Bio-protective effects of Lactobacillus plantarum subsp. plantarum against aflatoxin b1 genotoxicity on human blood lymphocytes: a native probiotic strain isolated from Iranian camel milk

Document Type : Original Articles


1 Department of Food Science and Technology, Ayatollah Amoli Branch, Islamic Azad University, Amol, Iran

2 Research and Development Unit of Referral Laboratory, Deputy of Health Management, Mazandaran University of Medical Sciences, Sari, Iran

3 Department of Food Science and Technology, Nour Branch, Islamic Azad University, Nour, Iran

4 Shams Bavaran Salamat Nour Company, Tehran, Iran.

5 Department of Toxicology and Pharmacology, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran

6 Pharmaceutical Sciences Research Center, Faculty of Pharmacy, Mazandaran University of Medical Sciences, Sari, Iran



Background and Purpose: Aflatoxin B1 is one of the main poisonous substances in certain kinds of fungi all over the world. The toxin is a serious health threat to humans and livestock, particularly via DNA damage, and induces multiple cancers. Probiotic agents have confirmed positive beneficial effects in DNA protection against various toxic compounds. In this regard, the present study aimed to investigate the bio-protective effects of a native Lactobacillus plantarum subsp. plantarumNIMBB003 strain isolated from Iranian one-humped camel milk against AflatoxinB1 (AFB1)-induced genotoxicity damage, based on the micronucleus test as a genotoxicity monitoring method.
Materials and Methods: In this study, a human male blood sample was treated and incubated with107, 109, and 1011CFU/mL of viable L. plantarum and IC50 dose ofAFB1alone and in combination. Afterward, assessed the rate of production of the micronucleus in bi-nucleated lymphocytes. It must be noted that a p-value of less than0.05 was considered significantly significant.
Results: Based on the findings, the combined treatment of the L. plantarum at 1011 and109CFU/mL dose with 5.33±0.57% of the micronuclei fragments had protective effects and significantly decreased the genotoxicity of AFB1 by 76%.
Conclusion: According to the findings, it can be concluded that L. plantarum in 109 CFU/mL had high protective potency against AFB1 genotoxicity. Consequently, the use of local, natural, and native protected compounds with antioxidant effects, such as probiotics agents, is one of the objectives of developing a green strategy in macro-management policies for the discovery and production of new medicines and functional foods with protective/therapeutic effects against nutritional and endogenous DNA toxins


