Toxicity assessment of reference and natural freshwater sediments with the LuminoTox assay

We examined the possibility of adapting the LuminoTox, a recently-commercialized bioanalytical testing procedure initially developed for aqueous samples, to assess the toxic potential of sediments. This portable fluorescent biosensor uses photosynthetic enzyme complexes (PECs) to rapidly measure photosynthetic efficiency. LuminoTox testing of 14 CRM (Certified Reference Material) sediments was first undertaken with (1) a "solid phase assay" (Lum-SPA) in which PECs are in intimate contact with sediment slurries for a 15 min exposure period and (2) an elutriate assay (Lum-ELU) in which PECs are exposed for 15 min to sediment water elutriates. CRM sediment toxicity data were then compared with those generated with the Microtox Solid Phase Assay (Mic-SPA). A significant correlation (P < 0.05) was shown to exist between Lum-SPA and Mic-SPA, indicating that both tests display a similar toxicity response pattern for CRM sediments having differing contaminant profiles. The sediment elutriate Lum-ELU assay displayed toxicity responses (i.e. measurable IC20s) for eight of the 14 CRM sediments, suggesting that it is capable of determining the presence of sediment contaminants that are readily soluble in an aqueous elutriate. Lum-SPA and Mic-SPA bioassays were further conducted on 12 natural freshwater sediments and their toxicity responses were more weakly, yet significantly, correlated. Finally, Lum-SPA testing undertaken with increasing mixtures of kaolin clay confirmed that its toxicity responses, in a manner similar to those reported for the Mic-SPA assay, are also subject to the influence of grain size. While further studies will be required to more fully understand the relationship between Lum-SPA assay responses and the physicochemical makeup of sediments (e.g., grain size, combined presence of natural and anthropogenic contaminants), these preliminary results suggest that LuminoTox testing could be a useful screen to assess the toxic potential of solid media.     

Toxicity assessment of sediments containing tributyltin around Hong Kong harbour

Rather high levels of tributyltin (TBT) in sediments were found in certain areas around the Hong Kong coastline. Sediments from two contaminated sites (Tsing Yi and Yam O) and two clean sites were used to study the release of TBT contaminants from resuspended sediment to the water column by elutriate tests. The chemical analysis data of elutriates were compared with the effective or lethal concentrations from the toxicity tests of spiked solution using marine microalga (Dunaliella tertiolecta) and benthic invertebrate (Melita koreana) to assess the potential ecotoxicological impacts arising from the dredging work. Concentrations of TBT in filtered sediment elutriates were mostly below 0.002 microg/l which suggested that very little of TBT was released in soluble form. The lethal toxicity of TBT for the invertebrate expressing as 10 day-LC(50) was 2.31 microg/l for TBT and the alga expressing as 48 h-EC(50) was 0.46 microg/l. The results of this evaluation demonstrated that the release of TBT from two contaminated sites during dredging was unlikely to attain the effective or lethal concentrations on the two test species.     

Toxicity assessment of peptaibols and contaminated sediments on Crassostrea gigas embryos

Peptaibols are known membrane-modifying peptides that were recently detected in marine sediments and mussels collected from a shellfish farming area (Fier d'Ars, Atlantic coast, France). In this investigation, embryotoxicity bioassays with oysters (Crassostrea gigas) were performed to assess acute toxicity of alamethicin and different groups of peptaibols produced by a Trichoderma longibrachiatum strain isolated from marine environment. C. gigas embryos appeared very sensitive to all the metabolites examined with higher toxic effects for long-sequence peptides (EC50 ranging from 10 to 64 nM). D-shaped larvae with mantle abnormality were particularly noticed when peptaibol concentrations increased. Disturbances of embryogenesis were also observed following exposure to organic and aqueous extract of sediments from Fier d'Ars (EC50=42.4 and 6.6 g L(-1) dry weight, respectively). Although peptaibol concentrations measured in these sediments could explain only a part of the toxic effects observed, this study suggests that these mycotoxins can induce larval abnormalities in a population of exposed animals at environmentally realistic concentrations. Their detection in coastal areas devoted to bivalve culture should be taken into account.     

