Holistic evaluation of the environmental impacts of shipping in the sensitive region of Ria de Aveiro.

Shipping activity can be a substantial source of pollution and impact on the environment, including air, water and ecosystems, as well as adverse health and climatic effects. Due to the distribution of maritime transport activity routes in the EU, a large portion of the population is exposed to shipping pollution throughout Europe. The ongoing European project EMERGE aims to investigate and quantify these impacts over Europe, and in more detail, in specific case studies regions. The Aveiro lagoon region in Portugal is one of these case studies. This region is a Natura 2000 area, and also includes a medium-sized port. Both air quality and water modelling tools were applied to assess the impact of the emissions and discharges from shipping (to air and water) in the region in 2018. Additionally, ecotoxicological impacts were determined by bioassays to evaluate the impact of scrubber-water discharges on the most sensitive stages of marine invertebrates, and on the post-exposure feeding inhibition of crustacean and bivalve species. The results show that there was a substantial increase in atmospheric pollutant concentrations due to emissions attributed to shipping, which was most relevant for NOx and SO2 (up to a 30 % shipping contribution). There was no significant degradation of the water quality, mainly as the ships operating in this area did not have scrubber equipment. The ecotoxicological tests were performed with three samples of scrubber water, including one artificial sample and two samples collected on-board ships. If scrubber water would have been discharged in this area, the results indicated that the majority of the tested species would be exposed to lowest observed effect concentration (LOEC) for the different scrubber-water samples, as well as to substantial concentrations of metals, PAHs, and alkylated PAHs.

Gene expression analysis of Chironomus riparius in response to acute exposure to tire rubber microparticles and leachates.

Tire rubber microparticles (TRPs) entering aquatic ecosystems through stormwater runoffs is a significant challenge. TRPs are formed by the abrasion of tires with the road surface and include chemical additives that are an additional cause for concern. Currently, information on the molecular effects of TRPs, or especially its additives, in freshwater organisms is scarce. To address this problem, an array covering different cellular processes has been designed for the freshwater midge Chironomus riparius. Fourth-instar larvae were exposed to two concentrations of TRPs (1 mg L-1, 10 mg L-1) and tire rubber leachates (TRLs) (0,0125 %, 5 %) to evaluate the transcriptional activity by Real-Time PCR. To assess acute toxicity, larvae were exposed for 24 h and genes related to the endocrine system, stress response, DNA repair mechanisms, immune system, oxidative stress, and detoxification mechanisms were evaluated. The activity of the enzymes: glutathione S-transferase (GST) and catalase was also examined. The main pathway affected was the stress response showing overexpression of HSPs (HSC70.3, HSC70.4, HSC70.5, HSP60). Moreover, there was a reduction of the GSTd3 and catalase disrupting the antioxidant system. The upregulation of InR indicates a potential disturbance in the insulin pathway and ABCB6 activation only in TRPs exposure suggests its potential implication in their transport. However, most of these alterations are caused by TRLs, showing higher toxicity than TRPs. The results obtained in this work provide the first approach at the molecular and cellular levels to elucidate the impact of TRLs in freshwater organisms. To perform a realistic evaluation of the TR effects, additional research is required to assess the TR's long-term effects at the molecular level.

Cross cultural adaptation and psychometric properties of the Finnish version of Western Ontario shoulder instability index (WOSI).

