Copper and nanocopper toxicity using integrated biomarker response in Pangasianodon hypophthalmus.

The current study focused on assessing the toxicological effects of copper (Cu) and copper nanoparticles (Cu-NPs) in acute condition on Pangasianodon hypophthalmus. The median lethal concentration (LC50 ) for Cu and Cu-NPs were determined as 8.04 and 3.85 mg L-1 , respectively. For the subsequent definitive test, varying concentrations were selected: 7.0, 7.5, 8.0, 8.5, and 9.0 mg L-1 for Cu, and 3.0, 3.3, 3.6, 3.9, and 4.2 mg L-1 for Cu-NPs. To encompass these concentration levels and assess their toxic effects, biomarkers associated with toxicological studies like oxidative stress, neurotransmission, and cellular metabolism were measured in the liver, kidney, and gill tissues. Notably, during the acute test, the activities of catalase, superoxide dismutase, glutathione-s-transferase, glutathione peroxidase, and lipid peroxide in the liver, gill, and kidney tissues were significantly increased due to exposure to Cu and Cu-NPs. Similarly, acetylcholinesterase activity in the brain was notably inhibited in the presence of Cu and Cu-NPs when compared to the control group. Cellular metabolic stress was greatly influenced by the exposure to Cu and Cu-NPs, evident from the considerable elevation of cortisol, HSP 70, and blood glucose levels in the treated groups. Furthermore, integrated biomarker response, genotoxicity, DNA damage in gill tissue, karyotyping in kidney tissue, and histopathology in gill and liver were investigated, revealing tissue damage attributed to exposure to Cu and Cu-NPs. In conclusion, this study determined that elevated concentrations of essential trace elements, namely Cu and Cu-NPs, induce toxicity and disrupt cellular metabolic activities in fish.

The invasive crayfish Cherax quadricarinatus facing chlordecone in Martinique: Bioaccumulation and depuration study.

The redclaw crayfish, Cherax quadricarinatus, was introduced to Martinique Island for aquaculture purposes at the beginning of the 21st century, in an attempt to revitalize the freshwater crustacean aquaculture sector. Mainly due to its high economical value, it was intentionally released in the wild and was caught and sold by fishermen. Martinican rivers are polluted by chlordecone, considered as one of the worst Persistant Organic Pollutants (POP). Despite its dangerousness, it was used until 1993 in the French West Indies against a banana pest and was always found in the ecosystems. This study aimed to investigate the level of contamination in the muscle of crayfish caught in the wild, as well as the potential of bioconcentration and depuration in the C. quadricarinatus muscle. This study could allow us to quantify the risk for consumers but also, to evaluate a depuration process to reduce the risk related to its consumption. Using both in-vitro and in-situ experiments, results highlighted the importance of the chlordecone concentration in the water and the time of exposure to the pollutant. The bioconcentration seems to be very quick and continuous in crayfish muscle, as chlordecone can be detectable as early as 6 h of exposure, whatever the concentration tested. Finally, it appears that, even after 20 days of depuration in chlordecone-free water, chlordecone concentrations remained higher to the residual maximum limit (i.e. 20 ng/g wet weight), concluding that the decontamination of the muscle seems not very efficient, and the risk for the Martinican people could be serious.

Inorganic mercury effects on biomarker gene expressions of a freshwater amphipod at two temperatures.

