Abstract-The risks associated with exposure to engineered nanomaterials (ENM) will be determined in part by the processes that control their environmental fate and transformation. These processes act not only on ENM that might be released directly into the environment, but more importantly also on ENM in consumer products and those that have been released from the product. The environmental fate and transformation are likely to differ significantly for each of these cases. The ENM released from actual direct use or from nanomaterial-containing products are much more relevant for ecotoxicological studies and risk assessment than pristine ENM. Released ENM may have a greater or lesser environmental impact than the starting materials, depending on the transformation reactions and the material. Almost nothing is known about the environmental behavior and the effects of released and transformed ENM, although these are the materials that are actually present in the environment. Further research is needed to determine whether the release and transformation processes result in a similar or more diverse set of ENM and ultimately how this affects environmental behavior. This article addresses these questions, using four hypothetical case studies that cover a wide range of ENM, their direct use or product applications, and their likely fate in the environment. Furthermore, a more definitive classification scheme for ENM should be adopted that reflects their surface condition, which is a result of both industrial and environmental processes acting on the ENM. The authors conclude that it is not possible to assess the risks associated with the use of ENM by investigating only the pristine form of the ENM, without considering alterations and transformation processes. Environ. Toxicol. Chem. 2012;31:50-59. # 2011 SETAC
The unique or enhanced properties of manufactured nanomaterials (MNs) suggest that their use in nanoenabled products will continue to increase. This will result in increased potential for human and environmental exposure to MNs during manufacturing, use, and disposal of nanoenabled products. Scientifically based risk assessment for MNs necessitates the development of reproducible, standardized hazard testing methods such as those provided by the Organisation of Economic Cooperation and Development (OECD). Currently, there is no comprehensive guidance on how best to address testing issues specific to MN particulate, fibrous, or colloidal properties. This paper summarizes the findings from an expert workshop convened to develop a guidance document that addresses the difficulties encountered when testing MNs using OECD aquatic and sediment test guidelines. Critical components were identified by workshop participants that require specific guidance for MN testing: preparation of dispersions, dose metrics, the importance and challenges associated with maintaining and monitoring exposure levels, and the need for reliable methods to quantify MNs in complex media. To facilitate a scientific advance in the consistency of nanoecotoxicology test results, we identify and discuss critical considerations where expert consensus recommendations were and were not achieved and provide specific research recommendations to resolve issues for which consensus was not reached. This process will enable the development of prescriptive testing guidance for MNs. Critically, we highlight the need to quantify and properly interpret and express exposure during the bioassays used to determine hazard values.
One target of development and application of TiO(2) nanoparticles (nano-TiO(2) ) is photochemical degradation of contaminants and photo-killing of microbes and fouling organisms. However, few ecotoxicological studies have focused on this aspect of nano-TiO(2) , specifically whether this photoreactivity might significantly increase hazard and risk of the materials in the natural environment. In the present study, we evaluated acute phototoxicity of nano-TiO(2) under simulated solar radiation (SSR) to two aquatic species-Daphnia magna and Japanese medaka, using 48-h and 96-h assays, respectively. A thorough characterization of the exposure system was performed by measuring particle agglomeration and TiO(2) concentration in suspension in a time-course manner. Sedimentation and loss of bulk concentration of nano-TiO(2) particles occurred at all concentrations above 2 mg/L and was more significant as concentration increased. Phototoxicity of nano-TiO(2) under SSR was enhanced by two to four orders of magnitude as compared to toxicity under ambient laboratory light, with a 48-h median lethal concentration (LC50) of 29.8 µg/L in D. magna and a 96-h LC50 of 2.2 mg/L in medaka. Our results also indicate that these effects are dependent on simultaneous exposure of the organisms to nanoparticles and SSR. This dramatic increase in toxicity of nano-TiO(2) at environmentally realistic levels of SSR indicates the need to incorporate this mode of action into risk assessment for nano-TiO(2) and other photoreactive nanomaterials.