BACKGROUND: Western Ontario shoulder instability index (WOSI) is a widely used disease-specific self-assessment measurement tool for patients with shoulder instability. The main aim of this study was to translate and cross culturally adapt the WOSI into Finnish language and to test its measurement properties. METHODS: WOSI was translated in Finnish and adapted into an electronic user interface. 62 male patients with traumatic anteroinferior shoulder instability, programmed for stabilizing shoulder surgery, answered the questionnaire twice preoperatively (2 and 0 weeks), and twice postoperatively (3 and 12 months). Additional scoring tools, such as satisfaction to treatment outcome, subjective shoulder value (SSV), Oxford shoulder instability index (OSIS) and Constant score (CS), were used as comparators. The reliability, validity and responsiveness of WOSI were investigated through statistical analysis. RESULTS: Preoperative test-retest results were available for 49 patients, and 54 patients were available at final follow up. The mean WOSI was 57.8 (SD 20.3), 70.4 (SD 18.9), and 85.9 (SD 15.5), at baseline, 3, and 12 months, respectively. There was a statistically significant mean improvement of 28.8 (SD 24.5) in WOSI between baseline and 12 months (p < 0.0001). The intraclass correlation coefficient for the preoperative WOSI was excellent 0.91. At 12 months WOSI had an excellent Pearson's correlation coefficient both with SSV (0.69), OSIS (-0.81), and poor with CS (0.25) scores, confirming our a priori hypothesis. There were no detected floor nor ceiling effects for WOSI pre- or postoperatively. The calculated minimal detectable change was 9.2 and the estimated minimal clinically important difference 13.4 to 18.1. CONCLUSION: Finnish version of WOSI is a reliable and valid tool for assessing health state and improvement after operative treatment of shoulder instability in young male patients.

Tracing the fate of microplastic carbon in the aquatic food web by compound-specific isotope analysis.

Increasing abundance of microplastics (MP) in marine and freshwaters is currently one of the greatest environmental concerns. Since plastics are fairly resistant to chemical decomposition, breakdown and reutilization of MP carbon complexes requires microbial activity. Currently, only a few microbial isolates have been shown to degrade MPs, and direct measurements of the fate of the MP carbon are still lacking. We used compound-specific isotope analysis to track the fate of fully labelled 13C-polyethylene (PE) MP carbon across the aquatic microbial-animal interface. Isotopic values of respired CO2 and membrane lipids showed that MP carbon was partly mineralized and partly used for cell growth. Microbial mineralization and assimilation of PE-MP carbon was most active when inoculated microbes were obtained from highly humic waters, which contain recalcitrant substrate sources. Mixotrophic algae (Cryptomonas sp.) and herbivorous zooplankton (Daphnia magna) used microbial mediated PE-MP carbon in their cell membrane fatty acids. Moreover, heteronanoflagellates and mixotrophic algae sequestered MP carbon for synthesizing essential ω-6 and ω-3 polyunsaturated fatty acids. Thus, this study demonstrates that aquatic micro-organisms can produce, biochemically upgrade, and trophically transfer nutritionally important biomolecules from PE-MP.

Perioperative platelet rich plasma (PRP) in total hip arthroplasty through the Hardinge approach: protocol to study the effectiveness for gluteus medius healing.

BACKGROUND: Platelet-rich plasma (PRP) has been used to support tendon regeneration mainly in sports medicine. PRP is a concentrate of platelet-rich plasma proteins derived from whole blood by centrifugation to remove erythrocytes and leukocytes. PRP has high amounts of platelets which may promote healing tendons affected by degenerative conditions. These platelets contain growth factors and are known to facilitate the regeneration of injured tendon structures. Total hip arthroplasty (THA) through the Hardinge approach may leave the patient with impaired gait and poor regeneration of the gluteus medius tendon if the tendon is not reattached properly after closure of the surgical wound. METHODS: The study will be a multicenter, double-blinded and randomized study enrolling 90 patients based on power calculations. The efficacy of perioperative PRP treatment will be assessed by subjective and objective outcome variables. The participants will be randomized (sealed envelope) into either a placebo (saline) or a PRP group (1:1). For subjective outcomes, the Oxford Hip Score (OHS) will be collected before surgery and 3 and 12 months after surgery. The objective measures are findings at magnetic resonance imaging and plain radiographs and recorded values of measured strength. DISCUSSION: We present the perioperative use and the ways to measure the clinical efficacy of PRP. As PRP may have benefits regarding degenerative tendon regeneration, studies on the use of PRP in hip arthroplasty are warranted to facilitate postoperative recovery. TRIAL REGISTRATION: This study has been approved by the ethics committee of the Hospital District of Southwest Finland and approved by the local institutional research board. The study has been registered in ClinicalTrials.gov ( NCT02607462 ).