Mercury (Hg) is a global contaminant resulting of both natural processes and human activities. In aquatic environments, studies conducted on vertebrates highlighted changes of gene expression or activity of antitoxic and oxidative enzymes. However, although Hg is a highly toxic compound in aquatic environments, only a few studies have evaluated the lethal and sublethal effects of inorganic Hg on Gammarus sp. Therefore, this study aimed at evaluating the effects of inorganic Hg (HgCl2) on the expression of 17 genes involved in crucial biological functions or mechanisms for organisms, namely respiration, osmoregulation, apoptosis, immune and endocrine system, and antioxidative and antitoxic defence systems. The study was performed in males of the freshwater amphipod Gammarus pulex exposed to two environmentally relevant concentrations (50 and 500 ng/L) at two temperature regime fluctuations (16 °C and 20 °C +/-2 °C) for 7 and 21 days. Results showed that G. pulex mortality was dependent on Hg concentration and temperature; the higher the concentration and temperature, the higher the mortality rate. In addition, the Integrated Biomarker Response emphasized that HgCl2 toxicity was dependent on the concentration, time and temperature of exposure. Overall, antioxidant and antitoxic defences, as well as the endocrine and immune systems, were the biological functions most impacted by Hg exposure (based on the concentration, duration, and temperature tested). Conversely, osmoregulation was the least affected biological function. The results also demonstrated a possible adaptation of G. pulex after 21 days at 500 ng/L, regardless of the exposure temperature. This study allowed us to show that Hg deregulates many crucial biological functions after a short exposure, but that during a long exposure, an adaptation phenomenon could occur, regardless of temperature.

First assessment of persistent organic pollutants in the Greater rhea (Rhea americana), a near-threatened flightless herbivorous bird of the Pampas grasslands.

Persistent organic pollutants (POPs) are still globally distributed and can exert different effects on ecosystems. Little is known about the occurrence of these contaminants in terrestrial birds from South America. In this study, POPs were assessed for the first time in a flightless herbivorous species from the Pampas grasslands, the Greater rhea (Rhea americana). Concentrations of polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and organochlorine pesticides (OCPs) were determined in 18 samples of feathers from free-ranging and captive individuals inhabiting four sites with different land uses in central Argentina. Among the 16 POPs tested in those feathers, 6 PCBs (28, 52, 101, 138, 153, and 180) and 8 OCPs (α-HCH, ß-HCH, γ-HCH, p,p'-DDE, p,p'-DDD, o,p'-DDT, p,p'-DDT, and HCB) were quantified. No PBDEs were detected. The total concentration of POPs was higher in populations living in an intensive crop production area (agriculture 159 ng g -1 and farm: 97.53 ng g-1) compared with the population in an urban area (zoo 45.86 ng g-1) and an agroecosystem with extensive rearing of livestock (cattle rearing 36.77 ng g-1). PCBs were the most abundant pollutants in all the populations studied. Lower chlorinated CB 52 and CB 101 were the principal PCB congeners detected, representing at least 70% of the total quantified. All populations studied showed a DDE + DDD/DDT ratio > 1, indicating a historical application of this insecticide. This study provides a new contribution to the scarce data on POP concentrations in South American bird species. Further investigations are needed to evaluate their potential effects on the health of individuals and populations.

Preliminary study of oxidative stress biomarkers and trace elements in North Sea Harbour Seals.

This preliminary study investigated the potential correlations between trace elements (mercury, zinc, cadmium, copper, selenium, lead, nickel, chromium, lithium and vanadium) concentrations, measured in red blood cells, and oxidative stress biomarkers (total thiols, total glutathione, total and selenium-dependent glutathione peroxidases, triglycerides, malondialdehyde) assessed in the respective serum, in males and females P. vitulina, sampled in the Wadden Sea in spring and autumn 2015. Only concentrations of total mercury and zinc showed significant differences by sex, and only lipid peroxidation was different by season. Moreover, significant positive and negative correlations were observed between biomarkers (triglycerides, thiols, malondialdehyde, glutathione) and trace element concentrations (copper, lead, mercury, nickel, zinc). These findings suggest that the studied biomarkers could be useful for the assessment of oxidative stress in harbour seals exposed to trace elements, but further research with larger sample sizes is needed to better understand their specific associations.

Roundup® disrupts chitinolytic enzyme activity and ecdysteroid concentration in Macrobrachium potiuna.

