Earthworm sublethal responses to titanium dioxide nanomaterial in soil detected by ¹H NMR metabolomics.

¹H NMR-based metabolomics was used to examine the response of Eisenia fetida earthworms raised from juveniles for 20-23 weeks in soil spiked with either 20 or 200 mg/kg of a commercially available uncoated titanium dioxide (TiO(2)) nanomaterial (nominal diameter of 5 nm). To distinguish responses specific to particle size, soil treatments spiked with a micrometer-sized TiO(2) material (nominal diameter, <45 μm) at the same concentrations (20 and 200 mg/kg) were also included in addition to an unspiked control soil. Multivariate statistical analysis of the (1)H NMR spectra for aqueous extracts of E. fetida tissue suggested that earthworms exhibited significant changes in their metabolic profile following TiO(2) exposure for both particle sizes. The observed earthworm metabolic changes appeared to be consistent with oxidative stress, a proposed mechanism of toxicity for nanosized TiO(2). In contrast, a prior study had observed no impairment of E. fetida survival, reproduction, or growth following exposure to the same TiO(2) spiked soils. This suggests that (1)H NMR-based metabolomics provides a more sensitive measure of earthworm response to TiO(2) materials in soil and that further targeted assays to detect specific cellular or molecular level damage to earthworms caused by chronic exposure to TiO(2) are warranted.

[1]  Jianmeng Chen,et al.  Effects of phenanthrene on the mortality, growth, and anti-oxidant system of earthworms (Eisenia fetida) under laboratory conditions. , 2011, Chemosphere.

[2]  B. Nowack,et al.  Exposure modeling of engineered nanoparticles in the environment. , 2008, Environmental science & technology.

[3]  Timothy M. D. Ebbels,et al.  Validation of metabolomics for toxic mechanism of action screening with the earthworm Lumbricus rubellus , 2009, Metabolomics.

[4]  Jing Chen,et al.  Toxicological effects of TiO2 and ZnO nanoparticles in soil on earthworm Eisenia fetida. , 2010 .

[5]  M. Simpson,et al.  Evaluation of sample preparation methods for nuclear magnetic resonance metabolic profiling studies with Eisenia fetida , 2008, Environmental toxicology and chemistry.

[6]  Yonglong Lu,et al.  Antioxidant and metabolic responses induced by cadmium and pyrene in the earthworm Eisenia fetida in two different systems: contact and soil tests , 2009 .

[7]  P. Kille,et al.  Metabolic profile biomarkers of metal contamination in a sentinel terrestrial species are applicable across multiple sites. , 2007, Environmental science & technology.

[8]  Ralf J M Weber,et al.  Discriminating between different acute chemical toxicities via changes in the daphnid metabolome. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[9]  Douglas M. Hawkins,et al.  Assessing Model Fit by Cross-Validation , 2003, J. Chem. Inf. Comput. Sci..

[10]  M. Simpson,et al.  1H NMR metabolomics of earthworm responses to sub-lethal PAH exposure , 2009 .

[11]  T. Ebbels,et al.  Metabolic profiling, metabolomic and metabonomic procedures for NMR spectroscopy of urine, plasma, serum and tissue extracts , 2007, Nature Protocols.

[12]  M. Simpson,et al.  Correlations of Eisenia fetida metabolic responses to extractable phenanthrene concentrations through time. , 2010, Environmental pollution.

[13]  G. Oberdörster,et al.  Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles , 2005, Environmental health perspectives.

[14]  J. Lindon,et al.  Scaling and normalization effects in NMR spectroscopic metabonomic data sets. , 2006, Analytical chemistry.

[15]  D. Spurgeon,et al.  Effects of metal-contaminated soils on the growth, sexual development, and early cocoon production of the earthworm Eisenia fetida, with particular reference to zinc. , 1996, Ecotoxicology and environmental safety.

[16]  Pratim Biswas,et al.  Does nanoparticle activity depend upon size and crystal phase? , 2008, Nanotoxicology.

[17]  S. Stürzenbaum,et al.  'Systems toxicology' approach identifies coordinated metabolic responses to copper in a terrestrial non-model invertebrate, the earthworm Lumbricus rubellus , 2008, BMC Biology.

[18]  Age K. Smilde,et al.  Exploring the analysis of structured metabolomics data , 2009 .

[19]  D. Oughton,et al.  Ecotoxicological effects of an aged TiO2 nanocomposite measured as apoptosis in the anecic earthworm Lumbricus terrestris after exposure through water, food and soil. , 2011, Environment international.

[20]  Douglas B. Kell,et al.  Statistical strategies for avoiding false discoveries in metabolomics and related experiments , 2007, Metabolomics.

