An algal probe for copper speciation in marine waters: laboratory method development

Laboratory-based algal assays were developed to explore the bioavailability of copper to the marine alga Thalassiosira weissflogii. A calibration strategy was developed that avoided use of the synthetic ligand ethylenediaminetetraacetic acid (EDTA) in the Aquil growth medium, thereby allowing ambient metal speciation. In a comparison of T. weissflogii cells grown in Aquil medium with EDTA to medium containing no added copper, zinc, and less than 0.003 nM of EDTA, no significant growth differences were observed after 8 d, indicating adequate stored nutrients. A 30-h assay was selected as the optimal time frame after examination of data from concentration-response experiments. Using 65Cu stable isotope additions, parameters examined included growth, chlorophyll a, copper uptake, phytochelatin production, and dissolved organic carbon excretion. The T. weissflogii specific growth rates decreased from 1.36 d(-1)( at pCu (i.e., the negative logarithmic concentration of free Cu) = 8.8 to 0.56 d(-1) at pCu = 7.8, whereas intercellular copper concentrations increased from 13.6 to 70.1 fg/cell, respectively. Calculated values of the copper concentration that caused a 50% reduction in algal growth of pCu = 7.7 and copper per algal mass of 625 microg/g were established. Using an algal assay based on EDTA-free culture medium, along with trace-metal clean techniques, the effect of copper on T. weissflogii and the speciation of copper in marine waters can be studied.     

Water pollution by Cu and Pb can adversely affect mallard embryonic development

The effects of heavy metal pollutants on aquatic birds have been widely studied in ecotoxicological investigations; however, the predominant focus has been on the postnatal period of life. Limited information on the adverse effects of metals to bird eggs is available. The possible toxic effects of lead and copper were studied in mallard eggs. After the accidental severe heavy metal pollution of the Tisa river (Hungary) in March 2000, these metals were detected in the highest concentration in both the water and the sediment, reaching far beyond acceptable concentrations. Pb treatment (2.9 mg/L) significantly increased the rate of mortality after a single immersion of the eggs into polluted water for 30 min. The rate of dead embryos significantly increased after the combined exposure to Cu and Pb (0.86 and 2.9 mg/L, respectively) both in the single- (once for 30 min) and in the multiple- (10s daily during first trimester of incubation) immersion groups. It was concluded that elevated metal concentrations similar to those found in the Tisa river after the tailing dam failure may cause toxic effects (mortality and teratogenicity) upon exposure of mallard eggs.     

Evaluation of Lethality and Genotoxicity in the Freshwater Mussel <Emphasis Type="Italic">Utterbackia imbecillis</Emphasis> (Bivalvia: Unionidae) Exposed Singly and in Combination to Chemicals Used in Lawn Care

Many chemicals, including fertilizers, herbicides, and insecticides, are routinely applied to turf in the care and maintenance of lawns. These chemicals have the potential to leach into nearby surface waters and adversely affect aquatic biota. In this study, we evaluated the lethal and genotoxic effects of chemicals used in lawn care on an early life stage of freshwater mussels (Utterbackia imbecillis). The chemicals tested were copper and commercial formulations of atrazine, glyphosate, carbaryl, and diazinon. Mussel glochidia were exposed to chemicals singly or in combination (equitoxic and environmentally realistic mixtures) for 24 h and toxic interactions were evaluated with Markings additive index. Genotoxicity was quantified with the alkaline single-cell gel electrophoresis assay (Comet assay). In acute tests, copper was the most toxic of all chemicals evaluated (LC50 = 37.4 µg/L) and carbaryl was the most toxic of all pesticides evaluated (LC50 = 7.9 mg/L). In comparison to other aquatic organisms commonly used in toxicity tests (e.g., amphipods, cladocerans, and chironomids), mussel glochidia were as or more sensitive to the chemicals evaluated with the exception of diazinon, where mussels were observed to be less sensitive. The combined toxicity of equitoxic and environmentally realistic mixtures to mussels was additive. Genotoxic responses were observed in mussels exposed to copper, atrazine and diazinon at levels below their respective no-observed-effect concentrations. Together, these data indicate that freshwater mussels are among the most sensitive aquatic organisms tested for some chemicals commonly used in lawn care and that DNA damage may be useful as a screening tool to evaluate potential sublethal effects of lawn care products on non-target aquatic organisms.     

