Water chemistry influences the toxicity of silver to the green-lipped mussel Perna viridis

The study determined the influence and relative importance of water chemistry parameters (pH, alkalinity, hardness) on the acute toxicity of silver to the green mussel Perna viridis. A preliminary bioassay revealed that 4 mg L^???1 of silver caused 50% mortality (LC₅₀) in 96 h for mussels placed in seawater with pH 8.5, hardness 1,872 mg L^???1, and alkalinity 172 mg L^???1. Mortality of mussels increased with decreasing pH and increasing hardness and alkalinity variables. In contrast the mortality decreased with increasing pH and decreasing hardness and alkalinity values. The water chemistry also affected the concentration of sliver in experimental seawater and bioaccumulation of silver in mussels. The results revealed that the chemical properties of seawater must be considered while conducting toxicity tests with metals like silver. The possible explanations for the influence of water chemistry on silver toxicity to P. viridis are discussed.     

Investigations of the impact of effluent from the Ok Tedi copper mine on the fisheries resource in the Fly River,papua new guinea

The Ok Tedi copper mine currently discharges overburden and partially treated ore residues into the Ok Tedi, a major tributary of the Fly River. At peak production these discharges will result in an increase in the suspended solids levels in the Fly River from a background of 76 mg L^-1 to approximately 800 mg L^-1, and an increase in median particulate copper levels from 90 g g^-1 to 1220 g g^-1. The dissolved copper levels are not expected to exceed 10 g L^-1. The Strickland River, a tributary of the Fly River, has natural suspended solids levels in excess of 500 mg L^-1 and provided a natural bioassay for the impacts of elevated suspended solids levels. The fish communities of the Strickland River were dominated by the sediment-tolerant Ariidae catfish, but the levels of biomass caught per standard sampling effort were comparable to those in the Fly River. Laboratory bioassays were performed to assess the impact of particulate associated copper. The fish fauna of the Fly River does not include any standard test species, and it is unlikely that standard species would be unaffected by the high suspended solids levels of the Fly River. Test species were selected on the basis of ecological importance. Two freshwater prawns,Macrobrachium rosenbergii andMacrobrachium sp., a catfish,Neosilurus ater, and a cladoceran,Ceriodaphnia dubia, were tested for acute toxicity and bioaccumulation of copper from particulates. There was no evidence of acute toxicity to prawns or fish, nor for bioaccumulation by prawns. Acute toxicity toC. dubia and bioaccumulation byN. ater were probably due to dissolved copper in the test environments. The test results indicated that provided dissolved copper levels did not exceed the predicted levels, there should not be toxic affects of particulate-associated copper in the Fly River.     

The importance of adult Hediste (Nereis) diversicolor in managing heavy metal pollution in shores and estuaries

The main aim of the toxicity test is to be able to measure toxicity in organisms, and therefore to assess whether a nationally or internationally preset standard is met. LC₅₀ values (median lethal concentration that kills 50% of the population at a given time) or LT₅₀ values (median lethal time in which 50% of the test species died) provide useful information for risk assessment. However, extrapolating laboratory data to actual field conditions is difficult because of changing environmental factors like temperature and salinity. LC₅₀ copper data for Hediste diversicolor treated multifactorially show the plasticity of the worm to environmental variables. Increasing temperature, 12 to 22°C, and increasing salinity, 7.6 to 30.5 reduced toxicity of copper to the worms without sediment. In the presence of sediment increasing temperature and increasing salinity increased toxicity of copper to the worms.     

