Multibiomarker assessment of cerium dioxide nanoparticle (nCeO2) sublethal effects on two freshwater invertebrates, Dreissena polymorpha and Gammarus roeseli.

Cerium nanoparticles (nCeO2) are widely used in everyday products, as fuel and paint additives. Meanwhile, very few studies on nCeO2 sublethal effects on aquatic organisms are available. We tried to fill this knowledge gap by investigating short-term effects of nCeO2 at environmentally realistic concentrations on two freshwater invertebrates; the amphipod Gammarus roeseli and the bivalve Dreissena polymorpha, using an integrated multibiomarker approach to detect early adverse effects of nCeO2 on organism biology. Differences in the behaviour of the organisms and of nanoparticles in the water column led to differential nCeO2 bioaccumulations, G. roeseli accumulating more cerium than D. polymorpha. Exposure to nCeO2 led to decreases in the size of the lysosomal system, catalase activity and lipoperoxidation in mussel digestive glands that could result from nCeO2 antioxidant properties, but also negatively impacted haemolymph ion concentrations. At the same time, no strong adverse effects of nCeO2 could be observed on G. roeseli. Further experiments will be necessary to confirm the absence of severe nCeO2 adverse effects in long-term environmentally realistic conditions.

Community structure and nutrient level control the tolerance of autotrophic biofilm to silver contamination.

Autotrophic biofilms are complex and fundamental biological compartments of many aquatic ecosystems. Since microbial species differ in their sensitivity to stressors, biofilms have long been proposed for assessing the quality of aquatic ecosystems. Among the many stressors impacting aquatic ecosystems, eutrophication and metal pollution are certainly the most common. Despite that these stressors often occur together, their effects on biofilms have been far much studied separately than interactively. In this study, we evaluated the interactive effects of silver (Ag), a reemerging contaminant, and phosphorus (P), a nutrient often associated with freshwater eutrophication, on the structure and functioning of two types of autotrophic biofilms, one dominated by diatoms and another one dominated by cyanobacteria. We hypothesized that P would alleviate the toxic effects of Ag, either directly, through the contribution of P in metal detoxification processes, or indirectly, through P-mediated shifts in biofilm community compositions and associated divergences in metal tolerance. Results showed that Ag impacted biofilm community structure and functioning but only at unrealistic concentrations (50 µg/L). P availability led to significant shifts in biofilm community composition, these changes being more pronounced in diatom- than those in cyanobacteria-dominated biofilm. In addition, P tended to reduce the impact of Ag but only for the cyanobacteria-dominated biofilm. More generally, our results highlight the preponderant role of the initial community structure and nutrient level on biofilm response to metallic pollutants.

Phosphorus availability modulates the toxic effect of silver on aquatic fungi and leaf litter decomposition.

The functioning of forested headwater streams is intimately linked to the decomposition of leaf litter by decomposers, mainly aquatic hyphomycetes, which enables the transfer of allochthonous carbon to higher trophic levels. Evaluation of this process is being increasingly used as an indicator of ecosystem health and ecological integrity. Yet, even though the individual impacts of contaminants and nutrient availability on decomposition have been well studied, the understanding of their combined effects remains limited. In the current study, we investigated whether the toxic effects of a reemerging contaminant, silver (Ag), on leaf litter decomposition could be partly overcome in situations where microorganisms were benefitting from high phosphorus (P) availability, the latter being a key chemical element that often limits detritus decomposition. We also investigated whether these interactive effects were mediated by changes in the structure of the aquatic hyphomycete community. To verify these hypotheses, leaf litter decomposition by a consortium of ten aquatic hyphomycete species was followed in a microcosm experiment combining five Ag contamination levels and three P concentrations. Indirect effects of Ag and P on the consumption of leaf litter by the detritivorous crustacean, Gammarus fossarum, were also evaluated. Ag significantly reduced decomposition but only at the highest concentration tested, independently of P level. By contrast, P and Ag interactively affected fungal biomass. Both P level and Ag concentrations shaped microbial communities without significantly affecting the overall species richness. Finally, the levels of P and Ag interacted significantly on G. fossarum feeding rates, high [Ag] reducing litter consumption and low P availability tending to intensify the feeding rate. Given the high level of contaminant needed to impair the decomposition process, it is unlikely that a direct effect of Ag on leaf litter decomposition could be observed in situ. However, subtle Ag effects in relation to nutrient levels in ecosystems could be expected. In particular, owing to higher consumption of low P leaf litter, shredding invertebrates could increase the ingestion of contaminated resources, which could, in turn, represent an important threat to headwater stream ecosystems.

Behavioural and physiological responses of Gammarus fossarum (Crustacea Amphipoda) exposed to silver.

