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Original Articles

Evaluation of interactive effects of phosphorus-32 and copper on marine and freshwater bivalve mollusks

, , & ORCID Icon
Received 24 Apr 2020, Accepted 09 Sep 2020, Accepted author version posted online: 24 Sep 2020, Published online: 06 Oct 2020

Abstract

Purpose

Contaminants seldom occur in isolation in the aquatic environment. While pollution of coastal and inland water bodies has received considerable attention to date, there is limited information on potential interactive effects between radionuclides and metals. Whether by accidental or controlled release, such contaminants co-exist in aquatic ecosystems and can pose an enhanced threat to biota. Using a range of biological responses, the study aimed to evaluate relative interactive effects on representative freshwater and marine bivalve species.

Methods

An integrated, multi-biomarker approach was adopted to investigate response to copper (Cu, 18 μg L−1), a known environmentally relevant genotoxic metal and differing concentrations of phosphorus-32 (32P; 0.1 and 1 mGy d−1), alone and in combination in marine (Mytilus galloprovincialis) and freshwater (Dreissena polymorpha) mussels. Genetic and molecular biomarkers were determined post-exposure and included DNA damage (as measured by the comet assay), micronuclei (MN) formation, γ-H2AX foci induction and the expression of key stress-related genes (i.e. hsp70/90, sod, cat, gst).

Results

Overall, using a tissue-specific (i.e. gill and digestive gland) approach, genotoxic response was reflective of exposures where Cu had a slight additive effect on 32P-induced damage across the species (but not all), cell types and dose rates. Multivariate analysis found significant correlations between comet and γ-H2AX assays, across both the tissues. Transcriptional expression of selected genes were generally unaltered in response to contaminant exposures, independent of species or tissues.

Conclusions

Our study is the first to explore the interactive effects of ionizing radiation (IR) and Cu on two bivalve species representing two ecological habitats. The complexity of IR-metal interactions demonstrate that extrapolation of findings obtained from single stressor studies into field conditions could be misrepresentative of real-world environments. In turn, environmental protective strategies deemed suitable in protecting biota from a single, isolated stressor may not be wholly adequate.

Highlights

  • Adoption of an integrated, multi-biomarker approach in two bivalve species.

  • Toxicity of combined mixtures of 32P and Cu compared.

  • Cu induced additive effects with 32P in the tissues.

  • DNA damage and DDR showed strong correlations.

  • Multiple stressors should be considered in assessing the impact of ionizing radiations.

Abbreviations: μg: microgram; γ-H2AX: Gamma Histone 2AX; 60Co: Cobalt 60; 137Cs: Cesium 137; 3H: Tritium; 32P: Phosphorus 32; 32S: Sulfur 32; 90Sr: Strontium 90; ACT: Actin; ATP: Adenosine triphosphate; Bq: Becquerel; BHA: Butylated hydroxyanisole; CAT: Catalase; cDna: Complimentary DNA; CPM: Counts per minute; Cq: Threshold cycle; Cu: Copper; DAPI: 4′:6-diamidino-2-phenylindole dihydrochloride; DDR: DNA damage response; DG: Digestive gland; DI: Deionized water; DO: Dissolved oxygen; DOC: Dissolved organic carbon; DP: Dreissena polymorpha; DSB: Double strand break; EF1: Elongation factor 1; ERICA: Environmental Risk from Ionizing Contaminants: Assessment and Management; GST: Glutathione-S-Transferase; HCl: Hydrochloric acid; HSP: Heat Shock Protein; ICP-MS: Inductively coupled plasma mass spectrometry; IR: Ionizing radiation; LET: Linear energy transfer; LMA: Low melting point agarose; LSC: Liquid Scintillation Counting; MBq: Mega Becquerel; MG: Mytilus galloprovincialis; mGy: Milligray; MN: Micronuclei; MOA: Mechanism of action; MT-10: Metallothionein 10; NPP: Nuclear power plant; PCA: Principal component analysis; PCR: Polymerase chain reaction; qPCR: Quantitative PCR; RER: Relative expression ratio; ROS: Reactive oxygen species; SOD: Superoxide Dismutase; SSB: Single strand break; TBHQ: Tertiary-butyl hydroquinone

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Acknowledgements

The authors thank Ms Charlotte Crowther, Mr Nick Crocker, Dr Alex Taylor and Professor William Blake, University of Plymouth, for their technical assistance and radiological protection advices.

Disclosure statement

The authors declare no conflict of interest.

Additional information

Funding

This work was funded by the Natural Environment Research Council (NERC), the Environment Agency (EA) and Radioactive Waste Management Limited (RWM) under the Radioactivity and the Environment (RATE) programme [Grant no.: NE/L000393/1].

Notes on contributors

Emily L. Vernon

Dr Emily L. Vernon is a post-doctoral research fellow working in the areas of ecotoxicology and environmental radioactivity in the School of Biological and Marine Sciences, University of Plymouth, UK.

Michael N. Moore

Professor Michael N. Moore is a Consultant Environmental Toxicologist, Cell Pathologist and Systems Biologist. He is an Honorary Professor at the European Center for Environment and Human Health (ECEHH), University of Exeter Medical School, Truro; and at the School of Biological & Marine Sciences, University of Plymouth; and also an Emeritus PML Fellow at Plymouth Marine Laboratory.

Tim P. Bean

Dr Tim P. Bean, is a Career Track Fellow at the Roslin Institute, University of Edinburgh. His research group uses molecular techniques to study aspects of fundamental bivalve biology which have direct implications for aquaculture industry and environmental policy.

Awadhesh N. Jha

Professor Awadhesh N. Jha, is a Professor of Genetic Toxicology and Ecotoxicoloy in the School of Biological and Marine Sciences, University of Plymouth, UK. His research interest involves elucidation of interactions of environmental stressors including anthropogenic chemicals and radiations with the genomes and its potential impact on the health of humans and wild species.
 

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