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Updated: Jul 18, 2026

In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage in Rat Liver
Published on: May 4, 2016
DNA damage by ochratoxin A in rat kidney assessed by the alkaline comet assay
D Zeljezić1, A-M Domijan, M Peraica
1Institute for Medical Research and Occupational Health, Ksaverska c.2, 10001 Zagreb, Croatia.
Abstract:
There are few studies of ochratoxin A (OTA) genotoxicity in experimental animals and the results obtained with cell cultures are inconsistent, although the carcinogenic potential of OTA for the kidney of experimental animals has been well established. We studied the genotoxic potential of OTA in the kidney of adult female Wistar rats (5 in each group) treated intraperitoneally with OTA (0.5 mg kg body weight-1 day-1 for 7, 14, and 21 days) measuring DNA mobility on agarose gel stained with ethidium-bromide using standard alkaline single-cell gel electrophoresis (comet assay). Negative control animals were treated with solvent (Tris buffer, 1.0 mg/kg) and positive control animals were treated with methyl methanesulfonate (40 mg/kg) according to the same schedule. OTA concentrations in plasma and kidney homogenates in 7-, 14-, and 21-day treated animals were 4.86 +/- 0.53, 7.52 +/- 3.32, 7.85 +/- 2.24 microg/mL, and 0.87 +/- 0.09, 0.99 +/- 0.06, 1.09 +/- 0.15 microg/g, respectively. In all OTA-treated groups, the tail length, tail intensity, and tail moment in kidney tissue were significantly higher than in controls (P < 0.05). The tail length and tail moment were higher after 14 days than after 7 days of treatment (P < 0.05), and still higher after 21 days (P < 0.05). The highest tail intensity was observed in animals treated for 21 days, and it differed significantly from animals treated for 7 and 14 days (P < 0.05). OTA concentrations in plasma and kidney tissue increased steadily and OTA concentration in kidney tissue strongly correlated with tail intensity and tail moment values. These results confirm the genotoxic potential of OTA, and show that the severity of DNA lesions in kidney correlates with OTA concentration.
Insights
Ochratoxin A (OTA) exposure causes significant DNA damage in rat kidneys, with damage severity increasing with exposure duration. This study confirms OTA
Area of Science:
- Toxicology
- Genetics
- Animal Studies
Background:
- Ochratoxin A (OTA) is a mycotoxin with established carcinogenic potential in animal kidneys.
- Previous studies on OTA genotoxicity show inconsistent results, particularly in cell cultures.
- Limited data exists on OTA genotoxicity in experimental animals.
Purpose of the Study:
- To investigate the genotoxic potential of Ochratoxin A (OTA) in the kidneys of adult female Wistar rats.
- To evaluate the dose-dependent and time-dependent effects of OTA exposure on DNA damage.
- To correlate OTA concentrations in plasma and kidney tissue with observed genotoxicity.
Main Methods:
- Adult female Wistar rats were administered OTA intraperitoneally (0.5 mg/kg/day) for 7, 14, and 21 days.
- DNA damage was assessed in kidney tissue using the alkaline single-cell gel electrophoresis (comet assay).
- OTA concentrations were measured in plasma and kidney homogenates.
Main Results:
- All OTA-treated groups exhibited significantly higher DNA migration (tail length, tail intensity, tail moment) compared to controls.
- Genotoxicity markers increased progressively with prolonged OTA exposure (7, 14, and 21 days).
- Kidney OTA concentrations strongly correlated with the severity of DNA lesions.
Conclusions:
- Ochratoxin A (OTA) demonstrates significant genotoxic potential in the kidneys of Wistar rats.
- The extent of DNA damage in kidney tissue is directly related to OTA concentration and exposure duration.
- These findings support the established carcinogenic risk of OTA to the kidney.

