A sensitive screening bioassay technique for the toxicological assessment of small quantities of contaminated bottom or suspended sediments

Biological impact assessment of sediment-bound contaminants is currently considered essential in addition to routine chemical characterization. Conventional methods of sediment bioassessment require relatively large quantities of the sample. When sample sizes are limited however, these methods become ineffective. To overcome this problem, the Limited Sample Bioassay (LSB) technique was developed. Bioassays conducted with bottom sediments and suspended particulates from western, central and Arctic regions of Canada indicated that the newly developed LSB technique successfully evaluated the contaminant/nutrient impact on laboratory-grown cultures of algae. The LSB method produced results which were comparable to those from standard elutriate bioassays conducted with natural phytoplankton size assemblages. The data suggest that the LSB can play an important role in providing a quick, simple, sensitive, and inexpensive screening technique for assessing the bioavailability of contaminants (or nutrients) from sediment of limited quantity.     

Comparative ecotoxicity of suspended sediment in the lower Rhone River using algal fractionation,Microtox® and Daphnia magna bioassays

The toxicity associated with suspended sediments from the Rhone River (Switzerland-France) was determined with three acute bioassays. Large volume water samples were centrifuged for recovery of suspended solids in November 1989; one sample was taken as a control upstream from Lake Geneva and the 9 remainder downstream from Geneva to the Mediterranean Sea, with a single sample of the major tributary the Saône at Lyon. Heavy metals (Hg, Cd, Cr, Cu, Ni, Pb, Zn) and organic contaminants (OCs, PCBs, PAHs) bound to sediment were analysed and extracted by elutriation with filtered lake water and by organic solvent (dichloromethane). Sediment water elutriates were tested with algal fractionation bioassays (AFB) using Lake Geneva ambient phytoplankton, with Daphnia magna and Microtox® acute toxicity tests, whereas organic extracts were utilized in the latter two bioassays to evaluate the potential sediment toxicity.The bulk analyses of the sediment together with elutriate metal concentrations indicated the highest contamination of sediment downstream of Lyon. Medium contamination appeared for the stations downstream of Geneva, in the Saône River and at the Rhone River mouth. The station upstream of Lyon had low concentrations, comparable to the values in the Upper Rhone. Organic contaminants are mainly observed downstream of Lyon and their concentrations decline onwards to the sea. The bioassays Microtox® on organic extracts and AFB on the elutriates show a similar toxicity trend, but differ in that Microtox was more sensitive to organics whereas the algal test responded predominantly to metals. This difference is believed to be due to the different extraction procedures used, rather than to the tests themselves. Daphnia magna was the least sensitive and appeared to give a broader band response to the observed contaminants in the sediment. The bioassay results when integrated confirm that the biotoxicity trends relate well to the composition of the sediment, a factor which emphasizes the need for battery testing in ecotoxicological assessment.     

Phytoplankton bioassays for evaluating toxicity of in situ sediment contaminants

Routine bulk chemical characterization of sediments does not provide useful information on toxicity of sediment bound contaminants. This study reviewed and evaluated the utility of phytoplankton bioassays for evaluation of toxicity of sediment bound contaminants, including state-of-the-art techniques. Several techniques such as Algal Fractionation Bioassays, microcomputer-based toxicity testing and in situ bioassays including plankton cages have been developed and successfully applied in our research at various contaminated sites in the St. Lawrence Great Lakes. These bioassay techniques are sensitive, rapid and inexpensive for screening contaminants. The use and application of such techniques, based on bioavailability and physiological response of micro-organisms, are essential for the detection of environmental perturbations of an ecosystem. Such an early warning system will facilitate the preservation and rehabilitation of the Great Lakes.     

Functional bioassays utilizing zooplankton: a comparison

Functional zooplankton bioassays based on ingestion, reproduction and respiration are described, with methods for a new ingestion bioassay included. All bioassays are compared using three indices, including the variability of controls, the range of experimental responses, and a listing of contaminants causing inhibition/stimulation of response. The ingestion bioassay showed the greatest range of response, and was sensitive to pesticides, PCBs and heavy metals. It was also commonly characterized by a hormesis response. The reproduction bioassay showed the lowest variability, illustrated a reduced range of response, and was sensitive to nutrients and heavy metals. In one study, the respiration bioassay was sensitive only to PCBs.     

