Berta Bonet

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.

Discharge and the response of biofilms to metal exposure in Mediterranean rivers

The expected response of fluvial biofilms to the environment and metal pollution prevailing under different discharge conditions was investigated. The relationship between inter-annual hydrological variability and metal concentration in water and sediments was explored in Mediterranean rivers (Catalonia, NE Spain) affected by low but chronic metal pollution, using monitoring data provided by the Catalan Water Agency (ACA). During the period investigated (2000–2006), metal pollution was characterized by low water concentrations and high concentrations in sediments. The most consistent pattern was observed for sediment cadmium (Cd) concentrations, showing a positive relationship with annual discharge, reaching values of environmental concern (above ecotoxicological benchmarks). A different pattern was observed for Cu, Zn, and As increasing with flow in some sites and decreasing in others. While Cd seems to proceed from diffuse sources being washed by surface runoff, Zn, Pb, and As may proceed from either diffuse or point-sources in the different river sites investigated. The relevance of diffuse metal pollution in the area of study indicates that polluted landfills runoff might be an important source of metals causing repetitive pulses of high metal concentration in the receiving water courses. The experimental results presented demonstrate that metal effects in fluvial biofilms may be accumulative, increasing the toxicity after repetitive pulse exposures. Since draughts and extreme rain events are expected to increase at higher latitudes due to global change, the sources of metal pollution, its final concentration and potential effects on the fluvial ecosystem may also change following the patterns expected for human-impacted Mediterranean rivers.

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.

Metal Ecotoxicology in Fluvial Biofilms: Potential Influence of Water Scarcity

Human activity is responsible for the entrance of toxic substances into aquatic ecosystems. These substances entail a risk for the components of the ecosystem (toxicological stress). As a result of global change, aquatic ecosystems are under strong environmental stress due to changes in water flow or nutrient concentration among others. This chapter presents a review of experimental and field studies addressing metal effects on fluvial biofilms and their implications for understanding the potential influence of water scarcity on the fate and effects of metals in fluvial systems. Water scarcity might increase metal exposure (due to low dilution), uptake (due to higher retention under low flow), toxicity and/or accumulation (depending on the dose and time of exposure) but may also cause opposite effects depending on the source of pollution. In addition, the influence that water scarcity might have on nutrient loads will also modulate the fate and effects of metals. Future studies addressing the role of environmental stress on the effects of toxicants at a community scale will be fundamental to predict the impact of toxicants in the aquatic ecosystems.

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.

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.

Cumulative Stressors Trigger Increased Vulnerability of Diatom Communities to Additional Disturbances

Chronic, non-lethal stressors occurring gradually (in space or time) can result in cumulative impacts that are more dramatic than higher intensities or occasional critical levels of any single one of these stressors. The negative effects of the chronic stressors trigger lasting impacts that may grow in intensity and become problematic over time and/or to higher trophic levels. In rivers, aquatic organisms experience this type of cumulative stress along the up- to downstream gradient in natural and anthropogenic contaminants generally observed in inhabited watersheds. Diatoms are a major component of the periphyton in rivers; their richness and diversity in natural communities are directly related to their varied ecological preferences and sensitivity to disturbance. In this study, we monitored from 2003 to 2008 the changes in the diversity of taxonomic and non-taxonomic features along a small river (Riou-Mort, South West France), at three sites: one site upstream considered as a reference for this watershed, one intermediate site with high nutrient load, and one downstream site exposed to both nutrient and metal pollution. The cumulative impacts of nutrients plus metals led to a gradual decrease in species richness and diversity, and in a potential capacity to cope with additional stresses, e.g., climate change-related ones. This is reflected by a decrease in species richness downstream, more dramatic in the hot summer of 2003 than in cooler summers. With the increasingly protective environmental regulations (e.g., Water Framework Directive in Europe), accumulation of stresses on aquatic resources are recommended to receive increasing attention, in particular considering the expected changes in climate.

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.