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.

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.

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.

Adverse effects of fullerenes (nC60) spiked to sediments on Lumbriculus variegatus (Oligochaeta).

Effects of fullerene-spiked sediment on a benthic organism, Lumbriculus variegatus (Oligochaeta), were investigated. Survival, growth, reproduction, and feeding rates were measured to assess possible adverse effects of fullerene agglomerates produced by water stirring and then spiked to a natural sediment. L. variegatus were exposed to 10 and 50 mg fullerenes/kg sediment dry mass for 28 d. These concentrations did not impact worm survival or reproduction compared to the control. Feeding activities were slightly decreased for both concentrations indicating fullerenes' disruptive effect on feeding. Depuration efficiency decreased in the high concentration only. Electron and light microscopy and extraction of the worm fecal pellets revealed fullerene agglomerates in the gut tract but not absorption into gut epithelial cells. Micrographs also indicated that 16% of the epidermal cuticle fibers of the worms were not present in the 50 mg/kg exposures, which may make worms susceptible to other contaminants.

Sublethal toxicity and biotransformation of pyrene in Lumbriculus variegatus (Oligochaeta).

The aim of this work was to study the toxicity and biotransformation of polyaromatic hydrocarbon (PAH) pyrene in the oligochaete aquatic worm, Lumbriculus variegatus. PAHs are ubiquitous environmental pollutants that pose a hazard to aquatic organisms, and metabolizing capability is poorly known in the case of many invertebrate species. To study the toxicity and biotransformation of pyrene, the worm was exposed for 15 days to various concentrations of water-borne pyrene. The dorsal blood vessel pulse rate was used as a sublethal endpoint. Pyrene biotransformation by L. variegatus was studied and the critical body residues (CBR) were estimated for pyrene toxicity. The toxicokinetics of pyrene uptake was evaluated. A combination of radiolabeled (14C) and nonlabeled pyrene was used in the exposures, and liquid scintillation counting (LSC) and high-pressure liquid chromatography were employed in both water and tissue residue analyses. The results showed that L. variegatus was moderately able to metabolize pyrene to 1-hydroxypyrene (1-HP), thus demonstrating that the phase-I-like oxidizing enzyme system metabolizes pyrene in L. variegatus. The amount of the 1-HP was 1-2% of the amount of pyrene in the worm tissues. The exposure to pyrene reduced the blood vessel pulse rate significantly (p<0.05), showing that pyrene had a narcotic effect. The estimated CBRs remained constant during the exposure time, varying from 0.120 to 0.174 mmol pyrene/kg worm wet weight. The bioconcentration factors (BCF) decreased as exposure concentration increased. It was suggested that the increased toxicity of pyrene accounted for the decrease in BCFs by lowering the activity of the organism.

Examining the role of temperature and sediment-chemical contact time on desorption and bioavailability of sediment-associated tetrabromo diphenyl ether and benzo(a)pyrene.

The effects of temperature and sediment-chemical contact time on desorption and bioaccumulation of sediment-spiked (14)C-labelled 2,2',4,4'-tetrabromo diphenyl ether (BDE-47) and benzo(a)pyrene (BaP) were examined. Experiments were performed after 2 or 6 weeks and 23 or 24 months sediment-chemical contact time at 10 and 20 degrees C. Desorption was measured in a sediment-water suspension using Tenax extraction, and bioaccumulation was measured by exposing Lumbriculus variegatus (Oligochaeta) to BDE-47 and BaP-spiked sediments in a 10d kinetic study. Biota-sediment accumulation factors (BSAFs) ranged between 2.9 and 4.3 for BDE-47 and between 0.5 and 0.9 for BaP. Thus, temperature and aging had a minor effect on bioavailability estimates. On the other hand, the difference between the chemicals was clear and could not be interpreted solely by reference to the size of the desorbing fractions, although the rapidly desorbing fraction-revised estimate clearly reduced the difference. The remaining discrepancy may be related to methodological (Tenax extraction vs. worm exposure) and/or biological (digestive extraction) causes. However, the data support the role of diffusional forces in the bioavailability of sediment-associated organic contaminants. Therefore, desorption-revised bioavailability estimates would lead to more precise bioavailability estimates than the traditional sediment organic carbon-organisms' lipids-based equilibrium partitioning approach.

Bioaccumulation, sublethal toxicity, and biotransformation of sediment-associated pentachlorophenol in Lumbriculus variegatus (Oligochaeta).

The xenobiotics accumulated in sediments represent a hazard to organisms. In order to study the toxic effects of xenobiotics in organisms, body residue has been proposed as a more relevant dose-metric than the environmental concentration of the chemical. In this study, the benthic oligochaetes Lumbriculus variegatus were exposed to sediment-spiked pentachlorophenol (PCP) in a chronic study at different exposure concentrations. The aim was to examine sublethal toxic effects in sediment-dwelling and sediment-ingesting organisms, and to link the effects with chemical body residues. Growth, reproduction, and egestion rate were used as sublethal endpoints. Bioaccumulation, sublethal toxic effects, and biotransformation of PCP were investigated by exposing organisms to both artificial and natural sediments with similar organic carbon content. Sediment characteristics were assumed to have an effect on toxicity since PCP retarded both growth and reproduction in L. variegatus in the artificial sediment. In natural sediment, growth, and reproduction was also reduced in control treatments, probably indicating poor nutritional quality. Most of the extracted chemicals in L. variegatus tissues were water-soluble metabolites, indicating that L. variegatus was capable of biotransforming PCP. The extractable parent PCP body residues (CBR(50)) for L. variegatus growth and reproduction were in agreement with the values estimated for respiratory uncouplers in the literature.

Feeding activity of midge larvae (Chironomus riparius meigen) in metal-polluted river sediments.

A method was developed for monitoring the feeding activity of larvae of the midge Chironomus riparius. The egestion rate (mg dry wt feces/mg dry wt larva/h) of the deposit-feeding larvae was measured and used as an indication of the feeding activity. Both the egestion rate and survival of several metal-adapted and reference larvae were measured in five test sediments with various Cd and Zn concentrations. The reference larvae suffered increased mortality in two contaminated sediments by comparison with field-exposed larvae. The feeding of all individuals decreased in the most polluted test sediment but also in another supposedly clean reference sediment. No differences were found in egestion rates between the reference and field-exposed larvae. This observation, however, does not reliably answer the hypothesis whether the feeding activity of the adapted and nonadapted midge larvae differs in metal-contaminated sediments because the data are based on total egestion during the short (96-h) experiment and the progress of feeding was not followed.