, Volume 27, Issue 4, pp 430–439 | Cite as

Protein profiling as early detection biomarkers for TiO2 nanoparticle toxicity in Daphnia magna

  • Paula Sá-Pereira
  • Mário S. Diniz
  • Liliana Moita
  • Teresa Pinheiro
  • Elsa Mendonça
  • Susana M. Paixão
  • Ana Picado


The mode of action for nanoparticle (NP) toxicity in aquatic organisms is not yet fully understood. In this work, a strategy other than toxicity testing was applied to Daphnia magna exposed to TiO2-NPs: the use of nuclear microscopy and the assessment of protein profile. D. magna is a keystone species broadly used as a model system in ecotoxicology. Titanium (Ti) was found in the D. magna digestive tract, mainly in the gut. The penetration of Ti into the epithelial region was greater at higher exposure levels and also observed in eggs in the brood pouch. The protein profile of individuals exposed to different concentrations showed that 2.8 and 5.6 mg/L TiO2-NP concentrations induced an over-expression of the majority of proteins, in particular proteins with molecular weight of ∼120, 85 and 15 kDa, while 11.2 mg/L TiO2-NP had an inhibitory effect on protein expression. The Matrix-assisted laser desorption ionization with tandem time of flight mass spectrometry (MALDI-TOF/TOF MS) analysis of these proteins consistently identified them as vitellogenin (Vtg)-like proteins, associated with enzymes involved in redox balance. These results indicate that Vtg-like proteins are up-regulated in D. magna exposed to TiO2-NPs. Vitellogenesis is associated with the reproduction system, suggesting that TiO2-NP exposure can impair reproduction by affecting this process. The precise mode of action of TiO2-NPs is still unclear and the results from this study are a first attempt to identify specific proteins as potential markers of TiO2-NP toxicity in D. magna, providing useful information for future research.


TiO2 nanoparticles Daphnia magna 1D-electrophoresis Protein profiling Early biomarkers 



Project funded by Fundação para a Ciência e Tecnologia, PTDC/CTM/099446/2008 and PEst-C/EQB/LA0006/2011 granted to Requimte. This work was also financed by national funds through FCT–Foundation for Science and Technology, under the project UID/BIO/04565/2013 and Programa Operacional Regional de Lisboa (LISBOA-01-0145-FEDER-007317). In addition, the authors would like to acknowledge Dr. José Carlos Roseiro, senior researcher from LNEG, and Dr. Andrew Wilbey, professor from University of Reading at UK, for carefully proof reading of the English in this article.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.INIAV—Instituto Nacional de Investigação Agrária e Veterinária, IPOeirasPortugal
  2. 2.REQUIMTE, Departamento de Química, Centro de Química Fina e Biotecnologia, Faculdade de Ciências e TecnologiaUniversidade Nova de LisboaCaparicaPortugal
  3. 3.LNEG–Laboratório Nacional de Energia e Geologia, IPAmadoraPortugal
  4. 4.Instituto de Bioengenharia e Biociências, Departamento de Engenharia e Ciências Nucleares, Instituto Superior TécnicoUniversidade de LisboaLisboaPortugal
  5. 5.APA-Agência Portuguesa do AmbienteAmadoraPortugal

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