The study aims at investigating the effects of silver (Ag), a re-emerging contaminant, on physiological and behavioural responses in Gammarus fossarum. In a first experiment, G. fossarum Ag LC50s were evaluated during 96 h under semi-static mode of exposure. Juveniles appeared to be more sensitive to Ag (LC5096h: 1.01 µg L(-1)) than ovigerous females (LC5096h: 1.9 µg L(-1)) and adult males (LC5096h: 2.2 µg L(-1)). In a second experiment, the physiological (osmo-/ionoregulation; antioxidant enzymes; lipid peroxidation (LPO)) and behavioural (locomotor activity and ventilation) responses of male G. fossarum exposed to Ag (0, 0.5, 1, 2, and 4 µg L(-1)) were investigated. The mortality and Ag bioconcentration of gammarids exposed to Ag were significantly higher than controls. Concerning physiological responses, a 48 h-exposure to Ag had no impact on catalase activity but led to a significant decrease of haemolymph osmolality and [Na(+)]. On the contrary, LPO, Se-GPx and Na(+)/K(+)-ATPase activity were significantly increased. Behavioural responses, such as locomotor and ventilatory activities, were also significantly reduced in Ag exposed gammarids. After 96 h-exposure, especially to 0.5 µg Ag L(-1), most responses (ventilation, locomotor activity, haemolymph osmolality and [Na(+)]) were even more pronounced and haemolymph [Cl(-)] was significantly decreased but, contrary to observations after 48 h-exposure, Na(+)/K(+)-ATPase activity was significantly reduced. Our results demonstrate the drastic effects of realistic [Ag] concentration (0.5 µg Ag L(-1)) on an ubiquitous and functionally important freshwater invertebrate (implied in detritus breakdown), but also strongly suggest an energetic reallocation to the detriment of locomotor activity and in favour of maintenance functions (i.e., osmoregulation and detoxification). These results highlight the risk represented by Ag and the need to perform integrated studies (at different scales, from individual to ecosystem).

Effects of deltamethrin (pyrethroid insecticide) on growth, reproduction, embryonic development and sex differentiation in two strains of Daphnia magna (Crustacea, Cladocera).

Acute and different chronic ecotoxic effects of deltamethrin have been investigated on two strains (coming from two different laboratories) of Daphnia magna. The effective concentrations immobilizing 50% of daphnids (EC50s) after 24 h and 48 h were 9.40 and 0.32 µg L(-1), 8.86 and 0.63 µg L(-1) for first strain (strain 1) and second strain (strain 2), respectively. Thus, there was an increase of deltamethrin ecotoxicity with time of exposure as confirmed by chronic studies. After 21 days of exposure to deltamethrin, daphnids have showed significant effects on survival at deltamethrin concentrations of 0.16 µg L(-1) and 0.31 µg L(-1) for strains 1 and 2, respectively. Eleven other endpoints were examined: body length, population growth rate and various reproductive parameters (days to first brood, number of broods, number of cumulative molts and number of neonates), embryotoxicity and appearance of males. IC10 values related to the number of juveniles per live adult were 11 and 46 ng L(-1) for strains 1 and 2, respectively. Furthermore, an increase in embryo deformities was observed at the highest concentrations tested for both strains. Following deltamethrin exposure, undeveloped second antennae, curved or unextended shell spines, and curved post abdomen spines were observed in live neonates. The production of male juveniles was only registered with strain 1 at 0.16 µg L(-1). Results suggest that deltamethrin could act as an endocrine disruptor in D. magna as it interferes with sex determination and development abnormality but there is a difference in sensitivity between the two tested strains.

Physiological and behavioural responses of Gammarus pulex (Crustacea: Amphipoda) exposed to cadmium.

The aim of this study was to investigate the effects of cadmium on physiological and behavioural responses in Gammarus pulex. In a first experiment, cadmium LC50s for different times were evaluated in 264 h experiment under continuous mode of exposure (LC50(96 h)=82.1 microgL(-1), LC50(120 h)=37.1 microgL(-1), LC50(168 h)=21.6 microgL(-1), LC50(264 h)=10.5 microgL(-1)). In a second experiment, the physiological and behavioural responses of the amphipod exposed to cadmium (0, 7.5 and 15 microgL(-1)) were investigated under laboratory conditions. The mortality and the whole body cadmium concentration of organisms exposed to cadmium were significantly higher than in controls. Concerning physiological responses, cadmium exposure exerted a significant decrease on osmolality and haemolymph Ca(2+) concentration, but not on haemolymph Na(+) and Cl(-) concentrations, whereas the Na(+)/K(+)-ATPase activity was significantly increased. Behavioural responses, such as feeding rate, locomotor and ventilatory activities, were significantly reduced in Cd exposed organisms. Mechanism of cadmium action and consequent energetic reallocation in favour of maintenance functions (i.e., osmoregulation) are discussed. The results of this study indicate that osmolality and locomotor activity in G. pulex could be effective ecophysiological/behavioural markers to monitor freshwater ecosystem and to assess the health of organisms.

Elevated aluminium concentration in acidified headwater streams lowers aquatic hyphomycete diversity and impairs leaf-litter breakdown.

