Natàlia Corcoll

Copper accumulation and toxicity in fluvial periphyton: the influence of exposure history.

Periphyton communities have a good bioaccumulation capacity and can be used to monitor metal pollution in fluvial ecosystems. Depending on the dose and exposure time, metals may produce changes in the structure and function of these communities, thus it is expected that the kinetics of metal accumulation and metal sensitivity will also be influenced by the exposure history. In this study, the effects of pulsed and continuous Cu exposures during the colonization of the communities were investigated under controlled conditions. This investigation includes the study of metal accumulation kinetics and the evaluation of community tolerance. Pulsed copper exposure did not affect the community structure but influenced the accumulation kinetics (decreasing intracellular copper uptake). On the other hand, continuous copper exposure caused a huge increase in metal content (both total and intracellular) and modified the structure of the community (increasing the percentage of cyanobacteria and diatom diversity). Both pulsed and continuous periphyton metal exposure may have negative repercussions for the fluvial ecosystem. While Cu pulsed exposure may be toxic to periphyton communities, continuous exposures may lead to community adaptation, which is often related to changes in species composition and higher metal contents being transferred to higher trophic levels of the stream food chain.

Consistency in Diatom Response to Metal-Contaminated Environments

Diatoms play a key role in the functioning of streams, and their sensitivity to many environmental factors has led to the development of numerous diatom-based indices used in water quality assessment. Although diatom-based monitoring of metal contamination is not currently included in water quality monitoring programs, the effects of metals on diatom communities have been studied in many polluted watersheds as well as in laboratory experiments, underlying their high potential for metal contamination assessment. Here, we review the response of diatoms to metal pollution from individual level (e.g. size, growth form, and morphological abnormalities) to community structure (replacement of sensitive species by tolerant ones). These potential effects are then tested using a large, multi-country database combining diatom and metal information. Metal contamination proved to be a strong driver of the community structure, and enabled for the identification of tolerant species like Cocconeis placentula var. euglypta, Eolimna minima, Fragilaria gracilis, Nitzschia sociabilis, Pinnularia parvulissima, and Surirella angusta. Among the traits tested, diatom cell size and the occurrence of diatom deformities were found to be good indicators of high metal contamination. This work provides a basis for further use of diatoms as indicators of metal pollution.

Catalase in fluvial biofilms: a comparison between different extraction methods and example of application in a metal-polluted river

Antioxidant enzymes are involved in important processes of cell detoxification during oxidative stress and have, therefore, been used as biomarkers in algae. Nevertheless, their limited use in fluvial biofilms may be due to the complexity of such communities. Here, a comparison between different extraction methods was performed to obtain a reliable method for catalase extraction from fluvial biofilms. Homogenization followed by glass bead disruption appeared to be the best compromise for catalase extraction. This method was then applied to a field study in a metal-polluted stream (Riou Mort, France). The most polluted sites were characterized by a catalase activity 4–6 times lower than in the low-polluted site. Results of the comparison process and its application are promising for the use of catalase activity as an early warning biomarker of toxicity using biofilms in the laboratory and in the field.

Antioxidant enzyme activities as biomarkers of Zn pollution in fluvial biofilms

The potential of the antioxidant enzyme catalase (CAT) and ascorbate peroxidase (APX) as molecular biomarkers of Zn toxicity in freshwater biofilms has been explored in this study jointly with other classical functional and structural endpoints (photosynthetic parameters, algal group composition and bioaccumulation). Biofilms were colonized in an indoor microcosm system for 5 weeks and then exposed to Zn for 5 weeks. To evaluate Zn effects, biofilms were sampled 5 and 3 days before exposure, just before exposure (time 0), and after 6h, 1, 3, 7, 21 and 35 days of metal exposure. Most endpoints measured were affected by Zn exposure (320 μg Zn L(-1)) during both periods of exposure. APX was the only functional parameter responding after a few hours of Zn exposure, highlighting its use as an early toxicity biomarker. Structural changes began after 3 days of exposure, starting with a decrease in algal biomass and an increase in the OD 430:665 ratio. Structural changes in biofilm communities were observed after 1 week, leading to a shift from diatoms to cyanobacteria and green algae-dominated communities. CAT activity was thereafter enhanced (after three weeks of exposure) and attributed not only to a direct effect of Zn bioaccumulation but also to an indirect effect of the community composition changes driven by chronic metal exposure. It can be concluded that biofilm antioxidant enzyme activities may provide evidence of early stress caused by metal exposure and also provide information about the mechanism of community adaptation. This information can be of great interest to improve current tools used for risk assessment.

The Use of Photosynthetic Fluorescence Parameters from Autotrophic Biofilms for Monitoring the Effect of Chemicals in River Ecosystems

Photosynthetic processes play a key role in aquatic ecosystems. These processes are highly sensitive to the presence of toxicants, leading to an increase in their use as ecotoxicological endpoints. The use of chlorophyll-a fluorescence techniques to assess the impact of toxicants on the photosynthesis of the autotrophic component of fluvial biofilms has increased in the last decades. However, these photosynthetic endpoints are not currently used in water quality monitoring programs.

