Biological Trace Element Research

, Volume 147, Issue 1–3, pp 206–216 | Cite as

Modified Natural Clinoptilolite Detoxifies Small Mammal’s Organism Loaded with Lead II: Genetic, Cell, and Physiological Effects

  • Margarita Topashka-Ancheva
  • Michaela BeltchevaEmail author
  • Roumiana Metcheva
  • J. Antonio Heredia Rojas
  • Abraham O. Rodriguez-De la Fuente
  • Tsvetelina Gerasimova
  • Laura E. Rodríguez-Flores
  • Svetla E. Teodorova


The detoxification capacity of the clinoptilolite modification KLS-10-MA used as food additive in small mammals, chronically lead-exposed, was proven for the first time. The modified clinoptilolite was prepared based on natural Bulgarian clinoptilolite deposits. As a powder, it was mechanically mixed at 12.5% concentration with the conventional forage for small rodents. Lead in the form of aqueous solution of Pb(NO3)2 was diluted in the drinking water. In the ecotoxicological experiment covering 90 days, imprinting control region laboratory mice were used. They were allocated into four groups: group 1, (control): animals fed with conventional food for small rodents and water; group 2: animals fed with conventional food + clinosorbent KLS-10-MA and water; group 3: animals fed with conventional food and water + Pb(NO3)2; and group 4: animals fed with conventional food + KLS-10-MA and water + Pb(NO3)2. A group of non-exposed healthy animals was fed with conventional forage mixed with KLS-10-MA to prove eventual toxicity of the sorbent and influence on growth performance. The changes in the chromosome structure, mitotic index, erythrocyte form, erythropoiesis, and body weight gain were recorded. On day 90, the following relations were established: Pb-exposed and clinoptilolite-supplemented mice exhibited 2.3-fold lower chromosome aberrations frequency, 2.5-fold higher mitotic index, and 1.5-fold higher percentage normal erythrocytes 1.3-fold higher body weight compared to Pb-exposed and unsupplemented animals. The obtained data showed that the sorbent is practically non-toxic. The results of the present study encourage a further elaboration of a reliable drug based on the tested substance in the cases of chronic lead intoxication.


Lead Chronic exposure Clinoptilolite Detoxification Laboratory mice Chromosome aberrations Mitotic index Erythrocyte form Body weight 



Authors express their special thanks to “Mineral agro Z” LTD—Bulgaria for the total financial support of this work.


  1. 1.
    Tsuchia K (1979) Lead. In: Friberg L (ed) Handbook on the toxicology of metals. Elsevier/North-Holland Biomedical, Amsterdam, pp 451–484Google Scholar
  2. 2.
    Goyer RA (1993) Lead toxicity: current concerns. Environ Health Perspect 100:177–187PubMedCrossRefGoogle Scholar
  3. 3.
    Jin Y, Liao Y, Lu C, Li G, Yu F, Zhi X, Xu J, Liu S, Liu M, Yang J (2006) Health effects of children aged 3–6 years induced by environmental lead exposure. Ecotoxicol Environ Saf 63:313–317PubMedCrossRefGoogle Scholar
  4. 4.
    Patil AJ, Bhagwat VR, Patil JA, Dongre NN, Ambekar JG, Das KK (2006) Biochemical aspects of lead exposure in silver jewelry workers in western Maharashtra (India). J Basic Clin Physiol Pharmacol 17:213–229PubMedCrossRefGoogle Scholar
  5. 5.
    Milton A, Cooke JA, Johnson MS (2003) Accumulation of lead, zinc and cadmium in a wild population of Clethrionomys glareolus from an abandoned lead mine. Arch Environ Contam Toxicol 44:405–411PubMedCrossRefGoogle Scholar
  6. 6.
    Ma WC (1996) Lead in mammals. In: Beyer WN, Heinz GH, Redmon-Norwood AW (eds) Environmental contaminants in wildlife interpreting tissue concentrations. Lewis, Boca Raton, pp 281–296Google Scholar
  7. 7.
    Robles HV, Romo E, Sanchez-Mendoza A, Rios A, Soto V, Avila-Casado MC, Medina A, Escalante B (2007) Lead exposure effect on angiotensin II renal vasoconstriction. Hum Exp Toxicol 26:499–507PubMedCrossRefGoogle Scholar
  8. 8.