Generation of reactive oxygen species (ROS) by titanium dioxide nanoparticles (nano‐TiO2) and its consequent phototoxicity to Daphnia magna were measured under different solar ultraviolet (UV) spectra by applying a series of optical filters in a solar simulator. Removing UV‐B (280–320 nm) from solar radiation had no significant impact on photocatalytic ROS production of nano‐TiO2, whereas removal of UV‐A (320–400 nm) decreased ROS production remarkably. Removal of wavelengths below 400 nm resulted in negligible ROS production. A linear correlation between ROS production and D. magna immobilization suggests that photocatalytic ROS production may be a predictor of phototoxicity for nano‐TiO2. Intracellular ROS production within D. magna was consistent with the immobilization of the organism under different solar UV spectra, indicating that oxidative stress was involved in phototoxicity. The dependence of nano‐TiO2 phototoxicity on environmentally realistic variations in solar radiation suggests that risk assessment of these nanomaterials requires careful evaluation of exposure conditions in the environment. Environ. Toxicol. Chem. 2012; 31: 2099–2107. © 2012 SETAC
Abstract-Recently a suite of relatively specific hindlimb deformities have been observed in several anuran species in North America. These deformities include ectopic and supernumerary limbs and missing limbs, limb segments, or digits. The objective of this study was to assess two stressors hypothesized as responsible for limb malformations in amphibians: methoprene, an insect growth regulator that, through interaction with the retinoic acid signaling system, could possibly cause limb deformities, and ultraviolet (UV) light. Northern leopard frogs (Rana pipiens) were exposed to several different concentrations of methoprene both in the absence and presence of UV light designed to mimic the UV wavelength spectrum present in sunlight. Exposures were initiated at early embryonic stages (newly fertilized eggs) and continued through emergence of the forelimbs of the frogs. At the highest methoprene concentration tested, both in the absence and presence of UV light, severe developmental effects were observed, with all organisms dying within 12 to 16 d of test initiation. However, exposure to the pesticide did not cause limb malformations. Irrespective of methoprene treatment, a very high percentage (ϳ50%) of animals held under the UV light for 24 d developed hindlimb malformations. These malformations usually were bilateral and sometimes completely symmetrical, and consisted of missing limb segments and missing or reduced digits. A complete proximal to distal representation of the deficiencies occurred, ranging from missing or malformed femurs to the absence of single digits or digit segments. The developmental period of greatest sensitivity to UV light occurred during very early limb bud development, corresponding with formation of the apical ectodermal ridge. The significance of these findings in terms of deformed frogs in the field is uncertain. Although the deformity types observed (i.e., missing limb segments and digits) were similar to those seen in some field specimens, the UV light treatment did not cause the full range of malformations observed in animals from the field (e.g., supernumerary limbs, nonbilateral deformities). Furthermore, although the artificial light spectrum utilized mimicked the relative UV spectrum present in sunlight, it did not match full sunlight intensity, and did not accurately mimic visible wavelengths. Finally, the relationship of the UV light dose used in the laboratory to that actually experienced by amphibians in the field is uncertain. Despite these questions, our findings suggest that UV light should be further considered as a plausible factor contributing to amphibian malformations in field settings.
A number of environmental stressors have been hypothesized as responsible for recent increases in limb malformations in several species of North American amphibians. The purpose of this study was to generate dose-response data suitable for assessing the potential role of solar ultraviolet (UV) radiation in causing limb malformations in a species in which this phenomenon seemingly is particularly prevalent, the northern leopard frog (Rana pipiens). Frogs were exposed from early embryonic stages through complete metamorphosis to varying natural sunlight regimes, including unaltered (100%) sunlight, sunlight subjected to neutral density filtration to achieve relative intensities of 85%, 75%, 65%, 50%, and 25% of unaltered sunlight, and sunlight filtered with glass or acrylamide to attenuate, respectively, the UVB (290-320 nm) and UVB plus UVA (290-380 nm) portions of the spectrum. The experiments were conducted in a controlled setting, with continual monitoring of UVB, UVA, and visible light to support a robust exposure assessment. Full sunlight caused approximately 50% mortality of the frogs during early larval development; no significant treatment-related mortality occurred under any of the other exposure regimes, including 100% sunlight with glass or acrylamide filtration. There was a dose-dependent (p < 0.0001) induction of hindlimb malformations in the frogs, with the percentage of affected animals ranging from about 97% under unaltered sunlight to 0% in the 25% neutral density treatment. Malformations were comprised mostly of missing or truncated digits, and generally were bilateral as well as symmetrical. Filtration of sunlight with either glass or acrylamide both significantly reduced the incidence of malformed limbs. The estimated sunlight dose resulting in a 50% limb malformation rate (ED50) was 63.5%. The limb ED50 values based on measured sunlight intensities corresponded to average daily doses of 4.5 and 100 Wh x m(-2) for UVB and UVA, respectively. Exposure to sunlight also resulted in increased eye malformations in R. pipiens, however, the dose-response relationship for this endpoint was not monotonic. The results of this study, in conjunction with measured or predicted exposure data from natural settings, provide a basis for quantitative prediction of the risk of solar UV radiation to amphibians.
Genetic plasticity in a mosquitofish (Gambusia affinis Baird and Girard) population was examined relative to acute mercury toxicity. Genotypes at eight loci (isocitrate dehydrogenase‐1 and ‐2, mannosephosphate isomerase, glucosephosphate isomerase‐2, fumarate hydratase, malate dehydrogenase‐1, leucylglycylglycine peptidase and phenylalanylproline peptidase) were scored using starch gel electrophoresis. Two null hypotheses were tested: (a) time to death does not differ between genotypes at individual loci and (b) time to death does not differ with multiple‐locus heterozygosity. Genotypes at three of the eight loci displayed significant effects on mosquitofish time to death. Multiple‐locus heterozygosity also had a significant effect on time to death. Significant amounts of genetic plasticity were found in a population of mosquitofish with no previous exposure to inorganic mercury.
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