Detection of Genotoxic Insult as Dna Strand Breaks in Fish Blood Cells by Agarose Gel Electrophoresis

Theodorakis, Christopher W.; D'Surney, S.J.; Shugart, Lee R.

doi:10.1897/1552-8618(1994)13[1023:dogiad]2.0.co;2

Cited by 12 publications

(18 citation statements)

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“…Our results show that both hypoxic and hyperoxic conditions have considerable impact on the level of DNA strand breaks in fish erythrocytes. These results are consistent with previous studies where fish erythrocytes have been shown to be sensitive for measuring the genotoxic effects either in laboratory or in field studies (Belpaeme et al 1998;Frenzilli et al 2004;Theodorakis et al 1994;Li et al 2005;Jha 2004Jha , 2008.…”

Section: Discussionsupporting

confidence: 93%

Hypoxia-induced oxidative DNA damage links with higher level biological effects including specific growth rate in common carp, Cyprinus carpio L.

et al. 2011

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Both hypoxia and hyperoxia, albeit in different magnitude, are known stressors in the aquatic environment. Adopting an integrated approach, mirror carp (Cyprinus carpio L.), were exposed chronically (i.e. 30 days) to hypoxic (1.8 ± 1.1 mg O(2) l(-1)) and hyperoxic (12.3 ± 0.5 mg O(2) l(-1)) conditions and resultant biological responses or biomarkers were compared between these two treatments as well as with fish held under normoxic conditions (7.1 ± 1.04 mg O(2) l(-1)). The biomarkers determined included the activities of glutathione peroxidase (GPx), measurement of oxidative DNA damage (using modified Comet assay employing bacterial enzymes: Fpg and Endo-III), haematological parameters, histopathological and ultrastructural examination of liver and gills. Specific growth rate (SGR) of the fish, as an important ecotoxicological parameter was also determined over the exposure period. The study suggested that while the levels of hepatic GPx were unaffected, there was a significant difference in activity in the blood plasma under different exposure conditions; the hyperoxic group showed increased GPx activity by approximately 37% compared to normoxic group and the hypoxic group showed a decrease by approximately 38% than the normoxic group. Interestingly, oxidative DNA damage was significantly higher in both hypoxic and hyperoxic by approximately 25% compared to normoxic conditions, Fpg showing enhanced level of damage compared to the Endo-III treatment (P< 0.001). The haematological parameters showed enhanced values under hypoxic conditions. Transmission electron microscopic (TEM) studies revealed damage to liver and gill tissues for both the treatments. Interestingly, SGR of fish was significantly lowered in hypoxic by approx. 30% compared to normoxic condition and this was found to be correlated with DNA damage (R = -0.82; P = 0.02). Taken together, these results indicate that prolonged exposure to both hypoxic and hyperoxic conditions induce oxidative stress responses at both DNA and tissue levels, and hypoxia can result in compensatory changes in haematological and growth parameters which could influence Darwinian fitness of the biota with wider ecological implications.

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Section: Discussionsupporting

confidence: 93%

Hypoxia-induced oxidative DNA damage links with higher level biological effects including specific growth rate in common carp, Cyprinus carpio L.

et al. 2011

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“…These results were similar to DNA strand breakage seen in other species exposed to complex mixtures of genotoxins. The MMLs measured for erythrocytes isolated from our herring gulls are in the same range as those measured by the same technique in bluegill sunfish (Leopomis macrochirus) exposed to sediments containing high levels of PAHs, PCBs and heavy metals (10.9 ± 10.7 kb) and in unexposed fish (31.9 ± 7.8 kb) (Theodorakis et al, 1994). Extensive inter-animal heterogenetity (dispersion) in DNA migration was also observed among golden mice (Ochrotomys nuttalli) live trapped from both a control and hazardous waste site in North Carolina and damage was increased in animals from the hazardous waste site (Tice, 1995).…”

Section: Origin Of the Exposures And Nature Of The Genotoxinssupporting

confidence: 58%

“…We assessed DNA integrity by quantifying the relative length of nuclear DNA strands isolated from the packed erythrocytes using the agarose gel electrophoresis technique of Theodorakis et al (1994). Methods for controlling DNA shearing during isolation, controlling the temperature, and recirculation of the buffer to dissipate pH gradients at the electrodes during electrophoresis were as described in Theodorakis et al (1999).…”

Section: Study Area and Methodsmentioning

confidence: 99%

“…In situ assays used to detect effects of PAHs and complex mixtures of genotoxins include measurements of DNA integrity including sister chromatid exchanges, chromosome aberrations, unscheduled DNA synthesis, variability in DNA content, micronuclei, and DNA adducts. Another measure of DNA integrity is the relative length of nuclear DNA strands extracted from erythrocytes and separated under alkaline conditions using the agarose gel electrophoresis technique (Theodorakis et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

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DNA Strand Length and EROD Activity in Relation to Two Screening Measures of Genotoxic Exposure in Great Lakes Herring Gulls

et al. 2005

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We collected tissues from herring gulls (Larus argentatus) nesting within and outside of the Great Lakes basin. Genotoxin exposure was assessed as fluorescent aromatic compounds (FACs) in bile and SOS Chromotest-inducing activity in muscle extracts. We determined whether these exposures were associated with decreased erythrocyte DNA strand length and/or induction of hepatic ethoxyresorufin-O-deethylase (EROD) activity. FACs were detected in all bile samples. Most muscle extracts produced a positive or marginal SOS response in the presence of S9. SOS induction potentials were strongly associated with dietary trophic level. The median molecular length of DNA isolated from erythrocytes for 14 of 17 adult and 10 of 11 prefledgling collections was reduced compared to the modal class for their respective age group suggesting widespread DNA damage. DNA damage was greatest in gulls from Saginaw Bay, Lake Huron. Median EROD activity in both adults and prefledglings from remote locations was significantly lower than that of gulls from the lower Great Lakes and was not associated with concentrations of benzo[a]pyrene (B[a]P)-like FACs. Our results indicate Great Lakes herring gulls were exposed to genotoxins and Ah-receptor activating agents in biologically significant concentrations in the early 1990s. These agents appear to be persistent bioaccumulative compounds and/or their metabolites.

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“…The latter two of these responses have been used as relatively quick and sensitive biomarker assays for exposure to genotoxic compounds (e.g. Stein et al, 1993Stein et al, , 1994Theodorakis et al, 1994;.…”

Section: Introduction and Conceptual Modelmentioning

confidence: 99%

Introduction and Conceptual Model

Lawrence¹,

Elliott²

2003

Effects of Pollution on Fish

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Detection of Genotoxic Insult as Dna Strand Breaks in Fish Blood Cells by Agarose Gel Electrophoresis

Cited by 12 publications

References 0 publications

Hypoxia-induced oxidative DNA damage links with higher level biological effects including specific growth rate in common carp, Cyprinus carpio L.

Hypoxia-induced oxidative DNA damage links with higher level biological effects including specific growth rate in common carp, Cyprinus carpio L.

DNA Strand Length and EROD Activity in Relation to Two Screening Measures of Genotoxic Exposure in Great Lakes Herring Gulls

Introduction and Conceptual Model

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