Mysids in toxicity testing — a review

The use of mysid shrimp, particularly the genusMysidopsis, along with specific testing procedures, has become accepted in aquatic toxicology. Investigators have developed methodologies for both culture and testing of these organisms. Acute and chronic (life cycle) toxicity tests in addition to dredge spoil and effluent tests with mysids are now becoming common. Attempts have been made to use mysids as test organisms in behavioral, physiological, nutritional, and food-chain studies. In general,Mysidopsis spp. have been shown to be as sensitive or more sensitive to toxic substances than other marine species tested. The ease of handling and culture, relative sensitivity to toxicants, short life cycle, small size and direct larval development make these organisms desirable for research purposes. Continued research using mysid species will probably demonstrate even greater usefulness of these organisms in assessment of pollutant impacts on estuarine or marine communities.     

An Evaluation of a Point Source Brine Discharge into a Riverine System and Implications for TDS Limitations

The North Fork Holston River (NFHR) below Saltville, Virginia, has been the subject of many studies due to years of contamination from a closed chloroalkali plant. In addition, a point source brine discharge high in total dissolved solids (TDS) may also be adversely affecting aquatic biota. This study assessed the toxicity of the brine discharge to selected test organisms, including two freshwater mussel species and also discusses implications for development of a TDS limit based on brine toxicity. Ceriodaphnia dubia was the most sensitive organism in acute and chronic brine toxicity tests. Bivariate analysis of the brine conductivity and C. dubia LC₅₀ values showed no relationship (p = .1756); however, LC₅₀ values were strongly related (r² = 0.8818) to sodium concentrations in the brine discharge. A comparison of the results from acute tests with the brine effluent and the reference toxicant sodium chloride (NaCl) demonstrated that organisms were more sensitive to NaCl. Brine discharge toxicity to C. dubia was variable during the study duration with toxicity determined to be dependent on specific TDS constituents rather than conductivity or TDS. These results suggest the need for site-specific development of TDS criteria that are based on the specific constituents in a discharge.     

Toxicity tests with Daphnids: Its application in the management of water quality

The possible role of Daphnia species in toxicity tests is discussed. In screening tests, effluent monitoring tests and legal tests daphnids are recommended as useful test organisms. They also may help to establish concentration limits for chemical water pollution (water quality criteria). As biological indicator organisms their use is limited.     

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.     

Use of Bivalve Metrics to Quantify Influences of Coal-related Activities in the Clinch River Watershed, Virginia

Previously, we reported that survivorship and growth of field-caged Asian clams (Corbicula fluminea [Müller]) were significantly reduced downstream of the wastewater effluent discharge of the Clinch River Plant (CRP), a coal-fired electric power-generating facility in Russell County, VA (USA). These findings warranted the present study, which investigated whether reduced survivorship and growth of transplanted C. fluminea were predictive of adverse effects on resident bivalves – most notably the Clinch River’s (CR’s) federally protected freshwater mussel fauna (Unionoidea). Thus, surveys of unionoid assemblages and C. fluminea population densities and age distributions were conducted to assess ecotoxicological effects on bivalve assemblages naturally occurring in the CR. Results of these surveys suggested that field bioassays with transplanted C. fluminea were predictive of in-stream C. fluminea densities and the presence/absence of indigenous unionoid assemblages. Relative to C. fluminea, additional field bioassays with a juvenile unionoid species (Villosa iris) native to the CR were less predictive of adverse effects on resident bivalves. Concurrence of transplant studies and in-stream surveys was evident at most study sites, particularly within 0.6 km downstream of the CRP effluent where resident bivalves were virtually non-existent relative to other stream sites having similar habitat conditions. Our findings indicate that in situ field bioassays with transplanted C. fluminea are valuable tools for predicting in stream occurrence and distribution of resident freshwater bivalves, and that field studies integrating multiple bivalve metrics may be useful for prioritizing source-reduction efforts in similar watersheds. Discrepancies observed for transplanted C. fluminea and V. iris have important implications to selection of bivalve species used in field bioassays and warrant further study.     