A sediment suspension system for bioassays with small aquatic organisms

Exposure of aquatic organisms to suspended sediments can impair growth and survival and increase bioaccumulation of sediment-associated contaminants. However, evaluation of the effects of suspended sediments and their associated contaminants on aquatic organisms has been hampered by the lack of a practical and inexpensive exposure system for conducting bioassays. We present a cost-effective system for assessing the effects of suspended sediments and associated contaminants on small aquatic organisms. A 7-day suspension test was conducted with nominal sediment concentrations ranging from 0.0 to 5.0 g 1^–1. The system maintained relatively constant suspended sediment concentrations, as measured by turbidity, and caused minimal mortality to test organisms.     

Assessing toxicity of Lake Diefenbaker (Saskatchewan,Canada) sediments using algal and nematode bioassays

Lake Diefenbaker, on the South Saskatchewan River, Saskatchewan, Canada, receives, on average, 90% of its inflow from snowmelt and rainfall in the Rocky Mountains. The inflowing rivers also receive irrigation return flows and municipal and industrial effluents which may result in the contamination of lake sediments. The sediments were assessed by nematode and algal bioassays.The toxicity of five chemical fractions of the sediment was determined using the nematode Panagrellus redivivus as the test organisms. The results suggest that the sediment chemical fractions frequently inhibit growth and maturation, while lethality was observed at 4 of 12 sites.Samples from 3 of these sites were further evaluated using conventional elutriate Algal Fractionation Bioassays (AFB) with both natural Lake Diefenbaker phytoplankton and a mixed laboratory grown algal culture. The natural phytoplankton showed inhibition at sediment: water ratios of 10: 1; whereas the algal cultures showed both enhancement and inhibition. Evidently, the sediments are frequently toxic to the species tested except for the algal culture. The AFB assesses the mitigative and synergistic effects of contaminants and nutrients and being a conventional elutriate, is more realistic and potentially more acceptable than the chemical fractionation/nematode bioassay technique which essentially considers potential trace organic contaminant effects.     

Assessing the impact of sewage effluent on the ecosystem health of the Toronto Waterfront (Ashbridges Bay), Lake Ontario

The ecosystem health of the Toronto Waterfront (Ashbridges Bay), Lake Ontario which receives treated sewage effluent was investigated during 1987 and 1988 by means of a functional and structural battery of tests. The functional tests included in situ size-fractionated primary productivity, Algal Fractionation Bioassays (AFBs), unfiltered and filtered bioassays, and sediment assays with Daphnia magna and Hyalella azteca. The structural evaluation involved the biomonitoring of the components of the microbial loop, such as bacteria, autotrophic picoplankton, heterotrophic nanoflagellates, and protozoa. The experimental results reveal a diversity of physiological responses to the complex nutrient and contaminant regimes by the indigenous phytoplankton. There was no evidence of the impact of chlorination on the primary productivity of the Bay. The overall productivity was higher during the post-chlorination period than the pre-chlorination phase. The high rates of microplankton + netplankton productivity near the outfall have been attributed to the bioavailability of nutrients which, quite possibly, exert ameliorating effects on metal toxicity. In contrast, the low ultraplankton rates have been interpreted to be due to their well-known sensitivity to contaminants. The Effluent Receiving Water Bioassays (ERWB) with filtered and unfiltered experiments provided interesting insight and appear to be a potentially useful assessment tool. Generally, the unfiltered water compared to the filtered was toxic to the offshore test phytoplankton. This demonstrates a unique ecological adaptation to the prevailing in situ conditions by the Bay community which might be important from the restoration point of view. However, the offshore population was found to be sensitive to the particulate-bound toxicity as indicated by the unfiltered bioassays. Consequently, it is essential to probe the complexity of nutrient-contaminant interactions which ultimately appear to determine the toxicity and the resulting health of the biota. Furthermore, our experiments have shown that the particulate-matter is an important carrier of both nutrients and contaminants in Ashbridges Bay. The sediment bioassays for Station 419 indicated that sediments were toxic during both the pre- and post-chlorination phases. Both solid and liquid phase testing indicated toxicity of sediment to the acute Daphnia test. The Hyalella chronic assay showed good survival during the 4-wk period of the experiment, in contrast to the toxicity observed for phytoplankton and Daphnia. This may be due to large mounts of organic matter available in the Bay. The invertebrate bioassays confirmed the lack of impact of chlorination. Finally, the microbial loop seems to be a sensitive, rapid, and an early warning bioindicator of anthropogenic stress. The multi-trophic battery of structural and functional strategy adopted in our laboratory appear to be holistic and effective. The strategy has a considerable potential for developing eco-technology for a badly needed assessment and restoration of ecosystem health of the Great Lakes as well as other perturbed environments in the world.     