Toxicity of elemental sulfur in sediments

Elemental sulfur occurs naturally in marine and limnic sediments. Elemental sulfur, brought in solution in aqueous media by using organic solvents such as methanol as carrier solvent, was toxic in a bacterial luminescence test, known as the Microtox test. Previously, it has been shown that the toxicity in the luminescence test of whole sediments also was correlated to i.a. elemental sulfur using multivariate statistical analysis. Organic solvent extracts of sediments obtained in receiving waters of effluents from a pulp and paper mill was toxic in the luminescence test, and using a toxicity evaluation procedure, the toxic substance was identified as octameric cyclic sulfur, S₈. The substance dominated the toxicity in extracts of both a contaminated sediment and a sediment from a control area. Since the toxicity in the Microtox test of aqueous solutions of S₈ decreased upon storage, a conversion process of the toxic form was indicated. Acute toxicity of S₈ was not limited to the luminescent bacteria in the Microtox test, but was observed in tests with fish larvae if tested with the transient form of elemental sulfur. Tests of acute toxicity with zebra fish and perch larvae were responsive to elemental sulfur. Probably, the toxic form of elemental sulfur is the single cyclic octamer, that due to low aqueous solubility, binding to particulate sediment material or aggregation is converted into a nontoxic form. Acute toxic effects may occur in sulfur containing sediments of varying redox potentials or where elemental sulfur deposits are turbated. © 1998 John Wiley & Sons, Inc. Environ Toxicol Water Qual 13: 217–224, 1998     

Toxicity assessment of wastewaters, river waters, and sediments in Austria using cost-effective microbiotests

The toxicity and chemical quality of surface water and sediment in the River Traun in Austria were studied because of recurrent fish mortality in some alpine rivers over the last few years. The analyses were carried out on samples collected during winter and summer upstream and downstream of two municipal wastewater treatment plants (WWTPs) and on effluents taken at the points of discharge of these two plants. Toxicity tests were performed on 20 samples of surface water, effluent, and sediment pore water. The test battery was composed of microbiotests with protozoans (Protoxkit F), microalgae (Algaltoxkit F), crustaceans (Daphtoxkit F magna and Thamnotoxkit F), and a higher plant (seed germination and root elongation assay on cress). Direct contact tests were performed on whole sediment with crustaceans (Ostracodtoxkit F). The physical-chemical characteristics of the surface water, effluent, and sediment pore water samples analyzed were conductivity, total hardness, pH, O(2), BOD(5), TOC, DOC, AOX, NH(4), NH(3), NO(2), PO(4)--P, Cd, Pb, Cu, and Zn. The toxicity data were expressed as percentage mortality or percentage inhibition, depending on the effect criterion of the respective assay. None of the water samples collected upstream and downstream of the WWTPs showed any significant (short-term) toxicity in either winter or in summer, but the effluents of the first municipal wastewater treatment plant were toxic to some of the test biota. All the sediment pore water samples induced serious inhibition of root growth of cress, and several pore waters were toxic to other test biota as well, particularly at the outlets of the WWTPs. The toxic character of some sediments was confirmed by direct contact tests with the ostracod crustacean. The chemical analyses did not reveal particularly high concentrations of any chemical that is very toxic. As a result no direct causal relationship could be established between the detected toxic effects and the chemical composition of the surface waters or sediment pore waters. The outcome of this preliminary study again highlights the need to complement chemical analyses with toxicity tests to determine the toxic hazard to aquatic environments that may be threatened by contamination. Furthermore, the investigations also confirmed the need to apply a battery of tests for an ecologically meaningful evaluation of the hazards of waters, sediments, and wastewaters. Finally, the results of the 360 bioassays performed show that culture-independent microbiotests are practical and reliable tools for low-cost toxicity monitoring of aquatic environments.     