An interlaboratory comparison of sediment elutriate preparation and toxicity test methods

Elutriate bioassays are among numerous methods that exist for assessing the potential toxicity of sediments in aquatic systems. In this study, interlaboratory results were compared from 96-h Ceriodaphnia dubia and Pimephales promelas static-renewal acute toxicity tests conducted independently by two laboratories using elutriate samples prepared from the same sediment. The goal of the study was to determine if the results from the elutriate tests were comparable between two U.S. Environmental Protection Agency (USEPA) laboratories when different elutriate preparation procedures were employed by each lab. Complete agreement in site characterization was attained in 22 of the 25 samples for both bioassays amongst each lab. Of the 25 samples analyzed, 10 were found to be toxic to at least one of the species tested by either laboratory. The C. dubia elutriate tests conducted by the National Exposure Research Laboratory (NERL) indicated that 7 of the 25 sediment samples were toxic, while 8 sediment samples were characterized as such in testing conducted by USEPA Region 6 (Region 6). The P. promelas elutriate tests conducted by NERL determined 8 samples as toxic, while Region 6 tests displayed toxicity in 5 of the samples. McNemar's test of symmetry for C. dubia (S?=?0.33, p?=?0.5637) and P. promelas (S?=?3.0, p?=?0.0833) tests indicated no significant differences in designating a site toxic between NERL and Region 6 laboratories. Likewise, Cohen's kappa test revealed significant agreement between NERL and Region 6 C. dubia (K?=?0.7148, p?<?0.01) and P. promelas (K?=?0.6939, p?<?0.01) elutriate tests. The authors conclude that differences in interlaboratory elutriate preparation procedures have no bearing on the ability of either the C. dubia or P. promelas bioassay testing methods to detect toxicity while yielding similar results.     

The interaction between concrete pavement and corrosion-induced copper runoff from buildings

Changes in chemical speciation of copper and the capacity of concrete pavement to retain copper in runoff water from external buildings have been investigated at urban field conditions, and in parallel laboratory experiments simulating outdoor scenarios. The research study showed the concrete surface to form a copper rich surface layer (≈50 μm thick) upon exposure, and a high capacity to significantly reduce the bioavailable fraction of released copper (20–95%). The retention capacity of copper varied between 5 and 20% during single runoff events in the laboratory, and between 10 and 40% of the total copper release during single natural runoff events. The capacity to retain and reduce the bioavailable fraction of non-retained copper increased with increasing wetness of the concrete surfaces, increasing pH of the runoff water and decreasing flow rates. Bioassay testing with bacterial and yeast bioreporters showed the bioavailable fraction of non-retained copper to be significantly lower than the total copper concentration in the runoff water, between 22 and 40% for bacteria and between 8 and 31% for yeast. The application of generated data to simulate a fictive outdoor scenario, suggests a significant reduction of bioavailable and total copper to background values during environmental entry as a result of dilution, and the interaction with solid surfaces, organic matter and complexing agents already in the drainage system.     

Stress ethylene: A bioassay for rhizosphere-applied phytotoxicants

A bioassay for rhizosphere-applied phytotoxicants was developed and evaluated with a broad range of chemicals. Test substances were applied to the rhizosphere of whole, intact bush bean plants (Phaseolus vulgaris L. cv. Bush Blue Lake 290) grown in a solid support medium and the resultant ethylene production was measured to detect the presence of phytotoxic materials. The beans were encapsulated in plastic bags for 2 hr following treatment and then incubated for 24 hr in the dark. Ethylene and ethane accumulating within the bags were quantified via gas-solid chromatography. The application of various concentrations of inorganic and organic chemicals induced various responses. No single equation adequately described the dose-response curves; therefore, a critical value (a statistically significant increase in stress ethylene) was computed for each test substance. A phytotoxic-response threshold for each test substance was defined as the test-substance concentration that caused ethylene production to exceed its respective critical-value concentration. Based on threshold concentrations determined by analysis of stress ethylene production, the relative phytotoxicity rankings of the inorganic test substances were: CdCl₂>CuCl₂>Pb(C₂H₃O₂)₂>LiCl, while those of the organic test substances were 2,4-D esters>paraquat dichloride>1-butanol>2-propanone>2-propanol. Both stress ethylene and ethane production were nonresponsive to hydrogen-ion concentration (measured before application) over a broad pH range. However, significant ethane production was detected at pH 2 and stress ethylene was produced at pH 1. The measurement of stress-induced ethylene provides a rapid and simple means to determine the relative phytoxicity of rhizosphere-applied substances.     

