Assessment of oil refinery wastewater and effluent integrating bioassays, mechanistic modelling and bioavailability evaluation.

Water is used in petroleum oil refineries in significant volumes for cooling, steam generation and processing of raw materials. Effective water management is required at refineries to ensure their efficient and responsible operation with respect to the water environment. However, ascertaining the potential environmental risks associated with discharge of refinery effluents to receiving waters is challenging because of their compositional complexity. Recent European research and regulatory initiatives propose a more holistic approach including biological effect methods to assess complex effluents and surface water quality. The study presented here investigated potential effects of effluent composition, particularly hydrocarbons, on aquatic toxicity and was a component of a larger study assessing contaminant removal during refinery wastewater treatment (Hjort et al 2021). The evaluation of effects utilised a novel combination of mechanistic toxicity modelling based on the exposure composition, measured bioavailable hydrocarbons using biomimetic solid phase microextraction (BE-SPME), and bioassays. The results indicate that in the refinery effluent assessments measured bioavailable hydrocarbons using BE-SPME was correlated with the responses in standard bioassays. It confirms that bioassays are providing relevant data and that BE-SPME measurement, combined with knowledge of other known non-hydrocarbon toxic constituents, provide key tools for toxicity identification. Overall, the results indicate that oil refinery effluents treated in accordance to the EU Industrial Emissions Directive requirements have low to negligible toxicity to aquatic organisms and their receiving environments. Low-cost, animal-free BE-SPME represents a compelling tool for rapid effluent characterization.

Solubility Constraints on Aquatic Ecotoxicity Testing of Anionic Surfactants.

In order to develop models that can predict the environmental behavior and effects of chemicals, reliable experimental data are needed. However, for anionic surfactants the number of ecotoxicity studies is still limited. The present study therefore aimed to determine the aquatic ecotoxicity of three classes of anionic surfactants. To this purpose we subjected daphnids (Daphnia magna) for 48 h to alkyl carboxylates (CxCO2-), alkyl sulfonates (CxSO3-), and alkyl sulfates (CxSO4-) with different carbon chain lengths (x). However, all surfactants with x > 11 showed less than 50% immobility at water solubility. Hence, EC50 values for only few surfactants could be gathered: C9CO2- (16 mg L-1), C11CO2- (0.8 mg L-1) and C11SO4- (13.5 mg L-1). Data from these compounds showed an increase in ecotoxicity with a factor 4.5 per addition of a hydrocarbon unit to the alkyl chain, and a factor 20 when replacing the sulfate head group by a carboxylate head group. Unfortunately, we could not test carboxylates with a broader variety of chain lengths because solubility limited the range of chain length that can be tested.

Environmental impact and recovery at two dumping sites for dredged material in the North Sea.

The environmental impact and recovery associated with the long and uninterrupted disposal of large volumes of moderately contaminated dredged material from the port of Rotterdam was studied at nearby dumping sites in the North Sea. Observations were made on sediment contamination, ecotoxicity, biomarker responses and benthic community changes shortly after dumping at the 'North' site had ceased and at the start of disposal at the new dumping site 'Northwest'. During the period of dumping, very few benthic invertebrates were found at the North site. Concentrations of cadmium, mercury, polychlorinated biphenyls (PCBs), polyaromatic hydrocarbons (PAHs) and tributyltin (TBT) in the fine sediment fraction (<63 microm) from this site were 2-3 times higher than at the reference site. In four different bioassays with marine invertebrates the sediments showed no acute toxic effects. In tissue (pyloric caeca) of resident starfish Asterias rubens, residual levels of mercury, zinc, PCBs and dioxin-like activity were never more than twice those at the reference site. Four different biomarkers (DNA integrity, cytochrome P450 content, benzo[a]pyrene hydroxylase activity and acetylcholinesterase inhibition) were used on the starfish tissues, but no significant differences were found between North and the reference site. Minor pathological effects were observed in resident dab Limanda limanda. One year after dumping had ceased at the North site, a significant increase in the species richness and abundance of benthic invertebrates and a concomitant decrease in the fine sediment fraction of the seabed were observed. After 8.2 million m3 of moderately contaminated dredged material had been dumped at the new dumping site Northwest, the species richness and abundance of benthic invertebrates declined over an area extending about 1-2 km eastwards. This correlated with a shift in sediment texture from sand to silt. The contamination of the fine sediment fraction at the Northwest location doubled. It is concluded that marine benthic resources at and around the dumping sites have been adversely affected by physical disturbance (burial, smothering). However, no causal link could be established with sediment-associated contaminants from the dredged spoils.