The effects of the interaction of outboard motors with the aquatic environment--a review

The effects of the compounds associated with outboard motor subsurface exhausts on water quality and aquatic biota are reviewed. The problems affiliated with water quality may include the formation of undesirable tastes and odors and the appearance of oily substances. It has been demonstrated that outboard motor exhaust water can exhibit a toxic effect in sufficiently high concentrations to fathead minnows and bluegills, taints the flesh of various fish, and may affect the reproduction of fish.A discussion of the current research related to the effects of outboard motors on the aquatic environment is presented. Recommendations are given for future research to broaden the understanding of the interaction of outboard motors with the aquatic environment.     

Influence of multiple water-quality characteristics on copper toxicity to fathead minnows (Pimephales promelas)

Water quality influences the bioavailability and toxicity of copper to aquatic organisms. Understanding the relationships between water-quality parameters and copper toxicity may facilitate the development of site-specific criteria for water quality and result in better protection of aquatic biota. Many studies have examined the influence of a single water-quality parameter on copper toxicity, but the interactions of several characteristics have not been well studied in low-hardness water. The goal of the present research was to examine the interactions among water-quality characteristics and their effects on copper toxicity to larval fathead minnows (Pimephales promelas). The effects of dissolved organic carbon (DOC) concentration, DOC source, pH, and hardness on acute copper toxicity were determined using a complete factorially designed experiment. Hardness, pH, DOC, and interaction of pH and DOC all significantly affected copper toxicity. A predictive model based on these data described 88% of the variability in copper toxicity. This model also explained 58% of the variability in copper toxicity for an independent dataset of South Carolina (USA) waters. The biotic ligand model underpredicted the acute copper toxicity to fathead minnows when compared with observed values.     

医院污水对水环境毒理学风险的研究进展

医院污水作为水环境中毒性风险的主要来源之一,其毒性污染物质进入水环境后,会对水生生态平衡和饮用水水源造成潜在的负面影响。作者介绍了医院污水中对水环境造成毒性风险的主要污染物质和这些污染物在水环境中造成潜在毒性影响的研究状况与进展。     

Ecotoxicology of glutaraldehyde: review of environmental fate and effects studies

Glutaraldehyde is a biocide used in many industrial applications with potential releases to the environment. This review discusses the environmental fate and effects data on this important biocide. Information drawn from this review indicates that glutaraldehyde is acutely toxic to aquatic organisms. Glutaraldehyde is equally toxic to warm water and cold water fish, but is slightly more toxic to freshwater fish than salt water fish. The acute toxicity of glutaraldehyde for avian species is comparable to that for mammalian species. The toxicity of glutaraldehyde is not appreciably increased with repeated long-term exposures. Results from environmental partitioning studies indicate that glutaraldehyde tends to remain in the aquatic compartment and has little tendency to bioaccumulate. Aqueous solutions of glutaraldehyde are stable at room temperature under acidic to neutral conditions, and to sunlight, but unstable at elevated temperatures, and under alkaline conditions. Glutaraldehyde is readily biodegradable in the freshwater environment and has the potential to biodegrade in the marine environment. Aquatic metabolism studies suggest that glutaraldehyde, under aerobic conditions, is metabolized to CO(2) via glutaric acid as an intermediate. Under anaerobic conditions, glutaraldehyde is metabolized to 1,5-pentanediol. Pretreatment with sodium bisulfite is the best method for inactivating glutaraldehyde prior to disposal to treatment systems.     

Acute toxicity of atrazine, endosulfan sulphate and chlorpyrifos to Vibrio fischeri, Thamnocephalus platyurus and Daphnia magna, relative to their concentrations in surface waters from the Alentejo region of Portugal

Ecotoxicological effects of the herbicide atrazine and the insecticides endosulfan sulphate and chlorpyrifos were evaluated using a test battery comprising aquatic organisms from different trophic levels. According to the categories established in the EU legislation, atrazine can be considered non-harmful for the species tested, while the insecticides can be considered very toxic for the crustaceans. The results of acute toxicity tests showed that the sensitivity of organisms were as follows: Thamnocephalus platyurus > Daphnia magna > Vibrio fischeri. Chlorpyrifos may act as a toxic compound in the aquatic environment of Guadiana River, as it may be detected in water at levels that promote toxic effects.     

