Effects of oil and bioremediation on mussel (Mytilus edulis L.) growth in mudflats.

Mussels (Mytilus edulis L.) were exposed to crude oil during a field experiment to evaluate two bioremediation strategies (nutrient addition and nutrient addition with tilling). The mussels were placed in 4 mesocosms: Control, Oil, Oil + Nutrients, and Oil + Nutrients + Tilled. Tilling appeared to be clearly detrimental to mussel growth. Additionally, this field experiment demonstrated that at temperatures below 5 degrees C, growth was reduced to rates undetectable by the laser diffraction method. The data on mussel shell length show that this technique does offer very sensitive and useful comparative measurements of physiological function. Measurement of shell growth has the advantage over other techniques in that it is non-invasive and non-destructive and thus may be used continuously without disturbing critical physiological and biochemical functions; however, bivalve physiology is strongly linked to environmental conditions, so it is important to include such measures (i.e. seawater temperature and turbidity) in the design of the biomonitoring program. Elevated polycyclic aromatic hydrocarbon (PAH) levels reflected bioaccumulation in mussels from all the oiled mesocosms. This correlated with reduction in growth rate. Maximum reduction in growth was observed in mussels from the tilled mesocosm which contained the lowest phenanthrene and dibenzothiophene concentrations. The tilling caused an increase in suspended solids which inhibited filter feeding activity, and resulted in suppressed growth and slower intake of PAH-laden sediment.

Biomarker responses and PAH uptake in Mya truncata following exposure to oil-contaminated sediment in an Arctic fjord (Svalbard).

Expanding industrial activity (notably oil and gas exploration) in the Arctic requires assessment of the potential impact of chemicals on marine organisms living in seawater at low temperature. The bivalve Mya truncata is common in Svalbard fjord (Norway) where it experiences low temperature throughout the year. To measure the impact of polycyclic aromatic hydrocarbons (PAH) on M. truncata, the responses of three biomarkers [total oxyradical scavenging capacity-assay (TOSC), plasma membrane stability of haemocytes and respiration rates] were investigated from bivalves exposed to sediment contaminated with a PAH mixture (crude oil). After two weeks of exposure to the contaminated sediment, TOSC showed no change. The high TOSC value (4010+/-1339 unit mg(-1) protein) of Mya truncata (control group) is thought to protect biomolecules with a low turnover rate efficiently in a low food availability environment. In the exposed bivalves, the haemocyte cellular membranes were significantly destabilised compared with controls (P<0.05). Respiration rate of control and PAH-exposed individuals (0.055+/-0.020 mg O(2) dw(-1) h(-1)) was similar and relatively low as is typical for polar bivalves, reflecting a strategy to minimise energy expenditure to cope with 9 months of starvation. Bioaccumulation of PAH by M. truncata was also low, due probably to a combination of low metabolic rate and reduced solubility of the oil compounds at low temperature. Data indicated an uptake of mainly low molecular weight compounds (two and three ring molecules). A good correlation of logBAF(lipid) (bioaccumulation factor) and logK(ow) (octanol/water partitioning coefficient) was shown (r(2)=0.87). Tissue sensitivity and/or functional differences (digestive gland vs. haemocytes), PAH uptake route (dietary vs. gills), the low metabolic rate of M. truncata and the low environmental temperature (reducing the bioavailability of PAH) are factors that help explain these findings.

Total oxyradical scavenging capacity and cell membrane stability of haemocytes of the Arctic scallop, Chlamys islandicus, following benzo(a)pyrene exposure.

Industrial activities, notably oil and gas industries, are expanding in the Arctic. Most of the biomarkers were developed using temperate organisms living at temperatures above 10 degrees C. Little is known about the biomarker responses of organisms living between -1.88 and 5 degrees C. Therefore, assessment of the toxicity of chemicals to cold-water adapted species is required. In this study, the Arctic scallop, Chlamys islandicus, was selected as a key species for bio-monitoring because of wide distribution in Arctic waters and its commercial value. Test animals, stored in seawater at 2 degrees C, were injected with benzo(a)pyrene (diluted in cod liver oil 5 mg ml(-1)) in the adductor muscle every 24 h for four days giving a final dose of 0, 74 and 90.6 mg kg(-1) wet weight for control, low and high dose, respectively. The biomarkers used were total oxyradical scavenging capacity (TOSC) in the digestive gland and cell membrane stability of haemocytes. TOSC values were significantly reduced (ca. 30%) in exposed groups (P < 0.05), indicating a depletion in oxyradical molecular scavengers. The antioxidant defences appeared to be overwhelmed by the reactive oxygen species as the plasma membranes of haemocytes were destabilised (P < 0.05) probably due to lipid peroxidation. These data indicate that reactive oxygen species (ROS) were produced by Arctic scallops via the metabolisation of benzo(a)pyrene at 2 degrees C.

Heart rate, respiration and total oxyradical scavenging capacity of the Arctic spider crab, Hyas araneus, following exposure to polycyclic aromatic compounds via sediment and injection.