Effects of temperature on the acute toxicity of cadmium to Corophium Insidiosum

This study aimed to optimise the methodology for the use of Corophium insidiosum in a bioassay. Taking into account that it would be suitable to execute the bioassay with organisms having a good sensitivity during the year and low mortality in control sediment, the influence of different temperatures (10–15–20 and 25°C) has been examined. C. insidiosum was collected during August, November 2005 and January 2006, in Mar Piccolo basin (Ionian sea). The results obtained show that this species mortality in the negative control sediment, ranged from 2.6 ± 0.6% at 10°C in August to 17 ± 2.2% at 20°C in November, at different temperatures tested. At 20°C there were significant differences in mortality among different months examined. Indeed no relationship among months was found at 15°C. Significant differences between August and November at 25°C, between November and January were not found at 10°C. The 96-h LC50 values found for cadmium at all temperature experimental conditions ranged from 2.11 mg/l (1.57–2.82) to 0.70 mg/l (0.54–0.93). The highest values were found at 10°C in November and January. The results showed that the optimal temperature for the bioassay seems to be between 15°C and 20°C. Even if, at 20°C the mortality differs significantly among organisms sampled.     

Coal-Mine Hollow Fill and Settling Pond Influences on Headwater Streams in Southern West Virginia, USA

The influences of coal-mine hollow fills and associated settling ponds in three headwater streams were assessed in southern West Virginia, USA. Fill drainage waters had elevated conductivities and metal concentrations, compared to a regional reference. Benthic macroinvertebrate richness was not affected consistently by the hollow fill drainages, relative to a regional reference, although a more tolerant community, lacking in Ephemeroptera taxa at most locations, was evident. Collector-filterer populations were elevated at monitoring stations directly below the settling ponds, indicating that the ponds’ presence influenced macroinvertebrate community structure by means of organic enrichment. Corbicula fluminea growth was enhanced in monitoring locations directly below the settling ponds, also an apparent result of organic enrichment. Results of acute water column toxicity testing with Ceriodaphnia dubia, sediment chronic toxicity testing with Daphnia magna, and in-situ ecotoxicological assessments with C. fluminea demonstrated no mortality or toxic influence at most of the sites tested below the ponds. The settling ponds appear to serve as sinks in collecting some, but not all, trace metals.     

Evaluation of the Fertilization Capability of Paracentrotus Lividus Sea Urchin Storaged Gametes by the Exposure to Different Aqueous Matrices

Several experiments were performed to evaluate the fertilization capability of Paracentrotus lividus sea urchin gametes, which are usually used in fertilization tests, but which, until now, were employed only for few hours after spawning.The aim of this work is to evaluate the possibility of performing the bioassay for longer periods, employing ‘conserved' gametes. As far as the method, the sperm was stored ‘dry' at 4 ^∘C; eggs were conserved in an antibiotic solution at 4 ^∘C, and they were exposed both to copper solutions as reference toxicant and to aqueous matrices.The use of conserved gametes in the fertilization bioassay lead to EC50 values which were acceptable for about 3 days. Moreover, these experiments permitted to identify the limiting factor: the male gamete.     

Effects of Copper Sulfate and Copper Nitrate in Aquatic Medium on the Restoration Potential and Accumulation of Copper in Stem Cuttings of the Terrestrial Medicinal Plant, Portulaca Oleracea Linn

The stem cuttings of the terrestrial, ornamental plant, Portulaca oleracea, grew well in distilled water by producing adventitious roots and leaves. However, when exposed to various concentrations of sulfate and nitrate salts of copper resulted in a suppression of root growth, increase in initiation time of roots and sprouts and decay of stem cuttings from the cut open end, decrease in number of leaves with an increase in concentration of copper in the growth medium. Accumulation of copper increased with increasing concentrations of both copper sulfate and copper nitrate. However, copper accumulation was greater in copper nitrate than in copper sulfate treatment. Hence, copper in the presence of nitrate is more toxic than in the presence of sulfate. The accumulation factors in all treatment concentrations were greater than 1, hence P. oleracea is a copper accumulator.     