Toxicity assessment of thiodiglycol

Sulfur mustard (HD) undergoes hydrolysis to form various products such as thiodiglycol (TG) in biological and environmental systems. TG is a precursor in the production of HD and it is also considered as a "Schedule 2" compound (dual-use chemicals with low to moderate commercial use and high-risk precursors). Several toxicological studies on TG were conducted to assess environmental and health effects. The oral LD(50) values were >5000 mg/kg in rats. It was a mild skin and moderate ocular irritant and was not a skin sensitizer in animals. It was not mutagenic in Ames Salmonella, Escherichia coli, mouse lymphoma, and in vivo mouse micronucleus assays, but it induced chromosomal aberrations in Chinese hamster ovarian (CHO) cells. A 90-day oral subchronic toxicity study with neat TG at doses of 0, 50, 500, and 5000 mg/kg/day (5 days/week) in Sprague-Dawley rats results show that there are no treatment-related changes in food consumption, hematology, and clinical chemistry in rats of either sex. The body weights of both sexes were significantly lower than controls at 5000 mg/kg/day. Significant changes were also noted in both sexes in absolute weights of kidneys, kidney to body weight ratios, and kidney to brain weight ratios, in the high-dose group. The no-observed-adverse-effect level (NOAEL) for oral toxicity was 500 mg/kg/day. The developmental toxicity conducted at 0, 430, 1290, and 3870 mg/kg by oral gavage showed maternal toxicity in dams receiving 3870 mg/kg. TG was not a developmental toxicant. The NOAEL for the developmental toxicity in rats was 1290 mg/kg. The provisional oral reference dose (RfD) of 0.4 mg/kg/day was calculated for health risk assessments. The fate of TG in the environment and soil showed biological formation of thiodiglycalic acid with formation of an intermediate ((2-hydroxyethyl)thio)acetic acid. It was slowly biodegraded under anaerobic conditions. It was not toxic to bluegill sunfish at 1000 mg/L and its metabolism and environmental and biochemical effects are summarized.     

Toxicity of nitrite to three species of freshwater invertebrates

Nitrite is a compound with a high toxicity to aquatic animals. Several anthropogenic pollution sources are increasing the concentrations of this component of the nitrogen cycle. Despite this toxicity, there is little available literature on its effects on freshwater invertebrates. Laboratory bioassays were performed to obtain data on the lethal effects of nitrite to three species of freshwater invertebrates: the planarian Polycelis felina and the amphipods Echinogammarus echinosetosus and Eulimnogammarus toletanus. The LC(50), LC(10), and LC(0.01) values (mg/L NO(2)--N) at 24, 48, 72, and 96 h were calculated for each species. E. toletanus and E. echinosetosus were the most sensitive species, with 96 h LC(50) values of 2.09 and 2.59 mg/L NO(2)--N, respectively. In contrast, the planarian P. felina showed a higher tolerance to nitrite, with a 96 h LC(50) value of 60.0 mg/L NO(2)--N. The obtained results were compared with the reported nitrite data for other freshwater invertebrates. This study may contribute to a more appropriate assessment of the ecological risk of this compound in freshwater ecosystems.     

Toxicity assessment of electronic cigarettes

Sale of electronic cigarette (e-cigarette) products has exponentially increased in the past decade, which raise concerns about its safety. This updated review provides the available toxicology profile of e-cigarettes, summarizing evidence from in vitro and in vivo studies. Data regarding which components in e-liquids exhibit potential toxicities are inconsistent. Some studies have reported that nicotine plays a significant role in inducing adverse outcomes and that solvents alone do not induce any adverse effects. However, other studies have suggested that nicotine is not associated with any adverse outcomes, whereas solvents and flavorings are the key components to elicit considerable deleterious effects on cells or animals. In addition, most of the studies that have compared the toxicity of e-cigarettes with tobacco cigarettes have suggested that e-cigarettes are less toxic than tobacco cigarettes. Nevertheless, scientific evidence regarding the toxicity profile of e-cigarette is insufficient owing to the lack of a standardized research approach. In the future, scientific toxicology data derived from standardized testing protocols including nicotine, ingredients analysis, the various e-cigarette devices made from different materials are urgently needed for thorough toxicology assessment. This review aims to update the toxicity profiles, identify knowledge gaps, and outline future directions for e-cigarettes research, which would greatly benefit public health professionals.     

Toxicity assessment of the antiparasitic ivermectin

The citotoxicity of ivermectin, a streptomycete-derived macrocyclic lactone active against a wide variety of nematode and arthropod parasites, was evaluated in pig kidney cells IB-RS-2. The cells were cultured in the presence of different levels of ivermectin for 72 h. The cell count decreased as ivermectin concentration in the medium was increased from 0 to 20 ?g/mL, and this decrease was time and dose dependent; at 40 ?g/mL cell death occurred within 24 h. Ivermectin (20 ?g/mL) decreased protein synthesis and glucose utilization. These effects became progressively less between 48 and 72 h when compared to untreated cells.     

