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MONONUCLEAR LEUKOCYTE DNA DAMAGE ON HIGHER CELLS CAUSED BY ECO-FRIENDLY PESTICIDES AND THEIR ANALYSIS USING CASP® PROGRAMME

Year 2010, Volume: 14 Issue: 4, 47 - 56, 21.02.2014

Abstract

The common use of pesticides for domestic and agricultural purpose is a major concern for public health. Since many in vitro studies revealed that the use of pesticides had genotoxic effects on human health and targeted organisms, the use of environmentally friendly pesticides aimed to prevent or reduce the negative effects of those chemicals on non-targeted organisms. In the present study, the genotoxicity of so called eco-friendly pesticides such as dimethoate (400-, 200 µg ml-1), methyl parathion (360-, 180 µg ml-1) and alphacypermethrin (100-, 50 µg ml-1) was evaluated with their corresponding advised and low doses, respectively, on the induction of DNA damage in human peripheral blood lymphocytes (PBL) using the single cell gel electrophoresis assay (SCGE assay or comet assay) immediately after 30 min of treatment for the mononuclear leukocyte DNA damage. The chemicals tested on whole blood samples showed that the advised or even lower doses of each pesticide caused extensive DNA damage. The expression of DNA damage was also compared with that of CASP programme. 

References

  • Au, W.W, Sierra-Torres, C.H., Cajas-Salazar, N., Shipp, B.K., Legator, M.S. 1999. Cytogenetic effects from exposure to mixed pesticides and the influence from genetic susceptibility. Environmental Health Perspectives 107:501–505.
  • Barthel, E. 1981. Increased risk of lung cancer in pesticide-exposed male agricultural workers. Journal of Toxicolology and Environmental Health 8:1027–1040.
  • Blair, A., Zahm, S.H. 1995. Agricultural exposures and cancer. Environmental Health Perspectives 103:205–208.
  • Cavas, T., Konen, S. 2007. Detection of cytogenetic and DNA damage in peripheral erythrocytes of goldfish (Carassius auratus) exposed to a glyphosate formulation using the micronucleus test and the comet assay. Mutagenesis 22:263–268.
  • Collins, A., Dussinska, M., Franklin, M. 1997. Comet assay in human biomonitoring studies: reliability validation and application, Environmental and Molecular Mutagenesis 30: 439-462.
  • Collins, A.R. 2002. The comet assay. Principles, applications and limitations. Methods in Molecular Biology 203:163– 177.
  • Das, P., John, G. 1999. Induction of sister chromatid exchanges and chromosome aberrations in vivo in Etroplus suratensis (Bloch) following exposure to organophosphorus pesticides. Toxicology Letters 4:111–116.
  • Dikilitas, M., Kocyigit, A. ve Yigit, F. 2009. A molecular-based fast method to determine the extent of DNA damages in higher plants and fungi. African Journal of Biotechnology, 8 (14): 3118-3127.
  • Dimitrov, B.D., Gadeva, P.G., Benova, D.K., Bineva, M.V. 2006. Comparative genotoxicity of the herbicides roundup, stomp and reglone in plant and mammalian test systems. Mutagenesis 21:375–382.
  • Ecobichon, D.J. 2001. Toxic effects of pesticides. In: Klaassen CD (ed). Casarett and Doull’s toxicology. The basic science of poisons. McGraw-Hill, New York, pp 763–810.
  • Falck, G.C.M., Hirvonen, A., Scarpato, R., Saarikoski, S.T., Migliore, L., Norppa, H. 1999. Micronuclei in blood lymphocytes and genetic polymorphism for GSTM1, GSTT1 and NAT2 in pesticide exposed greenhouse workers. Mutation Research 441:225–237.
  • Garaj-Vrhovac, V., Zeljezic, D. 2000. Evaluation of DNA damage in workers occupationally exposed to pesticides using single-cell gel electrophoresis (SCGE) assay. Pesticide genotoxicity revealed by comet assay. Mutation Research 469:279– 285.
  • Garrett, N.E., Stack, H.F., Waters, M.D. 1986. Evaluation of the genetic activity profiles of 65 pesticides. Mutation Research 168: 301–325.
  • Gichner, T., Znidar, I., Wagner, E.D. ve Plewa, M.J. 2009. The use of higher plants in the comet assay. In: The Comet Assay in Toxicology Edited by Alok Dhawan and Diana Anderson. Royal Society of Chemistry, p: 98-119.