    Sindhu RK, Sindhu KK, Roberts CK (2007) Role of lead in hypertension and chronic kidney disease. Environ Res J 1(3/4):345–359Google Scholar
  9. 9.
    Goyer RA (1996) Toxic effects of metals. In: McGraw-Hill (ed) Casarett and Doull’s toxicology. The basic science of poisons, 5th edn. Health Professions Division, New York, pp 691–735Google Scholar
  10. 10.
    Marques CC, Nunes AC, Pinheiro T, Lopes PA, Santos MC, Viegas-Crespo AM, Ramalhinho MG, Mathias ML (2006) An assessment of time-dependent effects of lead exposure in Algerian mice (Mus spretus) using different methodological approaches. Biol Trace Elem Res 109:75–90PubMedCrossRefGoogle Scholar
  11. 11.
    Nayak BN, Ray M, Persaud TVN (1989) Maternal and fetal chromosomal aberrations in mice following prenatal exposure to subembryotoxic doses of lead nitrate. Acta Anat 135:185–188PubMedCrossRefGoogle Scholar
  12. 12.
    Hartwig A, Schlepegrell R, Beyersmann D (1990) Indirect mechanism of lead-induced genotoxicity in cultured mammalian cells. Mutat Res 241:75–82PubMedCrossRefGoogle Scholar
  13. 13.
    Topashka-Ancheva M, Metcheva R, Teodorova SE (2003) Bioaccumulation and damaging action of polymetal industrial dust on laboratory mice Mus musculus alba II. Genetic, cell and metabolic disturbances. Environ Res 92:152–160PubMedCrossRefGoogle Scholar
  14. 14.
    Topashka-Ancheva M, Metcheva R, Teodorova SE (2003) A comparative analysis of the heavy metal loading of small mammals in different regions of Bulgaria II: chromosomal aberrations and blood pathology. Ecotoxicol Environ Saf 54:188–193PubMedCrossRefGoogle Scholar
  15. 15.
    Valverde M, Trejo C, Rojas E (2005) Is the capacity of lead acetate and cadmium chloride to induce genotoxic damage due to direct DNA–metal interaction? Toxicol Ind Health 21:243–248CrossRefGoogle Scholar
  16. 16.
    Shaik AP, Sankar S, Ready SC, Das PG, Jamil K (2006) Lead-induced genotoxicity in lymphocytes from peripheral blood samples of humans: in vitro studies. Drug Chem Toxicol 29:111–124CrossRefGoogle Scholar
  17. 17.
    Silbergeld EK, Waalkes M, Rice MJ (2000) Lead as a carcinogen: experimental evidence and mechanisms of action. Am J Ind Med 38:316–323PubMedCrossRefGoogle Scholar
  18. 18.
    Beyersmann D, Hartwig A (2008) Carcinogenic metal compounds: recent insight into molecular and cellular mechanisms. Arch Toxicol 82:493–512PubMedCrossRefGoogle Scholar
  19. 19.
    Roy NK, Rossman TG (1992) Mutagenesis and comutagenesis by lead compounds. Mutat Res 298:97–103PubMedCrossRefGoogle Scholar
  20. 20.
    Mumpton FA, Fishman PH (1977) The application of natural zeolites in animal science and agriculture. J Anim Sci 45:1188–1203Google Scholar
  21. 21.
    Pond WG (1995) Zeolite in animal nutrition and health: a review. In: Ming DW, Mumpton FA (eds) Natural zeolites ’93: occurrence, properties, use. Int. Comm. Natural Zeolites, Brockport New York, pp 449–457Google Scholar
  22. 22.
    Oguz H (2011) A review from experimental trials on detoxification of aflatoxin in poultry feed. Eurasia J Vet Sci 27(1):1–12Google Scholar
  23. 23.
    Pavelić K, Hadžija M, Bedrica L, Pavelić J, Dikić I, Katić M, Kralj M, Bosnar NJ, Kapitanović S, Poljak-Blaži M, Krizaneć S, Stojković R, Jurin M, Subotić B, Colić M (2001) Natural zeolite clinoptilolite: a new adjuvant in anticancer therapy. J Mol Med 78:708–720PubMedCrossRefGoogle Scholar
  24. 24.