Toxicity assessment of cyanobacterial toxin mixtures

A method is presented for the toxicity assessment of mixtures of cyanobacterial toxins with hepatotoxic, neurotoxic, and carcinogenic effects. The method is based on the derivation of toxicity equivalent factors obtained from toxicological data presented for cyanobacterial toxins in the literature and on the use of toxicity equivalents in analogy with the method used for polychlorinated dibenzo[p]dioxins and dibenzofurans. Because the method easily can include all toxicological data for cyanobacterial toxins at any time, the quality of the toxicity assessment of these toxins can be increased continuously depending on data availability. Generally, all toxicologically relevant cyanobacterial toxins for which data are available should be included in this assessment. The method is considered useful in the toxicity assessment of cyanobacterial toxin mixtures in surface water bodies and in drinking water. When using the total sum of the toxicity equivalents, the approach proposed here seems to lead to a more realistic toxicity assessment of cyanobacterial toxin mixtures than does the worst-case approach. However, the availability of data about acute and, especially, chronic toxicity must be increased significantly in the future in order to establish toxicologically validated exposure limit values for cyanobacterial toxins, especially those with hepatotoxic, neurotoxic, and carcinogenic effects.     

Toxicity assessment of urban marine sediments from Western Norway using a battery of stress-activated receptors and cell-based bioassays from fish

A luciferase reporter gene-based bioassay battery consisting of stress-activated receptors from fish, complemented with traditional fish cell-based bioassays, were used to assess the toxicity of marine sediment samples from the Byfjorden area around the city of Bergen (Norway). The reporter assays covered a wide range of cellular signalling and metabolic pathways, representing different molecular initiating events in the adverse outcome pathway framework. Cytotoxicity, generation of reactive oxygen-species, and induction of 7-ethoxyresorufin-O-deethylase activity were analysed using fish liver and gill cell lines. Chemical analyses of the sediment extracts revealed complex contamination profiles, especially at the innermost stations, which contained a wide array of persistent organic pollutants, polycyclic aromatic hydrocarbons, and metals. Sediment extracts from these sites were more potent in activating the stress-activated receptors than the other extracts, reflecting their toxicant profiles. Importantly, receptor- and cell-based bioassays complemented the chemical analyses and provided important data for future environmental risk assessments of urban marine sediments.     

Toxicity of soybean rust fungicides to freshwater algae and Daphnia magna

Soybeans are intensively grown over large swaths of land in the Midwestern US. Introduction of the pathogenic fungus responsible for Soybean Rust (Phakopsora pachyrhizi) will likely result in a significant increase in the environmental load of strobilurin and conazole fungicides. We determined the toxicity of six such fungicides to the unicellular algae Pseudokirchneriella subcapitata and the aquatic invertebrate, Daphnia magna. We found that levels of concern of some fungicides were lower than annual average runoff concentrations predicted for Indiana. Our results suggest that pyraclostrobin and propiconazole, and to a lesser extent tebuconazole, may cause impacts to algae and daphnids in areas where soybeans are intensively grown. More studies are needed to describe the ecological effects of sublethal exposures to these fungicides, as well as monitoring environmental concentrations in watersheds where these fungicides are applied to soybeans.     

Toxicity assessment of unintentional exposure to multiple chemicals

Typically exposure to environmental chemicals is unintentional, and often the exposure is to chemical mixtures, either simultaneously or sequentially. When exposure occurs, in public health practice, it is prudent to ascertain if thresholds for harmful health effects are exceeded, whether by individual chemicals or by chemicals in combination. Three alternative approaches are available for assessing the toxicity of chemical mixtures. Each approach, however, has shortcomings. As the procedures of each approach are described in this paper, at various steps research needs are identified. Recently, reliance has increased on computational toxicology methods for predicting toxicological effects when data are limited. Advances in molecular biology, identification of biomarkers, and availability of accurate and sensitive methods allow us to more precisely define the relationships between multiple chemical exposures and health effects, both qualitatively and quantitatively. Key research needs are best fulfilled through collaborative research. It is through such collaborations that resources are most effectively leveraged to further develop and apply toxicity assessment methods that advance public health practices in vulnerable communities.     

