periods. The impairment of lipid metabolism by cadmium (Cd; 1 mg kg -1 added in diet) was investigated on palmiped Cairina moschata. Expression levels of genes involved in lipid metabolism, mitochondrial metabolism and detoxification were investigated in liver and muscle of ducks. Lipid content in muscle and liver were analysed and plasma triglycerides were quantified. After 20 days, ducks exposed to Cd displayed a lower body weight and lower lipid content in liver than controls. In muscle, the increase of lipid content was only significant for control ducks but not for exposed ducks. Exposed ducks appeared unable to sufficiently transport and store lipids into peripheral tissues. Cd impairs lipid metabolism by several ways. First, Cd triggered the down-regulation of fatty acids synthesis in liver even if the NADPH production and the mitochondrial metabolism are enhanced, suggesting a stronger energy needs. Secondly, the associated decrease of plasma triglycerides and lipoprotein lipase activity with Cd are consistent with impairment of lipids storage in peripheral tissues.

BioMetals, 2006

Metallothionein (MT) response to cadmium (Cd) and zinc (Zn) bioaccumulation after single or combi... more Metallothionein (MT) response to cadmium (Cd) and zinc (Zn) bioaccumulation after single or combined direct exposure was compared in two freshwater bivalves, Dreissena polymorpha (zebra mussel) and Corbicula fluminea (Asiatic clam). Bivalves were exposed to 0.133 lM Cd and/or 15.3 lM Zn, with metal and MT concentrations analysed in the whole soft body after 1, 3, 10 and 24 days of exposure and compared with controls. Results showed significant increase in MT concentrations in both species exposed to Cd and Cd+Zn with a higher accumulation of the protein compared to the control in D. polymorpha for nevertheless similar Cd levels accumulated with time. Exposure to Zn alone led to a significant increase in MT concentrations only in C. fluminea, whereas there was a lack of MT gene induction in the zebra mussels which was confirmed by MT mRNA quantification in gills (RT-PCR). Mussel mortality after 10 days of exposure to Zn and Cd+Zn is discussed with regard to detoxification mechanisms, which include metallothioneins.

BioMetals, 2006

The effects of cadmium (Cd) on gene expression were examined in four organs (gills, liver, skelet... more The effects of cadmium (Cd) on gene expression were examined in four organs (gills, liver, skeletal muscles and brain) of the zebrafish. Adult male fish were subjected to three different water contamination pressures over periods of 7 and 21 days: control medium (C 0 : no Cd added) and two contaminated media (C 1 : 1.9 ± 0.6 lg Cd l )1 , and C 2 : 9.6 ± 2.9 lg Cd l )1 ). Fourteen genes involved in antioxidant defences, metal chelation, active efflux of organic compounds, mitochondrial metabolism, DNA repair and apoptosis were selected and their expression levels investigated by quantitative real-time PCR. Cadmium concentrations were determined in the four organs and metallothionein (MT) protein levels investigated in brain, liver and gills. Although skeletal muscle was a poor Cd-accumulating tissue, many genes were up-regulated at day 7: mt1, cyt, bax, gadd and rad51 genes. Three additional genes, c-jun, pyc and tap, were up-regulated in muscles at day 21 whereas bax, gadd and rad51 had returned to basal levels. Surprisingly, mt1 and c-jun were the only genes displaying a differential induction after 21 days in liver, although this organ accumulated the highest cadmium concentration. In brain, only mt1, mt2 and c-jun genes were up-regulated after 21 days. In gills, the highest response was observed after 7 days, featuring the differential expression of oxidative stress-response hsp70 and mitochondrial sod genes, along with genes involved in mitochondrial metabolism and metal detoxification. Then, after 21 days, the expression of almost every genes returned to basal levels while both mt1 and mt2 genes were up-regulated.