Subchronic effects of dipyrone on the fish species Rhamdia quelen

The use of the non-steroidal anti-inflammatory drugs such as dipyrone is so widespread that this drug and its metabolites have been detected in effluents and surface water. This study aimed to evaluate the potential toxic effects of dipyrone on the aquatic environment, using a native fish species, Rhamdia quelen. Fish were exposed to three concentrations of dipyrone, 0.5, 5 and 50 μg/L, in the water for 15 days, and hematological, biochemical, genetic and morphological biomarkers were evaluated. The glutathione S-transferase activity decreased in the highest concentration in relation to the control group. In addition, hematocrit, red blood cells and thrombocyte counts were decreased in all three exposed groups in relation to the control group. The comet assay showed DNA damage at the lowest concentration of dipyrone and significant kidney damage. Those results suggest that a constant exposure of aquatic organisms to dipyrone presents potential toxic effects.     

Copper toxicity in relation to surface water-dissolved organic matter: biological effects to Daphnia magna

Water quality standards for copper are usually stated in total element concentrations. It is known, however, that a major part of the copper can be bound in complexes that are biologically not available. Natural organic matter, such as humic and fulvic acids, are strong complexing agents that may affect the bioavailable copper (Cu2+) concentration. The aim of this study was to quantify the relation between the concentration of dissolved natural organic matter and free Cu2+ in surface waters, and the biological effect, as measured in a standardized ecotoxicological test (48 h-median effective concentration [EC50] Daphnia magna, mobility). Six typical Dutch surface waters and an artificial water, ranging from 0.1 to 22 mg/L dissolved organic carbon (DOC), were collected and analyzed quarterly. Chemical speciation modeling was used as supporting evidence to assess bioavailability. The results show clear evidence of a linear relation between the concentration of dissolved organic carbon (in milligrams DOC/L) and the ecotoxicological effect (as effect concentration, EC50, expressed as micrograms Cu/L): 48-h EC50 (Daphnia, mobility) = 17.2 x DOC + 30.2 (r2 = 0.80, n = 22). Except for a brook with atypical water quality characteristics, no differences were observed among water type or season. When ultraviolet (UV)-absorption (380 nm) was used to characterize the dissolved organic carbon, a linear correlation was found as well. The importance of the free copper concentration was demonstrated by speciation calculations: In humic-rich waters the free Cu2+ concentration was estimated at approximately 10(-11) M, whereas in medium to low dissolved organic carbon waters the [Cu2+] was approximately 10(-10) M. Speciation calculations performed for copper concentrations at the effective concentration level (where the biological effect is considered the same) resulted in very similar free copper concentrations (approximately 10(-8) M Cu) in these surface waters with different characteristics. These observations consistently show that the presence of organic matter decreases the bioavailability, uptake, and ecotoxicity of copper in the aquatic environment. It demonstrates that the DOC content must be included in site-specific environmental risk assessment for trace metals (at least for copper). It is the quantification of the effects described that allows policy makers to review the criteria for copper in surface waters.     

The Effect of Petrochemical Effluent on the Water Quality of Ubeji Creek in Niger Delta of Nigeria

Water samples containing petrochemical effluents were evaluated for elemental contaminants along a kilometer distance in Ubeji Creek, a tributary of the Ubeji River in the Niger Delta of Nigeria. Twenty water samples were collected from six sites at various times. The water samples were analyzed for several physico-chemical parameters. Results showed wide varieties in temperature, pH, BOD, COD, dissolved and suspended solids as well as conductivity. The entire environment starting from the end-of-pipe source point was coated with black oily residue. Water quality parameters were very poor. The absence of fish and other aquatic lives, the high levels of Zn (2.4), Cr (0.24), Fe (63.44), Hg (4.24), Mn (2.49), and Pb (0.76) level (μg/L) confirm the toxic nature of Ubeji Creek. At the lower reaches, the mixing of effluent with brackish waters was not enough to support aquatic life, partly because of diminishing oxygen and toxic shock. Nevertheless, the study provides evidence to suggest that the water in Ubeji Creek is toxic. It also provides graphic data to suggest point source where effluents could be held for treatment or neutralization before being discharged into the aquatic environment.     

