Phylogenetic analysis of museum specimens of houting Coregonus oxyrinchus shows the need for a revision of its extinct status.

According to the IUCN Red List the anadromous houting Coregonus oxyrinchus is categorized as 'extinct'. However, this extinct status might be incorrect because taxonomic difference between C. oxyrinchus and the closely related C. lavaretus is based on a disputable morphological comparison. Also, phylogenetic studies on mtDNA only focused on recent obtained coregonids. We are the first to perform a mtDNA analysis on both historic and recent specimens, including the syntype specimen which was used for species description by Linnaeus originally. Two primer pairs for mitochondrial CytB and ND3 were used to extract sequences for phylogenetic analysis. Sequences from 14 out of 21 C. oxyrinchus museum specimens were successfully obtained and compared with sequences from recent obtained C. lavaretus. The sequences were combined with GenBank data from a previous phylogenetic study on houting to create a phylogenetic tree and two minimum spanning haplotype networks. Results show that C. oxyrinchus and C. lavaretus form a clade with limited genetic variation. Low bootstrap values also show weak support for geographical patterns in distribution of mitochondrial haplotypes. Statistical analysis of the haplotype networks also shows that historic and recent specimens are similar species. Our results suggest that C. oxyrinchus is a junior synonym of C. lavaretus. A definitive taxonomic revision could not be made because only CytB sequencing was successful for the syntype specimen. We discuss taxonomic consequences and the species-specific focus in nature conservation. We propose a shift in nature conservation to a more functional approach based on traits rather than species.

Enlarging the Arsenal of Test Species for Sediment Quality Assessment.

Since only a few standard benthic test species are available for sediment quality, our study aimed to employ multiple test species representing different sensitivity categories in the quality assessment of contaminated sediments. To this end three macroinvertebrate species, Sericostoma personatum (caddisfly, sensitivity category 10), Asellus aquaticus (isopod, category 3) and Chironomus riparius (chironomid, category 2), were exposed to sediments originating from various contamination sources in whole sediment bioassays using intact sediment cores. The agricultural sediment caused insect mortality, the agricultural and urban sediment caused isopod growth reduction and the urban and Wastewater Treatment Plant (WWTP) sediment affected chironomid emergence time. It is concluded that the arsenal of standard species can be successfully expanded by non-standard species, reducing over- or underestimation of the risks of contaminated sediments.

Contribution of sediment contamination to multi-stress in lowland waters.

Water bodies in densely populated lowland areas are often impacted by multiple stressors. At these multi-stressed sites, it remains challenging to quantify the contribution of contaminated sediments. This study, therefore, aimed to elucidate the contribution of sediment contamination in 16 multi-stressed drainage ditches throughout the Netherlands. To this end an adjusted TRIAD framework was applied, where 1) contaminants and other variables in the sediment and the overlying water were measured, 2) whole-sediment laboratory bioassays were performed using larvae of the non-biting midge Chironomus riparius, and 3) the in situ benthic macroinvertebrate community composition was determined. It was hypothesized that the benthic macroinvertebrate community composition would respond to all jointly present stressors in both water and sediment, whereas the whole-sediment bioassays would only respond to the stressors present in the sediment. The benthic macroinvertebrate community composition was indeed related to multiple stressors in both water and sediment. Taxa richness was positively correlated with the presence of PO4-P in the water, macrophyte cover and some pesticides. Evenness, the number of Trichoptera families and the SPEARpesticides were positively correlated to the C:P ratios in the sediment, whilst negative correlations were observed with various contaminants in both the water and sediment. The whole-sediment bioassays with C. riparius positively related to the nutrient content of the sediment, whereas no negative relations to the sediment-associated contaminants were observed, even though the lowered SPEARpesticides index indicated contaminant effects in the field. Therefore, it was concluded that sediment contamination was identified as one of the various stressors that potentially drove the benthic macroinvertebrate community composition in the multi-stressed drainage ditches, but that nutrients may have masked the adverse effects caused by low and diverse sediment contaminants on C. riparius in the bioassays.

Advancements in effect-based surface water quality assessment.

Legally-prescribed chemical monitoring is unfit for determining the pollution status of surface waters, and there is a need for improved assessment methods that consider the aggregated risk of all bioavailable micropollutants present in the aquatic environment. Therefore, the present study aimed to advance effect-based water quality assessment by implementing methodological improvements and to gain insight into contamination source-specific bioanalytical responses. Passive sampling of non-polar and polar organic compounds and metals was applied at 14 surface water locations that were characterized by two major anthropogenic contamination sources, agriculture and wastewater treatment plant (WWTP) effluent, as well as reference locations with a low expected impact from micropollutants. Departing from the experience gained in previous studies, a battery of 20 in vivo and in vitro bioassays was composed and subsequently exposed to the passive sampler extracts. Next, the bioanalytical responses were divided by their respective effect-based trigger values to obtain effect-based risk quotients, which were summed per location. These cumulative ecotoxicological risks were lowest for reference locations (4.3-10.9), followed by agriculture locations (11.3-27.2) and the highest for WWTP locations (12.8-47.7), and were mainly driven by polar organic contaminants. The bioanalytical assessment of the joint risks of metals and (non-)polar organic compounds resulted in the successful identification of pollution source-specific ecotoxicological risk profiles: none of the bioassays were significantly associated with reference locations nor with multiple location types, while horticulture locations were significantly characterized by anti-AR and anti-PR activity and cytotoxicity, and WWTP sites by ERα activity and toxicity in the in vivo bioassays. It is concluded that the presently employed advanced effect-based methods can readily be applied in surface water quality assessment and that the integration of chemical- and effect-based monitoring approaches will foster future-proof water quality assessment strategies on the road to a non-toxic environment.