Biochimie, 2007

In order to demonstrate the in vivo antioxidant properties of metallothioneins (MTs), the bacteri... more In order to demonstrate the in vivo antioxidant properties of metallothioneins (MTs), the bacteria Escherichia coli was used as a cell reactor in which we compared the metal binding and antioxidative functions of MTs from different species, with different structures and polypeptide lengths. No protective effects of cytoplasmic MTs from cadmium (Cd) or zinc (Zn) contamination were observed in a wild-type E. coli strain, although these MTs can efficiently bind both Cd and Zn. To test their antioxidant properties, MTs were expressed within the cytoplasm of a sodA sodB deficient mutated strain (QC1726). However, a paradoxical MT toxicity was found when this strain was contaminated with Cd and Zn, suggesting that in a wild-type strain, superoxide dismutase counteracts MT toxicity. The most toxic MT was the one with the strongest Cd and Zn binding capacities. This toxic effect was linked to the generation of superoxide radicals, since a Cd-contaminated QC1726 strain expressing oyster MT isoforms produced 75e85% more O 2 À than the control QC1726 strain. Conversely, under anaerobiosis or in the presence of a copper chelator, MTs protected QC1726 strain from Cd and Zn contamination. A model is proposed to explain the observed MT toxicity.

Chemosphere, 2014

Since 2007 a remediation process was initiated on a mining site contaminated by metals (Cd and Zn... more Since 2007 a remediation process was initiated on a mining site contaminated by metals (Cd and Zn). Bivalves were exposed for two years to observe early remediation impacts on hydrosystem. Metal concentrations in river and bivalves remained very high in organisms exposed downstream. Bivalves were still sharply genetically impacted by metal contamination. Recovery of aquatic communities after remediation can only be expected in the long term.

Comparative biochemistry and physiology. Toxicology & pharmacology : CBP, Jan 20, 2015

The effect of methylmercury (MeHg) on glass eels' propensity to migrate, mitochondrial activi... more The effect of methylmercury (MeHg) on glass eels' propensity to migrate, mitochondrial activity and antioxidative defence systems was investigated. Marine glass eels were first sorted in an experimental flume according to their response to dusk. Fish responding to the decrease in light intensity by ascending in the water column and moving with or against the flow were considered as having a high propensity to migrate (migrant). Glass eels still sheltering at the end of the 24h catching period were considered as having a low propensity to migrate and were called non-migrant. Migrant and non-migrant glass eels were then individually tagged and exposed to isotopically enriched (201)MeHg (50ngL(-1)) for 11days. The effect of contamination was studied on muscle fibre structure, and the expression level of genes involved in mitochondrial activity and antioxidative defence systems. To investigate the effect of MeHg on glass eel behaviour, migrant and non-migrant glass eels were sorted ...

Environmental pollution (Barking, Essex : 1987), 2014

This study aimed to assess the recovery capacity of the freshwater bivalve Corbicula fluminea sub... more This study aimed to assess the recovery capacity of the freshwater bivalve Corbicula fluminea subjected to industrial metal discharges (Cd, Zn). After a 24-day exposure in a metal-contaminated river, bivalves were transferred and maintained in the laboratory for one year under metal-free conditions. Metal accumulation, metallothionein production and genetic expressions of genes involved in metal stress were studied. Results demonstrated the high persistence of Cd in tissues (only 73% eliminated after 365 days) whereas Zn was rapidly depurated. The Cd half-life was estimated around 240 days. Metallothioneins were strongly induced within the 28 first days of decontamination, then decreased by 45% after 365 days. The metal exposure of bivalves led to a significant gene induction. After 28 days, most of the genes were no longer overexpressed, suggesting that the bivalves may withstand small amounts of non-essential metals in their tissues without showing signs of detrimental effects on ...