  • Hayes, W.J., Laws, E.R. 1990. Handbook of Pesticide Toxicology, Vol. 3, Classes of Pesticides. Academic Press, Inc., NY.
  • Howard, P.H. 1989. Handbook of Environmental Fate and Exposure Data for Organic Chemicals, Vol. III: Pesticides. Lewis Publishers, Chelsea, MI.
  • Kobayashi, H., Sugiyama, C., Morikawa, Y., Hayashi, M.,. Sofuni, T. A. 1995. Comparison between manual and microscopic analysis and computerized image analysis in single cell electrophoresis assay. MMS Communication 3:103-115.
  • Kocyigit, A., Keles, H., Selek, S., Guzel, S., Celik, H. ve Erel, O. 2005. Increased DNA damage and oxidative stress in patients with Cutaneous leishmaniasis. Mutation Research, 585:71-78.
  • Konca, K., Lankoff, A., Banasik, A., Lisowska, H., Kuszewski, T., Gozdz S, Koza, Z. ve Wojcik, A. 2003. A cross-platform public domain PC image-analysis program for the comet assay. Mutation Research 534: 1520.
  • Lebailby, P., Vigreux, C., Lechevrel, C., Ledemeney, D., Godard, T., Sicheb, F., LeTalaer, J. Y. Henry-Amar, M, Gauduchon P. 1998. DNA Damage in Mononuclear Leukocytes of Farmers Measured Using the Alkaline Comet Assay: Discussion of Critical Parameters and Evaluation of Seasonal Variations in Relation to Pesticide Exposure. Cancer Epidemiology, Biomarkers & Prevention 7: 917-927.
  • Lucero, L., Pastor, S., Suarez, S., Durban, R., Gomez, C., Parron, T., Creus, A., Marcos, R. 2000. Cytogenetic biomonitoring of Spanish greenhouse workers exposed to pesticides: micronuclei analysis in peripheral blood lymphocytes and buccal epithelial cells. Mutation Research 464:255–262.
  • Meister, R.T. 1992. Farm Chemicals Handbook '92. Meister Publishing Company, Willoughby, OH.
  • Moretti, M., Marcarelli, M., Villarini, M., Fatigoni, C., Scassellati-Sforzolini, G., Pasquini, R. 2002. In vitro testing for genotoxicity of the herbicide terbutryn:cytogenetic and primary DNA damage. Toxicology in Vitro 16:81–88.
  • Ostling, O., Johanson, K.J. 1984. Microelectrophoretic study of radiation induced DNA damages in individual mammalian cells. Biochemical and Biophysical Research Communications 30: 291-298.
  • Rupa, D.S., Reddy, P.P., Reddi, O.S. 1989. Analysis of sister-chromatid exchanges, cell kinetics and mitotic index in lymphocytes of smoking pesticide sprayers. Mutation Research 223:253–258.
  • Singh, N.P. McCoy, M.T., Tice, R.R., Schneider, E.L. 1988. A simple technique for quantification of low levels of DNA damage in individual cells. Experimental Cell. Research 175: 184–191.
  • Sinha, S.P. 1989. Genotoxicity of pesticides. Perspectives in Cytology and Genetics 6:749–753.
  • Undeger, U., Basaran, N. 2002. Assessment of DNA damage in workers occupationally exposed to pesticide mixtures by the alkaline comet assay. Archives Toxicology 76:430–436.
  • Undeger, U., Basaran, N. 2005. Effects of pesticides on human peripheral lymphocytes in vitro: induction of DNA damage. Archives Toxicology 79: 169176.
  • Undeger, U., Institoris, L., Siroki, O., Nehez, M., Desi, I. 2000. Simultaneous geno- and immunotoxicological invesitgations for early detection of organophosphate toxicity in rats. Ecotoxicological Environmental Safety 45:43–48.
  • Velazquez, A., Xamena, N., Creus, A., Marcos, R. 1990. Mutagenic evaluation of the organophosphorus insecticides methyl parathion and triazophos in Drosophila melanogaster. Journal of Toxicological Environmental Health 31:313–325.
  • Viel, J.F., Chalier, B. 1995. Bladder cancer among French farmers: does exposure to pesticides in vineyards play a part? Occupational and Environmental Medicine 52:587–592.
  • Weisenburger, D.D. 1993. Human health effects of agrichemical use. Human Pathology 24, 571–576.
  • WHO, 1988. The WHO recommended classification of pesticides by hazard and guidelines to the classification 1988–1989. World Health Organisation, Geneva.
  • WHO, 1993. Environmental Health Criteria 145. Methyl parathion. World Health Organization, Geneva. 1993.