The chronic effects of fullereneC60-associated sediments in the midge Chironomus riparius - Responses in the first and the second generation.

The life cycle parameters of the benthic invertebrate Chironomus riparius make it a relevant organism for use in multi-generation chronic ecotoxicology tests. Since studies on chronic exposures with fullerene carbon nanoparticles have revealed adverse effects at lower concentration ranges, it is crucial to gain understanding of the consequences in following generations. The aims of this study were to investigate whether sediment-associated fullereneC60 impacts on C. riparius emergence and breeding, thus affecting the growth of the second generation. Larvae were exposed to fullerene-spiked sediment at concentrations of 0.5, 10 and 40 mg/kg sediment dw. Total emergence and breeding success were monitored after the first generation and the newly hatched larvae from the first generation exposure were transferred either to continuous exposure or to pristine sediment without fullerene. Findings indicate that the presence of fullerenes has major impacts on the first generation, mainly shown as delayed emergence time of females. Increased larval growth was observed in the second generation, and we conclude that the C. riparius response to fullerene exposure indicated significant signs of recovery in second-generation larval growth. The result shows the effects to be important for population dynamics, revealing delayed female emergence time, which leads to situation where adults' breeding is impaired.

Chironomus riparius exposure to fullerene-contaminated sediment results in oxidative stress and may impact life cycle parameters.

A key component of understanding the potential environmental risks of fullerenes (C60) is their potential effects on benthic invertebrates. Using the sediment dwelling invertebrate Chironomus riparius we explored the effects of acute (12h and 24h) and chronic (10d, 15d, and 28d) exposures of sediment associated fullerenes. The aims of this study were to assess the impact of exposure to C60 in the sediment top layer ((0.025, 0.18 and 0.48) C60 mg/cm2) on larval growth, oxidative stress and emergence rates and to quantify larval body burdens in similarly exposed organisms. Oxidative stress localization was observed in the tissues next to the microvilli and exoskeleton through a method for identifying oxidative stress reactions generated by reactive oxygen species. Rapid intake of fullerenes was shown in acute experiments, whereas body residues decreased after chronic exposure. Transmission electron microscopy analysis revealed oxidative damage and structural changes in cells located between the lipid droplets and next to the microvilli layer in fullerene exposed samples. Fullerene associated sediments also caused changes in the emergence rate of males and females, suggesting that the cellular interactions described above or other effects from the fullerenes may influence reproduction rates.

Biological responses of midge (Chironomus riparius) and lamprey (Lampetra fluviatilis) larvae in ecotoxicity assessment of PCDD/F-, PCB- and Hg-contaminated river sediments.

We evaluated the utility of chironomid and lamprey larval responses in ecotoxicity assessment of polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDD/F)-, polychlorinated biphenyls (PCB)- and mercury (Hg)-contaminated river sediments. Sediment samples were collected from the River Kymijoki with a known industrial pollution gradient. Sediment for the controls and lamprey larvae were obtained from an uncontaminated river nearby. Contamination levels were verified with sediment and tissue PCDD/F, PCB and Hg analyses. Behaviour of sediment-exposed chironomid and lamprey larvae were measured with Multispecies Freshwater Biomonitor© utilizing quadrupole impedance conversion technique. In addition, mortality, growth and head capsule deformity incidence of chironomids were used as ecotoxicity indicators. WHOPCDD/F+PCB-TEQ in the R. Kymijoki sediments ranged from the highest upstream 22.36 ng g(-1) dw to the lowest 1.50 ng g(-1) near the river mouth. The sum of PCDD/Fs and PCBs correlated strongly with Hg sediment concentrations, which ranged from <0.01 to 1.15 µg g(-1). Lamprey tissue concentrations of PCDD/Fs were two orders and PCBs one order of magnitude higher in the R. Kymijoki compared to the reference. Chironomid growth decreased in contaminated sediments and was negatively related to sediment ∑PCDD/Fs, WHOPCDD/F+PCB-TEQ and Hg. There were no significant differences in larval mortality or chironomid mentum deformity incidence between the sediment exposures. The distinct behavioural patterns of both species indicate overall applicability of behavioural MFB measurements of these species in sediment toxicity bioassays. Chironomids spent less and lampreys more time in locomotion in the most contaminated sediment compared to the reference, albeit statistically significant differences were not detected. Lamprey larvae had also a greater activity range in some of the contaminated sediments than in the reference. High pollutant levels in lamprey indicate risks for biomagnification in the food webs, with potential health risks to humans consuming fish.