International Journal of Molecular Sciences, 2012

Methylmercury (MeHg) is a potent neurotoxin, and human beings are mainly exposed to this pollutan... more Methylmercury (MeHg) is a potent neurotoxin, and human beings are mainly exposed to this pollutant through fish consumption. We addressed the question of whether a diet mimicking the fish consumption of Wayanas Amerindians from French Guiana could result in observable adverse effects in mice. Wayanas adult men are subjected to a mean mercurial dose of 7 g Hg/week/kg of body weight. We decided to supplement a vegetarian-based mice diet with 0.1% of lyophilized Hoplias aimara fish, which Wayanas OPEN ACCESS

Chemosphere, 2015

This study aimed to demonstrate the recovery potential of Danio rerio after Cd and Zn contaminati... more This study aimed to demonstrate the recovery potential of Danio rerio after Cd and Zn contaminations. The 75-day decontamination failed to completely eliminate Cd whereas Zn was quickly depurated. Cd induced over-expressions of genes involved in detoxification and against oxidative stress. Zn binary contamination with Cd provided protective effects on Cd-induced toxicity in Danio rerio. After 14-30 d of depuration, genes were no longer over expressed in response to Cd.

Environmental Toxicology and Chemistry, 2006

Biological indicators can be used to assess polluted sites, but their success depends on their sp... more Biological indicators can be used to assess polluted sites, but their success depends on their specificity. The aim of the present study was to identify a specific cadmium biomarker of the freshwater bivalve Corbicula fluminea. Differential display technique was used to identify new molecular indicators for cadmium exposure. Gene expression of C. fluminea transplanted into a cadmium-and zinc-contaminated river was compared to gene expression of bivalves from a reference site. One differentially expressed band was identified, and the entire cDNA sequence coding for the ribosomal protein S9 (rpS9) was cloned. The rpS9 gene expression was studied by real-time reverse transcription-polymerase chain reaction. After controlled cadmium (15 g/L) and/or zinc (1 mg/L) laboratory contaminations of the bivalves, this gene was shown to be up-regulated by cadmium and downregulated by zinc. A 48-d transplantation of animals into a polymetallic field contamination in the Lot river basin (Midi-Pyrénées region, France) showed that rpS9 gene regulation was correlated with the fluctuating cadmium and zinc water contents. To our knowledge, the existence of a gene that is up-regulated by cadmium and repressed by zinc is demonstrated here for the first time.

Environmental Toxicology and Chemistry, 2006

Quantitative real-time polymerase chain reaction (PCR) was used to compare for the first time the... more Quantitative real-time polymerase chain reaction (PCR) was used to compare for the first time the differential expression of metallothionein (MT) isoform genes, together with biosynthesis of the total MT proteins, in the gills of triploid and diploid juvenile Pacific oyster Crassostrea gigas in response to cadmium (Cd) and zinc (Zn) exposure. Oysters were exposed to Cd (0.133 M), Zn (15.3 M), and CdϩZn for 14 d. Results showed similar response capacities to metal exposures in the two populations. No significant difference was revealed in terms of MT gene expression, MT protein synthesis, and Cd accumulation. However, triploid oysters bioaccumulated Zn 30% less efficiently than diploid oysters. Among the three MT isoform genes, CgMT2 appeared to be more expressed than CgMT1, whereas CgMT3 appeared to be anecdotal (10 6 times lower than CgMT2). CgMT2 and CgMT1 gene expression levels were increased sevenfold in the presence of Cd, whereas Zn appeared to have no effect. A twofold increase in MT protein levels occurred in response to Cd exposure. Discrepancies between mRNA and protein levels suggest that in C. gigas MT are regulated at the transcriptional level, as well as at the translational level.

Environmental Toxicology, 2007

The toxic and genotoxic potential of Cadmium (CdCl 2 ) were evaluated by the micronucleus test (M... more The toxic and genotoxic potential of Cadmium (CdCl 2 ) were evaluated by the micronucleus test (MNT) and comet assay (CA) using amphibian larvae (Xenopus laevis and Pleurodeles waltl). Acute toxicity results showed that Cd is toxic to Xenopus larvae exposed from 2 to 50 mg/L and to Pleurodeles from 5 to 50 mg/L, depending on the nature of the water (reconstituted water containing mineral salts or mineral water MW (Volvic 1 )). The MNT results obtained in MW showed that Cd (2 mg/L) is genotoxic to Xenopus, whereas it was not genotoxic to Pleurodeles at all concentrations tested. The CA established that the genotoxicity of Cd to Xenopus and Pleurodeles larvae depends on the concentration, the exposure times, and the comet parameters (Tail DNA, ETM, OTM, and TL). The CA and MNT results were compared for their ability to detect genotoxic effects, considering the concentrations of Cd applied and the exposure time. The CA showed Cd to be genotoxic from the first day of exposure. In amphibians, the CA appears to be a sensitive and suitable method for detecting genotoxicity such as that caused by Cd. # 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 422-435, 2007.