    Zarkovic N, Zarkovic K, Kraij M, Borovic S, Sabolovic S, Poliakblazi M, Cipak A, Pacelic K (2003) Anticancer and antioxidative effects of micronized zeolite clinoptilolite. Anticancer Res 23:1589–1596PubMedGoogle Scholar
  25. 25.
    Sverko V, Sobočanec S, Balog T, Colić M, Marotti T (2004) Natural micronised clinoptilolite and clinoptilolite mixtures with Urtica dioica L. extract as possible antioxidants. Food Technol Biotechnol 42:189–192Google Scholar
  26. 26.
    Pavelić K, Katic M, Sverko V, Marroti T, Bosnjak B, Balog T, Stojkovic R, Radacic M, Colic M, Poljak-Blazi M (2002) Immunostimulatory effect of natural clinoptilolite as a possible mechanism of its antimetastatic ability. J Cancer Res Clin Oncol 128:37–44PubMedCrossRefGoogle Scholar
  27. 27.
    Jung BG, Toan NT, Cho SJ, Ko JH, Jung YK, Lee BG (2010) Dietary aluminosilicate supplement enhances immune activity in mice and reinforces clearance of porcine circovirus type 2 in experimentally infected pigs. Vet Microbiol 143:117–125PubMedCrossRefGoogle Scholar
  28. 28.
    Armbruster T (2001) Clinoptilolite–heulandite: applications and basic research. In: Galarnau A, Di Renzo F, Faujula F, Vedrine J (eds) Studies in surface science and catalysis, vol 135, Zeolites and mesoporous materials at the dawn of the 21st century. Elsevier Science, Montpellier, pp 101–200Google Scholar
  29. 29.
    Papaioannou D, Katsoulos PD, Panousis N, Karatzias H (2005) The role of natural and synthetic zeolites as feed additives on the prevention and/or the treatment of certain farm animal diseases: a review. Micropor Mesopor Mat 84:161–170CrossRefGoogle Scholar
  30. 30.
    Tiwari J (2007) Zeolite as natural feed additives to reduce environmental impacts of swine manure. Department of Bioresource Engineering, McGill University, MontrealGoogle Scholar
  31. 31.
    Tao YF, Qui Y, Fang SY, Liu ZY, Wang Y, Zhu JH (2010) Trapping the lead ion in multi-component aqueous solutions by natural clinoptilolite. J Hazard Mater 180:282–288PubMedCrossRefGoogle Scholar
  32. 32.
    Sampson R (1997) Application for the approval of clinoptilolite. Euremica Environmental, ClevelandGoogle Scholar
  33. 33.
    Beltcheva M, Metcheva R, Popov N, Teodorova SE, Heredia Rojas JA, Rodriguez De la Fuente AO, Rodríguez-Flores LE, Topashka-Ancheva M (2011) Natural clinoptilolite detoxifies small mammal’s organism loaded with lead I. Lead disposition and kinetic model for lead bioaccumulation. Biol Trace Elem Res (in press)Google Scholar
  34. 34.
    Fethiere R, Milles RD, Harms RH (1994) The utilization of sodium in sodium zeolite A by broilers. Poult Sci 73:118–121PubMedCrossRefGoogle Scholar
  35. 35.
    Vrzgula L, Bartko P, Blažovsky J, Kozač J (1982) The effect of feeding clinoptilolite on health status, blood picture and weight gain in pigs. Vet Med-Czesh 27:267–274Google Scholar
  36. 36.
    Tl W, Watkins KL, Southern LL, Hoyt PG, French DD (1991) Interactive effects of sodium zeolite A and copper in growing swine: growth, and bone and tissue mineral concentrations. J Anim Sci 69:726–733Google Scholar
  37. 37.
    Eady SJ, Pritchard DA, Martin MOJ (1980) The effect of sodium bentonite on zeolite on wool growth of sheep fed either mulga (Acacia aneura) or lucerne (Medicago sativa). Proc Aust Soc Anim Prod 18:188–191Google Scholar
  38. 38.