[21]  Age K. Smilde,et al.  UvA-DARE ( Digital Academic Repository ) Assessment of PLSDA cross validation , 2008 .

[22]  Qian Bu,et al.  NMR-based metabonomic study of the sub-acute toxicity of titanium dioxide nanoparticles in rats after oral administration , 2010, Nanotechnology.

[23]  Oliver A.H. Jones,et al.  A metabolomics based approach to assessing the toxicity of the polyaromatic hydrocarbon pyrene to the earthworm Lumbricus rubellus. , 2008, Chemosphere.

[24]  R. Cramer.  Partial Least Squares (PLS): Its strengths and limitations , 1993 .

[25]  Francesco Falciani,et al.  Discovery of metabolic signatures for predicting whole organism toxicology. , 2010, Toxicological sciences : an official journal of the Society of Toxicology.

[26]  J. Lindon,et al.  Environmental Metabonomics: Applying Combination Biomarker Analysis in Earthworms at a Metal Contaminated Site , 2004, Ecotoxicology.

[27]  H. Roche,et al.  Effects of physical (hypoxia, hyperoxia) and chemical (potassium dichromate, fenitrothion) stress on antioxidant enzyme activities in Chironomus riparius mg. (diptera, chironomidae) larvae: Potential biomarkers , 2000 .

[28]  Xiaoshan Zhu,et al.  Toxicity and bioaccumulation of TiO2 nanoparticle aggregates in Daphnia magna. , 2010, Chemosphere.

[29]  Y. Berthelot,et al.  Integration of toxicological and chemical tools to assess the bioavailability of metals and energetic compounds in contaminated soils. , 2008, Chemosphere.

[30]  Xuezhi Zhang,et al.  Trophic transfer of TiO(2) nanoparticles from Daphnia to zebrafish in a simplified freshwater food chain. , 2010, Chemosphere.

[31]  O. Tsyusko,et al.  Evidence for bioavailability of Au nanoparticles from soil and biodistribution within earthworms (Eisenia fetida). , 2010, Environmental science & technology.

[32]  Mark R Viant,et al.  Recent developments in environmental metabolomics. , 2008, Molecular bioSystems.

[33]  C. Delisle,et al.  Ecotoxicological evaluation of three deicers (NaCl, NaFo, CMA)-effect on terrestrial organisms. , 2001, Ecotoxicology and environmental safety.

[34]  G. Lowry,et al.  Towards a definition of inorganic nanoparticles from an environmental, health and safety perspective. , 2009, Nature nanotechnology.

[35]  Karluss Thomas,et al.  Research strategies for safety evaluation of nanomaterials, Part I: evaluating the human health implications of exposure to nanoscale materials. , 2005, Toxicological sciences : an official journal of the Society of Toxicology.

[36]  R. Scholz,et al.  Modeled environmental concentrations of engineered nanomaterials (TiO(2), ZnO, Ag, CNT, Fullerenes) for different regions. , 2009, Environmental science & technology.

[37]  Pedro J J Alvarez,et al.  Comparative eco-toxicity of nanoscale TiO2, SiO2, and ZnO water suspensions. , 2006, Water research.

[38]  D. Volk,et al.  1H NMR metabonomic study of rat response to tri-phenyl phosphate and tri-butyl phosphate exposure , 2010, Metabolomics.

[39]  E. Want,et al.  The metabolomic responses of Caenorhabditis elegans to cadmium are largely independent of metallothionein status, but dominated by changes in cystathionine and phytochelatins. , 2009, Journal of proteome research.

[40]  J G Bundy,et al.  An NMR‐based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress , 2001, FEBS letters.

[41]  Pratim Biswas,et al.  Characterization of size, surface charge, and agglomeration state of nanoparticle dispersions for toxicological studies , 2009 .

[42]  B. Rossaro,et al.  Increased alanine concentration is associated with exposure to fenitrothion but not carbamates in Chironomus riparius larvae. , 2007, Ecotoxicology and environmental safety.

[43]  John A Tomenson,et al.  Titanium dioxide: inhalation toxicology and epidemiology. , 2005, The Annals of occupational hygiene.

[44]  Nouna Kettaneh-Wold,et al.  Analysis of mixture data with partial least squares , 1992 .

[45]  Peter Filzmoser,et al.  Introduction to Multivariate Statistical Analysis in Chemometrics , 2009 .

[46]  M. Simpson,et al.  1H NMR metabolomics of earthworm exposure to sub-lethal concentrations of phenanthrene in soil. , 2010, Environmental pollution.