Solubilities of hydrophobic compounds in aqueous-organic solvent mixtures

Solubilities of several hydrophobic organic substances (paradichlorobenzene, endrin, naphthalene, and dibutyl phthalate) in aqueous solutions containing up to 0.10 mole fraction of common alcohols and ketones, were measured by gas chromatography. The solubilities are significantly increased by the alcohols and ketones. The results are interpreted in terms of the association of n molecules of alcohol or ketone with each hydrophobic organic molecule. Values of n and the equilibrium constant for this association are reported for each hydrophobic organic-alcohol and organic-ketone combination. The implications of these results for the disposal of toxic wastes by landfilling is discussed.     

Heavy metal contamination from mining sites in South Morocco: Monitoring metal content and toxicity of soil runoff and groundwater

The aim of the present work is the assessment of metal toxicity in runoff, in their contaminated soils and in the groundwater sampled from two mining areas in the region of Marrakech using a microbial bioassay MetPLATE™. This bioassay is based on the specific inhibition of the β-galactosidase enzyme of a mutant strain of Escherichia coli, by the metallic pollutants. The stream waters from all sampling stations in the two mines were all very toxic and displayed percent enzyme inhibition exceeding 87% except SWA₄ and SWB₁ stations in mine C. Their high concentrations of copper (Cu) and zinc (Zn) confirm the acute toxicity shown by MetPLATE. The pH of stream waters from mine B and C varied between 2.1 and 6.2 and was probably responsible for metal mobilization, suggesting a problem of acid mine drainage in these mining areas. The bioassay MetPLATE™ was also applied to mine tailings and to soils contaminated by the acidic waters. The results show that the high toxicity of these soils and tailings was mainly due to the relatively concentration of soluble Zn and Cu. The use of MetPLATE™ in groundwater toxicity testing shows that, most of the samples exhibited low metal toxicity (2.7–45.5% inhibition) except GW3 of the mine B (95.3% inhibition during the wet season and 82.9% inhibition during the dry season). This high toxicity is attributed to the higher than usual concentrations of Cu (189 μg Cu l⁻¹) and Zn (1505 μg Zn l⁻¹). These results show the potential risk of the contamination of different ecosystems situated to the vicinity of these two metalliferous sites. The general trend observed was an increase in metal toxicity measured by the MetPLATE with increasing total and mobile metal concentrations in the studied matrices. Therefore, the MetPLATE bioassay is a reliable and fast bioassay to estimate the metals toxicity in the aquatic and solids samples.     

A new <Emphasis Type="Italic">P. putida</Emphasis> instrumental toxicity bioassay

Here, we present a new toxicity bioassay (CO₂-TOX), able to detect toxic or inhibitory compounds in water samples, based on the quantification of Pseudomonas putida KT2440 CO₂ production. The metabolically produced CO₂ was measured continuously and directly in the liquid assay media, with a potentiometric gas electrode. The optimization studies were performed using as a model toxicant 3,5-DCP (3,5-dichlorophenol); later, heavy metals (Pb²⁺, Cu²⁺, or Zn²⁺) and a metalloid (As⁵⁺) were assayed. The response to toxics was evident after 15 min of incubation and at relatively low concentrations (e.g., 1.1 mg/L of 3,5-DCP), showing that the CO₂-TOX bioassay is fast and sensitive. The EC₅₀ values obtained were 4.93, 0.12, 6.05, 32.17, and 37.81 mg/L for 3,5-DCP, Cu²⁺, Zn²⁺, As⁵⁺, and Pb²⁺, respectively, at neutral pH. Additionally, the effect of the pH of the sample and the use of lyophilized bacteria were also analyzed showing that the bioassay can be implemented in different conditions. Moreover, highly turbid samples and samples with very low oxygen levels were measured successfully with the new instrumental bioassay described here. Finally, simulated samples containing 3,5-DCP or a heavy metal mixture were tested using the proposed bioassay and a standard ISO bioassay, showing that our test is more sensible to the phenol but less sensible to the metal mixtures. Therefore, we propose CO₂-TOX as a rapid, sensitive, low-cost, and robust instrumental bioassay that could perform as an industrial wastewater-process monitor among other applications.     