Predicting the bioaccumulation of polyaromatic hydrocarbons and polychlorinated biphenyls in benthic animals in sediments.

There were two main objectives in this study. The first was to compare the accuracy of different prediction methods for the chemical concentrations of polyaromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) in the organism, based on the measured chemical concentrations existing in sediment dry matter or pore water. The predicted tissue concentrations were compared to the measured ones after 28-day laboratory test using oligochaeta worms (Lumbriculus variegatus). The second objective was to compare the bioaccumulation of PAHs and PCBs in the laboratory test with the in situ bioaccumulation of these compounds. Using the traditional organic carbon-water partitioning model, tissue concentrations were greatly overestimated, based on the concentrations in the sediment dry matter. Use of an additional correction factor for black carbon with a two-carbon model, significantly improved the bioaccumulation predictions, thus confirming that black carbon was important in binding the chemicals and reducing their accumulation. The predicted PAH tissue concentrations were, however, high compared to the observed values. The chemical concentrations were most accurately predicted from their freely dissolved pore water concentrations, determined using equilibrium passive sampling. The patterns of PCB and PAH accumulation in sediments for laboratory-exposed L. variegatus were similar to those in field-collected Lumbriculidae worms. Field-collected benthic invertebrates and L. variegatus accumulated less PAHs than PCBs with similar lipophilicity. The biota to sediment accumulation factors of PAHs tended to decrease with increasing sediment organic carbon normalized concentrations. The presented data yields bioconcentration factors (BCF) describing the chemical water-lipid partition, which were found to be higher than the octanol-water partition coefficients, but on a similar level with BCFs drawn from relevant literature. In conclusion, using the two-carbon model method, or the measured freely dissolved pore water concentrations method is recommended for predicting the bioaccumulation of PAHs and PCBs.

Particulate matter air pollution components and risk for lung cancer.

BACKGROUND: Particulate matter (PM) air pollution is a human lung carcinogen; however, the components responsible have not been identified. We assessed the associations between PM components and lung cancer incidence. METHODS: We used data from 14 cohort studies in eight European countries. We geocoded baseline addresses and assessed air pollution with land-use regression models for eight elements (Cu, Fe, K, Ni, S, Si, V and Zn) in size fractions of PM2.5 and PM10. We used Cox regression models with adjustment for potential confounders for cohort-specific analyses and random effect models for meta-analysis. RESULTS: The 245,782 cohort members contributed 3,229,220 person-years at risk. During follow-up (mean, 13.1 years), 1878 incident cases of lung cancer were diagnosed. In the meta-analyses, elevated hazard ratios (HRs) for lung cancer were associated with all elements except V; none was statistically significant. In analyses restricted to participants who did not change residence during follow-up, statistically significant associations were found for PM2.5 Cu (HR, 1.25; 95% CI, 1.01-1.53 per 5 ng/m(3)), PM10 Zn (1.28; 1.02-1.59 per 20 ng/m(3)), PM10 S (1.58; 1.03-2.44 per 200 ng/m(3)), PM10 Ni (1.59; 1.12-2.26 per 2 ng/m(3)) and PM10 K (1.17; 1.02-1.33 per 100 ng/m(3)). In two-pollutant models, associations between PM10 and PM2.5 and lung cancer were largely explained by PM2.5 S. CONCLUSIONS: This study indicates that the association between PM in air pollution and lung cancer can be attributed to various PM components and sources. PM containing S and Ni might be particularly important.