The endocrine system of crustaceans regulates the molt cycle with ecdysteroid hormones, mainly the 20-hydroxyecdysone (20-HE). Moreover, the molt process requires the action of chitinolytic enzymes (e.g., chitinase, chitobiase) to break down the old cuticle. However, endocrine disrupting compounds (EDC) are capable of altering their normal functioning. Glyphosate-based herbicides (GBH), such as Roundup®, the most widely used herbicides, are found in freshwater environments and have been considered EDC for many aquatic organisms. Therefore, this study examined the effects of environmentally relevant GBH concentrations (0.0065, 0.065, and 0.28 mg L-1) on the 20-HE concentration and chitobiase activity in the decapod prawn Macrobrachium potiuna exposed for 14 days. Additionally, lipid peroxidation, a biomarker of membrane lipid degradation, was evaluated in hepatopancreas to assess cellular damage. Results showed that GBH decreased the 20-HE concentration in females at the two highest concentrations tested, while an increase was observed in males exposed to the highest GBH concentration. In addition, GBH also decreased chitobiase activity in males (all concentrations) and females (the two highest concentrations). Finally, GBH caused increased lipid peroxidation in males, indicating cellular damage in the hepatopancreas. In conclusion, this work suggests that GBH is an EDC for crustaceans by disrupting molting, which could lead to altered reproduction and thus population dynamics. Graphical abstract Decrease in the 20-HE concentration and chitobiase activity in muscle of males and females of the freshwater prawn Macrobrachium potiuna exposed to the herbicide Roundup® for 14 days.

Gammarid exposure to pyriproxyfen and/or cadmium: what effects on the methylfarnesoate signalling pathway?

Due to expected changes in climate, it is predicted that disease-carrying mosquitoes will expand their geographical range, resulting in increased use of insect growth regulators (IGRs) to face their proliferation. Among IGRs, pyriproxyfen (PXF) is widely used and has been shown to prevent larvae from developing into adults, rendering them unable to reproduce. However, because of the similarity of crustacean and insect endocrine systems, PXF could also impact aquatic crustaceans. In addition, when spreading in the environment, PXF is found in a mixture with other pollutants such as metallic trace elements, which could alter its effect. Consequently, the present work was devoted to analysing the effects of PXF on the methylfarnesoate (MF) hormonal pathway of the freshwater amphipod Gammarus pulex, as well as its combined binary effects with cadmium (Cd), by measuring MF concentration, as well as the relative transcriptional expression of the farnesoic acid O-methyltransferase (FAMeT) (enzyme limiting the MF production), the methoprene-tolerant receptor (Met), and the broad-complex (BrC) as a transcription factor. Results revealed that single exposures to PXF or Cd have mainly overexpressed FAMeT, Met, and BrC but did not significantly affect MF concentration. Conversely, the mixture exposures seemed to suppress these effects and even achieve antagonistic effects. This work confirmed that PXF single exposure could impact non-target organisms such as amphipods through changes in hormonal pathways of MF. In the same way, it highlighted that Cd could also impair the endocrine system of exposed organisms. However, antagonistic effects have been observed in exposure to mixtures, suggesting different long-term consequences on the growth of amphipods under realistic exposure conditions.

Modulation of antioxidant gene expressions by Roundup® exposure in the decapod Macrobrachium potiuna.

Glyphosate-based herbicides (GBH), including Roundup®, are the most used herbicides in agricultural and non-agricultural areas, which can reach aquatic environments through drift during application or surface runoff. Some studies, mostly in fish, demonstrated that GBH caused oxidative stress in non-target animals. However, only few information is available on the GBH effects in the antioxidant and stress proteins of many other organisms, such as freshwater crustaceans. Thus, we aimed to investigate the effects of environmentally relevant GBH concentrations on the relative transcript expression (RTE) of the superoxide dismutase (sod1), catalase (cat), selenium-dependent glutathione peroxidase (gpx), glutathione-S-transferase (gst), thioredoxin (txn), heat shock protein (hsp70 and hsp90) in the hepatopancreas of the ecologically important freshwater prawn Macrobrachium potiuna. Moreover, this study aimed to assess the gender-differences responses to GBH exposure. Male and female prawns were exposed to three Roundup WG® concentrations (0.0065, 0.065 and 0.28 mg of glyphosate/L) and a control group (0.0 mg/L) for 7 and 14 days. In general, males had an under-expression of the studied genes, indicating an oxidative stress and possible accumulation of ROS in the hepatopancreas. In the opposite, females had an overexpression of the same genes, indicating a more robust antioxidant system, in order to cope with the possible ROS increase after Roundup WG® exposure. Therefore, results confirmed that gender could be a confounding factor in ecotoxicological assessment of GBH effects. Additionally, this work highlights that sod1, cat, gpx, gst, txn, hsp70 and hsp90 gene expressions seem to be useful biomarkers to investigate the oxidative stress caused by Roundup WG® in Macrobrachium sp.

