Metallic nanoparticles are among the most widely used types of engineered nanomaterials; however, little is known about their environmental fate and effects. To assess potential environmental effects of engineered nanometals, it is important to determine which species are sensitive to adverse effects of various nanomaterials. In the present study, zebrafish, daphnids, and an algal species were used as models of various trophic levels and feeding strategies. To understand whether observed effects are caused by dissolution, particles were characterized before testing, and particle concentration and dissolution were determined during exposures. Organisms were exposed to silver, copper, aluminum, nickel, and cobalt as both nanoparticles and soluble salts as well as to titanium dioxide nanoparticles. Our results indicate that nanosilver and nanocopper cause toxicity in all organisms tested, with 48-h median lethal concentrations as low as 40 and 60 microg/L, respectively, in Daphnia pulex adults, whereas titanium dioxide did not cause toxicity in any of the tests. Susceptibility to nanometal toxicity differed among species, with filter-feeding invertebrates being markedly more susceptible to nanometal exposure compared with larger organisms (i.e., zebrafish). The role of dissolution in observed toxicity also varied, being minor for silver and copper but, apparently, accounting for most of the toxicity with nickel. Nanoparticulate forms of metals were less toxic than soluble forms based on mass added, but other dose metrics should be developed to accurately assess concentration-response relationships for nanoparticle exposures.
Increasing use of metallic nanomaterials is likely to result in release of these particles into aqueous environments; however, it is unclear if these materials present a hazard to aquatic organisms. Because some dissolution of metal particles will occur, it is important to distinguish effects of nanoparticulates from dissolved metals. To address this issue, acute toxicity of soluble copper and 80 nm copper nanoparticle suspensions were examined in zebrafish. The results demonstrate that nanocopper is acutely toxic to zebrafish, with a 48 h LC50 concentration of 1.5 mg/L. Rapid aggregation of copper nanoparticles occurred after suspension in water, resulting in 50–60% of added mass leaving the water column. While dissolution of particulate copper occurred, it was insufficient to explain the mortality in nanocopper exposures. Histological and biochemical analysis revealed that the gill was the primary target organ for nanocopper. To further investigate the effects of nanocopper on the gill, zebrafish were exposed to 100 µg/L of nanocopper or to the concentration of soluble copper matching that present due to dissolution of the particles. Under these conditions, nanocopper produced different morphological effects and global gene expression patterns in the gill than soluble copper, clearly demonstrating that the effects of nanocopper on gill are not mediated solely by dissolution.
Research has demonstrated that metallic nanoparticles produce toxicity in aquatic organisms that is due largely to effects of particulates as opposed to release of dissolved ions. The present research examined the interplay of nanoparticle composition and dissolution on response of the zebrafish gill following exposure to toxic (nanocopper or nanosilver) or nontoxic (nano-TiO2) nanometals. Female zebrafish were exposed to 48-h no observable effects concentration of nanocopper and nanosilver or to soluble Cu and Ag that matched the concentration of dissolved metals released during nanoparticle exposure. Both nanocopper and nanosilver exposures increased metal content associated with gill tissue, though silver concentrations were much higher following nanosilver exposures suggesting that intact silver nanoparticles are associated with the gill. Morphological and transcriptional responses of the gills differed among various nanomaterials and between nanoparticulate and soluble species. Nanocopper increased mean gill filament width by three to fourfold between 24 and 48 h, whereas nanosilver did not alter gill filament width at either time point. Global gene expression analysis demonstrates that the exposure to each nanometal or soluble metal produces a distinct gene expression profile at both 24 and 48 h, suggesting that each exposure is producing biological response by a different mechanism. The differences in responses among the exposures indicates that each particle is having a distinct biological effect that does not appear to be driven solely by release of soluble metal ions into the water column. Based on these results, care should be taken when inferring toxicity of nanomaterials from data on a different material.
The 2010 Deepwater Horizon oil spill resulted in the accidental release of approximately 700 million L of crude oil into the Gulf of Mexico. Photo-induced toxicity after co-exposure to ultraviolet (UV) radiation is 1 mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Blue crab are an important commercial and ecological resource in the Gulf of Mexico, and their largely transparent larvae may make them sensitive to PAH photo-induced toxicity. The goal of the present study was to examine the sensitivity of early lifestage blue crab (Callinectes sapidus) zoea to slick oil collected during the Deepwater Horizon spill. Blue crab zoea were exposed to 1 of several dilutions of water accommodated fractions from 1 of 2 sources of oil and gradations of natural sunlight in a factorial design. Two 7-h solar exposures were carried out with a recovery period (dark) in between. Survival was found to be UV- and PAH-dependent. Toxicity was observed within the range of surface PAH concentrations reported in the Gulf of Mexico during the Deepwater Horizon spill. These findings indicate that early lifestage blue crab are sensitive to photo-induced toxicity from Deepwater Horizon slick oil.
A novel custom microarray for largemouth bass (Micropterus salmoides) was designed with sequences obtained from a normalized cDNA library using the 454 Life Sciences GS-20 pyrosequencer. This approach yielded in excess of 58 million bases of high-quality sequence. The sequence information was combined with 2,616 reads obtained by traditional suppressive subtractive hybridizations to derive a total of 31,391 unique sequences. Annotation and coding sequences were predicted for these transcripts where possible. 16,350 annotated transcripts were selected as target sequences for the design of the custom largemouth bass oligonucleotide microarray. The microarray was validated by examining the transcriptomic response in male largemouth bass exposed to 17beta-oestradiol. Transcriptomic responses were assessed in liver and gonad, and indicated gene expression profiles typical of exposure to oestradiol. The results demonstrate the potential to rapidly create the tools necessary to assess large scale transcriptional responses in non-model species, paving the way for expanded impact of toxicogenomics in ecotoxicology.
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