Bioconcentration, biotransformation and elimination of pyrene in the arctic crustacean Gammarus setosus (Amphipoda) at two temperatures.

The influence of temperature on the bioaccumulation, toxicokinetics, biotransformation and depuration of pyrene was studied in the arctic marine amphipod Gammarus setosus. A two-compartment model was used to fit experimental values of total body burden, total metabolites and parent pyrene concentrations and to calculate toxicokinetic variables derived for two experimental treatments (2 and 8 °C). No statistically significant differences were observed with temperature for these toxicokinetic variables or bioconcentration factors. Contrarily, the Q10 values suggested that the toxicokinetic variables ke and km were temperature-dependent. This may be explained by the high standard deviation of the Q10 values. Q10 is the variation in the rate of a metabolic reaction with a 10 °C increase in temperature. Depuration rate constants were calculated from linear best fit equations applied to measured pyrene concentrations over time during the depuration phase of the experiment. During depuration, the parent pyrene was eliminated in two stages with faster elimination observed at 8 °C compared to 2 °C. This finding was also indicated by the Q10. No changes in total body burdens of metabolite concentrations were observed during the monitoring of depuration over a period of 96 h. The biotransformation pathway of pyrene in G. setosus was also investigated in this study with two main phase II biotransformation products discovered by liquid chromatography. These products are conditionally identified as the sulphate and glucose conjugates of 1-hydroxy-pyrene. Overall, the study contributes new knowledge to the understanding of the fate of PAHs in arctic biota. In particular, the study provides valuable insight into the bioaccumulation and biotransformation of an important PAH and its metabolites in a species that serves as both a predator and prey in the arctic ecosystem.

Treatment of non-traumatic rotator cuff tears: A randomised controlled trial with one-year clinical results.

We have compared three different methods of treating symptomatic non-traumatic tears of the supraspinatus tendon in patients above 55 years of age. A total of 180 shoulders (173 patients) with supraspinatus tendon tears were randomly allocated into one of three groups (each of 60 shoulders); physiotherapy (group 1), acromioplasty and physiotherapy (group 2) and rotator cuff repair, acromioplasty and physiotherapy (group 3). The Constant score was assessed and followed up by an independent observer pre-operatively and at three, six and twelve months after the intervention. Of these, 167 shoulders were available for assessment at one year (follow-up rate of 92.8%). There were 55 shoulders in group 1 (24 in males and 31 in females, mean age 65 years (55 to 79)), 57 in group 2 (29 male and 28 female, mean age 65 years (55 to 79)) and 55 shoulders in group 3 (26 male and 29 female, mean age 65 years (55 to 81)). There were no between-group differences in the Constant score at final follow-up: 74.1 (sd 14.2), 77.2 (sd 13.0) and 77.9 (sd 12.1) in groups 1, 2 and 3, respectively (p = 0.34). The mean change in the Constant score was 17.0, 17.5, and 19.8, respectively (p = 0.34). These results suggest that at one-year follow-up, operative treatment is no better than conservative treatment with regard to non-traumatic supraspinatus tears, and that conservative treatment should be considered as the primary method of treatment for this condition.

Smoking and operative treatment of rotator cuff tear.