ÇEVRE DOSTU PESTİSİTLERİN YÜKSEK YAPILI HÜCRELERDE NEDEN OLDUĞU TEK HÜCRE LÖKOSİT DNA HASARI VE CASP PROGRAMI İLE ANALİZİ

Year 2010, Volume: 14 Issue: 4, 47 - 56, 21.02.2014

Abstract

Tarımsal alanlarda ve evlerimizde yaygın olarak kullanılan pestisitler halk sağlığı için büyük önem arz etmektedir. Pestisit kullanımının hedef organizmalar ve insan sağlığı üzerine genotoksik etkilerinin olduğu birçok in vitro çalışma ile ortaya konulması sebebiyle, bu kimyasalların hedef olmayan organizmalar üzerinde meydana gelen olumsuz etkilerini önlemek ya da azaltmak için çevre dostu pestisitlerin kullanılması hedeflenmiştir. Bu çalışmada, çevre dostu olarak bilinen pestisitlerden dimethoate (400-, 200 µg ml-1), methyl parathion (360-, 180 µgml-1) and alphacypermethrin (100-, 50 µg ml-1 )’in tavsiye edilen ve düşük dozlarının insan periferal kan hücrelerindeki DNA hasar oluşumu üzerine genotoksik etkileri araştırılmıştır. Bu amaçla, kan örneklerine 30 dk süresince farklı dozlarda çevre dostu pestisitler uygulandıktan hemen sonra tek hücre jel elektroforezi (SCGE ya da comet analizi) kullanılarak, tek çekirdekli
lökosit DNA hasarı incelenmiştir. Sonuçlar bu kimyasalların tavsiye edilen ya da tavsiye edilenden daha düşük dozlarının bile büyük oranda DNA hasarına neden olduğunu göstermiştir. Elde edilen DNA hasarının durumu CASP programı kullanılarak kıyaslama yoluna gidilmiştir. 

