Advances on assessing nanotoxicity in marine fish – the pros and cons of combining an ex vivo approach and histopathological analysis in gills

C. L. Mieiro, M. Martins, M. da Silva, J. P. Coelho, C. B. Lopes, A. Alves da Silva, Joana Alves, E. Pereira, M. Pardal, M. H. Costa, M. Pacheco

Research output: Contribution to journalArticle

Abstract

The need to overcome logistic and ethical limitations of in vivo nanotoxicity evaluation in marine organisms is essential, mostly when dealing with fish. It is well established that medium/solvent conditions affect dispersion and agglomeration of nanoparticles (NPs), which represents a constraint towards a solid and realistic toxicity appraisal. In this way the pros and cons of an ex vivo approach, using a simplified exposure medium (seawater) and addressing gills histopathology, were explored. The nanotoxic potential of environmentally realistic concentrations of titanium dioxide NPs (TiO2 NPs) was also assessed, disclosing the morpho-functional effects on the gills and the possible uptake/elimination processes. Excised gills of the Senegalese sole (Solea senegalensis) were directly exposed in artificial seawater to 20 and 200 μg L−1 TiO2 NPs, for 2 h and 4 h. Semi-quantitative and quantitative histological analyses were applied. The normal morphology of the gill's epithelia was only slightly altered in the control, reflecting protective mechanisms against the artificiality of the experimental conditions, which, together with the absence of differences in the global histopathological index (Ih), corroborated that the gill's morpho-functional features were not compromised, thereby validating the proposed ex vivo approach. TiO2 NPs induced moderate severity and dissemination of histopathological lesions. After 2 h, a series of compensatory mechanisms occurred in NP treatments, implying an efficient response of the innate defense system (increasing number of goblet cells) and effective osmoregulatory ability (chloride cells proliferation). After 4 h, gills revealed signs of recovery (normalization of the number of chloride and goblet cells; similar Ih), highlighting the tissue viability and effective elimination and/or neutralization of NPs. The uptake of the TiO2 NPs seemed to be favored by the higher particle sizes. Overall, the proposed approach emerged as a high-throughput, reliable, accurate and ethically commendable methodology for nanotoxicity assessment in marine fish.

Original languageEnglish
Article number105322
JournalAquatic Toxicology
Volume217
DOIs
Publication statusPublished - 1 Dec 2019

Fingerprint

nanoparticles
marine fish
Nanoparticles
gills
Fishes
fish
Solea senegalensis
Goblet Cells
goblet cells
Seawater
Chlorides
chlorides
seawater
chloride
Flatfishes
titanium dioxide
uptake mechanisms
Tissue Survival
Aquatic Organisms
histopathology

Keywords

  • Ex vivo
  • Gills
  • Histopathology
  • Marine fish
  • Realistic concentrations
  • TiO nanoparticles

Cite this

Mieiro, C. L. ; Martins, M. ; da Silva, M. ; Coelho, J. P. ; Lopes, C. B. ; da Silva, A. Alves ; Alves, Joana ; Pereira, E. ; Pardal, M. ; Costa, M. H. ; Pacheco, M. / Advances on assessing nanotoxicity in marine fish – the pros and cons of combining an ex vivo approach and histopathological analysis in gills. In: Aquatic Toxicology. 2019 ; Vol. 217.
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Advances on assessing nanotoxicity in marine fish – the pros and cons of combining an ex vivo approach and histopathological analysis in gills. / Mieiro, C. L.; Martins, M.; da Silva, M.; Coelho, J. P.; Lopes, C. B.; da Silva, A. Alves; Alves, Joana; Pereira, E.; Pardal, M.; Costa, M. H.; Pacheco, M.

In: Aquatic Toxicology, Vol. 217, 105322, 01.12.2019.

Research output: Contribution to journalArticle

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AU - Mieiro, C. L.

AU - Martins, M.

AU - da Silva, M.

AU - Coelho, J. P.