This registry study was set up to evaluate the effect of smoking on the pre-operative status, intraoperative findings, and post-operative status after rotator cuff reconstruction. Five hundred seventy-six consecutive shoulders with primarily arthroscopically repaired penetrating rotator cuff tear were followed up. Tobacco consumption was recorded as pack-years. Age-adjusted Constant score was used as an outcome measure. Five hundred sixty-four patients were available for 1-year follow-up (dropout rate 2%). One hundred fourteen (20%) and 450 (80%) patients were pre-operatively recorded to be smokers and non-smokers, respectively. The gender distribution did not differ between the groups (P = 0.286). The mean age of all patients was 55 years in smokers (SD 9.1) and 61 years in non-smokers (SD 9.4) (P < 0.001). There was no statistically significant difference in pre-operative Constant score (P = 0.075) or mean size of intraoperatively measured tendon tear (P = 0.290) between the groups. At final follow-up, there was a statistically significant difference in Constant scores between smokers [71 (SE 1.4)] and non-smokers [75 (SE 0.7)] (P = 0.017). The pack-years of smoking correlated with neither the Constant score (P = 0.815) nor the size of the tear (P = 0.786). We conclude that operatively treated rotator cuff tear patients who smoked were significantly younger than non-smokers, and that smoking was associated with lower post-operative Constant score.

Orexin/hypocretin receptor signalling: a functional perspective.

Multiple homeostatic systems are regulated by orexin (hypocretin) peptides and their two known GPCRs. Activation of orexin receptors promotes waking and is essential for expression of normal sleep and waking behaviour, with the sleep disorder narcolepsy resulting from the absence of orexin signalling. Orexin receptors also influence systems regulating appetite/metabolism, stress and reward, and are found in several peripheral tissues. Nevertheless, much remains unknown about the signalling pathways and targets engaged by native receptors. In this review, we integrate knowledge about the orexin receptor signalling capabilities obtained from studies in expression systems and various native cell types (as presented in Kukkonen and Leonard, this issue of British Journal of Pharmacology) with knowledge of orexin signalling in different tissues. The tissues reviewed include the CNS, the gastrointestinal tract, the pituitary gland, pancreas, adrenal gland, adipose tissue and the male reproductive system. We also summarize the findings in different native and recombinant cell lines, especially focusing on the different cascades in CHO cells, which is the most investigated cell line. This reveals that while a substantial gap exists between what is known about orexin receptor signalling and effectors in recombinant systems and native systems, mounting evidence suggests that orexin receptor signalling is more diverse than originally thought. Moreover, rather than being restricted to orexin receptor 'overexpressing' cells, this signalling diversity may be utilized by native receptors in a site-specific manner.

Orexin/hypocretin receptor signalling cascades.

Orexin (hypocretin) peptides and their two known G-protein-coupled receptors play essential roles in sleep-wake control and powerfully influence other systems regulating appetite/metabolism, stress and reward. Consequently, drugs that influence signalling by these receptors may provide novel therapeutic opportunities for treating sleep disorders, obesity and addiction. It is therefore critical to understand how these receptors operate, the nature of the signalling cascades they engage and their physiological targets. In this review, we evaluate what is currently known about orexin receptor signalling cascades, while a sister review (Leonard & Kukkonen, this issue) focuses on tissue-specific responses. The evidence suggests that orexin receptor signalling is multifaceted and is substantially more diverse than originally thought. Indeed, orexin receptors are able to couple to members of at least three G-protein families and possibly other proteins, through which they regulate non-selective cation channels, phospholipases, adenylyl cyclase, and protein and lipid kinases. In the central nervous system, orexin receptors produce neuroexcitation by postsynaptic depolarization via activation of non-selective cation channels, inhibition of K⁺ channels and activation of Na⁺/Ca²âº exchange, but they also can stimulate the release of neurotransmitters by presynaptic actions and modulate synaptic plasticity. Ca²âº signalling is also prominently influenced by these receptors, both via the classical phospholipase C-Ca²âº release pathway and via Ca²âº influx, mediated by several pathways. Upon longer-lasting stimulation, plastic effects are observed in some cell types, while others, especially cancer cells, are stimulated to die. Thus, orexin receptor signals appear highly tunable, depending on the milieu in which they are operating.

A numerical model of birch pollen emission and dispersion in the atmosphere. Description of the emission module.