The Effects of Elevated Specific Conductivity on the Chronic Toxicity of Mining Influenced Streams Using Ceriodaphnia dubia

Salinization of freshwater ecosystems as a result of human activities has markedly increased in recent years. Much attention is currently directed at evaluating the effects of increased salinity on freshwater biota. In the Central Appalachian region of the eastern United States, specific conductance from alkaline discharges associated with mountain top mining practices has been implicated in macroinvertebrate community declines in streams receiving coal mining discharges. Whole effluent toxicity testing of receiving stream water was used to test the hypothesis that mine discharges are toxic to laboratory test organisms and further, that toxicity is related to ionic concentrations as indicated by conductivity. Chronic toxicity testing using Ceriodaphnia dubia was conducted by contract laboratories at 72 sites with a total of 129 tests over a 3.5 year period. The database was evaluated to determine the ionic composition of mine effluent dominated streams and whether discharge constituents were related to toxicity in C. dubia. As expected, sulfate was found to be the dominant anion in streams receiving mining discharges with bicarbonate variable and sometimes a substantial component of the dissolved solids. Overall, the temporal variability in conductance was low at each site which would indicate fairly stable water quality conditions. Results of the toxicity tests show no relationship between conductance and survival of C. dubia in the mining influenced streams with the traditional toxicity test endpoints. However, consideration of the entire dataset revealed a significant inverse relationship between conductivity and neonate production. While conductivity explained very little of the high variability in the offspring production (r² = 0.1304), the average numbers of offspring were consistently less than 20 neonates at the highest conductivities.     

The Need for Multiple Lines of Evidence for Predicting Site-Specific Ecological Effects

The goal of this paper is to illustrate the value and importance of the “weight of evidence” approach (use of multiple lines of evidence from field and laboratory data) to assess the occurrence or absence of ecological impairment in the aquatic environment. Single species toxicity tests, microcosms, and community metric approaches such as the Index of Biotic Integrity (IBI) are discussed. Single species toxicity tests or other single lines of evidence are valuable first tier assessments that should be used as screening tools to identify potentially toxic conditions in a effluent or the ambient environment but these tests should not be used as the final quantitative indicator of absolute ecological impairment that may result in regulatory action. Both false positive and false negative predictions of ecological effects can occur due to the inherent variability of measurement endpoints such as survival, growth and reproduction used in single species toxicity tests. A comparison of single species ambient toxicity test results with field data showed that false positives are common and likely related to experimental variability or toxicity to selected test species without measureable effects on the ecosystem. Results from microcosm studies have consistently demonstrated that chemical exposures exceeding the acute or chronic toxicity concentrations for highly sensitive species may cause little or no ecologically significant damage to an aquatic ecosystem. Sources of uncertainty identified when extrapolating from single species tests to ecological effects were: variability in individual response to pesticide exposure; variation among species in sensitivity to pesticides; effects of time varying and repeated exposures; and extrapolation from individual to population-level endpoints. Data sets from the Chesapeake Bay area (Maryland) were used to show the importance of using “multiple lines of evidence” when assessing biological impact due to conflicting results reported from ambient water column and sediment toxicity tests and biological indices (benthic and fish IBIs). Results from water column and sediment toxicity tests with multiple species in tidal areas showed that no single species was consistently the most sensitive. There was also a high degree of disagreement between benthic and fish IBI data for the various stations. The lack of agreement for these biological community indices is not surprising due to the differences in exposure among habitats occupied by these different taxonomic assemblages. Data from a fish IBI, benthic IBI and Maryland Physical Habitat Index (MPHI) were compared for approximately 1100 first through third-order Maryland non-tidal streams to show the complexity of data interpretation and the incidence of conflicting lines of evidence. A key finding from this non-tidal data set was the need for using more than one biological indicator to increase the discriminatory power of identifying impaired streams and reduce the possibility of “false negative results”. Based on historical data, temporal variability associated with an IBI in undisturbed areas was reported to be lower than the variability associated with single species toxicity tests.     

