Detecting genotoxicity using the Comet assay following chronic exposure of Manila clam Tapes semidecussatus to polluted estuarine sediments

Coughlan, B.M.; Hartl, Mark G. J.; O’Reilly, Sarah; Sheehan, David; Morthersill, C.; Pelt, Frank N.A.M. van; O’Halloran, John; O’Brien, Nora M.

doi:10.1016/s0025-326x(02)00254-0

Cited by 67 publications

(52 citation statements)

References 35 publications

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“…Heavy metal concentrations in samples of water, sediment and soft tissues of P. undulata were analysed according to the methods of Dybern (1983), Chevereuil et al (1996) and Coughlan (2002) …”

Section: Metal Analysismentioning

confidence: 99%

The effect of depuration on heavy metals, petroleum hydrocarbons, and microbial contamination levels in Paphia undulata (Bivalvia: Veneridae)

El-Gamal¹

2011

CZECH J ANIM SCI

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ABSTRACT:The depuration of 8 heavy metals (Zn, Pb, Ni, Mn, Cu, Cr, Co and Cd), total petroleum hydrocarbons and pathogenic bacteria of Paphia undulata was tested and the survival of depurated clams was evaluated. Investigated samples were collected from Ismailia, Egypt. The initial metal concentrations were significantly higher in the whole soft tissues than in water and sediment except for Mn. After 24 h depuration, Zn, Pb, Ni, Mn, Cu, Cr and Cd were significantly reduced to 44, 23, 25, 17, 61, 41, and 75%, respectively. After three days of depuration the reduction was significant only for Cu, Cr, Co and Cd (27, 15, 23 and 52%, respectively). The total petroleum hydrocarbons were reduced significantly to 72% after three days of depuration, while after 24 h they were reduced to 90% compared to their initial concentrations. Four pathogenic bacteria were identified in the soft tissues of P. undulata (Vibrio sp., Shigella sp., Escherichia coli and Salmonella sp.). After one-day depuration the results evidenced the mean microbial reduction to 75, 31, 68, and 36%, respectively, compared to their initial counts. After three days of depuration the counts of Vibrio sp. and Salmonella sp. were reduced to 3% and 8%, respectively, while Escherichia coli was not detected on the third day. Shigella sp. was increased by 22% compared to the first day of depuration. The viability and mortality were not influenced by the depurative treatment.

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Section: Metal Analysismentioning

confidence: 99%

The effect of depuration on heavy metals, petroleum hydrocarbons, and microbial contamination levels in Paphia undulata (Bivalvia: Veneridae)

El-Gamal¹

2011

CZECH J ANIM SCI

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“…The size and fluorescence intensity of the resulting Comet tail represents the degree of DNA damage in individual cells. Originally developed by Singh et al [1988] to measure genotoxic damage in cultured human lymphocytes, the Comet assay has since been developed further to include the examination of various cell types from a number of species, including bivalves [Rank, 1999;Coughlan et al, 2002;Hartl et al, 2004].…”

Section: Introductionmentioning

confidence: 99%

Maintenance of bivalve hemocytes for the purpose of delayed DNA strand break assessment using the comet assay

Hartl¹,

Grigson²,

Sinet³

2009

Environ and Mol Mutagen

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The lack of appropriate methods for storing intact and viable cells for the purpose of delayed DNA strand break analysis has hitherto limited the application of the Comet assay to in vitro or in vivo laboratory studies and restricted ecologically more relevant field-collected samples to sites in proximity to suitable laboratory facilities. In the present article, osmotically corrected cell culture media Hanks Balanced Salt Solution (HBSS) and Leibovitz Media (L-15) were assessed for their suitability as temporary storage media of blue mussel (Mytilus edulis) hemocytes. It was found that hemocytes maintained in either HBSS or L-15 could be stored for at least 7 days at 4 degrees C without any significant deterioration in cell viability (Trypan blue) or increase in DNA strand breaks, expressed as % tail DNA. This approach allows the acquisition and examination of samples from organisms exposed in situ at previously unsuitable remote sites, thereby greatly increasing the potential ecological relevance of Comet assay-derived genotoxicity data.

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“…In this study, an attempt was made to establish a uniform and reproducible protocol suitable for use with a variety of tissue types derived from turbot. The assay conditions used by Coughlan et al [2002], with some modifications, were found to be suitable for this study.…”

Section: Discussionmentioning

confidence: 99%

Genotoxicity of field‐collected inter‐tidal sediments from Cork Harbor, Ireland, to juvenile turbot (Scophthalmus maximus L.) as measured by the Comet assay

Kilemade

Hartl

Sheehan

et al. 2004

Environ and Mol Mutagen

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The alkaline single cell gel electrophoresis (SCGE) or Comet assay was employed to test the potential of surficial sediment collected from Cork Harbor, Ireland, to induce DNA damage in turbot (Scophthalmus maximus L.) in a laboratory exposure experiment. Turbot were exposed for 21 days to field-collected sediment from Cork Harbor and from a relatively clean reference site at Ballymacoda and sampled at 0, 7, 14, and 21 days. As a positive control for the sediment exposure experiment, a subsample of the turbot was exposed to cadmium chloride-spiked seawater. DNA damage analysis was performed on epidermal, gill, spleen, liver, and whole blood cell preparations. Liver, gill, and blood were the most sensitive tissues while a lower level of damage was detected in the epidermis and spleen. The blood was determined to be a suitable predictor of DNA damage in the whole organism. Chemical analysis of the sediment indicated that polycyclic aromatic hydrocarbons formed the bulk of the contaminants, with the harbor sites having almost double the levels of those from the reference site. The data indicated that turbot exposed to sediments from Cork Harbor elicited a significant increase in DNA damage in comparison with those exposed to sediment from the reference site and that exposure to the contaminated sediments caused a multi-organ genotoxic response. Results from the study indicate a relationship between the presence of genotoxicants in sediment and DNA damage. This finding was encouraging with regard to the potential use of the Comet assay as part of a marine biomonitoring strategy.

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Detecting genotoxicity using the Comet assay following chronic exposure of Manila clam Tapes semidecussatus to polluted estuarine sediments

Cited by 67 publications

References 35 publications

The effect of depuration on heavy metals, petroleum hydrocarbons, and microbial contamination levels in Paphia undulata (Bivalvia: Veneridae)

The effect of depuration on heavy metals, petroleum hydrocarbons, and microbial contamination levels in Paphia undulata (Bivalvia: Veneridae)

Maintenance of bivalve hemocytes for the purpose of delayed DNA strand break assessment using the comet assay

Genotoxicity of field‐collected inter‐tidal sediments from Cork Harbor, Ireland, to juvenile turbot (Scophthalmus maximus L.) as measured by the Comet assay

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