Aluminium Induced DNA-damage and Oxidative Stress in Cultures of the Marine Sponge Hymeniacidon perlevis

Rachael Ununuma Akpiri (School of Biosciences, University of Birmingham, United Kingdom)
Rosline Sonayee Konya (Department of Animal and Environmental Biology, University of Port Harcourt, Nigeria)
Nikolas John Hodges (School of Biosciences, University of Birmingham, United Kingdom)


Aluminium is the most abundant element in the earth crust, and has no known biological function. However, it is an established neurotoxicant in its trivalent oxidation state, with exposure resulting in neurodegenerative diseases like Parkinson’s disease and presenile dementia. Although, the potential genotoxic and carcinogenic effects of aluminium are established in mammalian and other model system, there is however very limited information on aluminium genotoxicity in aquatic invertebrates. Mechanism of aluminium toxicity is also largely unclear. With a concentration range between 0.001– 0.05mg/L in near neutral pH water, and up to 0.5-1mg/L in an acidic water , aluminium poses a potential threat to the marine ecosystem, however it is poorly studied. This study, therefore presents for the first time, aluminium-induced DNA damage using the comet assay and reactive oxygen Species (ROS) formation using 2’, 7’-dichlorodihydrofluorescein diacetate (H2DCF-DA) assay as biomarkers of genotoxicity and oxidative stress in the inter-tidal marine sponge Hymeniacidon perlevis, respectively. H. perlevis is widely distributed in the British Isles, Mediterranean and the Arctic sea and has been reported as a model for environmental biomonitoring in aquatic ecosystem and as a suitable alternative to bivalves. In this study, cryopreserved single sponge cells of H. perlevis were cultured as viable aggregates and were thereafter treated with 0.1, 0.2, 0.3 and 0.4mg/L aluminium chloride (AlCl3) for 12 hours. Cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Our results showed that non-cytotoxic concentrations of AlCl3 caused a statistically significant concentration-dependent increase in the level of DNA-strand break and reactive oxygen species formation single sponge cells of H. perlevis. There was also a statistically significant positive linear correlation between aluminium-induced DNA strand break and ROS formation suggesting the involvement of ROS in the causative mechanism of the aluminium induced DNA-strand breaks observed.


Aluminium; Comet assay; DNA Damage; ROS; Sponge; Hymeniacidon perlevis

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[1] Ferrante, M., et al.. In vivo exposure of the marine sponge Chondrilla nucula Schmidt, 1862 to cadmium (Cd), copper (Cu) and lead (Pb) and its potential use for bioremediation purposes. Chemosphere, 2018, 193: 1049-1057.

[2] Cebrian, E., M.J. Uriz, and X. Turon. Sponges as biomonitors of heavy metals in spatial and temporal surveys in northwestern Mediterranean: multispecies comparison. Environmental Toxicology and Chemistry, 2007, 26(11): 2430-2439.

[3] Zheng, L., et al.. Identification of norharman as the cytotoxic compound produced by the sponge (Hymeniacidon perleve)‐associated marine bacterium Pseudoalteromonas piscicida and its apoptotic effect on cancer cells. Biotechnology and applied biochemistry, 2006, 44(3): 135-142.

[4] Mahaut, M.-L., et al.. The porifera Hymeniacidon perlevis (Montagu, 1818) as a bioindicator for water quality monitoring. Environmental Science and Pollution Research, 2013, 20(5): 2984-2992.

[5] Longo, C., et al.. Bacterial accumulation by the Demospongiae Hymeniacidon perlevis: a tool for the bioremediation of polluted seawater. Marine pollution bulletin, 2010, 60(8): 1182-1187.

[6] Akpiri, R.U., R.S. Konya, and N.J. Hodges. Development of cultures of the marine sponge Hymeniacidon perleve for genotoxicity assessment using the alkaline comet assay. Environmental toxicology and chemistry, 2017.

[7] Batista, D., et al.. Marine sponges as bioindicators of oil and combustion derived PAH in coastal waters. Marine environmental research, 2013, 92: 234-243.

[8] Cai, X., et al.. Establishing primary cell cultures from Branchiostoma belcheri Japanese. In Vitro Cellular & Developmental Biology-Animal, 2013, 49(2): p. 97-102.

[9] Pomponi, S.A., R. Willoughby. Development of sponge cell cultures for biomedical application, 2000.

[10] Pomponi, S., R. Willoughby. Sponge cell culture for production of bioactive metabolites. Sponges in time and space. Balkema, Rotterdam, 1994: 395-400.

[11] de Caralt, S., M.J. Uriz, R.H. Wijffels. Cell culture from sponges: pluripotency and immortality. Trends in biotechnology, 2007, 25(10): 467-471.

[12] Schröder, H.C., et al.. Stress response in Baikalian sponges exposed to pollutants. Hydrobiologia, 2006, 568: 277-287.