Effects of roof and rainwater characteristics on copper concentrations in roof runoff

Copper sheeting is a common roofing material used in many parts of the world. However, copper dissolved from roof sheeting represents a source of copper ions to watersheds. Researchers have studied and recently developed a simple and efficient model to predict copper runoff rates. Important input parameters include precipitation amount, rain pH, and roof angle. We hypothesized that the length of a roof also positively correlates with copper concentration (thus, runoff rates) on the basis that runoff concentrations should positively correlate with contact time between acidic rain and the copper sheet. In this study, a novel system was designed to test and model the effects of roof length (length of roof from crown to the drip edge) on runoff copper concentrations relative to rain pH and roof angle. The system consisted of a flat-bottom copper trough mounted on an apparatus that allowed run length and slope to be varied. Water of known chemistry was trickled down the trough at a constant rate and sampled at the bottom. Consistent with other studies, as pH of the synthetic rainwater decreased, runoff copper concentrations increased. At all pH values tested, these results indicated that run length was more important in explaining variability in copper concentrations than was the roof slope. The regression equation with log-transformed data (R ² = 0.873) accounted for slightly more variability than the equation with untransformed data (R ² = 0.834). In log-transformed data, roof angle was not significant in predicting copper concentrations.     

The Asiatic clam,Corbicula spp., as a biological monitor in freshwater environments

Asiatic clams, Corbicula spp., are filter-feeding freshwater bivalves that are widely distributed, abundant, and fast growing with a lifespan of 1–3 yrs. A review of the existing literature demonstrates that Asiatic clams can concentrate organic pollutants from both water and sediment and heavy metals from water. In conjunction with these traits, they exhibit a high tolerance for the effects resulting from exposure to toxic substances. While an organism must possess these traits to serve as an effective biological monitor, they have also permitted the Asiatic clam to rapidly colonize natural and industrial environments resulting in purported ecological disturbances and severe economic repurcussions, respectively. Its invasive biofouling attributes therefore restrict the use of Asiatic clams for biomonitoring purposes from Corbicula-free drainage systems.     

Standardizing the Methodology of Sperm Cell Test with Paracentrotus Lividus

The sperm cell toxicity test with Paracentrotus lividus is widely used in Italy to assess the quality of complex aqueous matrices, but at present there is a shortage of standardized methodologies for the bioassay performance. In this work several critical points were considered, in order to verify the reliability of this bioassay and to improve its standardization.In particular, we have studied the differences in EC50 values at different sperm: egg ratios and with different quantities of gametes; the influence of sperm and eggs on fertilization rate and the influence of different recipients materials (plastic or glass). At the same time, it was evaluated if the obtained EC50 values were in accordance with those reported in literature.Experiments performed yielded EC50 values for copper ranging from 16.54 μ g/l to 69.59 μ g/l, with a total mean value ± SD of 39.79 μ g/l ± 11.17. These data fit with those found in literature for other echinoid species. According to the same authors, poor fertilization was probably due to the effects on sperm and not on eggs, and moreover the linear regression analysis performed on each experiment suggested that the best amount of eggs to use in the test was 1000.Results from the comparison between the performance of the test in plastic or glass tubes showed an higher success in fertilization rate using borosilicate glass tubes.     

Preliminary results of laboratory toxicity tests with the mayfly, Isonychia bicolor (Ephemeroptera: Isonychiidae) for development as a standard test organism for evaluating streams in the Appalachian coalfields of Virginia and West Virginia

Selecting the most appropriate test species for sediment and water column assays has been a primary goal for ecotoxicologists. Standard test organisms and established test guidelines exist, but the USEPA-recommended species may not be the most sensitive organisms to anthropogenic inputs. This paper describes preliminary results of toxicity tests with the mayfly, Isonychia bicolor (Ephemeroptera). Results suggested that Isonychia were moderately sensitive to NaCl after 96 h with an average LC₅₀ value of 3.10 g NaCl per liter. This value decreased after 7 days of exposure, resulting in a mean LC₅₀ value of 1.73 g NaCl per liter. When exposed to a coal-mine-processed effluent, Isonychia generated LC₅₀ values that ranged from 13% to 39% effluent. I. bicolor were more sensitive to the coal processing effluent than Ceriodaphnia dubia with conductivity lowest observable effects concentration (LOEC) values for mayfly survivorship that ranged from 1,508 to 4,101 μS/cm, while LOEC values for C. dubia reproduction ranged from 2,132 to 4,240 μS/cm.     