How to Link Field Observations with Causality? Field and Experimental Approaches Linking Chemical Pollution with Ecological Alterations

This chapter summarizes field and laboratory investigations dealing with metals and pesticides (90) and emerging compounds’ (10) effects on fluvial communities. The Arkansas River case study is a good example showing how field observations, together with long-term natural experiments and microcosm experiments, provide consistent evidence of metals effects on macroinvertebrate communities. In the case of biofilms, microcosm and mesocosm experiments confirm that metals and pesticides are responsible for the loss of sensitive species in the community, and that this influence is modulated by several biological and environmental factors. Information about the effects of emerging pollutants is very scarce, highlighting the existence of a missing gap requiring future investigations. The examples provided and the recommendations given are proposed as a general guide for studies aiming to link chemical pollution with ecological alterations.

Seasonal changes in antioxidant enzyme activities of freshwater biofilms in a metal polluted Mediterranean stream

While seasonal variations in fluvial communities have been extensively investigated, effects of seasonality on community responses to environmental and/or chemical stress are poorly documented. The aim of this study was to describe antioxidant enzyme activity (AEA) variability in fluvial biofilms over an annual cycle, under multi-stress scenarios due to environmental variability (e.g., light intensity, water flow, and temperature) and metal pollution (Zn, Mn and Fe). The annual monitoring study was performed at three sites according to their water and biofilm metal concentrations. Metal concentration was affected by water flow due to dilution. Low flow led to higher dissolved Zn concentrations, and thus to higher Zn accumulation in the biofilm. Water temperature, light intensity and phosphate concentration were the environmental factors which determined the seasonality of biofilm responses, whereas dissolved Zn and Zn accumulation in biofilms were the parameters linked to sites and periods of highest metal pollution. Community algal succession, from diatoms in cold conditions to green algae in warm conditions, was clearer in the non metal-polluted site than in those metal-polluted, presumably due to the selection pressure exerted by metals. Most AEA were related with seasonal environmental variability at the sites with low or no-metal pollution, except glutathione-S-transferase (GST) which was related with Zn (dissolved and accumulated in biofilm) pollution occurring at the most polluted site. We can conclude that seasonal variations of community composition and function are masked by metal pollution. From this study we suggest the use of a multi-biomarker approach, including AEA and a set of biological and physicochemical parameters as an effect-based field tool to assess metal pollution.

Light History Influences the Response of Fluvial Biofilms to Zn Exposure

Fluvial biofilms are subject to multistress situations in natural ecosystems, such as the co‐occurrence of light intensity changes and metal toxicity. However, studies simultaneously addressing both factors are rare. This study evaluated in microcosm conditions the relationship between short‐term light intensity changes and Zn toxicity on fluvial biofilms with long‐term photoacclimation to different light conditions. Biofilms that had long‐term photoacclimation to 25 μmol photons · m−2 · s−1 (low light [LL] biofilms), 100 μmol photons · m−2 · s−1 (medium light [ML] biofilms), and 500 μmol photons · m−2 · s−1 (high light [HL] biofilms) were characterized by different structural (Chlorophyll‐a [Chl‐a], total biomass‐AFDW, EPS, algal groups, and diatom taxonomy) and physiological attributes (ETR‐I curves and photosynthetic pigments). HL biofilms showed higher light saturation intensity and a higher production of xanthophylls than LL biofilms. In contrast, LL biofilms had many structural differences; a higher proportion of diatoms and lower AFDW and EPS contents than ML and HL biofilms. A clear effect of light intensity changes on Zn toxicity was also demonstrated. Zn toxicity was enhanced when a sudden increase in light intensity also occurred, mainly with LL biofilms, causing higher inhibition of both the Φ′PSII and the ΦPSII. A decoupling of NPQ from de‐epoxidation reaction (DR) processes was also observed, indicating substantial damage to photoprotective mechanisms functioning in biofilms (i.e., xanthophyll cycle of diatoms) due to Zn toxicity. This study highlights the need to take into account environmental stress (e.g., light intensity changes) to better assess the environmental risks of chemicals (e.g., metals).

Antioxidant enzyme activities in biofilms as biomarker of Zn pollution in a natural system: an active bio-monitoring study.

This study aimed to explore the use of antioxidant enzyme activities (AEA) and biofilm metal accumulation capacity in natural communities as effect-based indicator of metal exposure in fluvial systems. To achieve these objectives, an active biomonitoring using fluvial biofilm communities was performed during 5 weeks. Biofilm was colonized over artificial substrata in a non-polluted site. After 5 weeks, biofilms were translocated to four different sites with different metal pollution in the same stream. The evolution of environmental parameters as well as biofilm responses was analysed over time. Physicochemical parameters were different between sampling times as well as between the most polluted site and the less polluted ones, mainly due to Zn pollution. In contrast, AEA and metal accumulation in biofilms allowed us to discriminate the high and moderate metal pollution sites from the rest. Zn, the metal with the highest contribution to potential toxicity, presented a fast and high accumulation capacity in biofilms. According to the multivariate analysis, AEA showed different responses. While catalase (CAT) and ascorbate peroxidase (APX) variability was mainly attributed to environmental stress (pH, temperature and phosphate concentration), glutathione-S-transferase (GST) changes were related to metal pollution. Glutathione reductase (GR) and superoxide dismutase (SOD) responses were related to both stress factors. AEA and metal accumulation are proposed as sensitive effect-based field methods, to evaluate biofilm responses after acute metal exposure (e.g. an accidental spill) due to their capacity to respond after few hours, but also in routinely monitoring due to their persistent changes after few weeks of exposure. These tools could improve the Common Implementation Strategy (CIS) of the Water Framework Directive (WFD) as expert group request.