SNL  Vol.5 No.2 , April 2015
Efficacy of Organo-Modified Nano Montmorillonite to Protect against the Cumulative Health Risk of Aflatoxin B1 and Ochratoxin A in Rats
Abstract: The aim of the current study was to prepare organo-modified nano montmorillonite (OMNM) and to evaluate its chemopreventive effects against the hapatonephrotoxicity induced by aflatoxin B1 (AFB1) and ochratoxin A (OA) singly or in combination in rats. OMNM was prepared using Cetyltrimethylammoniumbromide (CTAB) as organic modifier. Eighty male Sprague Dawley were divided into 8 groups and treated for 8 weeks as follow: the control group; the group treated orally with AFB1 (80 μg/kg b.w.); the group treated with OA (100 μg/kg b.w.); the group treated with AFB1 plus OA, the group treated with OMNM (5 g/kg diet) and the groups treated with AFB1 and/or OA plus OMNM. At the end of treatment period, blood and tissue samples were collected from all animals for biochemical and histological analysis. The results revealed that the expansion in the basal spacing of the montmorillonite due to the intercalation of CTAB was 7.20 Å and the average particle size of OMNM was 120 nm. The in vivo results indicated that treatment with both AFB1 and OA singly or in combination resulted in a significant increase in liver and kidney function parameters, oxidative stress and tumor markers accompanied with a significant decrease in antioxidant enzyme activities and significant histological changes in liver and kidney tissues. These changes were severe in the group received the combined treatment of AFB1 and OA. OMNM alone did not show any toxic effect and it succeeded to prevent or at least diminish the toxic effects and the histological changes in liver and kidney. It can be concluded that treatment with AFB1 and OA has a synergistic toxic effects and OMNM is safe and it is a promise candidate as an additive to protect against the exposure to multi-mycotoxins in high risk population.
Cite this paper: Abdel-Wahhab, M. , El-Denshary, E. , El-Nekeety, A. , Abdel-Wahhab, K. , Hamzawy, M. , Elyamany, M. , Hassan, N. , Mannaa, F. , Shaiea, M. , Gado, R. and Zawrah, M. (2015) Efficacy of Organo-Modified Nano Montmorillonite to Protect against the Cumulative Health Risk of Aflatoxin B1 and Ochratoxin A in Rats. Soft Nanoscience Letters, 5, 21-35. doi: 10.4236/snl.2015.52004.

[1]   Stoev, S.D. (2013) Food Safety and Increasing Hazard of Mycotoxin Occurrence in Foods and Feeds. Critical Reviews in Food Science and Nutrition, 53, 887-901.

[2]   Kana, J.R., Gnonlonfin, B.G., Harvey, J., Wainaina, J., Wanjuki, I., Skilton, R.A. and Teguia, A. (2013) Assessment of Aflatoxin Contamination of Maize, Peanut Meal and Poultry Feed Mixtures from Different Agroecological Zones in Cameroon. Toxins (Basel), 5, 884-894.

[3]   Rodriguez-Carrasco, Y., Ruiz, M.J., Font, G. and Berrada, H. (2013) Exposure Estimates to Fusarium mycotoxins through Cereals Intake. Chemosphere, 93, 2297-2303.

[4]   van Egmond, H.P., Schothorst, R.C. and Jonker, M.A. (2007) Regulations Relating to Mycotoxins in Food: Perspectives in a Global and European Context. Analytical and Bioanalytical Chemistry, 389, 147-157.

[5]   IARC (1993) Monographs on the Evaluation of the Carcinogenic Risks to Human.

[6]   Liu, Y. and Wu, F. (2010) Global Burden of Aflatoxin-Induced Hepatocellular Carcinoma: A Risk Assessment. Environmental Health Perspectives, 118, 818-824.

[7]   Hassan, A.M., Abdel-Azeim, S.H., El-Nekeety, A.A. and Abdel-Wahhab, M.A. (2014) Panax ginseng Extract Modulates Oxidative Stress, DNA Fragmentation and Up-Regulate Gene Expression in Rats Sub Chronically Treated with Aflatoxin B1 and Fumonisin B1. Cytotechnology.

[8]   Joint FAO/WHO Expert Committee on Food Additives (JECFA) (2001) Safety Evaluation of Certain Mycotoxins in Food (WHO Food Additives Series 47). Geneva: International Programme on Chemical Safety. Food and Nutrition Paper 74.