References

  • Au, W.W, Sierra-Torres, C.H., Cajas-Salazar, N., Shipp, B.K., Legator, M.S. 1999. Cytogenetic effects from exposure to mixed pesticides and the influence from genetic susceptibility. Environmental Health Perspectives 107:501–505.
  • Barthel, E. 1981. Increased risk of lung cancer in pesticide-exposed male agricultural workers. Journal of Toxicolology and Environmental Health 8:1027–1040.
  • Blair, A., Zahm, S.H. 1995. Agricultural exposures and cancer. Environmental Health Perspectives 103:205–208.
  • Cavas, T., Konen, S. 2007. Detection of cytogenetic and DNA damage in peripheral erythrocytes of goldfish (Carassius auratus) exposed to a glyphosate formulation using the micronucleus test and the comet assay. Mutagenesis 22:263–268.
  • Collins, A., Dussinska, M., Franklin, M. 1997. Comet assay in human biomonitoring studies: reliability validation and application, Environmental and Molecular Mutagenesis 30: 439-462.
  • Collins, A.R. 2002. The comet assay. Principles, applications and limitations. Methods in Molecular Biology 203:163– 177.
  • Das, P., John, G. 1999. Induction of sister chromatid exchanges and chromosome aberrations in vivo in Etroplus suratensis (Bloch) following exposure to organophosphorus pesticides. Toxicology Letters 4:111–116.
  • Dikilitas, M., Kocyigit, A. ve Yigit, F. 2009. A molecular-based fast method to determine the extent of DNA damages in higher plants and fungi. African Journal of Biotechnology, 8 (14): 3118-3127.
  • Dimitrov, B.D., Gadeva, P.G., Benova, D.K., Bineva, M.V. 2006. Comparative genotoxicity of the herbicides roundup, stomp and reglone in plant and mammalian test systems. Mutagenesis 21:375–382.
  • Ecobichon, D.J. 2001. Toxic effects of pesticides. In: Klaassen CD (ed). Casarett and Doull’s toxicology. The basic science of poisons. McGraw-Hill, New York, pp 763–810.
  • Falck, G.C.M., Hirvonen, A., Scarpato, R., Saarikoski, S.T., Migliore, L., Norppa, H. 1999. Micronuclei in blood lymphocytes and genetic polymorphism for GSTM1, GSTT1 and NAT2 in pesticide exposed greenhouse workers. Mutation Research 441:225–237.
  • Garaj-Vrhovac, V., Zeljezic, D. 2000. Evaluation of DNA damage in workers occupationally exposed to pesticides using single-cell gel electrophoresis (SCGE) assay. Pesticide genotoxicity revealed by comet assay. Mutation Research 469:279– 285.
  • Garrett, N.E., Stack, H.F., Waters, M.D. 1986. Evaluation of the genetic activity profiles of 65 pesticides. Mutation Research 168: 301–325.
  • Gichner, T., Znidar, I., Wagner, E.D. ve Plewa, M.J. 2009. The use of higher plants in the comet assay. In: The Comet Assay in Toxicology Edited by Alok Dhawan and Diana Anderson. Royal Society of Chemistry, p: 98-119.
  • Hayes, W.J., Laws, E.R. 1990. Handbook of Pesticide Toxicology, Vol. 3, Classes of Pesticides. Academic Press, Inc., NY.
  • Howard, P.H. 1989. Handbook of Environmental Fate and Exposure Data for Organic Chemicals, Vol. III: Pesticides. Lewis Publishers, Chelsea, MI.
  • Kobayashi, H., Sugiyama, C., Morikawa, Y., Hayashi, M.,. Sofuni, T. A. 1995. Comparison between manual and microscopic analysis and computerized image analysis in single cell electrophoresis assay. MMS Communication 3:103-115.
  • Kocyigit, A., Keles, H., Selek, S., Guzel, S., Celik, H. ve Erel, O. 2005. Increased DNA damage and oxidative stress in patients with Cutaneous leishmaniasis. Mutation Research, 585:71-78.
  • Konca, K., Lankoff, A., Banasik, A., Lisowska, H., Kuszewski, T., Gozdz S, Koza, Z. ve Wojcik, A. 2003. A cross-platform public domain PC image-analysis program for the comet assay. Mutation Research 534: 1520.