Effects of natural sediment properties on test results in bioassays with oyster larvae (Crassostrea gigas) on sediment elutriates

Toxicity tests with oyster larvae are becoming increasingly important as sensitive bioassays for assessing water and sediment quality. In 1983 a procedure was presented specifically for the testing of seawater elutriates of sediment samples. Testing sediment elutriates with oyster larvae in a sediment-water system invokes specific problems related to natural sediment characteristics. Experiments were set up to study the effects of these sediment properties on the bioassay results. The effects of sediment grain size, suspended solids, elutriation period, as well as the possible effects of naturally occurring toxic nutrients, like ammonia, were studied. Also the effect of presence or absence of the elutriated sediment and the distribution of larvae in the test vessels were studied.It is concluded that natural sediment properties can have substantial effects on the test results in elutriate tests with sediment present in the test vessel. Recommendations are formulated to minimize these effects or to use other test fractions for predicting bulk sediment toxicity.     

Limnological and ecotoxicological studies in the cascade of reservoirs in the Tietê River (S?o Paulo, Brazil).

An evaluation was made of the quality of samples of water and sediment collected from a series of reservoirs in the Tietê River (SP), based on limnological and ecotoxicological analyses. The samples were collected during two periods (Feb and Jul 2000) from 15 sampling stations. Acute toxicity bioassays were performed using the test organism Daphnia similis, while chronic bioassays were carried out with Ceriodaphnia dubia and Danio rerio larvae. The water samples were analyzed for total nutrients, total suspended matter and total cadmium, chromium, copper and zinc concentrations, while the sediment samples were examined for organic matter, granulometry and potentially bioavailable metals (cadmium, chromium, copper and zinc). The results obtained for the limnological variable, revealed differences in the water quality, with high contribution of nutrients and metals for Tietê and Piracicaba rivers, besides the incorporation and sedimentation, consequently causing a reduction of materials in Barra Bonita reservoir, thus promoting the improvement of the water quality in the other reservoirs. The toxicity bioassays revealed acute toxicity for Daphnia similis only in the reservoirs located below Barra Bonita dam. On the other hand, chronic toxicity for Ceriodaphnia dubia and acute for Danio rerio showed a different pattern, decreasing in magnitude from Barra Bonita to Três Irm?os, demonstrating an environmental degradation gradient in the reservoirs.     

Background and overview of current sediment toxicity identification evaluation procedures

