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Evaluation of the genetic alterations in direct and indirect exposures of hexavalent chromium [Cr(VI)] in leather tanning industry workers North Arcot District, South India

  • Vellingiri BalachandarEmail author
  • Meyyazhagan Arun
  • Subramaniam Mohana Devi
  • Palanivel Velmurugan
  • Pappusamy Manikantan
  • Alagamuthu Karthick Kumar
  • Keshavarao Sasikala
  • Chinnakulandai Venkatesan
Original Article

Abstract

Purpose

The focal aim of the present study was to identify the genetic alterations occurring in the tannery workers and surrounding inhabitants chronically exposed to hexavalent chromium [Cr(VI)].

Methods

A total of 108 samples which includes 72 exposed subjects [36 directly exposed (DE) subjects and 36 indirectly exposed (IE) subjects] and 36 controls were recruited for this study. The exposed subjects and controls were selected based on the Cr level present in air and their urine. Directly exposed subjects were categorized based on their work duration in the tannery industries, whereas the indirectly exposed subjects were categorized based on their year of residence in the place adjacent to tannery industries for more than 3 decades. Controls were normal and healthy. Age was matched for the exposed subjects and controls. The exposed subjects as well as the controls were categorized based on their age (group I, <40 years; group II, >41 years). Cell cultures were established from blood samples (5 ml from each subject) collected from the subjects (exposed subjects and controls) after obtaining informed consent. G-banding (Giemsa staining) of the cultures, micronucleus (MN) assay and comet assay were used to identify the genetic alterations of individuals exposed to Cr(VI) in comparison with the controls.

Results

A higher degree of total CA [12 ± 8.49 (21–25 years)] and MN [18.69 ± 7.39 (11–15 years)] was found in DE subjects compared to other groups. In IE subjects, elevated levels of CA [5.67 ± 1.15 (51–60 years)] and MN [25 ± 9.89 (71–80 years)] were observed. As expected, controls exhibited minimal number of alterations. The overall CA frequency due to Cr exposure was significantly different from that of the controls for both chromatid and chromosome type aberrations (P < 0.05 by ANOVA). The MN/1,000 binucleated cells were significantly increased (P < 0.05) in the peripheral lymphocytes of DE and IE subjects in comparison with controls. The mean tail length of comet assay for DE, IE and controls were analyzed. The mean tail length of DE subjects [4.21 (3.21–10.98)] was higher compared to that of IE subjects [3.98 (2.98–11.27)] and controls [3.01 (2.68–9.40)].

Conclusion

In conclusion, this work shows a clear genotoxic effect associated with chromium exposure, both directly and indirectly. Our result reinforces the higher sensitivity of cytogenetic assays for the biomonitoring of occupationally exposed populations. There is a strong need to educate those who work with potentially hazardous heavy about its adverse effects and highlight the importance of using protective measures.

Keywords

Human lymphocytes Hexavalent chromium [(Cr(VI)] Chromosome aberrations (CA) Micronuclei (MN) Comet assay Occupational exposure 