Environmental Science and Pollution Research, 2010

Ecotoxicology and Environmental Safety, 2012

a b s t r a c t Metallothionein (MT) genes encode crucial metal-binding proteins ubiquitously exp... more a b s t r a c t Metallothionein (MT) genes encode crucial metal-binding proteins ubiquitously expressed in living organisms and which play important roles in homeostasis of essential metals and detoxification processes. Here, the molecular organization of the first metallothionein gene of the edible cockle Cerastoderma edule and its expression after cadmium (Cd) or mercury (Hg) exposures were determined. The resulting sequence (Cemt1) exhibits unusual features. The full length cDNA encodes a protein of 73 amino acids with nine classical Cys-X (1-3) -Cys motifs, but also one Cys-Cys not generally found in molluscan MT. Moreover, characterization of the molecular organization of the Cemt1 gene revealed two different alleles (A1 and A2) with length differences due to large deletion events in their intronic sequences involving direct Short Interspersed repeated Elements (SINE), while their exonic sequences were identical. To our knowledge, such large excision mechanisms have never before been reported in a bivalve gene sequence. After 10 days of Cd exposure at environmentally relevant doses, quantitative real-time PCR revealed a strong induction of Cemt1 in gills of C. edule. Surprisingly, neither induction of the Cemt1 gene nor of MT protein was shown after Hg exposure, despite the fact that this organism is able to bioaccumulate a high amount of this trace metal which is theoretically one of the most powerful inducers of MT biosynthesis.

Ecotoxicology and Environmental Safety, 2010

This study aimed to assess the effect of two dietary cadmium (Cd) levels (C1: 1 mg kg À 1 ; C10: ... more This study aimed to assess the effect of two dietary cadmium (Cd) levels (C1: 1 mg kg À 1 ; C10: 10 mg kg À 1 ) on bird Anas platyrhynchos exposed for 10, 20 and 40 days (5 animals per experimental condition). Ducks were able to accumulate high amounts of Cd, especially in kidneys (after 40 days: C1 8.1 7 1 mg kg À 1 , C10 37.7 7 4.3 mg kg À 1 ). After 40 days, the lowest Cd level triggered oxidative stress and stimulated mitochondrial metabolism. At the same time, highest amounts of Cd (C10 group) only triggered repression of genes encoding for catalase and acetyl-CoA carboxylase, with repression factors of 1/50 and 1/5, respectively. High dose exposures were then associated with the repression of genes encoding for antioxidant, whereas low dose exposure triggered their induction. In contrast, the onset of MT gene expression appeared quickly for the C10 group even if a time delay was observed between gene expression and protein accumulation. Through the comparison of A. platyrhynchos and Cairina moschata, the response to Cd toxicity appeared species-dependent. Discrepancies between species could be explained by differential utilization of MT. This pathway of detoxification seemed sufficient to counter Cd toxicity.