Laboratory bioassays can provide an integrated assessment of the potential toxicity of contaminated sediments to aquatic organisms; however, toxicity as a sole endpoint is not particularly useful in terms of identifying remedial options. To focus possible remediation (e.g., source control), it is essential to know which contaminants are responsible for toxicity. Unfortunately, contaminated sediments can contain literally thousands of potentially toxic compounds. Methods which rely solely on correlation to identify contaminants responsible for toxicity are limited in several aspects: (a) actual compounds causing toxicity might not be measured, (b) concentrations of potentially toxic compounds may covary, (c) it may be difficult to assess the bioavailability of contaminants measured in a sediment, and (d) interactions may not be accounted for among potential toxicants (e.g., additivity). Toxicity identification evaluation (TIE) procedures attempt to circumvent these problems by using toxicity-based fractionation procedures to implicate specific contaminants as causative toxicants. Phase I of TIE characterizes the general physio-chemical nature of sample toxicants. Phase II employs methods to measure toxicants via different analytical methods, and Phase III consists of techniques to confirm that the suspect toxicants identified in Phases I and II of the TIE actually are responsible for toxicity. These TIE procedures have been used to investigate the toxicity of a variety of samples, including sediments. Herein we present a brief conceptual overview of the TIE process, and discuss specific considerations associated with sediment TIE research. Points addressed include: (a) selection and preparation of appropriate test fractions, (b) use of benthic organisms for sediment TIE work, and (c) methods for the identification of common sediment contaminants.     

The role of bivalve molluscs as tools in estuarine sediment toxicity testing: a review

Estuarine sediments frequently are repositories and therefore potential sources of anthropogenic contaminants. Many organic and metallic chemical compounds released into aquatic systems bind to particulates and so accumulate in the sediments, thus, sediments become repositories of contaminants in estuaries. These may also cause contamination through diffusion of porewater, resuspension of particulates and dispersal of benthic fauna. There is a need to assess the biological affects of these anthropogenic contaminants because they may be toxic to infauna and bottomfish. Sediment toxicity bioassays are a means for carrying out such an assessment and primarily provide data on toxicity by measuring the effects on the test organism. Existing sediment toxicity bioassays rely on a battery of aquatic toxicity tests, which are based on the extraction of pore water, and elutriate from sediments and then subjecting these sediment phases to toxicity testing regimes. Two estuarine bivalve molluscs, Scrobicularia plana and Tapes semidecussatus were used to assess the ecotoxicity of field-collected sediments from estuarine and coastal areas around the Irish and English Coast over a 3-year study period. A variety of endpoints were measured during the study including survival in air, behaviour, animal condition, biochemistry, soft tissue metal concentrations, lysosomal membrane integrity and histopathology. Of these endpoints, the most sensitive were survival, survival in air, lysosomal membrane integrity, behaviour and histopathology.     

An improved elutriation technique for the bioassessment of sediment contaminants

An improved method is proposed for the preparation of sediment elutriates which permits relatively realistic determination of bioavailable contaminants. It suggests the use of rotary tumbling in a cycle of 3–4 rpm to achieve sediment-water mixing. Experiments were undertaken to evaluate the mixing efficiency of the rotary tumbler as compared to that of the compressed air, wrist-action shaker, and reciprocal shaker methods. Sediment to water ratios of 0 : 1, 1 : 20, 1 : 10, and 1 : 4 were tested over 0.5, 1.0, 24, and 48-h elution periods. Elutriate evaluations were based on chemical, physico-chemical and gravimetric determinations; and also on ¹⁴C-phytoplankton bioassays using Chlorella vulgaris (Beyerinck). Results indicated that rotary tumbling produced the most consistent bioassay-supportable data. It was also the most efficient procedure when used for 1 h with 1 : 4 sediment-water mixtures.     

Subsidy‐stress and multiple‐stressor effects along gradients of deposited fine sediment and dissolved nutrients in a regional set of streams and rivers