Aquatic hyphomycetes play an essential role in the decomposition of allochthonous organic matter which is a fundamental process driving the functioning of forested headwater streams. We studied the effect of anthropogenic acidification on aquatic hyphomycetes associated with decaying leaves of Fagus sylvatica in six forested headwater streams (pH range, 4.3-7.1). Non-metric multidimensional scaling revealed marked differences in aquatic hyphomycete assemblages between acidified and reference streams. We found strong relationships between aquatic hyphomycete richness and mean Al concentration (r = -0.998, p < 0.0001) and mean pH (r = 0.962, p < 0.002), meaning that fungal diversity was severely depleted in acidified streams. By contrast, mean fungal biomass was not related to acidity. Leaf breakdown rate was drastically reduced under acidic conditions raising the issue of whether the functioning of headwater ecosystems could be impaired by a loss of aquatic hyphomycete species.

Physiological and behavioural responses of Gammarus pulex exposed to acid stress.

Physiological and behavioural responses of the acid-sensitive amphipod Gammarus pulex exposed to a wide range of acid conditions (pH 4.1, 5.1, 6.0) under laboratory conditions were investigated. An exposure of 38 h to acid conditions caused significant decreases in survival rate, osmolality, haemolymph Na+ concentration, ventilatory and locomotor activity compared to organisms exposed to a circumneutral medium (pH 7.9). We highlighted the interest of using individual response distribution, since drastic effects can be detected in organisms exposed to pH 6.0, in particular for osmolality: The response can be divided into two groups, unimpacted and impacted organisms. Moreover this representation permitted to evaluate the health level of individual organisms through the determination of threshold values. Significant correlations between mean pH and mean physiological/behavioural responses were observed. The relationships between individual responses permitted not only to compare endpoints, but also to show that affected organisms were impacted by ionoregulation failure, hypoventilation and low locomotor activity. The energetic reallocation in favour of maintenance functions, such as osmoregulation, is discussed. The results of this study indicate that the values of haemolymph Na+ concentration, osmolality and locomotor activity in G. pulex could be effective ecophysiological/behavioural markers to monitor freshwater ecosystems and to assess the health of organisms or populations.

Physiological recovery from episodic acid stress does not mean population recovery of Gammarus fossarum.

The physiological responses of the acid-sensitive amphipod Gammarus fossarum exposed in situ to acid stress (pH4.5 and 5.5) and then transferred back to neutral water were investigated. Survival rate and haemolymph [Cl(-)] and [Na(+)] were assessed after 24, 48 and 72h of exposure in acidic streams and after a recovery period of 12, 24, 36, 48 and 60h. After 24h, exposure to slightly acidic (pH5.5) and strongly acidic water (pH4.5) led to a severe and significant depletion in haemolymph [Na(+)] and [Cl(-)] compared to organisms exposed in circumneutral water (pH7.3). However, after only a 12h-period of transfer back in neutral water and whatever the previous exposure time (24, 48 and 72h) in both slightly and strongly acidic water, haemolymph [Na(+)] and [Cl(-)] were equal or superior to the control level without associated mortality. In spite of this fast physiological recovery capacity, populations of G. fossarum living in streams undergoing episodic acid stresses were drastically affected thus, demonstrating the high acid-sensitivity of this species. We discuss the possible reasons of population regression and the absence of population recovery.

Short-term physiological responses to a severe acid stress in three macroinvertebrate species: a comparative study.

The present study focuses on the sensitivity among freshwater invertebrate species to acidic stress. Three common macroinvertebrate species in the Vosges Mountains (North-Eastern France), Gammarus fossarum (Amphipoda), Hydropsyche pellucidula (Trichoptera) and Dinocras cephalotes (Plecoptera) were exposed for 24, 72 and 120 h to natural acidified water (pH=4.73+/-0.08, [Ca2+]=39.1+/-0.6 micromol l(-1), [Al(tot)]=28.4+/-1 micromol l(-1)). Short-term exposure to acid stress caused significant decreases both in survival rate and haemolymph ions ([Cl-] and [Na+]). The relative sensitivity to a natural acidic stress slightly differed among the species and was in the following order: G. fossarum, as the most sensitive, then H. pellucidula and D. cephalotes. Results of this study confirm the interest of in situ tests to assess the toxicity of short-term acid exposure. Finally, our results reinforce the hypothesis that transient acidification can offset the recovery of sensitive species of macroinvertebrates in streams chemically recovering from acidification either through liming or declining deposition.

Hyperventilation and loss of hemolymph Na+ and Cl- in the freshwater amphipod Gammarus fossarum exposed to acid stress: a preliminary study.

The effect of acidification on the acid-sensitive species Gammarus fossarum was investigated in the laboratory. The results showed that as mortality increased, mean hemolymph chloride and sodium concentrations decreased rapidly. Concomitantly, organisms hyperventilated during the first 24 h and then started to hypoventilate. These results demonstrated that exposure to acid stress in the acid-sensitive species G. fossarum led to ion-regulatory and respiratory failure as previously reported in fish and crayfish exposed to acid stress.