Ecotoxicology and Environmental Safety, 2012

Because of their high capacity to accumulate contaminants such as persistent organic pollutants a... more Because of their high capacity to accumulate contaminants such as persistent organic pollutants and heavy metals, aquatic sediments are considered as a long-term source of contamination for aquatic organisms. In compliance with the increasing interest both for sediment quality evaluation and the use of fish early life stage (ELS) toxicity assays, we proposed an embryo-larval test to evaluate embryotoxicity and genotoxicity of sediment-bound contaminants. Pre-blastula stage medaka (Oryzias latipes) embryos were exposed by static sediment contact to two model heavy metals (cadmium and copper) at environmental concentrations during the whole 10-day embryonic development. Lethal and sub-lethal effects were recorded in both embryos and larvae for 20 days post fertilisation (dpf) using several global toxicity and phenotypic endpoints. The comet assay was also performed on medaka prolarvae to evaluate genotoxic effects of the tested chemicals. Environmental concentrations of cadmium (Cd) and copper (Cu) did not affect embryo and larval survival. However, both heavy metals significantly induced morphological abnormalities, particularly spinal and cardiovascular deformities. Cd but not Cu induced tachycardia. Both heavy metals induced a significant increase in DNA damage at all tested concentrations. Resulting LOEC values for Cd and Cu corresponded to 1.9 and 8.5 mg/g d.w. sediment, respectively. Although metal bioavailability is probably lower for naturally contaminated sediments, the relatively low toxicity thresholds for both Cd and Cu raise the question of possible risk for fish embryos developing in direct contact to sediments. This study demonstrates the applicability, sensitivity and relevance of the Japanese medaka embryo-larval assay (MELA) to evaluate sediment hazardous potency at environmental concentrations of heavy metals.

Ecotoxicology and Environmental Safety, 2011

Ecotoxicological studies aim to assess the potential environmental risks of various products. Thi... more Ecotoxicological studies aim to assess the potential environmental risks of various products. This implies the use of various biological models and tests on live animals. In case of handling fish and mammals, ethical rules have to be respected. The use of anaesthesia is considered to be the best way to ensure animal welfare. Eugenol and electro-narcosis are among the most popular chemical and physical anaesthetics used in fisheries and by field biologists. In this study, the genetic and endocrine impacts of these anaesthetics were assessed in order to establish whether the use of such methods could skew the results of ecotoxicological studies. Twenty yellow European eels (Anguilla anguilla) were submitted to Eugenol (50 mg/L) and electro-narcosis until they reached a level of deep anaesthesia, while 20 other eels were kept aware. Five anaesthetized and five unanaesthetized eels were sacrificed and analysed directly after treatment and after 1, 7 and 21 days of recovery. At the brain level, Eugenol triggered an increase in the transcription level of genes encoding proteins involved in oxidative stress responses (catalase expression 2.5-fold, mitochondrial superoxide dismutase expression 3-fold), probably due to a hypoxic event during anaesthesia. Later impacts were detected in muscles 21 days after anaesthesia (ATP synthase subunit 6 3-fold, NADH deshydrogenase subunit 5 4-fold and mitochondrial superoxide dismutase 3-fold increased) revealing oxidative stress from an accrued mitochondrial respiratory metabolism. Hormone dosages showed that the use of Eugenol reduced the release of plasma cortisol during anaesthesia. However, this impact seemed to be reversible within one day. In case of electro-narcosis, no significant variation in transcriptional levels could be detected between anaesthetized and unanaesthetized eels. Our results suggest that the use of Eugenol as an aesthetic in ecotoxicological studies measuring gene expression or plasma cortisol concentration is not appropriate, while electro-narcosis does not seem to have any impact, at least on the parameters taken into consideration in this study.

Ecotoxicology, 2010

In environment, birds often fast in connection with breeding, migration or drastic climatic condi... more In environment, birds often fast in connection with breeding, migration or drastic climatic conditions and need to mobilize lipid reserves during these periods. The impairment of lipid metabolism by cadmium (Cd; 1 mg kg -1 added in diet) was investigated on palmiped Cairina moschata. Expression levels of genes involved in lipid metabolism, mitochondrial metabolism and detoxification were investigated in liver and muscle of ducks. Lipid content in muscle and liver were analysed and plasma triglycerides were quantified. After 20 days, ducks exposed to Cd displayed a lower body weight and lower lipid content in liver than controls. In muscle, the increase of lipid content was only significant for control ducks but not for exposed ducks. Exposed ducks appeared unable to sufficiently transport and store lipids into peripheral tissues. Cd impairs lipid metabolism by several ways. First, Cd triggered the down-regulation of fatty acids synthesis in liver even if the NADPH production and the mitochondrial metabolism are enhanced, suggesting a stronger energy needs. Secondly, the associated decrease of plasma triglycerides and lipoprotein lipase activity with Cd are consistent with impairment of lipids storage in peripheral tissues.