[13] Perez, T., et al.. Marine sponges as biomonitor of polychlorobiphenyl contamination: concentration and fate of 24 congeners. Environmental science & technology, 2003. 37(10): 2152-2158.

[14] Zahn, R., et al.. The effect of benzo [a] pyrene on sponges as model organisms in marine pollution. Chemico-biological interactions, 1982, 39(2): 205-220.

[15] Pan, K., et al.. Sponges and sediments as monitoring tools of metal contamination in the eastern coast of the Red Sea, Saudi Arabia. Mar Pollut Bull, 2011, 62(5): 1140-6.

[16] Rao, J.V., et al.. Environmental contamination using accumulation of metals in marine sponge, Sigmadocia fibulata inhabiting the coastal waters of Gulf of Mannar, India. Toxicological & Environmental Chemistry, 2007, 89(3): 487-498.

[17] Hill, M., et al.. Toxic effects of endocrine disrupters on freshwater sponges: common developmental abnormalities. Environmental Pollution, 2002, 117(2): 295-300.

[18] Olesen, T. and J. Weeks, Accumulation of Cd by the marine sponge Halichondria panicea Pallas: effects upon filtration rate and its relevance for biomonitoring. Bulletin of environmental contamination and toxicology, 1994, 52(5): 722-728.

[19] Hansen, I.V., J.M. Weeks, and M.H. Depledge, Accumulation of copper, zinc, cadmium and chromium by the marine sponge Halichondria panicea Pallas and the implications for biomonitoring. Marine Pollution Bulletin, 1995, 31(1-3): 133-138.

[20] Ahmed, M., et al.. Chromium (VI) induced acute toxicity and genotoxicity in freshwater stinging catfish, Heteropneustes fossilis. Ecotoxicology and environmental safety, 2013, 92: 64-70.

[21] Martins, M. P.M. Costa. The comet assay in environmental risk assessment of marine pollutants: applications, assets and handicaps of surveying genotoxicity in non-model organisms. Mutagenesis, 2014, 30(1): 89-106.

[22] Sarkar, A., et al.. Genotoxicity of cadmium chloride in the marine gastropod Nerita chamaeleon using comet assay and alkaline unwinding assay. Environmental toxicology, 2015, 30(2): 177-187.

[23] Schröder, H.C., et al.. Induction of DNA strand breaks and expression of HSP70 and GRP78 homolog by cadmium in the marine sponge Suberites domuncula. Arch Environ Contam Toxicol, 1999, 36(1): 47-55.

[24] Stohs, S.J. D. Bagchi. Oxidative mechanisms in the toxicity of metal ions. Free radical biology and medicine, 1995, 18(2): 321-336.

[25] Henkler, F., J. Brinkmann, A. Luch. The role of oxidative stress in carcinogenesis induced by metals and xenobiotics. Cancers, 2010, 2(2): 376-396.

[26] IARC, IARC Monographs on the Evaluation of Carcinogenic Risks to Humans: Chromium, Nickel and Welding. IARC Working Group on the Evaluation of Carcinogenic Risks to Humans. 1990: International Agency for Research on Cancer.

[27] Deb, S., T. Fukushima. Metals in aquatic ecosystems: mechanisms of uptake, accumulation and release‐Ecotoxicological perspectives. International Journal of Environmental Studies, 1999, 56(3): 385-417.

[28] Rainbow, P.S., The biology of heavy metals in the sea. International Journal of Environmental Studies, 1985, 25(3): 195-211.

[29] Krewski, D., et al., Human health risk assessment for aluminium, aluminium oxide, and aluminium hydroxide. Journal of Toxicology and Environmental Health, Part B, 2007, 10(S1): 1-269.

[30] Alexopoulos, E., et al.. Bioavailability and toxicity of freshly neutralized aluminium to the freshwater crayfish Pacifastacus leniusculus. Archives of Environmental Contamination and Toxicology, 2003, 45(4): 509-514.

[31] Lankoff, A., et al.. A comet assay study reveals that aluminium induces DNA damage and inhibits the repair of radiation-induced lesions in human peripheral blood lymphocytes. Toxicology letters, 2006. 161(1): 27-36.

[32] Exley, C., et al.. Kinetic constraints in acute aluminium toxicity in the rainbow trout (Oncorhynchus mykiss). Journal of theoretical biology, 1996, 179(1): 25-31.

[33] Yousef, M.I., Aluminium-induced changes in hemato-biochemical parameters, lipid peroxidation and enzyme activities of male rabbits: protective role of ascorbic acid. Toxicology, 2004, 199(1): 47-57.

[34] Banasik, A., et al.. Aluminum‐induced micronuclei and apoptosis in human peripheral‐blood lymphocytes treated during different phases of the cell cycle. Environmental Toxicology: An International Journal, 2005, 20(4): 402-406.