[9]   Almeida, M.I., Almeida, N.G., Carvalho, K.L., Goncalves, G.A., Silva, C.N., Santos, E.A., Garcia, J.C. and Vargas, E.A. (2012) Co-Occurrence of Aflatoxins B1, B2, G1 and G2, Ochratoxin A, Zearalenone, Deoxynivalenol, and Citreoviridin in Rice in Brazil. Food Additives and Contaminants: Part A, Chemistry, Analysis, Control, Exposure and Risk Assessment, 29, 694-703.

[10]   Pfohl-Leszkowicz, A., Petk-Bocharova, T., Cherozemsky, I.N. and Castegnaro, M. (2002) Balkan Endemic Nephropathy and Associated Urinary Tract Tumors: A Review on Etiological Causes and the Potential Role of Mycotoxins. Food Additives and Contaminants, 19, 282-302.

[11]   Abdel-Wahhab, M.A., Ibrahim, A.A., El-Nekeety, A.A., Hassan, N.S. and Mohamed, A.A. (2012) Panax ginseng C.A. Meyer Extract Counteracts the Oxidative Stress in Rats Fed Multi-Mycotoxins-Contaminated Diet. Comunicata Scientiae, 3, 143-153.

[12]   Phillips, T.D. (1999) Dietary Clay in the Chemoprevention of Aflatoxin-Induced Disease. Toxicological Sciences, 52, 118-126.

[13]   Jordá-Beneyto, M., Alonso, J., Salas, J., Gallur, M., Aucejo, S., Clegg, F. and Breen, C. (2008) Processed Biopolymer Films Filled with Modified Montmorillonite for Food Packaging Applications. Proceedings of the Polymer Processing Society 24th Annual Meeting, PPS-24, Salerno (Italy), 15-19 June 2008, 15-19.

[14]   Bitinis, N., Hernández, M., Verdejo, R., Kenny, J.M. and López-Machado, M.A. (2011) Recent Advances in Clay/Polymer Nanocomposites. Advanced Materials, 23, 5229-5236.

[15]   Zawrah, M.F., Khattab, R.M., Saad, E.M. and Gado, R.A. (2014) Effect of Surfactant Types and Their Concentration on the Structural Characteristics of Nanoclay. Spectrochimica Acta, Part A: Molecular and Biomolecular Spectroscopy, 122, 616-623.

[16]   Mansour, M.A., Ginawi, O.T., El-Hadiyah, T., El-Khatib, A.S., Al-Shabanah, O.A. and Al-Sawaf, H.A. (2001) Effects of Volatile Oil Constituents of Nigella sativa on Carbon Tetrachloride-Induced Hepatotoxicity in Mice: Evidence for Antioxidant Effects of Thymoquinone. Research Communications in Molecular Pathology and Pharmacology, 110, 239-251.

[17]   Drury, R.A., Wallington, E.A. and Cancerson, R., Eds. (1976) Carlton’s Histopathological Techniques. 4th Edition, Oxford University Press, Oxford, London, New York.

[18]   Osman, M.A., Ploetze, M. and Skrabal, P. (2004) Structure and Properties of Alkylammonium Monolayers Self-Assembled on Montmorillonite Platelets. Journal of Physical Chemistry B, 108, 2580-2588.

[19]   Klaric, M.S. (2012) Adverse Effects of Combined Mycotoxins. Archives of Industrial Hygiene and Toxicology, 63, 519-530.

[20]   Speijers, G.J.A. and Speijers, M.H.M. (2004) Combined Toxic Effects of Mycotoxins. Toxicology Letters, 153, 91-98.

[21]   Diamond, J. (1999) Evolutionary Biology: Dirty Eating for Healthy Living. Nature, 400, 120-121.

[22]   Wingenfelder, U., Nowack, B., Furrer, G. and Schulin, R. (2005) Adsorption of Pb and Cd by Amine-Modified Zeolite. Water Research, 39, 3287-3292.

[23]   Abdel-Wahhab, M.A., Hassan, N.S., El-Kady, A.A., Mohamed, Y.A., El-Nekeety, A.A., Mohamed, S.R., Sharaf, H.A. and Mannaa, F.A. (2010) Red Ginseng Protects against Aflatoxin B1 and Fumonisin-Induced Hepatic Pre-Cancerous Lesions in Rats. Food and Chemical Toxicology, 48, 733-742.

[24]   Abdel-Wahhab, M.A., Abdel-Galil, M.M. and Ellithey, M.M. (2005) Melatonin Counteracts Oxidative Stress in Rats Fed an Ochratoxin A Contaminated Diet. Journal Pineal Research, 38, 130-135.

