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Monitoring of chemicals in the aquatic environment by chemical analysis alone cannot completely assess and predict the effects of chemicals on aquatic species and ecosystems. This is primarily because of the increasing number of (unknown) chemical stressors and mixture effects present in the environment. In addition, the ability of ecological indices to identify underlying stressors causing negative ecological effects is limited. Therefore, additional complementary methods are needed that can address the biological effects in a direct manner and provide a link to chemical exposure, i.e. (eco)toxicological tests. (Eco)toxicological tests are defined as test systems that expose biological components (cells, individuals, populations, communities) to (environmental mixtures of) chemicals to register biological effects. These tests measure responses at the sub-organismal (biomarkers and in vitro bioassays), whole-organismal, population, or community level. We performed a literature search to obtain a state-of-the-art overview of ecotoxicological tests available for assessing impacts of chemicals to aquatic biota and to reveal datagaps. In total, we included 509 biomarkers, 207 in vitro bioassays, 422 tests measuring biological effects at the whole-organismal level, and 78 tests at the population- community- and ecosystem-level. Tests at the whole-organismal level and biomarkers were most abundant for invertebrates and fish, whilst in vitro bioassays are mostly based on mammalian cell lines. Tests at the community- and ecosystem-level were almost missing for organisms other than microorganisms and algae. In addition, we provide an overview of the various extrapolation challenges faced in using data from these tests and suggest some forward looking perspectives. Although extrapolating the measured responses to relevant protection goals remains challenging, the combination of ecotoxicological experiments and models is key for a more comprehensive assessment of the effects o

Benthic macroinvertebrate communities are used as indicators for anthropogenic stress in freshwater ecosystems. To better understand the relationship between anthropogenic stress and changes in macroinvertebrate community composition, it is important to understand how different stressors and species traits are associated, and how these associations influence variation in species occurrence and abundances. Here, we show the capacity of the multivariate technique of double constrained correspondence analysis (dc-CA) to analyse trait-environment relationships, and we compare it with the redundancy analysis method on community weighted mean values of traits (CWM-RDA), which is frequently used for this type of analysis. The analyses were based on available biomonitoring data for macroinvertebrate communities from the Danube River. Results from forward selection of traits and environmental variables using dc-CA analyses showed that aquatic stages, reproduction techniques, dispersal tactics, locomotion and substrate relations, altitude, longitudinal and transversal distribution, and substrate preferendum were significantly related to habitat characteristics, hydromorphological alterations and water quality measurements such as physico-chemical parameters, heavy metals, pesticides and pharmaceuticals. Environmental variables significantly associated with traits using the CWM-RDA method were generally consistent with those found in dc-CA analysis. However, the CWM-RDA does neither test nor explicitly select traits, while dc-CA tests and selects both traits and environmental variables. Moreover, the dc-CA analysis revealed that the set of environmental variables was much better in explaining the community data than the available trait set, a kind of information that can neither be obtained from CWM-RDA nor from RLQ (Environment, Link and Trait data), which is a close cousin of dc-CA but not regression-based. Our results suggest that trait-based analysis based on dc-CA may be

Urban rivers often function as sinks for various contaminants potentially placing the benthic communities at risk of exposure. We performed a comprehensive biological survey of the benthic macroinvertebrate and bacterial community compositions in six rivers from the suburb to the central urban area of Guangzhou city (South China), and evaluated their correlations with emerging organic contaminants, heavy metals and nutrients. Overall, the benthic macroinvertebrate community shifted from molluscs to oligochaete from the suburban to the central urban rivers that receive treated and untreated sewage. An exception was the site in the Sha River where chironomids were most abundant. The differences in macroinvertebrate community assemblages were significantly associated with chromium, total phosphorus, galaxolide, triclosan and sand content in the sediment. There was no significant difference in benthic macroinvertebrate composition between the dry and wet season. As assessed by double constrained ordination, sexual reproduction was the only trait of benthic macroinvertebrates that showed a significant correlation with pollution variables, as it was significantly positively correlated with chromium and total phosphorus. This suggests that r-strategist occurs in polluted sampling sites. The benthic bacterial community composition showed a significant difference between seasons and among the Liuxi River, Zhujiang River and central urban rivers. The differences in community composition of the benthic bacteria were significantly correlated with galaxolide, total phosphorus, lead and triclosan. These results suggest that input of treated and untreated sewage significantly altered the benthic macroinvertebrate and bacterial community compositions in urban rivers.

Triclosan (TCS, 5-chloro-2-(2,4-dichlorophenoxy) phenol) is a broad-spectrum antimicrobial compound. Owing to its wide use, TCS has been frequently detected in river systems, especially in the (sub-)tropics. However, little information on its interaction with river biofilm in the (sub)tropics is currently available. In the present study, subtropical river biofilms were chronically exposed to TCS for 14 d at concentrations of 0.1–100 μg/L in artificial river water, which was followed by a 7 d recovery period. The results show that 100 μg/L TCS inhibited the growth of river biofilms and the no-observed-effect concentration (NOEC) of TCS on river biofilms was 10 μg/L. The affected biofilms did not completely recover within the 7 d of recovery period due to the adsorbed TCS which was not removed together with dissolved TCS. Exposure to TCS caused significant changes in prokaryotic species composition of river biofilms but no significant effects on eukaryotic species composition. In particular, the relative abundance of several TCS-tolerant bacterial species (e.g., Pseudoxanthomonas mexicana, Sphingopyxis alaskensis and Sphingomonas wittichii) in river biofilms increased following exposure to 10 and 100 μg/L TCS. River biofilm efficiently removed TCS from the liquid phase and the pH values of the aquatic system significantly affected the removal efficiency of TCS (from 36% at pH 6.5 to 60% at pH 8.5). No degradation products were detected in the liquid phase after 5 days of exposure, possibly due to strong adsorption of the hydrophobic degradation products to river biofilms and through biodegradation by bacteria utilizing TCS and its degradation products as source of carbon and energy for growth, such as Methyloversalitis universalis and Methylobacterium aquaticum.