Notes

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Albertini RJ, Anderson D, Douglas GR, Hagmar L, Hemminki K, Merlo F, Nataraja AT, Norppa H, Shuker DE, Tice R, Waters MD, Aition A (2000) IPCS guidelines for the monitoring of genotoxic effects of carcinogens in human. Mutat Res 463(2):111–172CrossRefGoogle Scholar
  2. Anderson D, Yu TW, Phillips BJ, Schmezer P (1994) The effects of various antioxidants and other modifying agents on oxygen radical-generated DNA damage in human lymphocytes in the comet assay. Mutat Res 307:261–271Google Scholar
  3. Appelo CAJ, Postma D (1996) Geochemistry ground-waters and pollution. Rotterdam. Balkema, 536Google Scholar
  4. Bajpayee M, Dhawan A, Parmar D, Pandey AK, Mathur N, Seth PK (2002) Gender related differences in basal DNA damage in lymphocytes of a healthy Indian population using the alkaline comet assay. Mutat Res 520:83–91Google Scholar
  5. Bonassi S, Fenech M, Lando C, Lin YP, Ceppi M, Chang WP, Holland N, Kirsch-Volders M, Zeiger E, Ban S, Barale R, Bigatti MP, Bolognesi C, Jia C, Di Giorgio M, Ferguson LR, Fucic A, Lima OG, Hrelia P, Krishnaja AP, Lee TK, Migliore L, Mikhalevich L, Mirkova E, Mosesso P, Muller WU, Odagiri Y, Scarffi MR, Szabova E, Vorobtsova I, Vral A, Zijno A (2001) Human micronucleus project: international database comparison for results with the cytokinesis-block micronucleus assay in human lymphocytes: I. Effect of laboratory protocol, scoring criteria, and host factors on the frequency of micronuclei. Environ Mol Mutagen 37:31–45CrossRefGoogle Scholar
  6. Bonassi S, Ugolini D, Kirsch-Volders M, Stromberg U, Vermeulen R, Tucker JD (2005) Human population studies with cytogenetic biomarkers: review of the literature and future prospectives. Environ Mol Mutagen 45:258–270CrossRefGoogle Scholar
  7. Bryant HE, Ying S, Helleday T (2006) Homologous recombinationis involved in repair of chromium-induced DNA damage in mammalian cells. Mutat Res 599:116–123Google Scholar
  8. Cid MG, Loria D, Vilensky M, Miotti JL, Matos E (1991) Leather tanning workers: chromosomal aberrations in peripheral lymphocytes and micronuclei in exfoliated cells in urine. Mutat Res 259:197–201CrossRefGoogle Scholar
  9. Collins AR (2004) The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol 26:249–261CrossRefGoogle Scholar
  10. Coogan TP, Squibb KS, Motz J, Kinney PL, Costa M (1991) Distribution of chromium within cells of the blood. Toxicol Appl Pharmacol 108:157–166CrossRefGoogle Scholar
  11. Countryman P, Heddle J (1976) The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes. Mutat Res 41(2–3):321–332Google Scholar
  12. De Flora S (2000) Threshold mechanisms and site specificity in chromium (VI) carcinogenesis. Carcinogenesis 21:533–541CrossRefGoogle Scholar
  13. De Flora S, Bagnasco M, Serra D, Zanacchi P (1990) Genotoxicity of chromium compounds. Mutat Res 238:99–172Google Scholar
  14. Fenech M (2000) The in vitro micronucleus technique. Mutat Res 455:81–95Google Scholar
  15. Fenech M, Morley AA (1985) Measurement of micronuclei in lymphocytes. Mutat Res 147:29–36Google Scholar
  16. Fenech M, Holland N, Chang WP, Zeiger E, Bonassi S (1999) The Human Micronucleus Project—an international collaborative study on the use of the micronucleus technique for measuring DNA damage in humans. Mutat Res 428:271–283Google Scholar
  17. Fenech M, Chang WP, Kirsch-Volders M, Holland N, Bonassi S, Zeiger E (2003) HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. Mutat Res 534(1–2):65–75Google Scholar
  18. Fracassoa ME, Doriaa D, Carrieri M, Bartolucci GB, Quintavalle S, Rosa ED (2009) DNA single- and double-strand breaks by alkaline- and immuno-comet assay in lymphocytes of workers exposed to styrene. Toxicol Lett 185:9–15CrossRefGoogle Scholar
  19. Gibb HJ, Lees PS, Pinsky PF, Rooneym BC (2000) Lung cancer among workers in chromium chemical production. Am J Ind Med 38:115–126CrossRefGoogle Scholar
  20. Govil PK, Reddy GLN, Krishna AK, Seshu CLVNS, Satya Priya V, Sujatha D (2004) Inventorization of contaminated sites in India, NGRI technical report no: NGRI-2004-EG 421:54–66Google Scholar
  21. Gowd SS, Govil PK (2008) Distribution of heavy metals in surface water of Ranipet industrial area in Tamil Nadu, India. Environ Monit Assess 136:197–207CrossRefGoogle Scholar