Confounding factors in bioassays with freshwater and marine organisms.

The use of bioassays in ecological risk assessments often raises questions about the causative factors, and insight into the possibility that confounding factors, such as pH or increased ammonia concentrations, might be responsible for the observed toxicity is needed. It was decided to develop a practical approach for the Dutch situation, in which a first screening is carried out based on provisional criteria. In collecting the required data, dozens of experiments were performed, while the scientific literature was searched for additional information. It is concluded that the provisional criteria specified are at present useful tools in interpreting results of bioassays. Depending on the outcome and the aim of the research, it might be necessary to further reduce uncertainties in the interpretation. This might require some additional experiments, using alternative controls or test procedures or altering the composition of the original sample.

Bioaccumulation and biomarkers in the sea star Asterias rubens (Echinodermata: Asteroidea): a North Sea field study.

During two North Sea field trips in March 1995 and September 1996 sea stars, Asterias rubens, were collected at various stations along pollution gradients in order to study the relation between biochemical markers and levels of accumulated contaminants. Biomarkers measured were: cytochrome P450 level, benzo[a]pyrene hydroxylase (BPH) activity, acetylcholinesterase (AChE) activity and DNA integrity. Accumulation levels of heavy metals, polychlorinated biphenyls, organochlorine pesticides and polycyclic aromatic hydrocarbons (PAHs) in the pyloric caeca of sea stars indicate different pollution gradients, influenced by rivers in The Netherlands, UK and Germany. For some contaminants, especially PAHs, relatively high levels were found in the central part of the North Sea (Dogger Bank). On the basis of multivariate statistics, stations near the mouth of the Elbe and the Rhine/Meuse were shown to have different patterns of biomarker responses. Sea stars from stations in coastal zones showed relatively high levels of cytochrome P450 and 'P418', another haemoprotein that is present in most marine invertebrates. The station nearest to the Elbe Estuary showed the lowest BPH and AChE activity. DNA integrity was lower especially in stations near the Dutch coast and in a station near the Tees/Tyne estuaries. Using these biomarkers as early warning signals of exposure and/or adverse effects, this type of monitoring can be used also in the future to study the spatial and temporal trends in the quality of coastal waters.

Comparison of laboratory and in situ sediment bioassays using Corophium volutator.

Bioassays with the marine amphipod Corophium volutator were performed simultaneously in situ and in the laboratory using sediments sampled from the in situ locations. In most cases, the in situ response was significantly higher compared to the laboratory response. This difference was not caused by direct influence of the use of the field chamber on Corophium sp., nor was the difference caused by the overlying water used. Experiments showed homogenization can affect the toxicity of a sediment, but not in such a way that it can completely explain the difference between the response in situ and in the laboratory. Possible explanatory factors are harbor activity, storms, and temperature. To reduce the influence of some of these factors, the best period of the year to perform in situ bioassays with C. volutator is May, June, or September.

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.

Tolerance induction and life cycle changes in cadmium-exposed Chironomus riparius (Diptera) during consecutive generations.

