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Due to their specific effect on photosynthesis, herbicides pose a potential threat to coastal and estuarine microalgae. However, comprehensive understanding of the hazard and risk of these contaminants is currently lacking. Therefore the aim of the present study was to investigate the toxic effects of four ubiquitous herbicides (atrazine, diuron, Irgarol(®)1051 and isoproturon) and herbicide mixtures on marine microalgae. Using a Pulse Amplitude Modulation (PAM) fluorometry based bioassay we demonstrated a clear species and herbicide specific toxicity and showed that the current environmental legislation does not protect algae sufficiently against diuron and isoproturon. Although a low actual risk of herbicides in the field was demonstrated, monitoring data revealed that concentrations occasionally reach potential effect levels. Hence it cannot be excluded that herbicides contribute to observed changes in phytoplankton species composition in coastal waters, but this is likely to occur only occasionally., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Toxicants potentially decouple links between biodiversity and ecosystem processes. This study aimed to evaluate how toxicants affect invertebrate bioturbation and decomposition. Effects of copper on functionally distinct macrofaunal species (Asellus aquaticus and Tubifex spp.), decomposition (release of dissolved organic carbon, DOC) and Average Metabolic Response (AMR) and Community Metabolic Diversity (CMD) of bacteria were determined in 5-day microcosm experiments. Bioturbation was assessed as sediment redox potential (Eh) profiles. Concentration-response curves of the functional parameters DOC, and the faunal mediated AMR and CMD in the presence of Tubifex spp. depended on Tubifex spp. survival, i.e. similar EC50 values for both endpoints. In contrast, functional parameters in the presence of A. aquaticus were more sensitive than survival. Sediment Eh-profiles showed that reduced decomposition was caused by reduced sediment reworking by A. aquaticus at sub-lethal copper concentrations. These observations hint at a decoupling of invertebrate community structure and ecosystem functioning upon stress., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

The aim of the present study was to improve the cause-effect relationship between toxicant exposure and chironomid mouthpart deformities, by linking induction of mouthpart deformities to contaminated field sediments, metal mixtures and a mutagenic polycyclic aromatic compound metabolite (acridone). Mouthpart deformities in Chironomus riparius larvae were induced by both the heavy metal mixture and by acridone. A clear correlation between metal concentrations in the sediment and deformities incidence was only observed when the contaminated field sediments were left out of the analysis, probably because these natural sediments contained other toxic compounds, which could be responsible for a higher incidence of deformities than predicted by the measured metal concentrations only. The present study clearly improved the cause-effect relationship between toxicant exposure and the induction of mouthpart deformities. It is concluded that the incidence of mouthpart deformities may better reflect the potential toxicity of contaminated sediments than chemical analysis., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

During acute exposure, polycyclic aromatic compounds (PACs) act mainly by narcosis, but during chronic exposure the same compounds may exert sublethal life cycle effects. The aim of this study was therefore to evaluate the chronic effects of sediment spiked PACs on the emergence of the midge Chironomus riparius. Three isomer pairs were selected, and 28-day LC50 values and 50% emergence times (EMt50) were determined. Concentration-response relationships were observed for phenanthrene, acridine, phenanthridine and acridone. Anthracene and phenanthridone had no effect on total emergence, but did cause a delay in emergence. Calculated porewater LC50 values correlated well with logKow values, suggesting narcosis as mode of action. In contrast, effect concentrations for delay in emergence (EMt50) deviated from narcosis, suggesting a specific mode of action during chronic exposure. It is concluded that emergence is a powerful endpoint to detect life cycle effects of PACs on a key sediment inhabiting invertebrate.

The aim of the present study was to investigate algal-bacterial interactions in a gradient of metal contaminated natural sediments. By means of multivariate techniques, we related the genetic structure (denaturing gradient gel electrophoresis, DGGE) and the physiological structure (community-level physiological profiling, CLPP) of the bacterial communities to the species composition of the algal communities and to the abiotic environmental variables, including metal contamination. The results revealed that genetic and physiological structure of the bacterial communities correlated with the species composition of the algal community, but hardly to the level of metal pollution. This must be interpreted as an indication for a strong and species-specific linkage of algal and bacterial species in floodplain sediments. Metals were, however, not proven to affect either the algal or the bacterial communities of the Dutch river floodplains.