  22. Hamamy HA, Al-Hakkak ZS, Hussain AF (1987) Chromosome aberrations in workers at a tannery in Iraq. Mutat Res 189:395–398CrossRefGoogle Scholar
  23. Heuser VD, Silva J, Moriske HJ, Dias JF, Yoneama ML, Freitas TRO (2002) Genotoxicity biomonitoring in regions exposed to vehicle emissions using the comet assay and the micronucleus test in native rodent Ctenomys minutus. Environ Mol Mutagen 40:227–235CrossRefGoogle Scholar
  24. Heuser VD, Erdtmann B, Kvitko K, Rohr P, Da Silva J (2007) Evaluation of genetic damage in Brazilian footwear-workers: biomarkers of exposure, effect, and susceptibility. Toxicology 232:235–247CrossRefGoogle Scholar
  25. IARC (1990) Monographs on the evaluation of carcinogenic risk to humans. chromium, nickel and welding. Geneva, Switzerland, vol 49. International Agency for Research on Cancer, Lyon, pp 49–256Google Scholar
  26. Keshava N, Ong T (1999) Occupational exposure to genotoxic agents. Mutat Res 155:117–120Google Scholar
  27. Kornhauser C, Wrobel K, Wrobel K, Malacara JM, Nava LE, Gomez L, Gonzalez R (2002) Possible adverse effect of chromium in occupational exposure of tannery workers. Ind Health 40:207–213CrossRefGoogle Scholar
  28. Kumaravel TS, Jha AN (2006) Comet assay measurements for detecting DNA damage induced by ionizing radiation and chemicals. Mutat Res 605:7–16Google Scholar
  29. Lam TH, Zhu CQ, Jiang CQ (2002) Lymphocyte DNA damage in elevator manufacturing workers in Guangzhou, China. Mutat Res 515:147–157Google Scholar
  30. Langard S (1990) One hundred years of chromium and cancer: a review of epidemiological evidence and selected case reports. Am J Ind Med 17:189–215CrossRefGoogle Scholar
  31. Linderberg E, Vesterberg O (1983) Monitoring exposure to chromic acid in chrome plating by measuring chromium in urine. Scand J Work Environ Health 9:333–340Google Scholar
  32. Luo H, Lu YD, Shi X, Mao Y, Dalal NS (1996) Chromium IV rH O Fenton-like reaction causes DNA 22 damage: implications to chromate’s genotoxicity. Ann Clin Lab Sci 26:1185–1191Google Scholar
  33. Mahimairaja S, Sakthivel S, Divakaran J, Naidu R, Ramasamy K (2000) Extent and severity of contamination around tanning industries in Vellore district. In: Naidu R et al (eds) Towards better management of soils contaminated with tannery wastes. ACIAR Proceedings No 88, pp 75–82Google Scholar
  34. Marzin D (1999) New approaches to estimating the mutagenic potential of chemicals. Cell Biol Toxicol 15:359–365CrossRefGoogle Scholar
  35. McAughey JJ, Samuel AM, Baxter PJ, Smith NJ (1988) Biological monitoring of occupational exposure in the chromate pigment production industry. Sci Total Environ 71(3):317–322CrossRefGoogle Scholar
  36. Medeiros MG, Rodrigues AS, Batoreu MC (2003) Elevated levels of DNA–protein cross links and micronuclei in peripheral lymphocytes of tannery workers exposed to trivalent chromium. Mutagenesis 18:19–24CrossRefGoogle Scholar
  37. Migliore L, Parrini M, Sbrana I, Biagini C, Battaglia A, Loprieno N (1991) Micronucleated lymphocytes in people occupationally exposed to potential environmental contaminants: the age effect. Mutat Res 256:13–20Google Scholar
  38. Moller P (2006) Assessment of reference values for DNA damage detected by the comet assay in human blood cell DNA. Mutat Res 612:84–104CrossRefGoogle Scholar
  39. Mondal NC, Saxena VK, Singh VS (2005) Assessment of groundwater pollution due to tannery industries in and around Dindigul, Tamilnadu, India. Environ Geol 48:149–157CrossRefGoogle Scholar
  40. Moorhead PS, Novell PC, Mellman WJ, Battips DM, Hungerford DA (1960) Chromosome preparation of leukocyte culture from peripheral blood. Exp Cell Res 20:613–615CrossRefGoogle Scholar
  41. Nordenson I, Beckman G (1978) Occupational and environmental risks in and around a smelter in northern Sweden. Hereditas 88:37–50Google Scholar
  42. Nriagu JO (1988) Production and uses of chromium. In: Nriagu, Niebner E (eds) Chromium in the natural and human environments. Wiley, New York, pp 81–104Google Scholar
  43. Perera F (1987) The potential usefulness of biological markers in risk assessment. Environ Health Perspect 76:141–145CrossRefGoogle Scholar