Effects of low glyphosate-based herbicide concentrations on endocrine-related gene expression in the decapoda Macrobrachium potiuna.

Glyphosate-based herbicides (GBH) are the most used herbicides worldwide and are considered as endocrine-disrupting compounds (EDC) for non-target organisms. However, effects of GBH on their endocrine systems remain poorly understood. Thus, the aim of this study was to assess the effects of low concentrations of Roundup WG® on growth and reproduction process molecules in both males and females of the decapod crustacean Macrobrachium potiuna, by the relative transcript expression levels of the ecdysteroid receptor (EcR), the molt-inhibiting hormone (MIH), and the vitellogenin (Vg) genes. Prawns were exposed to three concentrations of GBH (0.0065, 0.065, and 0.28 mg L-1) for 7 and 14 days. The results revealed that only in males the three genes transcript levels were influenced by the GBH concentration, time of exposure, and the interaction between the concentrations and time of exposure, suggesting that males were more sensitive to GBH than females. For males, after 7 days of exposure at 0.065 mg L-1, EcR and MIH were over-expressed, while the Vg expression was only over-expressed after 14 days. The present study highlighted that GBH impacted endocrine systems of M. potiuna. Moreover, EcR and MIH gene expressions could be promising EDC biomarkers of exposure in crustaceans. These results also indicate that GBH concentrations, considered secure by regulatory agencies, should be reviewed to minimize the effects on non-target organisms. Potential effects of glyphosate-based herbicides on the endocrine system of decapods Macrobrachium sp.

Relative expression of three key genes involved in the hormonal cycle of the freshwater amphipod, Gammarus pulex.

Ecdysteroids and sesquiterpenoids are the two groups of hormones controlling molt and growth processes in amphipods, and are mostly represented by ecdysone (i.e., 20-hydroxyecdysone) and methyl farnesoate (MF), respectively. The endocrine system responsible for their syntheses is located in the cephalothorax and is composed of three main organs: the X-organ/sinus gland complex (XO), the Y-organ (YO) and the mandibular organ. Ecdysone synthesis is negatively controlled by the molt-inhibiting hormone (MIH) produced by XO, and its signal is mediated through the ecdysteroid receptor (EcR); whereas the MF production is limited by the farnesoic acid O-methyltransferase enzyme (FAMeT). As little is known on MIH, EcR, and FAMeT in amphipods, this study focused on the evaluation of the expression variations of the gene of these three proteins in Gammarus pulex, during the embryonic development and the molt cycle of females. Results highlighted the presence of ecr and famet genes from the first stages of the embryogenesis, suggesting key roles in the embryo development. The mih gene was only measured from Stage 3 of embryogenesis, probably related to the gastrulation and the cephalothorax development. Regardless of the gene, a strong overexpression was also measured at the hatch time. Besides, differential expression levels of mih, ecr, and famet genes through the molt cycle were observed. The highest expressions of the three genes were measured at the premolt stage, confirming key roles of MIH and EcR in the ecdysteroid pathways, and also suggesting the involvement of the FAMeT enzyme during the ecdysis in G. pulex.

Identification of molt-inhibiting hormone and ecdysteroid receptor cDNA sequences in Gammarus pulex, and variations after endocrine disruptor exposures.