Effect-based nationwide surface water quality assessment to identify ecotoxicological risks.

A large portion of the toxic effects observed in surface waters cannot be attributed to compounds regularly measured by water authorities. Hence, there is an urgent need for an effect-based monitoring strategy that employs bioassays to identify environmental risks. The aim of the present study was to perform an effect-based nationwide water quality assessment to identify ecotoxicological risks in a wide variety of surface waters. At 45 locations silicone rubbers and polar organic chemical integrative samplers were exposed to surface water for 6 weeks. Alongside the passive samplers an in-situ daphnid test was performed. Subsequent to field exposure, accumulated compounds were extracted from the passive samplers after which a battery of in vivo and in vitro bioassays was exposed to the extracts. The bioassay battery was selected such that it could identify the risks posed by a wide range of chemical pollutants and their transformation products, while simultaneously allowing for targeted identification of groups of compounds that cause specific effects. Bioassay responses were compared to effect-based trigger values to identify potential ecotoxicological risks at the investigated locations. Responses were observed in all bioassays, and trigger values were exceeded in 9 out of the 21 applied assays, allowing for ranking of the investigated locations based on ecotoxicological risks. No relationship between land use and the identification of ecotoxicological risks was observed. Based on the results, considerations regarding future improvements of effect-based monitoring are given. It is concluded that effect-based water quality assessment allowed prioritization of sites based on ecotoxicological risks, identified the presence of hazardous compounds regardless of being listed as priority substances, and meanwhile could prevent costly chemical analysis at sites with low ecotoxicological risks.

The significance of refuge heterogeneity for lowland stream caddisfly larvae to escape from drift.

The process of macroinvertebrate drift in freshwater lowland streams is characterized by dislodgement, drift distance and subsequent return to the bottom. Refuges are important to all drift phases, since they may help larvae to avoid dislodgement and to escape from drift, even more so if the refuge structure is complex and heterogeneous. The aim of the present study was therefore to determine the influence of refuge heterogeneity on the ability of caddisfly larvae to return to the bottom from drift and to avoid secondary dislodgement. To this purpose a series of indoor flume experiments were undertaken, testing six Limnephilidae (Trichoptera) species, that occur on a gradient from lotic to lentic environments. Bed morphology (plain, refuges with or without leaf patches) and flow velocity (low (0.1 m/s), intermediate (0.3 m/s) and high (0.5 m/s) were manipulated. We showed that all species were favoured by refuges and that especially for species on the lentic end of the gradient (L. lunatus, L. rhombicus and A. nervosa), the ability to escape from drift and to avoid secondary dislodgement was increased. Moreover, we showed that all species spent more time in refuges than in open channel parts and more time in heterogeneous refuges (leaf patches) than in bare refuges, the latter being especially the case for larvae of the lotic species. For lentic species, not well adapted to high flow velocity, refuges are thus crucial to escape from drift, while for the lotic species, better adapted to high flow velocity, the structure of the refuge becomes increasingly important. It is concluded that refuges may play a crucial role in restoring and maintaining biodiversity in widened, channelized and flashy lowland streams.

Nationwide screening of surface water toxicity to algae.

According to the European Water Framework Directive (WFD), chemical water quality is assessed by monitoring 45 priority substances. However, observed toxic effects can often not be attributed to these priority substances, and therefore there is an urgent need for an effect-based monitoring strategy that employs bioassays to identify environmental risk. Algal photosynthesis is a sensitive process that can be applied to identify the presence of hazardous herbicides in surface water. Therefore, the aim of this study was to employ an algal photosynthesis bioassay to assess surface water toxicity to algae and to identify the compounds causing the observed effects. To this purpose, Raphidocelis subcapitata was exposed to surface water samples and after 4.5 h photosynthetic efficiency was determined using PAM fluorometry. In this rapid high throughput bioassay, algal photosynthesis was affected by surface water from only one of 39 locations. Single compounds toxicity confirmation elucidated that the observed effect could be solely attributed to the herbicide linuron, which occurred at 110 times the EQS concentration and which is not included in the WFD priority substances list. In conclusion, applying the algal photosynthesis bioassay enables more efficient and effective assessment of toxicity to primary producers because it: (i) identifies the presence of herbicides that would be overlooked by routine chemical WFD monitoring, and (ii) avoids redundant chemical analyses by focusing only on (non-)target screening in samples with demonstrated effects.