  44. Perera FP, Hemminki K, Gryzbowska E, Motykiewicz G, Michalska J, Santella RM (1992) Molecular and genetic damage in humans from environmental pollution in Poland. Nature 360:256–258CrossRefGoogle Scholar
  45. Rahkonen E, Junttila ML, Kalliomaki PL, Olkinouora M, Koponen M, Kalliomaki K (1983) Evaluation of biological monitoring among stainless steel welders. Int Arch Occup Environ Health 52:243–255CrossRefGoogle Scholar
  46. Rajaram R, Nair BU, Ramasami T (1995) Chromium(III) induced abnormalities in human lymphocyte cell proliferation: evidence for apoptosis. Biochem Biophys Res Commun 210:434–440CrossRefGoogle Scholar
  47. Randall JA, Gibson RS (1987) Serum and urine chromium as indices of chromium status in tannery workers. Proc Soc Exp Biol Med 185:16–23Google Scholar
  48. Rosenman KD, Stanbury M (1996) Risk of lung cancer among former chromium smelter workers. Am J Ind Med 29:491–500CrossRefGoogle Scholar
  49. Sbrana I, Carett S, Battaglia A (1991) Chromosomal aberration analysis of workers in tannery industries. Mutat Res 260:331–336CrossRefGoogle Scholar
  50. Schmid W (1975) The micronucleus test. Mutat Res 31(1):9–15Google Scholar
  51. Selvakumar M, Manoharan R (2002) Effect of tannery effluent in groundwater and its control—a case study at Dindigul. In: Proc. IGC-2002, DindigulGoogle Scholar
  52. Shi X, Mao Y, Knapton AD, Ding M, Rojanasakul Y, Gannett PM, Dalal NS, Liu K (1994) Reaction of Cr VI with ascorbate and hydrogen peroxide generates hydroxyl radicals and causes DNA damage: role of Cr VI-mediated Fenton-like reaction. Carcinogenesis 15:2475–2478CrossRefGoogle Scholar
  53. Silva J, Freitas TRO, Heuser VD, Marinho JR, Erdtmann B (2000) Genotoxicity biomonitoring in coal regions using wild rodent Ctenomys torquatus by comet assay and micronucleus test. Environ Mol Mutagen 35:270–278CrossRefGoogle Scholar
  54. Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res 175(1):184–191CrossRefGoogle Scholar
  55. Stupar J, Vrtovec M, Kocijancic A, Gantar A (1999) Chromium status of tannery workers in relation to metabolic disorders. J Appl Toxicol 19:437–446CrossRefGoogle Scholar
  56. Tariq SR, Shah MH, Shaheen N, Khalique A, Manzoor S, Jaffar M (2006) Multivariate analysis of trace metal levels in tannery effluents in relation to soil and water: a case study from Peshawar, Pakistan. J Environ Manag 79(1):20–29CrossRefGoogle Scholar
  57. Thangarajan M (1999) Modelling pollutant migration in the upper Palar River Basin, Tamil Nadu, India. Environ Geol 38:209–222CrossRefGoogle Scholar
  58. Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H (2000) Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen 35:206–221CrossRefGoogle Scholar
  59. Ueno S, Kashimoto T, Susa N, Furukawa Y, Ishii M, Yokoi K, Yasuno M, Sasaki YF, Ueda J, Nishimura Y, Sugiyama M (2001) Detection of dichromate(VI)-induced DNA strand breaks and formation of paramagnetic chromium in multiple mouse organs. Toxicol Appl Pharmacol 170:56–62CrossRefGoogle Scholar
  60. Welinder H, Littorin M, Gullberg B, Skerfving S (1983) Elimination of chromium in urine after stainless steel welding. Scand J Work Environ Health 9:397–403Google Scholar
  61. Werfel U, Langen V, Eickhoff I, Schoonbrood J, Vahrenholz C, Brauksiepe A (1998) Elevated DNA single strand breakage frequencies in lymphocytes of welders exposed to chromium and nickel. Carcinogenesis 19:413–418CrossRefGoogle Scholar
  62. WHO (1990) Chromium World Health Organisation. (Environmental Health Criteria 61). International Programme on Chemical Safety, Geneva, SwitzerlandGoogle Scholar

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Vellingiri Balachandar
    • 1
    Email author
  • Meyyazhagan Arun
    • 1
  • Subramaniam Mohana Devi
    • 1
  • Palanivel Velmurugan
    • 2
  • Pappusamy Manikantan
    • 1
  • Alagamuthu Karthick Kumar
    • 1
  • Keshavarao Sasikala
    • 1
  • Chinnakulandai Venkatesan
    • 3
  1. 1.Human Genetics Laboratory, School of Life SciencesBharathiar UniversityCoimbatoreIndia
  2. 2.Division of Biotechnology, College of Environmental and Bioresource SciencesChonbuk National UniversityJeonjuSouth Korea
  3. 3.Aquaculture Biotechnology Laboratory, Abdul Hakeem CollegeThiruvalluvar UniversityVellore DistrictIndia

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