1. Stream managers need to understand relationships between multiple stressors and ecological responses. We examined responses of benthic invertebrates and algae along two land‐use‐related stressor gradients of concern in running waters. Our correlative study of the consequences of augmented deposited fine sediment and nutrient concentrations was conducted in a regional set of streams and rivers monitored by a water management authority in New Zealand and incorporated a wide range of catchment geologies and stream orders. 2. We used multiple linear regression analysis and an information‐theoretic approach to select the best predictive models for our biological response variables by testing multiple competing hypotheses that include nonlinear subsidy‐stress relationships and interactive effects between the two stressors. 3. Patterns consistent with a subsidy‐stress response to increasing dissolved inorganic nitrogen concentration were found for the relative abundances of the common invertebrate genera Pycnocentrodes and Deleatidium and for the relative abundance of total individuals in the EPT orders (Ephemeroptera, Plecoptera, Trichoptera). 4. Fine sediment seemed the more pervasive stressor, apparently counteracting and overwhelming any initial subsidy effect of increased nutrients, and accounting for more of the variance in biological response variables. Relationships with high nutrient concentrations were weaker and modelled with less certainty, probably reflecting the indirect modes of action of nutrients compared to those underlying sediment effects. Nevertheless, in several cases, the models indicated that nutrients interacted synergistically with fine sediment, lending further weight to the conclusion that managers need to address both stressors to achieve the best outcomes.     

Short- and Long-Term Exposure of Lumbriculus variegatus (Oligochaete) to Metal Lead: Ecotoxicological and Behavioral Effects

Metals are naturally occurring constituents of the environment and although many are essential nutrients for living organisms, at higher concentrations they can be toxic. Some aquatic species can help understand and even predict the impact of those contaminants. Lumbriculus variegatus is a recommended species for use in sediment toxicity tests and is known to have a remarkable ability of segmental regeneration. Short- (10-day) and long-term (28-day) toxicity tests were used to test the effects of a metal on the survival, growth, and behavior of L. variegatus. This work aims to investigate and validate the use of behavior as a new parameter in standard toxicity tests. Worms were exposed to sediments contaminated with different levels of lead and the results indicated a positive relation between lead concentrations and mortality and growth: higher lead concentrations resulted in higher mortalities and strong inhibition of growth. An inhibition of behavior was observed and results suggested that although behavior could not be used in sediment toxicity tests, it proved useful as an addition to short-term tests and helps select sediments. Thus, exposure to sediments contaminated with lead affects the presence of this species in nature, because it interferes with growth and survival.     

Acute toxicity testing with the European estuarine amphipod <Emphasis Type="Italic">Corophium multisetosum</Emphasis>

This study reports on the use of the estuarine amphipod Corophium multisetosum in acute toxicity testing. The species was successfully acclimated to the laboratory and was used in a water-only whole effluent 96 h acute bioassay and in a 10 days whole estuarine sediment test. C. multisetosum response was compared to other species in 96 h bioassays, testing boiling cork effluent and iron filings lixiviates. The amphipod showed high sensitivity and the results were similar to those obtained with others species namely, the freshwater cladoceran Daphnia magna, the estuarine amphipod Gammarus chevreuxi and the seawater annelid Sabellaria alveolata. In a 10 days static exposure to natural impacted estuarine sediments, the response of the species was compared to a fertilization bioassay with the sea-urchin Paracentrotus lividus. Both species indicated the same sediment samples as the most potentially toxic. These samples were collected in sites where the resident macrofauna benthic community is also the most affected, with strong reduction of the species richness, abundance and biomass. The results revealed that C. multisetosum presents high potential to be used in routine acute toxicity testing in the estuarine environment.     

Ecotoxicological characterisation of sedimentation in the Kis-Balaton Water Protection System

The main function of the Kis-Balaton Water Protection System is to retain nutrients and total suspended solids, thus protecting the water quality of Lake Balaton. In this paper, the toxic nature of the sediment in the 2nd reservoir of the KBWPS has been characterised, using a battery of tests: Vibrio fischeri acute bioassay on whole sediment samples, and V. fischeri bioassay on pore water and elutriate samples. The latest version of the V. fischeri bioluminescence inhibition was applied, the Flash assay which uses a kinetic mode and is able to detect the toxicity of solid, turbid/coloured samples. Whole sediment toxicity showed a clear spatial distribution of toxicity, in parallel with elutriate toxicity. However, no pore water toxicity was detected, leading to the conclusion that contaminants are not water soluble.     