Increasing industrial activity in the European Arctic has raised concerns of the potential anthropogenic impact of chemicals on this polar marine ecosystem. For the past 20 years or so, biomarkers have been developed to provide early-warning signals of detrimental impacts of chemicals on the marine ecosystem, however, most biomarker methods have been established for organisms living in temperate rather than polar waters. Little is known about biomarker responses in organisms living within the temperature range of -1.88 to +5 degrees C. In this study, established biomarkers from temperate studies were tested on the Arctic spider crab Hyas araneus to validate their use in polar ecosystems. H. araneus is common in Svalbard fjord (Norway), although it is a temperate water species occurring from northern Spain to Svalbard at depths from 10 to 1200 m. In this paper, the effects of oil were investigated at 2 degrees C via two routes: (i) injection and (ii) contaminated sediment. After 2 weeks of exposure, heart rate, oxygen consumption and total oxyradical scavenging capacity (TOSC) were measured in the same individuals. In both methods of contaminant exposure, heart rate showed a significant increase compared with the control (P < 0.0001, n = 7); mean heart rate values (+/- S.D.) of H. araneus were 49.06 (+/- 13.72), 57.56 (+/- 7.28) and 63.30 (+/- 6.57) beats per minute in control, injected and sediment-treated groups, respectively. Respiration of H. araneus was not affected significantly by either oil treatment (P > 0.05), but two individuals (n = 8) showed a marked increase in oxygen uptake in the sediment-exposed group. The basal oxygen consumption of control H. araneus was lower (0.025 mg O(2) g wet wt.(-1) h(-1)) than reported for H. araneus living in temperate water. Although TOSC of H. araneus was not affected significantly by either exposure treatment (P > 0.05) the mean TOSC value in the sediment-exposed group was lower than the control, indicating some saturation of the oxyradical scavenging system. Results indicate that although low temperature appears to be the main factor reducing the bioavailability of polycyclic-aromatic hydrocarbons, the relatively low metabolic rate of Arctic H. araneus is also implicated in decreased uptake and metabolism of oil compounds into reactive oxygen species (ROS).

Bioaccumulation of polycyclic aromatic compounds: 1. Bioconcentration in two marine species and in semipermeable membrane devices during chronic exposure to dispersed crude oil.

Assessing the fate in marine biota of hydrocarbons derived from oil particles that are discharged during exploration and production is of relevant environmental concern. However, a rather complex experimental setup is required to carry out such investigations. In this study, a sophisticated tool, the continuous-flow system (CFS), was used to mimic dispersed oil exposure to marine biota. Polycyclic aromatic hydrocarbon (PAH) uptake was studied in two species, the blue mussel Mytilus edulis and juvenile of the turbot Scophthalmus maximus, and in semipermeable membrane devices (SPMD) exposed to crude oil dispersed in a flow-through system. After an exposure period of 8 to 21 d, elimination in organisms and devices was analyzed for 9 to 10 d following transfer to PAH-free seawater. Principal component analysis (PCA) revealed different PAH patterns. In mussel and SPMD, the PAH profiles were very close to that analyzed in seawater. Slight differences were, however, indicated for large molecules with log Kow above six. Nonachievement of steady-state concentration and bioavailability of PAH in oil droplets may account for these differences. The PAH composition in fish revealed only congeners with two to three aromatic rings. A combination of bioavailability and efficient metabolism of the larger PAH molecules may explain this pattern. The CFS made possible a better understanding of some critical factors governing bioconcentration in marine biota from dispersed oil. Yet the results illustrate that uptake of PAH from exposure to oil particles is complex and that different species may bioconcentrate different molecules depending on factors like life style and metabolic capability to degrade the potential harmful substances. Hence, risk assessment of the actual impact of discharges to marine biota should consider these essential biological and ecological factors.

Bioaccumulation of polycyclic aromatic compounds: 2. Modeling bioaccumulation in marine organisms chronically exposed to dispersed oil.

Within the frame of a large environmental study, we report on a research program that investigated the potential for bioaccumulation and subsequent effect responses in several marine organisms exposed to chronic levels of dispersed crude oil. Body burden can be estimated from kinetic parameters (rate constants for uptake and elimination), and appropriate body burden-effect relationships may improve assessments of environmental risks or the potential for such outcomes following chronic discharges at sea. We conducted a series of experiments in a flow-through system to describe the bioaccumulation kinetics of polycyclic aromatic hydrocarbons (PAH) at low concentrations of dispersed crude oils. Mussels (Mytilus edulis) and juvenile turbot (Scophthalmus maximus) were exposed for periods ranging from 8 to 21 d. Postexposure, the organisms were kept for a period of 9 to 10 d in running seawater to study elimination processes. Rate constants of uptake (k1) and elimination (k2) of the PAHs during and following exposure were calculated using a first-order kinetic model that assumed a decrease of the substances in the environment over time. The estimated bioconcentration factor was calculated from the ratio of k1/k2. The kinetic parameters of two-, three-, and four-ring PAHs in mussel and fish are compared with estimates based on hydrophobicity alone, expressed by the octanol-water partition coefficient, Kow (partitioning theory). A combination of reduced bioavailability of PAHs from oil droplets and degradation processes of PAHs in body tissues seems to explain discrepancies between kinetic rates based on Kow and actual kinetic rates measured in fish. Mussels showed a pattern more in compliance with the partitioning theory.