Use of bioassay-based whole effluent toxicity (WET) tests to predict benthic community response to a complex industrial effluent

Whole effluent toxicity (WET) tests are a usefulmonitoring tool because they provide a rapid andreplicable measure of the potential ecotoxicologicaleffect of effluents. Although WET tests have beenincorporated into toxicity-based effluent limits toprotect receiving systems from adverse effects, fewstudies have attempted to quantitativelyfield-validate laboratory-derived toxicity thresholds.In this study, we examine the ability of WET tests topredict response thresholds of an invertebratecommunity to a paper mill effluent discharged into theNicolet-SW River, Québec, Canada. We quantifiedinvertebrate community structure and density in theriver and detrended for the effects ofphysical/chemical variables. This allowed examinationof direct correlation between invertebrate communitystructure and effluent concentration. There was asignificant decrease in taxonomic richness at aneffluent concentration of 16%, but significantchanges in the density of invertebrates occurredbetween 0% and 2% effluent. This suggests thatalthough most taxa returned to the river downstream ofthe effluent, they did so at lower densities.Calculated field thresholds were compared tolaboratory thresholds for the effluent using chronicWET tests with algae, cladocerans and fish. The WETtests produced a mean MATC of 3.6%. Thus, standardWET tests overestimated response thresholds of theinvertebrate community in the receiving environmentand impacts were observed in areas where no impact wasexpected.     

Recent advances in the use of meiofaunal polychaetes for ecotoxicological assessments

While whole sediment toxicity tests with macrofaunal polychaetes are well developed and standardized, they are oftentimes not very sensitive to environmental contaminants. Meiofaunal polychaetes, however, are sensitive to contaminants, easy to culture, and representative of the interstitial habitat. These are desirable attributes for ecotoxicological assessments of marine and estuarine sediments. The meiofaunal polychaete, Dinophilus gyrociliatus Schmidt, 1857, is a cosmopolitan species that has become a useful tool for ecotoxicological assessments, particularly for its use in toxicity testing with sediment pore waters. Due to its short life cycle it is suitable for sublethal toxicity tests, with egg production by the females as a sensitive endpoint, which can be assessed in a 7-day exposure period. Toxicity assessments of pore waters from an industrialized bay in Texas, U.S.A., and of pore waters from sediments spiked with nitroaromatic explosives, demonstrated that the D. gyrociliatus reproduction endpoint was consistently among the most sensitive, when compared to early-life stage tests with several other marine species, including macro-algae, sea urchins and fish. There was also excellent agreement among the results of porewater tests with D. gyrociliatus, nauplii of the meiofaunal copepod, Longipedia americana, and embryos of the sea urchin, Arbacia punctulata in a survey with pore waters extracted from sediments collected in the vicinity of an oil platform in the Gulf of Mexico, where metals were the primary contaminants of concern.     

Spiked Sediment Toxicity Testing of Hydrophobic Organic Chemicals: Bioavailability,Technical Considerations,and Applications

Many evaluations estimating safe levels of hydrophobic organic chemicals in sediments do not account for confounding factors such as physical habitat quality or covariance among chemicals. Controlled experiments demonstrating cause and effect can be conducted with spiked sediment toxicity tests, but application of this methodology has been limited in part by concerns about chemical bioavailability and challenges in achieving target concentrations. Relevant literature was reviewed to assess the utility of standardizing sediment equilibration times; hydrophobicity, complex sediment characteristics, and temperature were identified as potentially equally important factors. Disequilibrium appears likely following limited equilibration time but should yield conservative toxicity test results relative to aged field sediments. Nominal and measured concentrations in over 20 published studies were compared to assess spiked chemical recovery (i.e., measured concentration/nominal concentration). Recovery varied substantially among studies and was not readily predictable based on spiking or extraction method, chemical properties, or measured sediment characteristics, although unmeasured differences between sediments appeared to be important. Factors affecting specific studies included chemical adsorption to glassware, biodegradation, and volatilization. Pre- and post-toxicity test analyses are recommended to confirm exposure concentrations. Studies with 2,3,7,8-tetrachloro-dibenzo-p-dioxin (2,3,7,8-TCDD) and hexachlorobenzene (HCB) exemplify the utility of verifying results of field studies using spiked sediment tests. Sediments spiked with these chemicals at concentrations greatly exceeding those in associated field studies caused no adverse effects in test organisms, demonstrating that other chemicals co-occurring in test sediment samples caused toxicity initially attributed to 2,3,7,8-TCDD and HCB in the field studies. Another key application of spiked sediment tests has been the investigation of TOC as the primary factor affecting bioavailability of hydrophobic organic chemicals. A review of LC50s for nine chemicals reported in 12 studies shows that comparable LC50s derived in different sediments generally agree within a factor of five when concentrations are normalized to a constant TOC. Additionally, use of spiked sediment toxicity testing to investigate toxicological interactions among chemicals provides a promising approach to improving the ability to predict sediment toxicity in the field.     