The response of sediment bacterial communities in subtropical freshwater benthic microcosms to sediment-associated triclosan (TCS; 28 d exposure) was analysed using Illumina high-throughput sequencing. This study highlights the interactive effects of TCS and the presence of benthic macroinvertebrates (Limnodrilus hoffmeisteri and Viviparidae bellamya) on sediment bacterial communities. Our results show that TCS alone significantly altered the taxonomic composition and decreased alpha diversity of sediment bacterial communities at concentrations ≥80 μg TCS/g dry weight (dw) sediment (sed). Regarding dominant phyla, TCS significantly reduced the relative abundance of Bacteroidetes and Firmicutes at these concentrations, whereas the relative abundance of Chloroflexi and Cyanobacteria increased. In the presence of benthic macroinvertebrates, the sediment bacterial community was affected by 8 μg TCS/g dw sed as well. However, the presence of benthic macroinvertebrates did not cause measurable changes to bacterial community in unspiked (i.e., control) sediment. These results indicate that TCS alone would not alter the sediment bacterial community at environmentally relevant concentrations (up till 8 μg/g dw sed), but may have an effect in combination with the presence of benthic macroinvertebrates. Therefore, we recommend to include benthic macroinvertebrates when assessing the response of sediment bacterial communities during exposure to environmental stress such as organic contaminants.

Galaxolide (HHCB) is used as a fragrance ingredient in household and personal care products, and has been ubiquitously detected in the environment. Here we investigated the fate of HHCB in subtropical freshwater microcosms, and evaluated effects of sediment-associated HHCB on a biological community consisting of algae, Daphnia, benthic macroinvertebrates and bacteria. The concentrations of sediment-associated HHCB did not change significantly during a 28 days exposure period, but HHCB accumulated in worms with biota-sediment accumulation-factor (BSAF) values in the range of 0.29–0.66 for Branchiura sowerbyi and 0.94–2.11 for Limnodrilus hoffmeisteri. There was no significant effects of HHCB (30 μg/g dry weight (dw) sediment) on chlorophyll-a content, sediment bacterial community composition, and survival and growth of benthic macroinvertebrates. However, the presence of benthic macroinvertebrates altered the sediment bacterial community structure relative to microcosms without introduced organisms. The findings of this study suggest that a single high-dose of HHCB, over 28 days, at environmentally relevant concentrations would not impose direct toxicological risks to aquatic organisms such as benthic macroinvertebrates.

Personal care products (PCPs) are ubiquitous in the environment due to their wide use in daily life. However, there are insufficient sediment toxicity data of PCPs under ecologically relevant conditions. Here we used Fourier transform infrared spectroscopy (FTIR) to investigate the sediment toxicity of triclosan (TCS) and galaxolide (HHCB) to two freshwater benthic macroinvertebrates, Limnodrilus hoffmeisteri and Branchiura sowerbyi, in microcosms containing a diverse biological community. Exposure to 8 µg TCS/g and 100 µg HHCB/g dry weight (dw) sediment induced significant biochemical alterations in the L. hoffmeisteri tissue. 8 µg TCS/g primarily affected proteins and nucleic acid while 100 µg HHCB/g mainly affected proteins and lipids of L. hoffmeisteri. However, 0.8 µg TCS/g and 30 µg HHCB/g did not cause significant subcellular toxicity to L. hoffmeisteri. In contrast, exposure of B. sowerbyi to 30 µg HHCB/g led to significant biochemical changes, including proteins, polysaccharides and lipids. Therefore, B. sowerbyi was more sensitive to sediment-associated HHCB than L. hoffmeisteri. Such effects were significantly enhanced when the HHCB concentration increased to 100 µg/g dw where death of B. sowerbyi occurred. These results demonstrate the application of FTIR spectroscopy to sediment toxicity testing of chemicals to benthic invertebrates with biochemical alterations as endpoints that are more sensitive than standard toxic endpoints (e.g., survival and growth).

Personal care products are widely used in our daily life in considerable quantities and discharged via the down-the-drain route to aquatic environments, resulting in potential risks to aquatic organisms. We investigated bioaccumulation and biotransformation of two widely used personal care products, triclosan (TCS) and galaxolide (HHCB) spiked to sediment, in the oligochaete worm Limnodrilus hoffmeisteri in water/sediment microcosms. After 7 days of sediment exposure to 3.1 μg of TCS or HHCB/g of dry weight sediment, the accumulation of TCS and HHCB in L. hoffmeisteri reached equilibrium, at which point the biota-sediment accumulation factors (BSAFs) were 2.07 and 2.50 for TCS and HHCB, respectively. The presence of L. hoffmeisteri significantly accelerated the dissipation of the levels of TCS and HHCB in the microcosms, with approximately 9.03 and 2.90% of TCS and HHCB, respectively, eliminated from the water/sediment systems after exposure for 14 days in the presence of worms. Two biotransformation products, methyl triclosan and triclosan O-sulfate, were identified for TCS in worm tissue, whereas only methyl triclosan was identified in the sediment. Unlike TCS, no evidence of biotransformation products was found for HHCB in either worm tissue or sediment. These experiments demonstrate that L. hoffmeisteri biotransformed TCS through methylation and sulfation, whereas HHCB biotransformation was undetectable.