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Assessment of cytotoxicity, genotoxicity and 7-ethoxyresorufin-O-deethylase (EROD) induction in sediment extracts from New Zealand urban estuaries

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Abstract

Sediments represent a major sink for contaminants resulting from industrial and agricultural activities — especially lipophilic substances. This study exclusively used in vitro methodologies to characterize specific toxicity effects of contaminants in sediment extracts from two urban New Zealand estuaries. Sediment extracts were prepared and tested for a range of biological endpoints. The micronucleus and comet assays in V79 cells were used to assess genotoxicity. Induction of 7-ethoxyresorufin-O-deethylase in piscine RTL-W1 cells was determined to estimate dioxin-like toxicity. Cytotoxic potentials were analyzed by neutral red uptake and MTT reduction. There was evidence of strong dioxin-like toxicity and moderate cytotoxicity. Genotoxicity was distinct in the micronucleus assay, but low in the comet assay. The results indicate the presence of chemicals in the sediments with the potential to pose a risk through multiple mechanisms of toxicity, the identities and amounts of which will be disclosed in a parallel study alongside with in vivo toxicity data.

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Abbreviations

ASE:

Accelerated solvent extraction

BNF:

β-Naphthoflavone

CP:

Cyclophosphamide

DAPI:

4′,6-Diamidin-2-phenylindol

DCP:

3,5-Dichlorophenol

DMSO:

Dimethyl sulfoxide

EDTA:

Ethylenediaminetetraacetic acid

EROD:

7-Ethoxyresorufin-O-deethylase

FBS:

Fetal bovine serum

FEL-TEQ:

Fixed-effect level toxicity equivalent

IPA:

Isopropyl alcohol

LMA:

Low melting agarose

MIC-EQ:

Maximally inducing concentration toxicity equivalent

MOA:

Mode of action

MTT:

3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; thiazolyl blue tetrazolium bromide

NADP:

Nicotinamide adenine dinucleotide phosphate

NMA:

Normal melting agarose

PAHs:

Polycyclic aromatic hydrocarbons

PBS:

Phosphate-buffered saline

S9:

Supernatant of rat-liver homogenate fractionated by centrifugation at 9000 rpm

SEQ:

Sediment equivalent

SPE:

Solid-phase extraction

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Acknowledgements

RTL-W1 cells were kindly provided by Dr. Niels C. Bols (Department of Biology, University of Waterloo, Canada) and Dr. Lucy Lee (Faculty of Science, University of the Fraser Valley, Canada).

Funding

Traveling cost associated with this research was partly funded by a New Zealand Ministry of Business, Innovation and Employment (MBIE) TTW grant (C09X0801), a Royal Society of New Zealand International Mobility Fund (IMF) NZ-Germany Science & Technology Programme: Contract Number: FRG11-26 and a German Federal Agency for Agriculture and Nutrition (‘Bundesanstalt für Landwirtschaft und Ernährung’, BLE) grant (530-06.01-NZL).

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Authors and Affiliations

  1. Aquatic Ecology and Toxicology Section, Centre for Organismal Studies, University of Heidelberg, Im Neuenheimer Feld 504, 69120, Heidelberg, Germany

    Patrick Heinrich, Lara L. Petschick & Thomas Braunbeck

  2. Northcott Research Consultants Ltd, 20 River Oaks Place, Hamilton, 3200, New Zealand

    Grant L. Northcott

  3. Cawthron Institute, 98 Halifax St E, The Wood, Nelson, 7010, New Zealand

    Louis A. Tremblay & James M. Ataria

  4. School of Biological Sciences, University of Auckland, Auckland, 1142, New Zealand

    Louis A. Tremblay

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  1. Patrick Heinrich
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  2. Lara L. Petschick
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Correspondence to Patrick Heinrich.

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Heinrich, P., Petschick, L.L., Northcott, G.L. et al. Assessment of cytotoxicity, genotoxicity and 7-ethoxyresorufin-O-deethylase (EROD) induction in sediment extracts from New Zealand urban estuaries. Ecotoxicology 26, 211–226 (2017). https://doi.org/10.1007/s10646-016-1756-1

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