In amphipods, growth, development and reproduction are mediated by the molt, which is a hormonally controlled process and which, therefore, could be impacted by endocrine disruption compounds (EDC). The molt process is controlled by both X-organ (XO) and Y-organ (YO) through a variety of hormones and receptors including the molt-inhibiting hormone (MIH) and the ecdysteroid receptor (EcR). However, although many studies were devoted to characterize MIH and EcR in crustaceans, only few works evaluated their variations under EDCs exposures. Consequently, the present work aimed to characterize MIH and EcR genes of the amphipod Gammarus pulex, as well as to study their relative expression variations after exposure to four EDCs, proved in vertebrates: ethinylestradiol (estrogen), 4-hydroxytamoxifen (anti-estrogen), 17α-methyltestosterone (androgen) and cyproterone acetate (anti-androgen). PCR amplification allowed to obtain 204 bp length and 255 bp length fragments, encoding for partial sequences of 68 amino acids and 85 amino acids, which correspond to EcR and MIH, respectively, and which are highly conserved in crustacean species. Results highlighted MIH and EcR expressions mainly in G. pulex head, which is the localization of XO and YO. Moreover, irrespective of the EDC exposure, increases of MIH and EcR relative expressions were observed, as it was observed after the exposure to 20-hydroxyecdysone (20HE), the natural molt hormone, used as positive control. Therefore, it appeared that tested EDCs behaved like 20HE, suggesting that their effects could occur through the ecdysteroids pathways, and so impact the molt process of G. pulex on the long term. Finally, the present study is a first step in the possibility of using MIH and EcR relative expressions as biomarkers of exposure for EDCs risk assessment. However additional studies must first be carried out to better characterize and understand their variations, and also better predicted consequences for the exposed amphipods.

Effects of cyproterone acetate and vertically transmitted microsporidia parasite on Gammarus pulex sperm production.

Endocrine disruption compounds (EDCs) and parasitism can both interfere with the reproduction process of organisms. The amphipod Gammarus pulex is the host of the vertically transmitted microsporidia Dictyocoela duebenum, and this work was devoted to the investigation of the effect of an exposure to the anti-androgen compound, cyproterone acetate (CPA), and/or of the presence of D. duebenum on the spermatozoa production and length. Significant reduction of the spermatozoa production was observed when G. pulex males were uninfected and exposed to CPA. There also appeared a lower number of spermatozoa when D. duebenum infects G. pulex, whatever the exposure condition. Moreover, we highlighted that CPA has no effect on spermatozoa production when males are infected by D. duebenum, and no treatment has impacted the spermatozoa length. Our results suggest CPA and D. duebenum could impact the endocrine system of G. pulex and especially processes close to the spermatozoa production (e.g., androgenic gland, androgen gland hormone released, gonad-inhibiting hormone synthesized by X-organ). However, as no mechanism of action was highlighted, further testing need to be performed to improve the understanding of their impacts. Finally, results confirm that vertically transmitted microsporidia could be a confounding factor in the endocrine disruption assessments in Gammaridae.

Bioaccumulation, distribution and elimination of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii: Field and laboratory studies.

Chlordecone is a persistent organochlorine pesticide that has been widely used in Guadeloupe (French West Indies) to control the banana weevil Cosmopolites sordidus from 1972 to 1993. A few years after its introduction, widespread contamination of soils, rivers, wild animals and aquatic organisms was reported. Although high chlordecone concentrations have been reported in several crustacean species, its uptake, internal distribution, and elimination in aquatic species have never been described. This study aimed at investigating the accumulation and tissue distribution of chlordecone in the giant freshwater prawn Macrobrachium rosenbergii, using both laboratory (30 days exposure) and field (8 months exposure) approaches. In addition, depuration in chlordecone-free water was studied. Results showed that chlordecone bioconcentration in prawns was dose-dependent and time-dependent. Moreover, females appeared to be less contaminated than males after 5 and 7 months of exposure, probably due to successive spawning leading in the elimination of chlordecone through the eggs. Chlordecone distribution in tissues of exposed prawns showed that cephalothorax organs, mainly represented by the hepatopancreas, was the most contaminated. Results also showed that chlordecone was accumulated in cuticle, up to levels of 40% of the chlordecone body burden, which could be considered as a depuration mechanism since chlordecone is eliminated with the exuviae during successive moults. Finally, this study underlined the similarity of results obtained in laboratory and field approaches, which highlights their complementarities in the chlordecone behaviour understanding in M. rosenbergii.