[25]   Abdel-Wahhab, M.A., Abdel-Azim, S.H. and El-Nekeety, A.A. (2008) Inula crithmoides Extract Protect against Ochratoxin A-Induced Oxidative Stress, Clastogenic and Mutagenic Alterations in Male Rats. Toxicon, 52, 566-573.

[26]   Kaneko, J.J. (1989) Clinical Chemistry of Domestic Animals. 4th Edition, Academic Press, San Diego.

[27]   Yang, C.F., Liu, J., Wasser, S., Shen, H.M., Tan, C.E. and Ong, C.N. (2000) Inhibition of Ebselen on Aflatoxin B1- Induced Hepatocarcinogenesis in Fisher 344 Rats. Carcinogenesis, 21, 2237-2243.

[28]   Abdel-Wahhab, M.A., Abdel-Galil, M.M., Hassan, A.M., Hassan, N.H., Nada, S.A., Saeed, A. and El-Sayed, M.M. (2007) Zizyphus spina-Christi Extract Protects against Aflatoxin B1-Intitiated Hepatic Carcinogenicity. African Journal of Traditional, Complementary and Alternative Medicines, 4, 248-256.

[29]   Sell, S., Xy, K.L., Huff, W.E., Kabena, L.F., Harvey, R.B. and Dunsford, H.A. (1998) Aflatoxin Exposure Produces Serum Alphafetoprotein Elevations and Marked oval Cell Proliferation in Young Male Pekin Ducklings. Pathology, 30, 34-39.

[30]   Moon, E.Y. and Pyo, S. (2000) Aflatoxin B1 Inhibits CD14-Mediated Nitric Oxide Production in Murine Peritoneal Macrophages. International Journal of Immunopharmacology, 22, 237-246.

[31]   Azeredo-Martins, A.K., Lortz, S., Lenzen, S., Curi, R., Eizirik, D.L. and Tiedge, M. (2003) Improvement of the Mitochondrial Antioxidant Defense Status Prevents Cytokine-Induced Nuclear Factor-Kappa B Activation in Insulin-Producing Cells. Diabetes, 52, 93-101.

[32]   Abdel-Azeim, S.H., Hassan, A.M. and Abdel-Wahhab, M.A. (2011) Dietary Supplementation with Whey Protein and Ginseng Extract Counteracts the Oxidative Stress and DNA Damage in Rats Fed Aflatoxins-Contaminated Diet. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 723, 65-71.

[33]   Abdel-Wahhab, M.A. and Aly, S.E. (2005) Antioxidant Property of Nagilia sativa (Black Cumin) and Syzygium aromaticum (Clove) in Rats during Aflatoxicosis. Journal of Applied Toxicology, 25, 218-223.

[34]   Gautier, J.C., Holzhaeuser, D., Markovic, J., Gremaud, E., Schilter, B. and Turesky, R.J. (2001) Oxidative Damage and Stress Response from Ochratoxin A Exposure in Rats. Free Radical Biology and Medicine 30, 1089-1098.

[35]   Gad, A.S., Khadrawy, Y.A., El-Nekeety, A.A., Mohamed, S.R., Hassan, N.S., Abdel-Wahhab, M.A. (2011) Antioxidant Activity and Hepatoprotective Effects of Whey Protein and Spirulina in Rats. Nutrition, 27, 582-589.

[36]   Schinella, G.R., Tournier, H.A., Prieto, J.M., Mordujovich de Buschiazzo, P. and Ríos, J.L. (2002) Antioxidant Activity of Anti-Inflammatory Plant Extracts. Life Science, 70, 1023-1033.

[37]   Pfohl-Leszkowicz, A., Grosse, Y., Kanr, A., Gharbi, A., Beaudrimont, I., Obrecht, S., Creppy, E.E. and Dirheimer, G. (1993) Is the Oxidative Pathway Implicated in the Genotoxicity of Ochratoxin A? In: Creppy, E.E., Castegnaro, M. and Direimer, G., Eds., Human Ochratoxicosis and Its Pathologies, John Libbey Eurotext Ltd., London, 177-187.

[38]   Busby, W.F. and Wogan, G.N. (1984) Aflatoxins. In: Searle, C.E., Ed., Chemical Carcinogenesis, II, American Chemical Society, Washington DC, 945-1136.

[39]   Hohler, D. (1998) Ochratoxin A in Food and Feed: Occurrence, Legislation and Mode of Action. Zeitschrift für Ernahrungswissenschaft, 37, 2-12.

[40]   Aly, S.E., Abdel-Galil, M.M. and Abdel-Wahhab, M.A. (2004) Application of Adsorbent Agents Technology in the Removal of Aflatoxin B1 and Fumonisin B1 from Malt Extract. Food and Chemical Toxicology, 42, 1825-1831.