Toxicity assessment of complex mixtures remains a goal

One of the initial steps in remediating contaminated environments is to assess the human and ecological health risk associated with exposure to contaminants in a specific medium. Presented here are the results of a five-year study investigating the toxicity of simple and complex mixtures. A series of model compounds and simple mixtures including polycyclic aromatic hydrocarbons (PAHs), pentachlorophenol (PCP), and halogenated aliphatic hydrocarbons (HAHs) were analyzed. Mixture toxicity was studied using microbial genotoxicity assays and cytotoxicity assays with renal and neural cells. The majority of binary mixtures described here induced additive responses. A limited number of samples were identified where binary mixtures induced inhibitory effects. For example, benzo(a)pyrene (BAP) alone induced 30% renal cell death, whereas an equimolar dose of chrysene and BAP only produced 1.6% cellular death. In none of the mixtures tested did the mixture toxicity results deviate from the predicted results by an order of magnitude. The results from testing binary mixtures in this study indicate that the results did not deviate significantly from additivity. Complex mixture results were more difficult to interpret. The toxicity of complex mixtures could not be accurately predicted based on chemical analysis. This could be due to chemical interactions or due to the presence of unidentified chemicals, such as alkyl PAHs or high molecular weight PAHs that are not included in the standard risk assessment procedure. Even though the results from these in vitro studies indicate that additive assumptions will generally be appropriate for binary mixtures similar to the ones tested here, the risk associated with complex mixtures remains a challenge to predict. Before the results of toxicity testing can be used to adjust risk assessment calculations, it is important to fully appreciate the chemical composition and to understand the mechanism of observed chemical interactions in animals chronically exposed to low doses of chemical mixtures. This research was supported by ATSDR Grant no. ATU684505 and NIEHS SBRP Grant no. P42 ES04917.     

Toxicity assessment of pramipexole in juvenile rhesus monkeys

Pramipexole (PPX) is a dopamine agonist approved for the treatment of the signs and symptoms of idiopathic Parkinson's disease as well as restless leg syndrome. The objective of this study was to investigate the toxicity of PPX when administered orally to juvenile rhesus monkeys once daily for 30 weeks, and to assess the reversibility of toxicity during a 12-week recovery. Rhesus monkeys (N = 4 males and 4 females/group; 22–24 months of age) were orally treated daily for 30 weeks with 0.0, 0.1, 0.5 or 2.0 mg/kg PPX, and subjects were assessed daily using the NCTR Operant Test Battery (OTB). Clinical chemistry, hematology, ophthalmology and other standard postmortem toxicological evaluations, including histopathology and neuropathology as well as toxicokinetics were performed. The systemic exposure to PPX was higher than that at therapeutic doses in man and AUC(0–24 h)-data increased proportionally to dose. Blood pressure significantly decreased over time in all groups including control. Near the end of treatment, there were statistically significant decreases in heart rate for the 0.5 and 2.0 mg/kg/day groups compared to control. After 4 weeks of dosing, serum prolactin was significantly decreased in all treatment groups compared to control. This decrease remained at the end of treatment in the 0.5 and 2.0 mg/kg/day groups. In summary, administration of PPX at doses of up to 2.0 mg/kg/day for 30 weeks to juvenile rhesus monkeys produced adverse findings which were attributable to its pharmacological properties, including hypoprolactinemia.     

Toxicity, bioaccumulation and metabolism of phenol in the freshwater fish Oreochromis mossambicus

The acute toxicity of phenol was determined to be 35.0 mg/l in Oreochromis mossambicus. The fish were exposed to two sublethal concentrations of phenol (2.3 and 3.5 mg/l) for 30 days. The effects of exposure were studied on the bioaccumulation and elimination of phenol from the kidney and biochemical parameters of liver, gill, and muscle at intervals of 10, 20, and 30 days. A statistically significant increase in phenol concentration was noted in tissues from all treated fish groups. Bioaccumulation and biochemical changes were dose and duration dependant. Recovery in fish after post exposure was observed after transferring these fish to normal tap water for 30 days. Elimination of phenol was noted, although the concentration of phenol remained significantly higher than the control after 30 days of the experiment. Total protein, total carbohydrate, and total lipids in the tissues of liver, gill, and muscle of fish decreased greatly. The longer the exposure time, the greater was the percentage reduction of organic matter of the fish exposed to the sublethal concentration of phenol.     