The effect of dissolved oxygen,sediment, and sewage treatment plant discharges upon growth,survival and density of Asiatic clams

The biology of Corbicula fluminea, the Asiatic clam, in the Vermilion River, Louisiana, as affected by sediment, dissolved oxygen (DO) levels, and sewage treatment plant (STP) effluents was investigated. A point source of high DO water to the Vermilion River established a gradient of DO that decreased as the river moved towards the Gulf of Mexico. Lowering DO levels were exacerbated by municipal sewage treatment plant discharges in the 20 km reach studied. Low dissolved oxygen was associated with reduced Corbicula density in the river and 30-day in-stream growth studies (weight and length) demonstrated that low DO inhibited growth. Generally, if DO was < 1.0 mg l^–1 in sediment pore water and/or < 3.0 mg l^–1 at the sediment-water interface, growth was significantly impaired (p < 0.05). Corbicula experienced substantial mortality near the STP discharges (up to 70% in 30 days) and laboratory toxicity tests with Ceriodaphnia dubia, a sensitive cladoceran, also strongly suggested discharges were chronically toxic at 6.25–25.0% effluent. Respiration experiments along with environmental measurements of DO, temperature, and STP discharge chemistry support a hypothesis that clam populations are adversely affected by the suite of environmental conditions present in the Vermilion River. Further, growth studies were consistent with observed population densities in situ.     

Mastax morphology under SEM and its usefulness in reconstructing rotifer phylogeny & systematics

Biological response could not be predicted based on chemical concentration of the sediment contaminants. Bioassays integrate the response of test organisms to contaminants and nutrients. Comparative results of five acute bioassays indicated that Neubauer phytoassay was the most sensitive. The microbial biomass and algal growth tests indicated a response to the availability of contaminants and nutrients. These results suggest the usefulness of a diversity of bioassays in toxicity testing of sediment contamination.     

Utility and relevance of aquatic oligochaetes in Ecological Risk Assessment

Ecological risk assessment (EcoRA) provides both a process and a framework to evaluate the potential for adverse ecological effects occurring as a result of exposure to contaminants or other stressors. EcoRA begins with problem formulation/hazard identification, progresses to effects and exposure assessment, and culminates with risk characterization (an estimate of the incidence and severity of any adverse effects likely to occur). Key components of EcoRA include determining: stressors/contaminants of concern; sensitive, exposed biota; and, appropriate tests and organisms for evaluating effects. Aquatic oligochaetes are not generally used directly in EcoRA because of three major perceptions. First, EcoRA personnel are generally not familiar with or comfortable using this group of organisms. Second, there is believed to be a paucity of widely accepted toxicity tests with these organisms. Third, their taxonomy is considered difficult and uncertain. In fact, aquatic oligochaetes potentially have great utility and relevance to EcoRAs because of factors including: their importance in the aquatic food chain (e.g. prey to fauna including fish and waterfowl; as a vector for contaminant movement through the food chain from bacteria); many species are widely distributed and well studied; representatives include fresh, estuarine and marine species; as a group, they range from sensitive to insensitive over a wide range of environmental insults; they have a long history of use in pollution monitoring and assessment; and, relevant toxicity and biaccumulation tests exist. Toxicity testing under defined conditions is appropriate for problem formulation while more realistic testing for effects assessment (e.g. microcosms) is logistically easier with this group of organisms than with others due to their relatively small size. The importance of aquatic oligochaetes for EcoRA, in particular of sediments, is particularly compelling.     

Investigation of the sublethal effects of some petroleum refinery effluents

In Canada, environmental regulations for protection of the biota from the adverse effects of effluents from petroleum refineries have tended to focus on acute toxicity. There is concern those effluents may have other subtle, but still deleterious, long-term effects on aquatic ecosystems. We have used a battery of toxicity tests to assess the acute toxicity, genotoxicity, and chronic toxicity of effluent samples from two Ontario refineries. The test organisms included representatives of the bacterial, algal, plant, cladoceran, and fish communities. The results of our preliminary study indicate that the effluent samples had little acute toxicity to the test organisms. There were indications of some sublethal toxicity to Ceriodaphnia dubia, Panagrellus redivivus, and Pimephales promelas. One of the effluents inhibited the growth of Selanastrum capricornutum (IC₅₀ of 59.9%) and Lemna gibba (IC₂₅ of 73.3%) and also caused a 15 percent reduction in the germination of Lactuca sativa seeds. The SOS-Chromotest, a commercially available test that measures the activity of a bacterial DNA repair system, detected genotoxic effects in a single effluent that had been concentrated ten fold. There was no apparent relationship between several chemical parameters and the observed sublethal effects. Further research is needed to establish whether or not the observed toxic effects are typical of effluents from Ontario refineries.     