This study reports on the development and application of a whole sediment toxicity test using a benthic cladoceran Chydorus sphaericus, as an alternative for the use of pelagic daphnids. A C. sphaericus laboratory culture was started and its performance under control conditions was optimised. The test was firstly validated by determining dose-response relationships for aqueous cadmium and copper and ammonia, showing a sensitivity of C. sphaericus (96 h LC(50) values of 594 microg Cd/L, 191 microg Cu/L and 46 mg ammonia/L at pH 8) similar to that of daphnids. Next, sediment was introduced into the test system and a series of contaminated sediments from polluted locations were tested. A significant negative correlation between survival and toxicant concentrations was observed. It is concluded that the test developed in the present study using the benthic cladoceran C. sphaericus is suitable for routine laboratory sediment toxicity testing.

Potential of bacterial communities in biofilms to recover after copper exposure was investigated. Biofilms grown outdoor in shallow water on glass dishes were exposed in the laboratory to 0.6, 2.1, 6.8 micromol/l copper amended surface water and a reference and subsequently to un-amended surface water. Transitions of bacterial communities were characterised with denaturing gradient gel electrophoresis (DGGE) and community-level physiological profiles (CLPP). Exposure to 6.8 micromol/l copper provoked distinct changes in DGGE profiles of bacterial consortia, which did not reverse upon copper depuration. Exposure to 2.1 and 6.8 micromol/l copper was found to induce marked changes in CLPP of bacterial communities that proved to be reversible during copper depuration. Furthermore, copper exposure induced the development of copper-tolerance, which was partially lost during depuration. It is concluded that bacterial communities exposed to copper contaminated water for a period of 26 days are capable to restore their metabolic attributes after introduction of unpolluted water in aquaria for 28 days.

The benthic diatom Gomphonema parvulum Kützing is a common species in both clean and metal contaminated rivers. Our aim was to investigate whether metal-induced tolerance could explain the persistance of this taxon under metal polluted conditions. G. parvulum strains were isolated from a Zn- and Cd-contaminated stream and from a relatively clean ("reference") stream. The strains were cultured in synthetic medium as mono-specific biofilms to maintain their specific benthic growth features. Moreover, the strain from the metal polluted stream was cultured in plain and Zn- and Cd-enriched synthetic medium. Short-term (5 h) toxicity experiments with Zn were performed with the strains using pulse amplitude modulated (PAM) fluorometry. Zn lowered significantly the minimal chlorophyll fluorescence (F0) and the photon yield (phi(p)) of the exposed strains after 5 h exposure. The actual Zn concentrations that caused a 50% reduction (EC50's) of the phi(p) of the strain from the metal polluted stream were significantly higher than those of the isolate from the unpolluted stream. The absence of tolerance to Cu of the "polluted" strain indicated that Zn tolerance resulted from specific induction by chronic exposure to Zn in the field. Observations on field biofilms confirmed a higher tolerance of the G. parvulum population from the polluted stream than of the G. parvulum population from the reference stream. A genetic nature of this metal adaptation was supported by the persistance of the Zn tolerance of the polluted strain 2 years after isolation.

Toxic and genotoxic effects of alachlor, metolachlor, amitraz, chlordimeform, their respective environmentally stable degradation products 2,6-diethylaniline, 2-ethyl-4-methylaniline, 2,4-dimethylaniline, and two other related compounds, 3,4-dichloroaniline and aniline were compared. Acute toxicity tests with Chironomus riparius (96 h) and Vibrio fischeri (Microtox) and genotoxicity tests with a dark mutant of V. fischeri (Mutato) were carried out. Our results demonstrate that toxicity and genotoxicity of the pesticides are retained upon degradation to their alkyl-aniline metabolites. In the case of the herbicides alachlor and metolachlor, the toxicity to V. fischeri was enhanced upon degradation. Narcosis alone explains toxicity of the compounds to the midge, but not so for the bacteria suggesting a disparity in the selectivity of the test systems. All compounds showed direct genotoxicity in the Vibrio test. but amitraz and its metabolite were genotoxic at concentrations 10(3)-10(5) lower than all the other compounds. The observations indicate that stable aniline degradation products of the pesticides may contribute considerably to environmental risks of pesticides application and that genotoxic effects may arise upon degradation of pesticides.

The ability of the non-biting midge Chironomus riparius to survive and reproduce in metal polluted lowland rivers facilitates the opportunity to study micro-evolutionary processes in situ. However, due to larval drift, adapted midge populations are subject to regular immigration of non-adapted specimens from clean upstream river reaches. To examine the influence of non-adapted genes in adapted midge populations on the level of metal adaptation, an upstream and downstream chironomid population were crossbred on eight separate occasions in the laboratory to mimic gene flow. Several life-history characteristics, indicating adaptation to metals, were followed seasonally in the parental strains as well as in the reciprocal crossings. Such crossings were done over a 14-month period and maternal effects were found to be absent, indicating a major genetic component for the increased metal tolerance in the exposed midge populations. Furthermore, results confirmed the presence of adaptation to metals in exposed chironomids. However, a rapid loss of metal adaptation in the first generation hybrid offspring was clearly demonstrated. Consequently, the large temporal variation in metal adaptation in midge populations from the river can be explained by the earlier reported seasonal variations in selection pressure and immigration rates from non-adapted sub-populations.