1. Moradi M, Tajik H, Mardani K, Ezati P. Efficacy of lyophilized cell-free supernatant of Lactobacillus salivarius (Ls-BU2) on Escherichia coli and shelf life of ground beef. Vet Res Forum. 2019;10(3):193-8.
2. World Health Organization. The state of food security and nutrition in the world 2018: building climate resilience for food security and nutrition. Food and Agriculture Organization. Geneva: World Health Organization; 2018.
3. Afshar P, Motamedzadegan A, Ahmady M, Alimi M, Shahidi SA, Golestan L, et al. Review on aflatoxins biodegradation strategies based on fungi. Clin Excell. 2017;7(1):36-48.
4. He XY, Tang L, Wang SL, Cai QS, Wang JS, Hong JY. Efficient activation of aflatoxin B1 by cytochrome P450 2A13, an enzyme predominantly expressed in human respiratory tract. Int J Cancer. 2006;118(11):2665-71.
5. Ráduly Z, Szabó L, Madar A, Pócsi I, Csernoch L. Toxicological and medical aspects of Aspergillus-derived mycotoxins entering the feed and food chain. Front Microbiol. 2019;10(1):2908.
6. Ostry V, Malir F, Toman J, Grosse Y. Mycotoxins as human carcinogens-the IARC Monographs classification. Mycotoxin Res. 2017;33(1):65-73.
7. Adam A, Adam M, Tabana YM, Musa KB, Sandai DA. Effects of different mycotoxins on humans, cell genome and their involvement in cancer. Oncol Rep. 2017;37(3):1321-36.
8. Moradi S, Azari H, Jafari Anarkooli I, Qasemi-Panahi B, Elhami S, Forouharmehr A. Effect of aflatoxin B1 on BRCA1 and BRCA2 genes expression under in vitro cultured cell line of normal Human Mammary Epithelial Cells (HMEC). J Police Med. 2015;3(4):211-20.
9. Afshar P, Shokrzadeh M, Kalhori S, Babaee Z, Saravi SS. Occurrence of ochratoxin A and aflatoxin M1 in human breast milk in Sari, Iran. Food Control. 2013;31(2):525-9.
10. Kurhan Ş, Cakir I. DNA-bioprotective effects of lactic acid bacteria against aflatoxin B1. Curr Res Nutr Food Sci. 2016;2016:87-91.
11. Horky P, Skalickova S, Baholet D, Skladanka J. Nanoparticles as a solution for eliminating the risk of mycotoxins. Nanomaterials. 2018;8(9):727.
12. Alpsoy L, Yildirim A, Agar G. The antioxidant effects of vitamin A, C, and E on aflatoxin B1-induced oxidative stress in human lymphocytes. Toxicol Ind Health. 2009;25(2):121-7.
13. Jama AM, Mitić-Ćulafić D, Kolarević S, Đurašević SF, Knežević-Vukčević J. Protective effect of probiotic bacteria against cadmium-induced genotoxicity in rat hepatocytes in vivo and in vitro. Arch Biol Sci. 2012;64(3):1197-206.
14. Afshar P, Shokrzadeh M, Raeisi SN, Ghorbani-HasanSaraei A, Nasiraii LR. Aflatoxins biodetoxification strategies based on
probiotic bacteria. Toxicon. 2020;178:50-8.
15. Walia S, Sood S, Kanwar SS. Exhibition of DNA-bioprotective activity by microflora of traditional fermented foods of North-Western Himalayas. Food Res Int. 2014;55(1):176-80.
16. Ghotbi M, Zenoozian MS, Roozbehnasiraii L. Survival of free and microencapsulated Lb. rhamnosus LC705 in simulated gastric and intestinal juice. Int J Biol Sci. 2015;6(3):330-7.
17. Deabes MM, Darwish HR, Abdel-Aziz KB, Farag IM, Nada SA, Tawfek NS. Protective effects of Lactobacillus rhamnosus GG on Aflatoxins-induced toxicities in male Albino Mice. J Environ Anal Toxicol. 2012;2(3):2-9.
18. Tukenmez U, Aktas B, Aslim B, Yavuz S. The relationship between the structural characteristics of lactobacilli-EPS and its ability to induce apoptosis in colon cancer cells in vitro. Sci Rep. 2019;9(1):8268.
19. Salehipour Z, Haghmorad D, Sankian M, Rastin M, Nosratabadi R, Dallal MM, et al. Bifidobacterium animalis in combination with human origin of Lactobacillus plantarum ameliorate neuroinflammation in experimental model of multiple sclerosis by altering CD4+ T cell subset balance. Biomed Pharmacother. 2017;95:1535-48.