Invertebrate biomarkers: links to toxicosis that predict population decline

The application of biochemical measurements that can be used as individual biomarkers of impaired biological function in invertebrates is reviewed to evaluate whether biochemical biomarkers of aquatic invertebrates can predict changes in natural populations. Biomarkers that measure toxic effects at the molecular level (e.g., the inhibition of brain acetylcholinesterase activity by organophosphorus pesticides) have been shown to provide rapid quantitative predictions of a toxic effect upon individuals in laboratory studies. Such biomarkers should not be used as a replacement for conventional aquatic monitoring techniques, but should be applied as supplementary approaches for demonstrating links between sublethal biochemical and adverse effects in natural populations in field studies. The research challenge for using biomarker measurements in aquatic invertebrates is to predict effects at the population level from effects at the individual level measured upon individuals collected in the field.     

Sublethal effects of copper on coho salmon: impacts on nonoverlapping receptor pathways in the peripheral olfactory nervous system

The sublethal effects of copper on the sensory physiology of juvenile coho salmon (Oncorhynchus kisutch) were evaluated. In vivo field potential recordings from the olfactory epithelium (electro-olfactograms) were used to measure the impacts of copper on the responses of olfactory receptor neurons to natural odorants (L-serine and taurocholic acid) and an odorant mixture (L-arginine, L-aspartic acid, L-leucine, and L-serine) over a range of stimulus concentrations. Increases in copper impaired the neurophysiological response to all odorants within 10 min of exposure. The inhibitory effects of copper (1.0-20.0 micrograms/L) were dose-dependent and they were not influenced by water hardness. Toxicity thresholds for the different receptor pathways were determined by using the benchmark dose method and found to be similar (a 2.3-3.0 micrograms/L increase in total dissolved copper over background). Collectively, examination of these data indicates that copper is broadly toxic to the salmon olfactory nervous system. Consequently, short-term influxes of copper to surface waters may interfere with olfactory-mediated behaviors that are critical for the survival and migratory success of wild salmonids.     

Sensitivity of mottled sculpins (Cottus bairdi) and rainbow trout (Onchorhynchus mykiss) to acute and chronic toxicity of cadmium, copper, and zinc

Studies of fish communities of streams draining mining areas suggest that sculpins (Cottus spp.) may be more sensitive than salmonids to adverse effects of metals. We compared the toxicity of zinc, copper, and cadmium to mottled sculpin (C. bairdi) and rainbow trout (Onchorhynchus mykiss) in laboratory toxicity tests. Acute (96-h) and early life-stage chronic (21- or 28-d) toxicity tests were conducted with rainbow trout and with mottled sculpins from populations in Minnesota and Missouri, USA, in diluted well water (hardness = 100 mg/L as CaCO3). Acute and chronic toxicity of metals to newly hatched and swim-up stages of mottled sculpins differed between the two source populations. Differences between populations were greatest for copper, with chronic toxicity values (ChV = geometric mean of lowest-observed-effect concentration and no-observed-effect concentration) of 4.4 microg/L for Missouri sculpins and 37 microg/L for Minnesota sculpins. Cadmium toxicity followed a similar trend, but differences between sculpin populations were less marked, with ChVs of 1.1 microg/L (Missouri) and 1.9 microg/L (Minnesota). Conversely, zinc was more toxic to Minnesota sculpins (ChV = 75 microg/L) than Missouri sculpins (chronic ChV = 219 microg/L). Species-average acute and chronic toxicity values for mottled sculpins were similar to or lower than those for rainbow trout and indicated that mottled sculpins were among the most sensitive aquatic species to toxicity of all three metals. Our results indicate that current acute and chronic water quality criteria for cadmium, copper, and zinc adequately protect rainbow trout but may not adequately protect some populations of mottled sculpins. Proposed water quality criteria for copper based on the biotic ligand model would be protective of both sculpin populations tested.     