A birch pollen emission model is described and its main features are discussed. The development of the model is based on a double-threshold temperature sum model that describes the propagation of the flowering season and naturally links to the thermal time models to predict the onset and duration of flowering. For the flowering season, the emission model considers ambient humidity and precipitation rate, both of which suppress the pollen release, as well as wind speed and turbulence intensity, which promote it. These dependencies are qualitatively evaluated using the aerobiological observations. Reflecting the probabilistic character of the flowering of an individual tree in a population, the model introduces relaxation functions at the start and end of the season. The physical basis of the suggested birch pollen emission model is compared with another comprehensive emission module reported in literature. The emission model has been implemented in the SILAM dispersion modelling system, the results of which are evaluated in a companion paper.

Trophic transfer of pyrene metabolites between aquatic invertebrates.

The trophic transfer of pyrene metabolites was studied using Gammarus setosus as a predator and the invertebrates Lumbriculus variegatus and Chironomus riparius as prey. The results obtained by liquid scintillation counting confirmed that the pyrene metabolites produced by the aquatic invertebrates L. variegatus and C. riparius were transferred to G. setosus through the diet. More detailed analyses by liquid chromatography discovered that two of the metabolites produced by C. riparius appeared in the chromatograms of G. setosus tissue extracts, proving their trophic transfer. These metabolites were not present in chromatograms of G. setosus exclusively exposed to pyrene. The present study supports the trophic transfer of PAH metabolites between benthic macroinvertebrates and common species of an arctic amphipod. As some PAH metabolites are more toxic than the parent compounds, the present study raises concerns about the consequences of their trophic transfer and the fate and effects of PAHs in natural environments.

Assessing the impact of chemical pollution on benthic invertebrates from three different European rivers using a weight-of-evidence approach.

The aim of this study was to combine different lines of evidence on the impact of chemical pollution on benthic invertebrate communities in three European river basins (Elbe, Scheldt, and Llobregat). The study integrates chemical analyses, a battery of different sediment toxicity tests, and field data from soft-sediment meio- and macrobenthic fauna within a sediment-quality triad in which chironomids, oligochaetes, and nematodes are identified on the species level. The use of TU (toxic units) and msPAF (multi-substance potentially affected fraction) in an approach assessing the chemical impact as well as the integration of sediment toxicity tests with bacteria (Vibrio fischeri), benthic invertebrates (Caenorhabditis elegans, Potamopyrgus antipodarum, Lumbriculus variegatus, Chironomus riparius), and fish embryos (Danio rerio), together with univariate and non-parametric multivariate statistical analyses of the biological data revealed significant differences between unpolluted and polluted sites in all three river basins. To combine the different results obtained in the sediment-quality triad, a scoring system was successfully developed based on a simple algorithm. This system provides an easily understandable scheme for non-experts among decision makers and water managers.

Trophic transfer of pyrene metabolites and nonextractable fraction from Oligochaete (Lumbriculus variegatus) to juvenile brown trout (Salmo trutta).

The trophic transfer of pyrene metabolites was evaluated by a 2-month exposure of the freshwater annelid Lumbriculus variegatus (Oligochaeta) to pyrene, followed by feeding to juvenile brown trout (Salmo trutta). The results obtained by scintillation counting (SC) proved that the pyrene metabolites produced by L. variegatus were transferred to juvenile S. trutta through diet. More detailed analyses by LC-FLD (liquid chromatography with fluorescence detection) showed that an unknown pyrene metabolite originating from L. variegatus was present in fish liver. This metabolite, although yet not properly identified, may be the glucose conjugate of 1-hydroxy-pyrene. This metabolite was not present in chromatograms of fish that were fed pyrene-spiked food pellets. In addition, the strongly bound tissue residue of L. variegatus, which was nonextractable neither by organic solvents nor by the proteolytic enzyme Proteinase K, was most likely not available for the fish through diet. Altogether, the present study shows that the metabolites of pyrene produced at low levels of the food chain may be potentially available for upper levels through diet, raising a concern about their potential toxicity to predators and supporting their inclusion in the risk assessment of PAHs.