  • Lebailby, P., Vigreux, C., Lechevrel, C., Ledemeney, D., Godard, T., Sicheb, F., LeTalaer, J. Y. Henry-Amar, M, Gauduchon P. 1998. DNA Damage in Mononuclear Leukocytes of Farmers Measured Using the Alkaline Comet Assay: Discussion of Critical Parameters and Evaluation of Seasonal Variations in Relation to Pesticide Exposure. Cancer Epidemiology, Biomarkers & Prevention 7: 917-927.
  • Lucero, L., Pastor, S., Suarez, S., Durban, R., Gomez, C., Parron, T., Creus, A., Marcos, R. 2000. Cytogenetic biomonitoring of Spanish greenhouse workers exposed to pesticides: micronuclei analysis in peripheral blood lymphocytes and buccal epithelial cells. Mutation Research 464:255–262.
  • Meister, R.T. 1992. Farm Chemicals Handbook '92. Meister Publishing Company, Willoughby, OH.
  • Moretti, M., Marcarelli, M., Villarini, M., Fatigoni, C., Scassellati-Sforzolini, G., Pasquini, R. 2002. In vitro testing for genotoxicity of the herbicide terbutryn:cytogenetic and primary DNA damage. Toxicology in Vitro 16:81–88.
  • Ostling, O., Johanson, K.J. 1984. Microelectrophoretic study of radiation induced DNA damages in individual mammalian cells. Biochemical and Biophysical Research Communications 30: 291-298.
  • Rupa, D.S., Reddy, P.P., Reddi, O.S. 1989. Analysis of sister-chromatid exchanges, cell kinetics and mitotic index in lymphocytes of smoking pesticide sprayers. Mutation Research 223:253–258.
  • Singh, N.P. McCoy, M.T., Tice, R.R., Schneider, E.L. 1988. A simple technique for quantification of low levels of DNA damage in individual cells. Experimental Cell. Research 175: 184–191.
  • Sinha, S.P. 1989. Genotoxicity of pesticides. Perspectives in Cytology and Genetics 6:749–753.
  • Undeger, U., Basaran, N. 2002. Assessment of DNA damage in workers occupationally exposed to pesticide mixtures by the alkaline comet assay. Archives Toxicology 76:430–436.
  • Undeger, U., Basaran, N. 2005. Effects of pesticides on human peripheral lymphocytes in vitro: induction of DNA damage. Archives Toxicology 79: 169176.
  • Undeger, U., Institoris, L., Siroki, O., Nehez, M., Desi, I. 2000. Simultaneous geno- and immunotoxicological invesitgations for early detection of organophosphate toxicity in rats. Ecotoxicological Environmental Safety 45:43–48.
  • Velazquez, A., Xamena, N., Creus, A., Marcos, R. 1990. Mutagenic evaluation of the organophosphorus insecticides methyl parathion and triazophos in Drosophila melanogaster. Journal of Toxicological Environmental Health 31:313–325.
  • Viel, J.F., Chalier, B. 1995. Bladder cancer among French farmers: does exposure to pesticides in vineyards play a part? Occupational and Environmental Medicine 52:587–592.
  • Weisenburger, D.D. 1993. Human health effects of agrichemical use. Human Pathology 24, 571–576.
  • WHO, 1988. The WHO recommended classification of pesticides by hazard and guidelines to the classification 1988–1989. World Health Organisation, Geneva.
  • WHO, 1993. Environmental Health Criteria 145. Methyl parathion. World Health Organization, Geneva. 1993.
There are 35 citations in total.

Details

Primary Language English
Subjects Food Engineering
Journal Section Araştırma Makaleleri
Authors

Murat Dikilitaş

Abdurrahim Koçyiğit This is me

Publication Date February 21, 2014
Submission Date February 21, 2014
Published in Issue Year 2010 Volume: 14 Issue: 4

Cite

APA Dikilitaş, M., & Koçyiğit, A. (2014). MONONUCLEAR LEUKOCYTE DNA DAMAGE ON HIGHER CELLS CAUSED BY ECO-FRIENDLY PESTICIDES AND THEIR ANALYSIS USING CASP® PROGRAMME. Harran Tarım Ve Gıda Bilimleri Dergisi, 14(4), 47-56.

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