Chemical contamination and toxicity of sediment from a coastal area receiving industrial effluents in Kuwait

The Shuaiba coastal area (12.5 × 1.5 km) was examined for contamination with total organic carbon, volatile organic matter, total petroleum hydrocarbons, polycyclic aromatic hydrocarbons, cadmium, chromium, copper, lead, nickel, vanadium, and zinc in sediment; their desorption by aqueous elution; and toxicity to aquatic biota. The pollutants were mainly accumulated in the upstream area facing Mina Al-Ahmadi refinery to Shuaiba harbour. Solid-phase Microtox assays showed severe toxicity, and the LC₅₀ was negatively correlated with most of the chemical parameters, suggesting that toxicity was the function of collective effects of the pollutants present in sediment. Sea water elutriation showed poor desorption of pollutants from sediment, and the elutriates were not found toxic to Microtox and brine shrimp larvae. Whole sediment suspension in sea water reduced the survival of fingerlings in fish bioassays. Action from Shuaiba Area Authority is required to reduce pollutant accumulation in identified depositional area on the Shuaiba coast by facilitating unrestricted water flow in the area and restricting pollutant discharge at source. Received: 15 November 2000/Accepted: 11 May 2001     

Effects of leachate from tree leaves and grass litter on tadpoles

Tree species composition can change as a result of succession, climate change, fire suppression, and invasive species. These changes clearly affect forests, but they can also affect aquatic ecosystems based on differences in the input quality of leaf litter, such as plant secondary compounds. These compounds vary in type and concentration depending on species and can be toxic to aquatic organisms. To examine toxic effects on Pseudacris maculata and Pseudacris crucifer tadpoles, we conducted 60-d laboratory experiments to compare leaf litter leachate from a dominant canopy species (red oak) and nonnative species (white pine) with an aquatic grass (prairie cordgrass) and plain water control. An additional experiment examined the effects of white pine on Bufo americanus, Hyla versicolor, and tannin concentrations in natural ponds. Compared with the control and grass, tree leaf extracts resulted in reduced tadpole survival. Leached compounds from pine reduced tadpole survival to 3 d or less. Tadpoles were able to metamorphose in significant numbers only from the two controls. The lowered survival with the red oak treatment might have been caused by lowered dissolved oxygen or high tannin concentrations. However, pine is known to have high concentrations of toxic monoterpenes, which should be investigated further. We found that tannin concentrations in natural ponds were much lower than the test concentrations, indicating that these results may represent worst-case scenarios or unrealistic concentrations.     

Integrated ecological risk assessment of pesticides in tropical ecosystems: a case study with carbofuran in Brazil

The aim of the present study is to contribute an ecologically relevant assessment of the ecotoxicological effects of pesticide applications in agricultural areas in the tropics, using an integrated approach with information gathered from soil and aquatic compartments. Carbofuran, an insecticide/nematicide used widely on sugarcane crops, was selected as a model substance. To evaluate the toxic effects of pesticide spraying for soil biota, as well as the potential indirect effects on aquatic biota resulting from surface runoff and/or leaching, field and laboratory (using a cost-effective simulator of pesticide applications) trials were performed. Standard ecotoxicological tests were performed with soil (Eisenia andrei, Folsomia candida, and Enchytraeus crypticus) and aquatic (Ceriodaphnia silvestrii) organisms, using serial dilutions of soil, eluate, leachate, and runoff samples. Among soil organisms, sensitivity was found to be E. crypticus < E. andrei < F. candida. Among the aqueous extracts, mortality of C. silvestrii was extreme in runoff samples, whereas eluates were by far the least toxic samples. A generally higher toxicity was found in the bioassays performed with samples from the field trial, indicating the need for improvements in the laboratory simulator. However, the tool developed proved to be valuable in evaluating the toxic effects of pesticide spraying in soils and the potential risks for aquatic compartments.     