Stability of lysosomal and cell membranes in haemocytes of the common mussel (Mytilus edulis): effect of low temperatures.

Expanding industrial activities in the Arctic require an urgent assessment of the toxicity of chemicals at low temperatures. Organisms acclimatized to low temperature exhibit specific adaptations. For example, the amount of unsaturated lipids is increased to maintain the fluidity of the cell membranes. It has been hypothesized that such temperature-induced alterations in membrane lipid composition may affect the stability of lysosomal and cell membranes in the common mussel, Mytilus edulis, an organism exposed to seasonal temperature extremes. As mussels may be exposed to petroleum compounds along industrialized coastlines, we tested the combined effects of exposure to low temperature and the petroleum compound, phenanthrene, on haemocyte membrane stability. Test animals, acclimated to either 0 or 10 degrees C, were exposed to phenanthrene (0 = control or 500 micrograms l-1) and haemocytes were examined using the neutral red retention assay (lysosomal stability) and a fluorescence assay (cell membrane stability). At 0 degree C, lysosomal and cell membranes from uncontaminated mussels were destabilized compared with 10 degrees C (P = 0.0005). No significant effects (P > 0.05) of phenanthrene were detected at either temperature. Possible mechanisms underlying membrane destabilization include a weaker physical resistance of the membrane due to a higher amount of unsaturated lipids, a potentially higher level of reactive oxygen radicals at low temperature and the higher susceptibility of unsaturated lipids to oxidative stress. More work is required to better understand the consequences of this membrane destabilization at low temperature on the susceptibility of the organism to pollutants.

Pollutant-induced depression of the transmembrane sodium gradient in muscles of mussels.

This study deals with the effects of chemical pollutants on the transmembrane potential difference for sodium (delta mu Na) in smooth muscle cells of Mytilus edulis. A method for indirect determination of extracellular space, intracellular ion concentrations and delta mu Na has been developed and is applied in the investigations. The determination is based on concentration data from haemolymph and muscle tissue samples. The precision of the method used was tested by direct measurements of the apparent intracellular concentration of sodium and the membrane potential. On the basis of these tests, the method was evaluated as reasonably good. The method was used to study the sensitivity of the transmembrane delta mu Na in Mytilus edulis to 96 h exposures to various sublethal concentrations of formaldehyde, methanol and mercury. Both formaldehyde and mercury induced a depression of delta mu Na. The observed depressions could be ascribed to a change in both the electrogenic and the chemical components of delta mu Na. A depression of delta mu Na was associated with subsequent clinical injury and death. Methanol did not cause death or any changes in delta mu Na. Because of the observed correlation between depression of delta mu Na and clinical injury, delta mu Na is suggested to have a potential as an indicator of toxicity.

Physiological parameters in ecotoxicology.

1. Regulated physiological parameters are normally maintained at a constant level by regulatory mechanisms. Acute toxic effects develop whenever a pollutant causes a regulated parameter to be displaced beyond tolerated limits, and thus, regulated parameters may be convenient toxicity parameters. The present study indicates that delta mu Na+ across the adductor muscle membrane of Mytilus edulis is a regulated parameter, and that injuries develop whenever this parameter drops below -700 J/mole. 2. Regulatory physiological parameters may display quick and substantial changes when regulatory mechanisms are activated to counteract variations in the regulated parameters. Thus, regulatory parameters may be used as sensitive alarm parameters in environmental monitoring. The present results indicate that the phosphate index [(ATP x P-arginine)/(Pi)2], metabolic rate and strombine may be used as alarm parameters. 3. The combined response of all parameters may provide a pollutant-specific fingerprint in environmental monitoring.

Measurements of oxygen consumption in Mytilus edulis during exposure to, and recovery from, high sublethal concentrations of formaldehyde, benzene and phenol.

1. The rate of oxygen consumption has been monitored continuously in M. edulis during acute exposure to high sublethal concentrations of formaldehyde, phenol and benzene and subsequent recovery periods of 96 hr. 2. The results are discussed in relation to changes in the electrochemical potential difference of sodium, the content of ATP and the tissue concentration of strombine. 3. After exposure to benzene and phenol, an increase in the rate of oxygen consumption that could not be explained by oxygen debt from the exposure period was observed. 4. Depression of the rate of oxygen consumption after exposure to formaldehyde may be explained by a reduced ability to extract oxygen from the water. 5. The pattern of oxygen consumption and behavioural responses, as well as the combined changes in the biochemical markers, were distinctly different in the three cases.