THE USE OF TOXICITY TESTING IN EFFLUENT REGULATION AND CONTROL: A NEW DIMENSION TO ASSESSMENT AND MANAGEMENT OF WATER QUALITY IN SOUTH AFRICA

Summary

Efficient management tools are continually required to cope with increasing demands placed on the quality and quantity of aquatic resources. Recent developments in environmental monitoring indicate the importance of incorporating biological indicators in assessment programmes. Aquatic toxicology has consequently become an important monitoring and regulatory science. Applications of aquatic toxicity testing include: deriving water quality criteria; toxicological evaluations of whole effluents and receiving waters; and the estimation of ecological risk. Toxicity testing can potentially play a significant role in improving water quality in years to come, especially through its application in effluent regulation. Currently however, few environmental laboratories in South Africa have the required expertise and facilities to carry out a representative range of toxicity tests. Training and funding are required to build the capacity for the necessary developmental research, before toxicity testing can routinely be implemented.

The significant problems we face cannot be solved at the same level of thinking we were at when we created them. Albert Einstein.     

Salt pollution in a Japanese stream and its effects on water chemistry and epilithic algal chlorophyll-a

Concentrations of major ions, total phosphorus, dissolved organic carbon, biological oxygen demand and chlorophyll-a of epilithic algae were determined weekly at nine sites in a Japanese stream receiving effluent from a groundwater treatment plant. The concentrations of four major cations (Na, K, Ca and Mg) and chloride ion increased significantly immediately at downstream sites of the effluent outfall. The ionic concentrations decreased with increasing dilution from merging tributaries but never reached the original concentrations and relative composition of stream water within a 10.7 km stream distance from the outfall. The changes in total ionic concentration and relative ionic proportion also changed the chlorophyll-a content of epilithic algae. The results also showed significantly higher chlorophyll-a content in epilithic algae under moderate salinity.     

Determining the influence of habitat and chemical factors on instream biotic integrity for a Southern Ohio watershed

The relative influences of water quality and in-streamhabitat on receiving water biology were investigatedin the Little Miami River, Ohio, a national and statescenic river located in the southwest corner of thestate. Monitored sample data for water chemistry,habitat, invertebrates and fish were obtained fromfederal and state sources and integrated via ageographical information system (GIS), using a uniquestream segmenting system. Locations and flowinformation for municipal wastewater treatment plants(WWTPs) were also integrated into the system. Therelative importance of water chemistry and habitat onbiological sample information was determined usingmultivariate forward stepwise regression. Theprincipal environmental factors that influencedbiological responses were in-stream habitat andpercent cumulative WWTP effluent at mean flow. Cumulative WWTP effluent at low flow dilution,nutrients and total toxic units from metals andammonia did not correlate with biological responses(e.g. number of mayfly species, invertebrate communityindex (ICI), number of fish species and index ofbiotic integrity (IBI)). We found the GIS-based datamethods to be highly useful for understanding theprincipal processes that guide receiving water health.     

A review of recent non-target toxicity testing of vertebrate pesticides: establishing generic guidelines

Abstract

An analysis of the range, extent and importance of New Zealand-based ecotoxicology studies has been completed to better understand the hazards and non-target risks associated with new toxins and baits. This review focuses on compounds that have been recently developed for incorporation into bait for terrestrial vertebrate pest control, namely cholecalciferol, para-aminopropiophenone (PAPP) and zinc phosphide and the effects of these toxins on non-target species. Testing of these compounds has included cage and pen toxicity and bait acceptance studies. Locally conducted acute toxicity studies clarify overseas data and enhance risk assessments. Consistency in approach and selection of surrogate species, and careful selection of native non-target species for acute toxicity testing in cage and pen trials are recommended as important steps before field trials. We propose a systematic approach to cage and pen trials and offer guidelines on species selection.