Long-Term Effects of Ammonia on the Behavioral Activity of the Aquatic Snail <Emphasis Type="Italic">Potamopyrgus antipodarum</Emphasis> (Hydrobiidae,Mollusca)

An appropriate approach to assess the effect of toxicants on aquatic animals is to monitor behavioral endpoints, as they are a link between physiological and ecological processes. A group that can be exposed long-term to low toxic concentrations is benthic macroinvertebrates, as their mobility in aquatic ecosystems is relatively limited. Therefore, the study of behavioral long-term effects in this group is suitable from an ecological point of view, as behavioral effects can appear before mortality. During the last decades there has been an increase in ammonia concentrations in freshwater ecosystems, threatening aquatic animals. The present study focuses on the long-term effects (40 days) of nonionized ammonia on the behavioral activity of the aquatic snail Potamopyrgus antipodarum. One control and three ammonia concentrations (0.02, 0.07, and 0.13 mg N–NH₃/L) were used in triplicate, and the activity of snails (as mean time to start normal movement) and immobility were recorded for each treatment after 0, 10, 20, 30, and 40 days of continuous exposure to nonionized ammonia. The results show that P. antipodarum presented a high tolerance to lethal long-term effects of nonionized ammonia, as no animal died during the bioassay. However, the behavioral activity of snails was a very sensitivity endpoint, as a mean nonionized ammonia concentration of 0.07 mg N–NH₃/L affected P. antipodarum. The results are discussed and compared with the available literature on long-term effects of ammonia on freshwater macroinvertebrates. Additionally, the ammonia water quality criteria, NOECs, LOECs, and long-term LCs are discussed on the basis of the current available data for freshwater macroinvertebrates.     

More than inorganic copper is bioavailable to aquatic mosses at environmentally relevant concentrations

The present study investigates how dissolved organic matter (DOM) alters copper bioavailability at environmentally relevant concentrations (1-5 microg/L of dissolved copper, 1-4 mg/L of dissolved organic copper). A methodology combining two biological endpoints (short-term and steady-state bioaccumulation of copper by the aquatic moss Fontinalis antipyretica) and a sampling of labile copper with diffusion gradient in thin films (DGT) is proposed for batch experiments conducted with mineral water and various DOM, ethylenediaminetetra-acetic acid (EDTA), humic acid, and natural Seine River (France) extracts (hydrophobic and transphilic fractions). All types of DOM reduce the bioavailability of copper to aquatic mosses, and this reduction was more pronounced for the short-term biological endpoint, which was taken as being representative for environmental exposure. Labile copper sampled with DGT made it possible to estimate short-term bioaccumulation in the case of EDTA and natural Seine River extracts. With humic acid solutions, however, labile copper was lower than bioavailable copper. This result suggests that at realistic metal concentrations and with certain types of natural DOM, bioavailable copper might comprise not only inorganic copper but also some weak organic complexes. Hence, labile copper, in situ sampled with DGT, might not systematically overestimate bioavailable copper, as suggested previously on the basis of in vitro toxicity studies.     

Immunology-Related Perturbations Induced by Copper and Chitosan in Carp (Cyprinus carpio L.)

Copper is used in treatment mixtures to control fungal diseases in vineyards. Its concentrations are relatively high in some aquatic ecosystems, and the main problem observed in this study was the antioxidant stress induced by this heavy metal. Copper toxicologic effects in aquatic organisms have prompted the demand for alternative use of low-toxicity molecules in culture treatments. Chitosan is a polymer with antifungal property similar to copper and may be an interesting biopesticide. Thus, it is necessary to investigate the potential toxicity of chitosan for aquatic animal health, either alone or in conjunction with copper. In this study, carp were exposed to two sublethal chitosan concentrations (75 and 150 mg/L) or to two sublethal copper concentrations (0.1 and 0.25 mg/L) or to a mixture of chitosan plus copper (75 mg/L and 0.1 mg/L, respectively). The results of the present study show that exposure to copper at environmentally realistic levels or to chitosan at sublethal concentrations may significantly stimulate various aspects of immune functions in carp such as nonspecific cellular immunity, represented by total immunoglobulin level, ceruloplasmin activity, and oxidative activity of phagocytes. This acute-phase inflammatory response induced separately by the two treatments was not observed, especially on phagocyte oxidative activity, when carp were exposed to the copper–chitosan mixture. This fact could be explained by a possible chelation of copper by chitosan decreasing the biodisponibility of the two products for immune cells. Thus, the immunotoxicologic impact of copper and chitosan on fish immune response would be less pronounced with the combined treatments than with separate treatments in an aquatic environment.