BioMetals, 2006

Metallothionein (MT) response to cadmium (Cd) and zinc (Zn) bioaccumulation after single or combi... more Metallothionein (MT) response to cadmium (Cd) and zinc (Zn) bioaccumulation after single or combined direct exposure was compared in two freshwater bivalves, Dreissena polymorpha (zebra mussel) and Corbicula fluminea (Asiatic clam). Bivalves were exposed to 0.133 lM Cd and/or 15.3 lM Zn, with metal and MT concentrations analysed in the whole soft body after 1, 3, 10 and 24 days of exposure and compared with controls. Results showed significant increase in MT concentrations in both species exposed to Cd and Cd+Zn with a higher accumulation of the protein compared to the control in D. polymorpha for nevertheless similar Cd levels accumulated with time. Exposure to Zn alone led to a significant increase in MT concentrations only in C. fluminea, whereas there was a lack of MT gene induction in the zebra mussels which was confirmed by MT mRNA quantification in gills (RT-PCR). Mussel mortality after 10 days of exposure to Zn and Cd+Zn is discussed with regard to detoxification mechanisms, which include metallothioneins.

BioMetals, 2006

The effects of cadmium (Cd) on gene expression were examined in four organs (gills, liver, skelet... more The effects of cadmium (Cd) on gene expression were examined in four organs (gills, liver, skeletal muscles and brain) of the zebrafish. Adult male fish were subjected to three different water contamination pressures over periods of 7 and 21 days: control medium (C 0 : no Cd added) and two contaminated media (C 1 : 1.9 ± 0.6 lg Cd l )1 , and C 2 : 9.6 ± 2.9 lg Cd l )1 ). Fourteen genes involved in antioxidant defences, metal chelation, active efflux of organic compounds, mitochondrial metabolism, DNA repair and apoptosis were selected and their expression levels investigated by quantitative real-time PCR. Cadmium concentrations were determined in the four organs and metallothionein (MT) protein levels investigated in brain, liver and gills. Although skeletal muscle was a poor Cd-accumulating tissue, many genes were up-regulated at day 7: mt1, cyt, bax, gadd and rad51 genes. Three additional genes, c-jun, pyc and tap, were up-regulated in muscles at day 21 whereas bax, gadd and rad51 had returned to basal levels. Surprisingly, mt1 and c-jun were the only genes displaying a differential induction after 21 days in liver, although this organ accumulated the highest cadmium concentration. In brain, only mt1, mt2 and c-jun genes were up-regulated after 21 days. In gills, the highest response was observed after 7 days, featuring the differential expression of oxidative stress-response hsp70 and mitochondrial sod genes, along with genes involved in mitochondrial metabolism and metal detoxification. Then, after 21 days, the expression of almost every genes returned to basal levels while both mt1 and mt2 genes were up-regulated.

Biochimie, 2007

In order to demonstrate the in vivo antioxidant properties of metallothioneins (MTs), the bacteri... more In order to demonstrate the in vivo antioxidant properties of metallothioneins (MTs), the bacteria Escherichia coli was used as a cell reactor in which we compared the metal binding and antioxidative functions of MTs from different species, with different structures and polypeptide lengths. No protective effects of cytoplasmic MTs from cadmium (Cd) or zinc (Zn) contamination were observed in a wild-type E. coli strain, although these MTs can efficiently bind both Cd and Zn. To test their antioxidant properties, MTs were expressed within the cytoplasm of a sodA sodB deficient mutated strain (QC1726). However, a paradoxical MT toxicity was found when this strain was contaminated with Cd and Zn, suggesting that in a wild-type strain, superoxide dismutase counteracts MT toxicity. The most toxic MT was the one with the strongest Cd and Zn binding capacities. This toxic effect was linked to the generation of superoxide radicals, since a Cd-contaminated QC1726 strain expressing oyster MT isoforms produced 75e85% more O 2 À than the control QC1726 strain. Conversely, under anaerobiosis or in the presence of a copper chelator, MTs protected QC1726 strain from Cd and Zn contamination. A model is proposed to explain the observed MT toxicity.