Use of a multiple addition bioassay to determine limiting nutrients in eagle lake,California

This investigation uses anin vitro enrichment bioassay (in which all essential nutrients except one are added to each culture) to determine which nutrients are important to algal growth in Eagle Lake. The technique was developed when it was discovered that addition of individual nutrients produced little if any growth response. Laboratory bioassays correlated well with comparative studies in the lake. A great deal of variation was found throughout the year but P, N, Fe, and S were found to be limiting at one time or another. The north and south basins of the lake, which differ in morphometry, were also found to differ in the intensity spectrum of limitation. While P was the most important nutrient in both basins, the other nutrients were more limiting in the north basin than the south, and Fe, which was least limiting in the south, was very important in the north. The multiple enrichment bioassay has several advantages over other bioassays. Supported in part by Research Corporation     

Contamination on sandflats and the decoupling of linked ecological functions

Benthic macrofauna can influence inputs and transformations of energy and matter in estuaries, affecting both the stocks of vital materials (e.g. carbon, oxygen) and the rates of key processes (e.g. organic matter decomposition, nutrient uptake). Although a number of studies have identified shifts in functional groups or biological traits in relation to anthropogenic stressors, there have been few field‐based assessments of changes in functioning associated with stress gradients. We used a comparative experimental approach to investigate functioning on two sandflats with differing exposures to urban contaminants. Apart from significant differences in sediment contaminant concentrations (43.2 ± 1.8 mg kg^?1 Zn and 15.6 ± 0.9 mg kg^?1 Pb at the Pollen site; 17.7 ± 0.7 mg kg^?1 Zn and 7.9 ± 0.9 mg kg^?1 Pb at the Waiheke site), the two sandflats were readily comparable: both had similar sediment grain size distributions and were dominated by the same macrofaunal species; and both were in non‐eutrophic New Zealand marine reserves with low ambient sediment organic matter content. To better understand the effects of contaminants on biologically mediated transformations of organic matter into inorganic nutrients, we manipulated sediment organic matter content and macrofaunal abundance in standardized treatments at each site. Fluxes of oxygen and ammonium, which are linked to key sandflat processes such as organic matter decomposition and benthic photosynthesis, were measured as response variables 1 week after the experimental manipulations. We predicted more efficient organic matter processing on the uncontaminated flat and thus expected to see elevated ammonium efflux in response to organic enrichment treatments at this site. Higher rates of benthic photosynthesis were predicted for plots with higher ammonium efflux, as ammonium is a readily utilizable form of limiting inorganic nitrogen. We documented significant positive relationships between ammonium uptake and benthic primary production on the uncontaminated flat, but weaker/insignificant relationships at the contaminated site. Our data were consistent with theories of increased variability and a decoupling of system processes with increasing amounts of stress.     

Assessment of sediment contamination at Great Lakes Areas of Concern: the ARCS Program Toxicity-Chemistry Work Group strategy

In response to a mandate in Section 118(c)(3) of the Water Quality Act of 1987, a program called Assessment and Remediation of Contaminated Sediments (ARCS) was established. Four technical work groups were formed. This paper details the research strategy of the Toxicity-Chemistry Work Group.The Work Group's general objectives are to develop survey methods and to map the degree of contamination and toxicity in bottom sediments at three study areas, which will serve as guidance for future surveys at other locations. A related objective is to use the data base that will be generated to calculate sediment quality concentrations by several methods. The information needed to achieve these goals will be collected in a series of field surveys at three areas: Saginaw Bay (MI), Grand Calumet River (IN), and Buffalo River (NY). Assessments of the extent of contamination and potential adverse effects of contaminants in sediment at each of these locations will be conducted by collecting samples for physical characterization, toxicity testing, mutagenicity testing, chemical analyses, and fish bioaccumulation assays. Fish populations will be assessed for tumors and external abnormalities, and benthic community structure will be analyzed. A mapping approach will use low-cost indicator parameters at a large number of stations, and will extrapolate by correlation from traditional chemical and biological studies at a smaller number of locations. Sediment toxicity testing includes elutriate, pore water and whole sediment bioassays in a three-tiered framework. In addition to the regular series of toxicity tests at primary mater stations, some stations are selected for a more extensive suite of tests.