Abiotic transformation of azaarenes in the environment has been analysed extensively, but metabolism is less well described. To further elucidate preliminary observations of interspecific differences in azaarene metabolism by aquatic organisms, phenanthridine biotransformation by midge larvae and carp was studied. In both experiments, 6(5H)-phenanthridinone (phenanthridone) was found as an important metabolite. The fish were clearly capable of metabolising phenanthridine, but in the midge experiment the metabolite was principally formed by bacteria growing on the food and not by the midges. Phenanthridone itself was further degraded to non-observed compounds in both experiments, due to bacteria and midges acting together in the midge experiment, and by carp in the fish experiment. Internal concentrations of phenanthridine and phenanthridone were non-detectable in the midge larvae, but concentrations of both compounds in carp organs suggested a major role of bile and liver. Since phenanthridone did not account for all phenanthridine loss, it was suggested that, apart from phenanthridone degradation, other metabolic pathways may play a role. This study clearly demonstrates the importance of interspecies differences in metabolism, which should not be neglected in risk assessment.

This study reports the effects of two model toxicants, copper and diazinon, on two characteristic riverine insect species, the caddisfly Cyrnus trimaculatus and the mayfly Ephoron virgo. It was demonstrated that these species are very sensitive to both compounds in comparison with aquatic insects traditionally used in ecotoxicity tests. For diazinon, the 96-h LC(50) value of Cyrnus trimaculatus (1.1 microg/l) is lower than for any other insect species known from the literature and for copper it was demonstrated that Ephoron virgo is among the most sensitive aquatic insect species. The observed low LC(50) values stress the importance of using these indigenous species in assessing the risk of environmental contaminants in large European rivers and in defining conditions for ecological recovery.

Invertebrate communities in polluted rivers are often exposed to a wide variety of compounds. Due to complex interactions, 'pollution tolerant' species are not necessarily the most tolerant species to toxicants tested under standard laboratory conditions. It was hypothesized that the distribution of species in polluted rivers is not only dependent on the tolerance of species to toxicants, but also on species-specific capacities to modify or compensate for negative effects of toxicants. To test this hypothesis, species-specific responses to metals in organically enriched river water were studied under controlled conditions. The zebra mussel Dreissena polymorpha and the midge Chironomus riparius were exposed to metal-polluted water from the River Dommel. Additionally, the (interactive) effects of metals and humic acids (HA) on both species were evaluated. In spite of a lower tolerance of Chironomus riparius to metals in laboratory studies, the midge was the most tolerant of the two test species to metal-polluted site water. The results indicated that the sensitivities of the two test species determined in laboratory tests were inversely related to their sensitivities to polluted river water. In accordance with these results, midge larvae were protected from copper (Cu) toxicity by HA, while metal toxicity was not reduced (Cu) or even amplified (cadmium) by HA for the zebra mussel. Thus, the presence of (naturally occurring) HA in site water may partly account for discrepancies between responses of species to bioassays and toxicity tests. It is suggested that these differences in responses to metals in site water are strongly influenced by species-specific preferences for organic compounds (like HA). It is concluded that the response to organic compounds present in site water largely determines whether a species is classified as 'pollution tolerant' or 'pollution sensitive'.

The toxicity of the nitrogen (N) heterocyclic polyaromatic hydrocarbon (PAH) acridine was tested for seven species of microalgae: Scenedesmus acuminatus, Selenastrum capricornutum, Chlamydomonas eugametos, Staurastrum chaetoceras, Staurastrum manfeldtii, Navicula salinarum and Nitzschia sigma. The effect of acridine on the algae was studied in a 96-h growth test, in which growth rates were determined using cell numbers and biovolume. The obtained EC50 values (for growth rates based on cell numbers) ranged from 0.08 mg litre(-1) for N. sigma to 0.78 mg litre(-1) for C. eugametos and N. salinarum. Effect concentrations based on biovolume were slightly higher for most species. Metabolism of acridine was observed for one species (S. capricornutum), but this capacity did not result in a very different tolerance. Acridine toxicity was neither related to taxonomical background (green algae versus diatoms) nor to original habitat of the species (planktonic or benthic, eutrophic or oligo-mesotrophic). The presence of near-UV radiation during the incubation might explain the higher toxicity of acridine than is expected on basis of QSAR derived narcotic toxicity.