Evaluation of genotoxic effects of lead in pottery-glaze workers using micronucleus assay, alkaline comet assay and DNA diffusion assay

  • V. KašubaEmail author
  • R. Rozgaj
  • M. Milić
  • D. Želježić
  • N. Kopjar
  • A. Pizent
  • Z. Kljaković-Gašpić
  • A. Jazbec
Original Article



We investigated genotoxic effects of occupational exposure to lead acetate in pottery-glaze ceramic workers.


The study was carried out in 30 exposed workers and 30 matched controls, to whom several biochemical parameters—the blood lead (B-Pb; range: exposed, 41.68–404.77; controls, 12–52) and cadmium (B-Cd) level, the activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), the level of vitamin B12 and folate in serum—were measured. The genotoxic effects were evaluated by the alkaline comet assay, the DNA diffusion assay and micronucleus test in peripheral blood lymphocytes.


Subjects exposed to lead had significantly higher B-Pb level and, consequently, increased values of tail intensity (TI), frequency of apoptotic and necrotic cells, and frequency of micronuclei (MN). In contrast, their activity of ALAD, the level of vitamin B12 and folate in serum were significantly lower compared to controls. Poisson regression analysis showed a significant correlation of profession, duration of exposure, smoking, level of cadmium in blood, ALAD and EP with primary DNA damage. A majority of primary damage repairs in a short period after exposure to a genotoxic agent. In addition, the influence of gender and level of vitamin B12 and folate in serum MN frequency in exposed group was observed.


In this study, DNA diffusion and micronucleus test showed higher influence of tested parameters to DNA damage. The results indicate a need for concomitant use of at least two different biomarkers of exposure when estimating a genetic risk of lead exposure.


Ceramic workers Lead exposure Biological markers Alkaline comet assay Micronucleus assay 



This investigation was supported by the Croatian Ministry of Science, Education and Sports as a part of the Projects No: 022-0222148-2137 and No: 022-0222411-2408. We thank Krešimir Nekić from Unit for analytical toxicology and mineral metabolism for performing ALAD and EP measurement.

Conflict of interest

None of authors had any personal or financial conflict of interest.


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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • V. Kašuba
    • 1
    Email author
  • R. Rozgaj
    • 1
  • M. Milić
    • 1
  • D. Želježić
    • 1
  • N. Kopjar
    • 1
  • A. Pizent
    • 2
  • Z. Kljaković-Gašpić
    • 2
  • A. Jazbec
    • 3
  1. 1.Mutagenesis UnitInstitute for Medical Research and Occupational HealthZagrebCroatia
  2. 2.Unit for Analytical Toxicology and Mineral MetabolismInstitute for Medical Research and Occupational HealthZagrebCroatia
  3. 3.Faculty of ForestryUniversity of ZagrebZagrebCroatia

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