Cultures of Chironomus riparius were exposed to cadmium during nine consecutive generations to determine whether cadmium tolerance could be induced. Selection for cadmium tolerance was assumed to influence the population dynamics of this species. Therefore, the responses and interactions of different population parameters (such as mortality, growth, and reproduction) were studied during the selection process. Exposure to cadmium during consecutive generations caused increasing effects on some life cycle parameters compared to a one-generation experiment. Tolerance to cadmium increased during exposure to 54.2 nM Cd and the tolerant population seemed to be stimulated by low cadmium concentrations (based on an acute growth experiment). Despite this tolerance development, mortality among cadmium-exposed tolerant chironomids remained high. These experiments illustrated that changes of the life cycle and tolerance can be expected as soon as single-generation NOEC values are exceeded, and in addition that "safe concentrations" based on a one-generation toxicity experiment could well underestimate the potential effects of a toxicant on midge populations.

Chronic toxicity of cadmium to Chironomus riparius (Diptera: Chironomidae) at different food levels.

The interacting effects of cadmium toxicity and food limitation on the midge, Chironomus riparius, were studied during chronic exposure in laboratory experiments. If the food was supplied ad libitum, both larval developmental time and mortality of the larvae were negatively affected by cadmium concentrations of 2.0-16.2 micrograms/L. The number of eggs deposited per female and the mean life span of the imagines were not affected by cadmium. Integration of these separate effects into a population growth rate showed a clear reduction with increasing cadmium concentrations. Food limitation of unexposed larvae at high population density reduced fitness, judged on all parameters studied and consequently reduced the population growth rate (up to 85%). The effects on larvae, which were exposed to both cadmium and food limitation, differed considerably from the response to the individual stress factors. Exposure to cadmium increased mortality among food-limited first and second instar larvae. Consequently, the amount of food available for each surviving larva increased. At the two lowest concentrations studied (2.0 and 5.6 micrograms Cd/L), these indirect positive effects of cadmium overruled the direct negative effects and caused an increase of the fitness of the food-limited exposed larvae compared to the food-limited, unexposed controls. At a concentration of 16.2 micrograms Cd/L, the negative effects of cadmium on food-limited midges balanced the positive effects of reduced food limitation. At this concentration, the population growth rate did not differ significantly from the food-limited control any more. It is concluded that the indirect positive effects of cadmium on food limitation could eliminate negative, direct effects of low cadmium concentrations on food-limited chironomid populations.

A comparison of test systems for assessing effects of metals on isopod ecological functions.

The effects of metals on the isopod Porcellio scaber and the implication of metal pollution for the functioning of isopods in soil decomposition processes were determined. The experiments were carried out using two test systems, a simple test container without natural substrate and a micro-ecosystem with leaf litter, the latter being more comparable to the field situation. Metals were offered to the isopods by feeding artificially contaminated litter and litter from contaminated field sites. The Cd concentrations in isopods in test containers with ground litter as food were similar to those in isopods in micro-ecosystems with intact leaves as food. Exposure to artificially contaminated litter with Cd alone or litter from contaminated field sites with Cd, Pb, Zn, and Cu resulted in comparable Cd concentrations in the isopods. Growth of the isopods was not affected by Cd or by the combination of Cd, Pb, Zn, and Cu, but differed between the test systems. Fragmentation activity of isopods decreased in litter containing 155 nmol/g Cd and 33500 nmol/g Zn, sampled at a contaminated field site. CO2 production from micro-ecosystems decreased at a concentration of 1010 nmol/g Cd, but NH4 and NO3 concentrations in the litter did not respond to Cd. Isopod growth was better in micro-ecosystems. These systems are therefore recommended for ecotoxicity tests using isopods. After comparison with literature data it was concluded that the bioavailability of Cd in laboratory tests is often higher than the bioavailability in the field at the same total concentrations. This may be due to environmental heterogeneity; this gives animals food choice and is absent in many laboratory test systems. For the quantification of a laboratory-field extrapolation factor for general use, more detailed information is needed than is available now.