    Keshavarz K, McCormick CC (1991) Effect of sodium aluminosilicates, oyster shell, and their combination on acid–base balance and eggshell quality. Poult Sci 70:313–325PubMedCrossRefGoogle Scholar
  39. 39.
    Preston RY, Dean LY, Galloway Sh, Helden H, Me Fee A (1987) Mammalian in vivo cytogenetic assays. Analysis of chromosome aberrations in bone marrow cells. Mutat Res 189:157–165PubMedCrossRefGoogle Scholar
  40. 40.
    Hoff J (2000) Methods of blood collection in the mouse. Lab Anim Tech 29:10–50Google Scholar
  41. 41.
    Environment, Health and Safety Committee [EHSC] (2001) Note On “LD50” [Lethal Dose 50%], version 2–31/7/01. Royal Society of Chemistry, LondonGoogle Scholar
  42. 42.
    Mumpton FA (1985) Using zeolites in agriculture in innovative biological technologies for lesser developed countries. Workshop Proceedings, Washington DCGoogle Scholar
  43. 43.
    Mumpton FA (1999) La Roca Magica: uses of natural zeolites in agriculture and industry. Proc Natl Acad Sci USA 96:3463–3470PubMedCrossRefGoogle Scholar
  44. 44.
    International Commission for Protection Against Environmental Mutagens and Carcinogens (1983) Report of Committee I. Mutat Res 114:120–177Google Scholar
  45. 45.
    World Health Organization (1985) Environmental health criteria 51, guide to short-term tests for detecting mutagenic and carcinogenic chemicals. WHO, Geneva, pp 100–114Google Scholar
  46. 46.
    Demarque D, Jouanny J, Poitevin B, Saint Jean Y (1995) Pharmacologie et Matiere Medicale Homeopathique. Boiron–C.E.D.H., FranceGoogle Scholar
  47. 47.
    Dhir H, Roy AK, Sharma A, Talukder G (1990) Modification of clastogenicity of lead and aluminum in mouse bone marrow cells by dietary ingestion of Phyllanthus emblica fruit extract. Mutat Res 241:305–312PubMedCrossRefGoogle Scholar
  48. 48.
    Sorsa M, Wilbourn J, Vainio H (1992) Human cytogenetic damage as a predictor of cancer risk. In: Vainio H, Magee PH, McGregor DB, McMichael AJ (eds) Mechanisms of carcinogenesis in risk identification. International Agency for Research on Cancer, Lyons, pp 543–554Google Scholar
  49. 49.
    Albertini RJ, Anderson D, Douglas GR, Hagmar L, Hemmimki K, Merlo F, Natarajan AT, Norppa H, Shuker DE, Tice R, Waters MD, Aitio A (2000) IPCS guidelines for the monitoring of genotoxic effects of carcinogens in humans. International Programme on Chemical Safety. Mutat Res 463:111–172PubMedCrossRefGoogle Scholar
  50. 50.
    Hagmar L, Stromberg U, Bonassi S, Hansteen I-L, Knudsen LE, Lindholm C, Norppa H (2004) Impact of types of lymphocyte chromosomal aberrations on human cancer risk: results from Nordic and Italian cohorts. Cancer Res 64:2258–2263PubMedCrossRefGoogle Scholar
  51. 51.
    Franco S, Alt FW, Manis JP (2006) Pathways that suppress programmed DNA breaks from progressing to chromosomal breaks and translocations. DNA Repair 5:1030–1041PubMedCrossRefGoogle Scholar
  52. 52.
    Mateuca R, Lombaert N, Aka PV, Decordier I, Kirsch-Volders M (2006) Chromosomal changes: induction, detection methods and applicability in human monitoring. Biochimie 88:1515–1531PubMedCrossRefGoogle Scholar
  53. 53.
    Privezentsev KV, Sirota NP, Gaziev AJ (1996) The genotoxic of cadmium studied in vivo. Tsitologia i genetika 30:45–51Google Scholar
  54. 54.
    Pfeiffer P, Goedecke W, Obe G (2000) Mechanisms of DNA double-strand repair and their potential to induce chromosomal aberrations. Mutagenesis 15:289–302PubMedCrossRefGoogle Scholar
  55. 55.