Direct and indirect effects of copper-contaminated sediments on the functions of model freshwater ecosystems

Ecotoxicology - Copper is acutely toxic to, and directly affects, primary producers and decomposers, which are key players in essential processes such as the nutrient cycle in freshwater...     

Toxicity assessment of cobalt ferrite nanoparticles on wheat plants

Cobalt ferrite nanoparticles (NPs) have received increasing attention due to their widespread therapeutic and agricultural applicability. In the environmental field, dry powder- and ferrofluid-suspended cobalt ferrite NPs were found to be useful for removing heavy metals and metalloids from water, while diluted suspensions of cobalt ferrite NP have been promisingly applied in medicine. However, the potential toxicological implications of widespread exposure are still unknown. Since cobalt ferrite NPs are considered residual wastes of environmental or medical applications, plants may serve as a point-of-entry for engineered nanomaterials as a result of consumption of these plants. Thus, the aim of this study was to assess the effects of dry powder and fresh cobalt ferrite NP on wheat plants. Seven-day assays were conducted, using quartz sand as the plant growth substrate. The toxicity end points measured were seed germination, root and shoot lengths, total cobalt (Co) and iron (Fe) accumulation, photosynthetic pigment production, protein (PRT) production, and activities of catalase (CAT), ascorbate peroxidase (APX), and guaiacol peroxidase (GPX). Increasing total Co and Fe in plant tissues indicated that wheat plants were exposed to cobalt ferrite NP. Seed germination and shoot length were not sufficiently sensitive toxicity end points. The effective concentration (EC₅₀) that diminished root length of plants by 50% was 1963 mg/kg for fresh ferrite NPs and 5023 mg/kg for powder ferrite NP. Hence, fresh ferrite NPs were more toxic than powder NP. Plant stress was indicated by a significant decrease in photosynthetic pigments. CAT, APX, and GPX antioxidant enzymatic activity suggested the generation of reactive oxygen species and oxidative damage induced by cobalt ferrite NP. More studies are thus necessary to determine whether the benefits of using these NPs outweigh the risks.     

Sensitivity assessment of freshwater macroinvertebrates to pesticides using biological traits

Assessing the sensitivity of different species to chemicals is one of the key points in predicting the effects of toxic compounds in the environment. Trait-based predicting methods have proved to be extremely efficient for assessing the sensitivity of macroinvertebrates toward compounds with non specific toxicity (narcotics). Nevertheless, predicting the sensitivity of organisms toward compounds with specific toxicity is much more complex, since it depends on the mode of action of the chemical. The aim of this work was to predict the sensitivity of several freshwater macroinvertebrates toward three classes of plant protection products: organophosphates, carbamates and pyrethroids. Two databases were built: one with sensitivity data (retrieved, evaluated and selected from the U.S. Environmental Protection Agency ECOTOX database) and the other with biological traits. Aside from the “traditional” traits usually considered in ecological analysis (i.e. body size, respiration technique, feeding habits, etc.), multivariate analysis was used to relate the sensitivity of organisms to some other characteristics which may be involved in the process of intoxication. Results confirmed that, besides traditional biological traits, related to uptake capability (e.g. body size and body shape) some traits more related to particular metabolic characteristics or patterns have a good predictive capacity on the sensitivity to these kinds of toxic substances. For example, behavioral complexity, assumed as an indicator of nervous system complexity, proved to be an important predictor of sensitivity towards these compounds. These results confirm the need for more complex traits to predict effects of highly specific substances. One key point for achieving a complete mechanistic understanding of the process is the choice of traits, whose role in the discrimination of sensitivity should be clearly interpretable, and not only statistically significant.     

Toxicity of the freshwater puffer fish Tetraodon fangi and T. palembangensis from Thailand

Toxicity of the freshwater puffer Tetraodon fangi and T. palembangensis from Amphur Chumpuang, Nakorn Ratchasima Province, Thailand (where food poisoning due to T. fangi recently occurred) was monitored from December 1988 through October 1989. The puffer were toxic throughout most of the year. There was marked variation in amounts of toxins among individuals and with the time of collection. The highest toxicity levels for T. fangi were found in skin (813 mouse units per g) following by eggs (336), muscle (331), liver (209) and intestine (159). Corresponding values for T. palembangensis were 907, 332, 282, 225 and 143 mouse units/g, respectively.