[47]  Anita Jemec,et al.  In vivo screening to determine hazards of nanoparticles: nanosized TiO2. , 2009, Environmental pollution.

[48]  X. Sima,et al.  Effects of nano-scale TiO2, ZnO and their bulk counterparts on zebrafish: acute toxicity, oxidative stress and oxidative damage. , 2011, The Science of the total environment.

[49]  T. Ebbels,et al.  Metabonomic investigations into the global biochemical sequelae of exposure to the pancreatic toxin 1‐cyano‐2‐hydroxy‐3‐butene in the rat , 2009, Magnetic resonance in chemistry : MRC.

[50]  Valery E Forbes,et al.  The use and misuse of biomarkers in ecotoxicology , 2006, Environmental toxicology and chemistry.

[51]  J. Lindon,et al.  Qualitative high field 1H-NMR spectroscopy for the characterization of endogenous metabolites in earthworms with biochemical biomarker potential , 2005, Metabolomics.

[52]  J. Oris,et al.  Anthracene photoinduced toxicity to plhc‐1 cell line (Poeciliopsis lucida) and the role of lipid peroxidation in toxicity , 2000 .

[53]  W. Hendershot,et al.  Ion Exchange and Exchangeable Cations , 2007 .

[54]  Rasmus Bro,et al.  Some common misunderstandings in chemometrics , 2010 .

[55]  Jamie R Lead,et al.  Nanomaterials in the environment: Behavior, fate, bioavailability, and effects , 2008, Environmental toxicology and chemistry.

[56]  Theodoros N. Arvanitis,et al.  A new approach to toxicity testing in Daphnia magna: application of high throughput FT-ICR mass spectrometry metabolomics , 2009, Metabolomics.

[57]  J. Morrow,et al.  Trophic transfer of nanoparticles in a simplified invertebrate food web. , 2008, Nature nanotechnology.

[58]  Ying Tang,et al.  Cellular Toxicity of TiO2 Nanoparticles in Anatase and Rutile Crystal Phase , 2011, Biological Trace Element Research.

[59]  Jaeweon Cho,et al.  Oxidative stress responses of Daphnia magna exposed to TiO(2) nanoparticles according to size fraction. , 2010, The Science of the total environment.

[60]  A. Simpson,et al.  Purge NMR: effective and easy solvent suppression. , 2005, Journal of magnetic resonance.

[61]  Baoshan Xing,et al.  Toxicity of nanoparticulate and bulk ZnO, Al2O3 and TiO2 to the nematode Caenorhabditis elegans. , 2009, Environmental pollution.

[62]  D. Larsson,et al.  Reproducible (1)H NMR-based metabolomic responses in fish exposed to different sewage effluents in two separate studies. , 2011, Environmental science & technology.

[63]  Julian Moger,et al.  Sublethal toxicity of nano-titanium dioxide and carbon nanotubes in a sediment dwelling marine polychaete. , 2010, Environmental pollution.

[64]  T. MacCormack,et al.  Identifying and Predicting Biological Risks Associated With Manufactured Nanoparticles in Aquatic Ecosystems , 2008 .

[65]  F. Besenbacher,et al.  Limit-test toxicity screening of selected inorganic nanoparticles to the earthworm Eisenia fetida , 2011, Ecotoxicology.

[66]  H. Senn,et al.  Probabilistic quotient normalization as robust method to account for dilution of complex biological mixtures. Application in 1H NMR metabonomics. , 2006, Analytical chemistry.

[67]  M. Simpson,et al.  1H NMR and GC/MS metabolomics of earthworm responses to sub-lethal DDT and endosulfan exposure , 2009, Metabolomics.

[68]  Clive A. Edwards,et al.  Biology and Ecology of Earthworms , 1995 .

[69]  J. West,et al.  Correlating nanoscale titania structure with toxicity: a cytotoxicity and inflammatory response study with human dermal fibroblasts and human lung epithelial cells. , 2006, Toxicological sciences : an official journal of the Society of Toxicology.

[70]  J. Whalen,et al.  Reproductive and behavioral responses of earthworms exposed to nano‐sized titanium dioxide in soil , 2012, Environmental toxicology and chemistry.

[71]  John L Markley,et al.  Metabolite identification via the Madison Metabolomics Consortium Database , 2008, Nature Biotechnology.

[72]  B. Xing,et al.  Effect of sub-acute exposure to TiO2 nanoparticles on oxidative stress and histopathological changes in Juvenile Carp (Cyprinus carpio). , 2009, Journal of environmental sciences.

[73]  T. W. Anderson,et al.  An Introduction to Multivariate Statistical Analysis , 1959 .