Proteomic response of Macrobrachium rosenbergii hepatopancreas exposed to chlordecone: Identification of endocrine disruption biomarkers?

The present work is the first study investigating the impacts of chlordecone, an organochlorine insecticide, on the proteome of the decapod crustacean Macrobrachium rosenbergii, by gel-free proteomic analysis. The hepatopancreas protein expression variations were analysed in organisms exposed to three environmental relevant concentrations of chlordecone (i.e. 0.2, 2 and 20µg/L). Results revealed that 62 proteins were significantly up- or down-regulated in exposed prawns compared to controls. Most of these proteins are involved in important physiological processes such as ion transport, defense mechanisms and immune system, cytoskeleton dynamics, or protein synthesis and degradation. Moreover, it appears that 6% of the deregulated protein are involved in the endocrine system and in the hormonal control of reproduction or development processes of M. rosenbergii (e.g. vitellogenin, farnesoic acid o-methyltransferase). These results indicate that chlordecone is potentially an endocrine disruptor compound for decapods, as already observed in vertebrates. These protein modifications could lead to disruptions of M. rosenbergii growth and reproduction, and therefore of the fitness population on the long-term. Besides, these disrupted proteins could be suggested as biomarkers of exposure for endocrine disruptions in invertebrates. However, further investigations are needed to complete understanding of action mechanisms of chlordecone on proteome and endocrine system of crustaceans.

Vitellogenin and vitellogenin receptor gene expression and 20-hydroxyecdysone concentration in Macrobrachium rosenbergii exposed to chlordecone.

Chlordecone is a persistent organochlorine pesticide widely used in Guadeloupe (French West Indies) to control the banana weevil Cosmopolites sordidus. Although it was previously highlighted that chlordecone may affect the reproduction and growth of vertebrate species, little information is available on the chlordecone effects in invertebrates. The present study investigated the effects of chlordecone on a hormone and a protein having key roles in reproduction and growth of the decapod crustacean Macrobrachium rosenbergii, by measuring the 20-hydroxyecdysone concentration, vitellogenin, and vitellogenin receptor gene expression, as well as the bioconcentration of chlordecone in exposed prawns. First, the results revealed that chlordecone was accumulated in M. rosenbergii. Then, it was found that Vg and VgR gene expression were increased in male and female M. rosenbergii exposed to chlordecone for 90 and 240 days, while the 20-hydroxyecdysone concentrations were decreased. This work suggests that chlordecone accumulates in prawn tissues and could affect key molecules involved in the reproduction and the growth of the invertebrate M. rosenbergii. However, many questions remain unresolved regarding the impacts of chlordecone on growth and reproduction and the signaling pathways responsible for these effects, as well as the potential role of confounding factors present in in situ studies.

Effects of chlordecone on 20-hydroxyecdysone concentration and chitobiase activity in a decapod crustacean, Macrobrachium rosenbergii.

Chlordecone (CLD) is an organochlorine insecticide abundant in aquatic environment of the French West Indies. However, few studies have investigated its impact on freshwater invertebrates. Whereas CLD is suspected of inducing endocrine disruption, this work aimed to study the effects of environmentally relevant concentrations of CLD on the 20-hydroxyecdysone (20-HE) hormone concentration and on the chitobiase activity, both having key roles in the molting process of crustaceans. In addition, the bioaccumulation of CLD was measured in the muscle tissue of Macrobrachium rosenbergii to underline potential dose-response relationship. The results have shown that CLD was bioaccumulated in exposed organisms according to a trend to a dose-response relationship. Moreover, it was observed that CLD decreased the 20-HE concentration in exposed prawns when compared to control, whatever the duration of exposure, as well as it inhibited the chitobiase activity after 30days of exposure. The present study indicates that CLD could interfere with molting process of M. rosenbergii by disturbing the 20-HE concentration and the activity of chitobiase, suggesting consequences at the long term on the shrimp development. This study also confirmed that CLD could be an endocrine disruptor in decapod crustaceans, as it was already observed in vertebrates.