[41]   Sposito, G., Skipper, N.T., Sutton, R., Park, S., Soper, A.K. and Greathouse, J.A. (1999) Surface Geochemistry of the Clay Minerals. Proceedings of the National Academy of Sciences of the United States of America, 96, 3358-3364.

[42]   Afriyie-Gyawu, E., Wang, Z., Ankrah, N.A., Xu, L., Johnson, N.M., Tang, L., Guan, H., Huebner, H.J., Jolly, P.E., Ellis, W.O., Taylor, R., Brattin, B., Ofori-Adjei, D., Williams, J.H., Wang, J.S. and Phillips, T.D. (2008) NovaSil Clay Does Not Affect the Concentrations of Vitamins A and E and Nutrient Minerals in Serum Samples from Ghanaians at High Risk for Aflatoxicosis. Food Additives and Contaminants. Part A, Chemistry, Analysis, Control, Exposure and Risk Assessment, 25, 872-84.

[43]   Abbès, S., Ben Salah-Abbès, J., Ouanes, Z., Houas, Z., Othman, O., Bacha, H., Abdel-Wahhab, M.A. and Oueslati, R. (2006) Preventive Role of Phyllosilicate Clay on the Immunological and Biochemical Toxicity of Zearalenone in Balb/c Mice. International Journal of Immunopharmacology, 6, 1251-1258.

[44]   Wang, J.S., Luo, H., Billam, M.,Wang, Z., Guan, H., Tang, L., Goldston, T., Afriyie-Gyawu, E., Lovett, C., Griswold, J., Brattin, B., Taylor, R.J., Huebner, H.J. and Phillips, T.D. (2005) Short-Term Safety Evaluation of Processed Calcium Montmorillonite Clay (NovaSil) in Humans. Food Additives and Contaminants, 22, 270-279.

[45]   Hassan, A.M., Kenawy, A.M., Abbas, W.T. and Abdel-Wahhab, M.A. (2010) Prevention of Cytogentic, Histochemical and Biochemical Alterations in Oreochromis niloticus by Dietary Supplement of Sorbent Materials. Ecotoxicology and Environmental Safety, 73, 1890-1895.

[46]   Maisanaba, S., Puerto, M., Pichardo, S., Jordá, M., Moreno, F.J., Aucejo, S. and Jos, A. (2013) In Vitro Toxicity Evaluation of Unmodified Commercial Clays on the Human Hepatic Cell Line HepG2. Food and Chemical Toxicology, 57, 266-275.

[47]   Houtman, J., Maisanaba, S., Puerto, M., Gutiérrez-Praena, D., Jordá, M., Aucejo, S. and Jos, A. (2014) Toxicity Assessment of Organomodified Clays Used in Food Contact Materials on Human Target Cell Lines. Applied Clay Science, 90, 150-158.

[48]   Sharma, A.K., Schmidt, B., Frandsen, H., Jacobsen, N.R., Larsen, E.H. and Binderup, M.L. (2010) Genotoxicity of Unmodified and Organo-Modified Montmorillonite. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 700, 18-25.

[49]   Silvestre, C., Duraccio, D. and Cimmino, S. (2011) Food Packaging Based on Polymer Nanomaterials. Progress in Polymer Science, 36, 1766-1782.

[50]   Baek, M., Lee, A.J. and Choi, S.J. (2012) Toxicological Effects of a Cationic Clay, Montmorillonite in Vitro and in Vivo. Molecular and Cellular Toxicology, 8, 95-101.

[51]   Liu, Q., Liu, Y., Xiang, S., Mo, X., Su, S. and Zhang, J. (2011) Apoptosis and Cytotoxicity of Oligo (Styrene-co-Acrylonitrile)-Modified Montmorillonite. Applied Clay Science, 51, 214-219.

[52]   Bouwmeester, H., Dekkers, S., Maryvon, Y., Noordam, M., Hagens, W., Bulder, A.S., de Heer, C., ten Voorde, S., Wijnhoven, S., Marvin, H. and Sips, A. (2009) Review of Health Safety Aspects of Nanotechnologies in Food Production. Regulatory Toxicology and Pharmacology, 53, 52-62.

[53]   Dougherty, S.H., Fiegel, V.D., Nelson, R.D., Rodeheaver, G.T. and Simmons, R.L. (1985) Effects of Soil Infection Potentiating Factors on Neutrophils in Vitro. American Journal Surgery, 150, 306-311.