20. Aryani DC, Den Besten HM, Zwietering MH. Quantifying variability in growth and thermal inactivation kinetics of Lactobacillus plantarum. Appl Environ Microbiol. 2016; 82(16):4896-908.
21. Shokrzadeh M, Emami S, Amirzadeh M, Modanloo M. Cytotoxic Effects of Dibromoisatin Derivatives on Hela and HepG2 Cell Lines Using MTT Assay. J Mazand Univ Med Sci. 2017;15(151):24-31.
22. Shokrzadeh M, Tasdighi A, Modanlo M, Shaki F. Evaluating and comparing genotoxicity mechanism of copper nanoparticles and copper sulfate. J Mazand Univ Med Sci. 2018;28(163):1-9.
23. Pizzolitto RP, Bueno DJ, Armando MR, Cavaglieri L, Dalcero AM, Salvano MA. Binding of aflatoxin B1 to lactic acid bacteria and Saccharomyces cerevisiae in vitro: a useful model to determine the most efficient microorganism. Aflatoxins-biochemistry and molecular biology.London:IntechOpen; 2011.
24. Gratz S, Täubel M, Juvonen R, Viluksela M, Turner P, Mykkänen H, et al. Lactobacillus rhamnosus strain GG modulates intestinal absorption, fecal excretion, and toxicity of aflatoxin B1 in rats. Appl Environ Microbiol. 2006;72(11): 7398-400.
25. Kirsch-Volders M, Decordier I, Elhajouji A, Plas G, Aardema MJ, Fenech M. In vitro genotoxicity testing using the micronucleus assay in cell lines, human lymphocytes and 3D human skin models. Mutagenesis. 2011;26(1):177-84.
26. Akyıl D, Özkara A, Erdoğmuş SF, Eren Y, Konuk M, Sağlam E. Micronucleus assay in human lymphocytes after exposure to alloxydim sodium herbicide in vitro. Cytotechnology. 2015;67(6):1059-66.
27. Corcuera LA, Vettorazzi A, Arbillaga L, Pérez N, Gil AG, Azqueta A, et al. Genotoxicity of Aflatoxin B1 and Ochratoxin A after simultaneous application of the in vivo micronucleus and comet assay. Food Chem Toxicol. 2015;76(1):116-24.
28. Türkez H, Şişman T. Anti-genotoxic effect of hydrated sodium calcium aluminosilicate on genotoxicity to human lymphocytes induced by aflatoxin B1. Toxicol Ind Health. 2007;23(2):83-9.
29. Ahmad S, Hoda A, Afzal M. Additive action of vitamins C and E against hydrocortisone-induced genotoxicity in human lymphocyte chromosomes. Int J Vitam Nutr Res. 2002; 72(4):204-9.
30. Peng X, Zhang S, Fang J, Cui H, Zuo Z, Deng J. Protective roles of sodium selenite against aflatoxin B1-induced apoptosis of jejunum in broilers. Int J Environ Res. 2014;11(12):13130-43.
31. Chen K, Shu G, Peng X, Fang J, Cui H, Chen J, et al. Protective role of sodium selenite on histopathological lesions, decreased T-cell subsets and increased apoptosis of thymus in broilers intoxicated with aflatoxin B1. Food Chem Toxicol. 2013; 59(1):446-54.
32. Jebali R, Salah-Abbès JB, Abbès S, Hassan AM, Abdel-Aziem SH, El-Nekeety AA, et al. Lactobacillus plantarum alleviate aflatoxins (B1 and M1) induced disturbances in the intestinal genes expression and DNA fragmentation in mice. Toxicon. 2018;146:13-23.
33. Huang L, Duan C, Zhao Y, Gao L, Niu C, Xu J, et al. Reduction of aflatoxin B1 toxicity by Lactobacillus plantarum C88: a potential probiotic strain isolated from Chinese traditional fermented food “Tofu”. PloS One. 2017;12(1):e0170109.
34. Prete R, Tofalo R, Federici E, Ciarrocchi A, Cenci G, Corsetti A. Food-associated Lactobacillus plantarum and yeasts inhibit the genotoxic effect of 4-nitroquinoline-1-oxide. Front Microbiol. 2017;8:2349.
35. Mohamed WF, Rabeea O, Abu Shady H, Youssef HA. Combining prebiotic and mixture of freeze dried probiotics to develop synbiotics that induce apoptosis in human colon tumor
cells. Egypt J Microbiol. 2017;52(1):101-11.
36. Adesulu-Dahunsi AT, Jeyaram K, Sanni AI, Banwo K. Production of exopolysaccharide by strains of Lactobacillus plantarum YO175 and OF101 isolated from traditional fermented cereal beverage. PeerJ. 2018;6:e5326.
37. Zhang L, Liu C, Li D, Zhao Y, Zhang X, Zeng X, et al. Antioxidant activity of an exopolysaccharide isolated from Lactobacillus plantarum C88. Int J Biol Macromol. 2013; 54(1):270-5.