    Kipling D, Wilson HE, Mitchell AR, Teylor BA, Cooke HJ (1994) Mouse centromeric mapping using oligonucleotide probe that detect variants of the minor satellite. Chromosoma 103:46–55PubMedCrossRefGoogle Scholar
  56. 56.
    Kelsey KT, Yano E, Liber HL, Little JB (1986) The in vitro genetic effects of fibrous erionite and crocidolite asbestos. Br J Cancer 54:107–114PubMedCrossRefGoogle Scholar
  57. 57.
    Paglia DE, Valentine WN, Dahlgner JG (1975) Effects of low-level lead exposure on pyrimidine-5-nucleosidase and other erythrocyte enzymes. J Clin Invest 56:1164–1169PubMedCrossRefGoogle Scholar
  58. 58.
    Ershov YA, Pleteneva TV (1989) Mechanisms of toxic action of inorganic compounds. Medizina, MoscowGoogle Scholar
  59. 59.
    Martin-Kleiner I, Flegar-Meštić Z, Zadro R, Breljak D, Stanović Janda S, Stojković R, Marušić M, Radačić M, Boranić M (2001) The effect of the zeolite clinoptilolite on serum chemistry and hematopoiesis in mice. Food Chem Toxicol 39:717–727PubMedCrossRefGoogle Scholar
  60. 60.
    Shurson GC, Ku PK, Miller ER, Yokoyama MT (1984) Effects of zeolite A or clinoptilolite in diets of growing swine. J Anim Sci 59:1536–1545PubMedGoogle Scholar
  61. 61.
    Kyriakis SC, Papaioannou DS, Alexopoulos C, Polizopoulou Z, Tzika ED, Kyriakis CS (2002) Experimental studies on safety and efficacy of the dietary use of a clinoptilolite-rich tuff in sows: a review of recent research in Greece. Micropor Mesopor Mat 51:65–74CrossRefGoogle Scholar
  62. 62.
    Pond WG, Yen JT (1988) Response of growing pigs to dietary clinoptilolite from two geographic sources. Nutr Rep Int 25:837–848Google Scholar
  63. 63.
    Yannakopoulos A, Tserveni-Gousi A, Kassoli-Four-Naraki A, Tsirambides A, Michailidis K, Filippidis A, Lutat U (2000) Effects of dietary clinoptilolite-rich tuff on the performance of growing-finishing pigs. In: Coela C, Mumpton FA (eds) Natural zeolites for the third millennium. De Frede Editore, Napoli, pp 471–481Google Scholar
  64. 64.
    Norouzian MA, Valizadeh R, Khadem AA, Afzalzadeh A, Nabipour A (2010) The effects of feeding clinoptilolite on hematology, performance, and health of newborn lambs. Biol Trace Elem Res 137:168–176PubMedCrossRefGoogle Scholar
  65. 65.
    Nestorov N (1984) Possible applications of natural zeolites in animal husbandry. In: Pond WG, Mumpton FA (eds) Zeo-agriculture: use of natural zeolite in agriculture and aquaculture. International Committee on Natural Zeolites. Westview, Boulder, pp 167–174Google Scholar
  66. 66.
    Kondo N, Wagai B (1968) Experimental use of clinoptilolite-tuff as a dietary supplement for pigs. Yotonkai, May 1968, 1–4Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Margarita Topashka-Ancheva
    • 1
  • Michaela Beltcheva
    • 1
    Email author
  • Roumiana Metcheva
    • 1
  • J. Antonio Heredia Rojas
    • 2
  • Abraham O. Rodriguez-De la Fuente
    • 2
  • Tsvetelina Gerasimova
    • 1
  • Laura E. Rodríguez-Flores
    • 3
  • Svetla E. Teodorova
    • 4
  1. 1.Institute of Biodiversity and Ecosystem ResearchBulgarian Academy of SciencesSofiaBulgaria
  2. 2.Departamento de Ciencias Exactas y Desarrollo Humano, Facultad de Ciencias BiológicasUniversidad Autónoma de Nuevo LeónSan Nicolás de los GarzaMexico
  3. 3.Departamento de Patología, Facultad de MedicinaUniversidad Autónoma de Nuevo LeónMonterreyMexico
  4. 4.Institute for Nuclear Research and Nuclear EnergyBulgarian Academy of SciencesSofiaBulgaria

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