AU - Lopes, C. B.

AU - da Silva, A. Alves

AU - Alves, Joana

AU - Pereira, E.

AU - Pardal, M.

AU - Costa, M. H.

AU - Pacheco, M.

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N2 - The need to overcome logistic and ethical limitations of in vivo nanotoxicity evaluation in marine organisms is essential, mostly when dealing with fish. It is well established that medium/solvent conditions affect dispersion and agglomeration of nanoparticles (NPs), which represents a constraint towards a solid and realistic toxicity appraisal. In this way the pros and cons of an ex vivo approach, using a simplified exposure medium (seawater) and addressing gills histopathology, were explored. The nanotoxic potential of environmentally realistic concentrations of titanium dioxide NPs (TiO2 NPs) was also assessed, disclosing the morpho-functional effects on the gills and the possible uptake/elimination processes. Excised gills of the Senegalese sole (Solea senegalensis) were directly exposed in artificial seawater to 20 and 200 μg L−1 TiO2 NPs, for 2 h and 4 h. Semi-quantitative and quantitative histological analyses were applied. The normal morphology of the gill's epithelia was only slightly altered in the control, reflecting protective mechanisms against the artificiality of the experimental conditions, which, together with the absence of differences in the global histopathological index (Ih), corroborated that the gill's morpho-functional features were not compromised, thereby validating the proposed ex vivo approach. TiO2 NPs induced moderate severity and dissemination of histopathological lesions. After 2 h, a series of compensatory mechanisms occurred in NP treatments, implying an efficient response of the innate defense system (increasing number of goblet cells) and effective osmoregulatory ability (chloride cells proliferation). After 4 h, gills revealed signs of recovery (normalization of the number of chloride and goblet cells; similar Ih), highlighting the tissue viability and effective elimination and/or neutralization of NPs. The uptake of the TiO2 NPs seemed to be favored by the higher particle sizes. Overall, the proposed approach emerged as a high-throughput, reliable, accurate and ethically commendable methodology for nanotoxicity assessment in marine fish.

AB - The need to overcome logistic and ethical limitations of in vivo nanotoxicity evaluation in marine organisms is essential, mostly when dealing with fish. It is well established that medium/solvent conditions affect dispersion and agglomeration of nanoparticles (NPs), which represents a constraint towards a solid and realistic toxicity appraisal. In this way the pros and cons of an ex vivo approach, using a simplified exposure medium (seawater) and addressing gills histopathology, were explored. The nanotoxic potential of environmentally realistic concentrations of titanium dioxide NPs (TiO2 NPs) was also assessed, disclosing the morpho-functional effects on the gills and the possible uptake/elimination processes. Excised gills of the Senegalese sole (Solea senegalensis) were directly exposed in artificial seawater to 20 and 200 μg L−1 TiO2 NPs, for 2 h and 4 h. Semi-quantitative and quantitative histological analyses were applied. The normal morphology of the gill's epithelia was only slightly altered in the control, reflecting protective mechanisms against the artificiality of the experimental conditions, which, together with the absence of differences in the global histopathological index (Ih), corroborated that the gill's morpho-functional features were not compromised, thereby validating the proposed ex vivo approach. TiO2 NPs induced moderate severity and dissemination of histopathological lesions. After 2 h, a series of compensatory mechanisms occurred in NP treatments, implying an efficient response of the innate defense system (increasing number of goblet cells) and effective osmoregulatory ability (chloride cells proliferation). After 4 h, gills revealed signs of recovery (normalization of the number of chloride and goblet cells; similar Ih), highlighting the tissue viability and effective elimination and/or neutralization of NPs. The uptake of the TiO2 NPs seemed to be favored by the higher particle sizes. Overall, the proposed approach emerged as a high-throughput, reliable, accurate and ethically commendable methodology for nanotoxicity assessment in marine fish.

KW - Ex vivo

KW - Gills

KW - Histopathology

KW - Marine fish

KW - Realistic concentrations

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