Changes in antitoxic defense systems of the freshwater amphipod Gammarus pulex exposed to BDE-47 and BDE-99.

Polybrominated diphenyl ethers (PBDEs) are emerging pollutants widely distributed in aquatic environment. Although the bioaccumulation of this compound has been well studied, few studies have investigated their impacts on antitoxic systems of invertebrates, considering both genders. Here, we have evaluated the effects of BDE-47 and BDE-99 congeners on the antitoxic defence systems of the freshwater amphipod Gammarus pulex, and especially on the multixenobiotic resistance (MXR) activity, as well as the activities of two antitoxic enzymes, the glutathione-S-transferase (GST) and the glutathione peroxidases (GPx). Results revealed that BDE-47 and BDE-99 have inhibited the MXR activity whatever the gammarid gender, which could lead to a reduction of the pollutant elimination from the organism. In addition, a gender-biased response and a congener-biased effect on the antitoxic enzymes activities were observed. Indeed, both BDE congener exposures increased the GST activity in males, whereas in females, only BDE-99 congener modified this activity by decreasing it. On the contrary, BDE exposures did not impact the GPx activity in females, while in males BDE-99 has increased it. Results of the present study highlight that a PBDE exposure at 0.1 µg L(-1) modify antitoxic enzymes activities differently according to gender, which could lead to a change in G. pulex sensitivity on the long term. Finally, this work confirms the ecotoxicological implication of gender in the pollutant toxicity assessment, in order to evaluate impact on populations.

Effects of two PBDE congeners on the moulting enzymes of the freshwater amphipod Gammarus pulex.

Polybrominated diphenyl ethers (PBDEs) are abundant in aquatic environment. However, only few studies have investigated their impacts on freshwater invertebrates. This work aimed to study the effects of BDE-47 and BDE-99 congeners on the chitobiase and chitinolytic enzymes activities of the freshwater amphipod Gammarus pulex, according to gender, PBDE concentration and time of exposure. In addition, the bioaccumulation of BDE-47 and BDE-99 were measured. Results revealed that females have bioaccumulated more PBDE than males, and BDE-99 was more accumulated than BDE-47. PBDE exposures for 96 h have caused chitobiase and chitinolytic enzymes inhibition. This study not only indicate the importance of taking into account various confounding factors (gender, congeners, concentration) to understand PBDE effects, but underline also disruptions of molting enzymes activities. These disturbances suggest effects on the gammarid development and reproduction, and consequently effects on the gammarid population, and on a larger scale, a dysfunction of the ecosystem.

Biochemical and morphological responses in Chironomus riparius (Diptera, Chironomidae) larvae exposed to lead-spiked sediment.

The aim of the present study was to assess the potential use of biochemical markers and mentum deformities as indicators of long-term exposure to lead (Pb) in Chironomus riparius larvae. To do this, the authors measured 3 biochemical markers (i.e., malondialdehyde level, metallothionein concentration, and energy reserve content) as well as larval growth and mentum deformities after 16-d exposure to sediment containing Pb. The concentrations studied ranged from 3.5 mg/kg to 505.5 mg/kg dry weight. Despite the bioaccumulation of Pb in C. riparius bodies, frequencies of both mentum deformities and the dry weight were not significantly different between the control and stressed groups. On the contrary, Pb exposure caused a significant increase of both malondialdehyde level and metallothionein concentration. The increase of body Pb concentrations did not significantly modify body copper and zinc concentrations. Moreover, we observed a decrease of total lipid content and an increase of glycogen content as a function of a dose-response relationship, while no variation in protein concentrations was observed. Despite the adverse effects observed at the biochemical level, larval development was not affected. These results suggested that measurements of malondialdehyde level, metallothionein concentration, and energy reserve content can be used as relevant biomarkers of long-term sublethal exposure to Pb in C. riparius larvae.

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.