[35] McLachlan, D., et al.. Risk for neuropathologically confirmed Alzheimer's disease and residual aluminum in municipal drinking water employing weighted residential histories. Neurology, 1996, 46(2): 401-405.

[36] Zatta, P., et al.. Aluminium (III) as a promoter of cellular oxidation. Coordination Chemistry Reviews, 2002, 228(2): 271-284.

[37] Oberholster, P.J., et al.. Bioaccumulation of aluminium and iron in the food chain of Lake Loskop, South Africa. Ecotoxicology and Environmental Safety, 2012. 75: 134-141.

[38] Mussino, F., et al.. Primmorphs cryopreservation: a new method for long-time storage of sponge cells. Marine biotechnology, 2013, 15(3): 357-367.

[39] Cold Spring Harbor Laboratory Protocols. Calcium- and magnesium-free artificial seawater (CMF-ASW). Cold Spring Harbor Protocols 2009 December 1, 2009 [cited 2009 12]; pdb.rec12053]. Available from:


[41] Duez, P., et al.. Statistics of the Comet assay: a key to discriminate between genotoxic effects. Mutagenesis, 2003.

[42] McKelvey-Martin, V., et al.. The single cell gel electrophoresis assay (comet assay): a European review. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 1993, 288(1): 47-63.

[43] Koppen, G., et al.. The next three decades of the comet assay: a report of the 11th International Comet Assay Workshop. 2017, Oxford University Press UK.

[44] Elmore, A.R.. Final report on the safety assessment of aluminum silicate, calcium silicate, magnesium aluminum silicate, magnesium silicate, magnesium trisilicate, sodium magnesium silicate, zirconium silicate, attapulgite, bentonite, Fuller's earth, hectorite, kaolin, lithium magnesium silicate, lithium magnesium sodium silicate, montmorillonite, pyrophyllite, and zeolite. International journal of toxicology, 2003, 22: 37-102.

[45] Synzynys, B., A. Sharetskiĭ, O. Kharlamova. Immunotoxicity of aluminum chloride. Gigiena i sanitariia, 2004(4): 70-72.

[46] Varella, S.D., et al.. Mutagenic activity in waste from an aluminum products factory in Salmonella/microsome assay. Toxicology in vitro, 2004. 18(6): 895-900.

[47] Ingersoll, C.G., et al.. Aluminum and acid toxicity to two strains of brook trout (Salvelinus fontinalis). Canadian Journal of Fisheries and Aquatic Sciences, 1990. 47(8): 641-1648.

[48] Roberts, D.A., E.L. Johnston, A.G. Poore. Contamination of marine biogenic habitats and effects upon associated epifauna. Marine Pollution Bulletin, 2008, 56(6): 1057-1065.

[49] Rainbow, P., S. Luoma. Metal toxicity, uptake and bioaccumulation in aquatic invertebrates—modelling zinc in crustaceans. Aquatic toxicology, 2011. 105(3-4): 455-465.

[50] Rosseland, B., T.D. Eldhuset, M. Staurnes. Environmental effects of aluminium. Environmental Geochemistry and Health, 1990. 12(1-2): 17-27.

[51] Wood, C., et al.. Physiological evidence of acclimation to acid/aluminum stress in adult brook trout (Salvelinus fontinalis). 1. Blood composition and net sodium fluxes. Canadian Journal of Fisheries and Aquatic Sciences, 1988, 45(9): 1587-1596.

[52] Bergman, H. and J. Mattice, Lake acidification and fisheries project: acclimation to low pH and elevated aluminum by trouts. Canadian Journal of Fisheries and Aquatic Sciences, 1991. 48(10): 1987-1988.

[53] Wang, Z., J.P. Meador, K.M. Leung. Metal toxicity to freshwater organisms as a function of pH: A meta-analysis. Chemosphere, 2016, 144: 1544-1552.

[54] Iwegbue, C.M., et al.. Distribution, sources and ecological risks of metals in surficial sediments of the Forcados River and its Estuary, Niger Delta, Nigeria. Environmental Earth Sciences, 2018. 77(6): 227.

[55] Ipeaiyeda, A., N. Umo, and G. Okojevoh, Environmental Pollution Induced By an Aluminium Smelting Plant in Nigeria. Glo. J. Sci. Front. Res. Chem, 2012. 12(1).

[56] Kádár, E., et al., Avoidance responses to aluminium in the freshwater bivalve Anodonta cygnea. Aquatic Toxicology, 2001. 55(3-4): 137-148.

[57] Bondy, S.C. and A. Campbell, Aluminum and neurodegenerative diseases, in Advances in Neurotoxicology, Elsevier, 2017: 131-156.

[58] Hartwig, A.. Current aspects in metal genotoxicity. Biometals, 1995, 8(1): 3-11.



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