Bottom sediments around the world store large amounts of legacy hydrophobic organic contaminants (HOCs), forming mixtures of unknown chemical composition. Primary emissions to the environment of many HOCs have been reduced as a consequence of regulation. However, HOCs may be released from the sediments to water and biota, and there is therefore a risk of negative effects on local ecosystems. The activity of benthic organisms can enhance the sediment-to-water flux of HOCs, a process called bioturbation. Few in situ assessments of the sediment-to-water flux are available in the scientific literature, and the effect of bioturbation on the sediment-to-water flux of HOCs has not been studied in the field. Thus, there is a need to improve in situ methods for direct determination of sediments as a source of HOCs to water, and thereby include the effect of bioturbation. In Paper I, a benthic flow-through chamber was developed for environmentally realistic in situ assessments of the sediment-to-water flux. In Paper II, the sediment-to-water flux of polycyclic aromatic hydrocarbons (PAHs) was assessed using the flow-through chamber at four sites on the Swedish Baltic Sea coast. The sediments at all four sites acted as sources of PAHs to water. In the same study, potential effects of bioturbation, with an increase of the sediment-to-water flux by up to one order of magnitude, were observed at sites with bioturbating organisms. In the past, assessing the toxicity of HOCs has been challenging due to difficulties in maintaining stable exposure concentrations of the test chemical. In Paper III, a passive dosing method, where the test chemical partitions from a polymer (silicone) to the aquatic exposure medium, was developed and tested for chronic exposure. A stable exposure concentration could be maintained, and the chronic toxicity to the sediment-dwelling harpacticoid Nitocra spinipes of chronic exposure to triclosan was assessed in a 6-week population development test. In Paper IV, a passive sampling and dosing method was developed and used to assess the toxicity of an environmental chemical mixture of bioavailable sediment-associated HOCs transferred from a contaminated sediment to the laboratory-based bioassay. The passive sampling and dosing method can be used to assess the toxicity of environmental mixtures of chemicals at environmentally realistic concentrations to which ecosystems are constantly exposed. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 2: Manuscript.</p>

Passive sampling, as opposed to the conventional spot or bottle water sampling technique, has shown to be reliable and efficient in monitoring the toxicologically relevant, freely dissolved (e.g. bioavaialable) concentrations of a wide range of organic contaminants in water. At the same time, partitioning controlled delivery (passive dosing; PD) techniques promise to overcome many of the challenges associated with toxicity testing of hydrophobic substances that may bias the interpretation of toxicity data. The present study investigated the feasibility of coupling silicone rubber passive sampling devices (SR-PSDs) with bioassay techniques for both chemical and ecotoxicological assessment of complex mixtures of organic contaminants in the aquatic environment. SR-PSDs were deployed in water at various locations within the Ythan catchment (north east, Scotland, UK), Forth estuary and the Firth of Forth (east coast of central Scotland, UK) for 7 to 9 weeks. Following retrieval, extracts from the SR-PSDs were analysed for dissolved concentrations of a variety of organic contaminants including PAHs and PCBs using GC-MS and GC-ECD respectively and were screened for a wide range of pesticides using GC-MS/MS and LC-MS/MS. The extracts were further evaluated for acute cytotoxicity (i.e. neutral red uptake assay) and EROD induction potential using rainbow trout liver cell line (Oncorhynchus mykiss; RTL-W1) and for phytotoxicity and developmental toxicity potential using algal growth inhibition test (with a marine phytoplankton, Diacronema lutheri) and fish embryo toxicity test (with embryos from zebrafish Danio rerio) respectively. Overall, the individual and total dissolved concentrations of PAHs (ΣPAH40; parent and branched) and PCBs (ΣPCB32; ortho and mono-ortho) measured in water from the Ythan, Forth estuary and Firth of Forth were relatively low compared with other studies using PSDs. A number and level of pesticides, including insecticides, herbicides and fungicides of varying hydrophobicity (log KOWs ~2.25 to ~5.31) were detected in the silicone rubber (SR) extracts from the Ythan catchment, the Forth estuary and the Firth of Forth, suggesting input mainly from agricultural run-off and possibly from direct discharges. No statistically significant (p<0.05) acute cytotoxicity was observed following 48 h exposure of RTL-W1 cells to SR extracts from the Ythan catchment. But, on a sublethal level, for every site, statistically significant EROD activity was observed to some degree following 72 h exposure. In addition, developmental and algal toxicities on embryos of D. rerio and D. lutheri respectively, were measured in all the deployed samples compared with the procedural controls (undeployed samples). Interestingly, extracts of SR-PSDs from the Forth estuary and the Firth of Forth exhibited growth inhibitions on D. lutheri that were similar to those of extracts from the Ythan, even though, fewer numbers of pesticides were detected in the Forth estuary and Firth of Forth than the Ythan. This suggests that pesticides were not solely responsible for the observed effects in the Ythan catchment. To further improve data from toxicity testing of hydrophobic substances, the study identified the use of SR O-rings as a suitable passive dosing format in in vitro toxicity tests and was partially validated through their use in dosing RTL-W1 cells with two individual PAHs and subsequently determining cytotoxicity and EROD-activity.