Bioluminescent enzymatic rapid assay of water integral toxicity

A bioluminescent rapid method was developed to estimate the integral toxicity of natural and wastewater. This method is based on registering the effect of the polluted water sample on the parameters of the bioluminescent reaction catalyzed by the multi-component reagent containing NADH:FMN oxidoreductase, luciferase, and their substrates co-immobilized in a starch carrier. Several ways to increase the method's sensitivity to toxic substances were suggested; conditions were selected to make it possible to determine, with maximum efficiency, the content of toxic substances corresponding to a certain maximum permissible concentration. The sensitivity of soluble and immobilized coupled enzymatic systems to a series of organic pollutants (phenols, quinones, and salts of heavy metals) was compared. It was shown that the reagent is the most sensitive to the effect of phenols and quinones. The method was tested during analysis of the wastewater from a pulp and paper plant and can be used for biotesting in both laboratory and field conditions.     

A modified toxicity testing method using tropical marine microalgae

Toxicity testing of four heavy metals(Cd, Cu, Mn and As) using four species of tropicalmarine phytoplankton, Chaetoceros calcitrans, Isochrysis galbana, Tetraselmis tetrahele and Tetraselmis sp., was carried out in multiwellplates with test volumes of 2 mL and the resultscompared to those of standard, large volume,shake-flasks. IC₅₀ values (concentrations of metalsestimated to inhibit 50% growth relative to thecontrol) were determined after 96 hours based onautomated O.D. readings measured in Elisa microplatesby a Multiskan spectrophotometer. Good agreement wasachieved between O.D. readings and cell countsindicating that this new method is a simple,economical, practical and rapid technique for toxicitytesting, and provides good reproducibility of IC₅₀ values. Results of the toxicity testsindicate that Cu was the most toxic metal (averageIC₅₀ values ranging from 0.04 to 0.37 mg L^-1), followed by Cd (0.06–5.7 mg L^-1), Mn (7.2–21.4 mg L^-1) and As (33.9–319.3 mg L^-1).Test species had different degrees of sensitivity tothe metals tested, with I.galbana and C. calcitrans the mostsensitive to Cu, Cd and Mn. Based on these findingsit is recommended that the existing Malaysian InterimStandards for Marine Water Quality for Cd and Cu be reviewed.     

A new system to bioassay pesticides present in the surface microlayer using floating propagules of an aero-aquatic hyphomycetous fungus Pseu doaegerita matsushimae

The surface microlayer often contains pesticides at levels which may be more than one hundred thousand times those sampled in the remainder of the water column. Standard bioassay systems cannot asses these pesticides since no system yet devised has made use of organisms inhabiting the surface microlayer. Using floating propagules of Pseudoaegerita matsushimae, an aeroaquatic hyphomycetous fungus, a bioassay system was devised in which the percentage germinations of such propagules after exposure to various levels of pesticides were plotted to produce a dosegermination scale onto which subsequent data could be fitted. The following pesticides and pesticide mixtures were used: PCP; DDT; Methoxychlor; Bis(tributyl)tin Oxide; Malathion; Captan; 1 part PCP/1 part DDT; 2 parts PCP/1 part DDT; 3 parts PCP/1 part DDT; 2 parts DDT/1 part PCP; 3 parts DDT/1 part PCP; 1 part PCP/1 part Methoxychlor; 2 parts PCP/1 part Methoxychlor; 3 parts PCP/1 part Methoxychlor; 2 parts Methoxychlor/1 part PCP; 3 parts Methoxychlor/1 part PCP. The bioassay system revealed increased sensitivity to pesticides and pesticide mixtures which affected respiratory metabolism. Some pesticide mixtures were more effective in inhibiting germination than their individual components while others appeared much less toxic to the propagules than their unassisted components.