توکسین زدایی آفلاتوکسین B1 توسط لاکتوباسیلوس رامنوسوس در مدل شبیه‌سازی شده دستگاه گوارش انسان
محورهای موضوعی : میکروب شناسی غذایی
1 - استادیار، گروه بهداشت و ایمنی مواد غذایی، دانشکده بهداشت، دانشگاه علوم پزشکی فسا، فارس، ایران
2 - استاد، گروه بهداشت موادغذایی، دانشکده دامپزشکی، دانشگاه ارومیه، ارومیه، ایران
کلید واژه: دستگاه گوارش, HPLC, آفلاتوکسین B1, سم‌زدایی, لاکتوباسیلوس رامنوسوس,
چکیده مقاله :
سابقه و هدف: بر اساس گزارش سازمان فائو سالیانه 10 درصد از محصولات غذایی تولید شده در دنیا توسط سموم قارچی آلوده می‌شوند که در این آلودگی آفلاتوکسین‌ها سهم بیشتری نسبت به سایر سموم دارند. این پژوهش با هدف ارزیابی توانایی اتصال آفلاتوکسین به لاکتوباسیلوس رامنوسوس زیرگونهGG در محیط شبیه‌سازی دستگاه گوارش انسان حاوی شیراستریلیزه به منظور کاهش سمیت آفلاتوکسینB1 انجام شد.مواد و روش‌ها: تعداد باکتری cfu/ml 1010×1 و غلظت آفلاتوکسین 5 میکروگرم بر میلی لیتر در نظر گرفته شد که در محیط شبیه سازی شده ترشحات مصنوعی دستگاه گوارش انسان تلقیح گردید. در این مطالعه 6 گروه تیمار در حضور و غیاب باکتری، شیراستریلیزه، سوسپانسیون شیره دستگاه گوارش مورد بررسی قرار گرفت. غلظت آفلاتوکسین باقیمانده توسط کروماتوگرافی مایع با کارایی بالا (HPLC) و تخلیص با ستون ایمونوافینیتی تعیین گردید.یافته ها: کاهش آفلاتوکسین B1 در تمام تیمارها با استفاده از کروماتوگرافی مایع با کارایی بالا با حد تشخیص 0.25 میکروگرم بر میلی‌لیتر و حد تعیین کمی 0.75 میکروگرم بر میلی‌لیتر اندازه‌گیری شد. مقادیر بازیافت مایکوتوکسین AFB1 بین 89 تا 94 درصد بود. منحنی درجه بندی آفلاتوکسین B1 با ضریب همبستگی 0.995 در گستره غلظتی 1 تا 10 نانوگرم بر میلی لیتر خطی بود. بیشترین و کمترین میانگین درصد حذف آفلاتوکسین B1 به ترتیب مربوط به تیمار5 و 1 ( 0.018 ±58.8 و 0.017± 13.86) بود که تفاوت معنی‌‌داری در بین شش گروه وجود داشت.نتیجه گیری: نتایج نشان داد که علاوه بر باکتری لاکتوباسیلوس رامنوسوس زیر گونه GG، شیره معده و روده کوچک نیز در کاهش یا حذف آفلاتوکسین B1 موثر می‌‌باشند.
Background & Objectives: According to the FAO annual report, 10 percent of the world's food products are contaminated with fungal toxins, among which aflatoxins have the most contribution as compared to others. This research was aimed to assess the ability of Lactobacillus rhamnosus strain GG in the reduction of aflatoxin B1 (AFB1) in a simulated human gastrointestinal tract containing sterilized milk. Materials & Methods: For this purpose, the bacterial count and aflatoxin concentration were adjusted to 1×1010 cfu/ml and 5 ppm, respectively, where artificial gastrointestinal tract discharges were inoculated in the simulated environment. In this study 6 treatment groups were assessed in the presence and absence of bacteria, sterilized milk, and gastrointestinal juice suspension. The concentration of residual aflatoxin was determined using high-performance liquid chromatography (HPLC) and purification by Immunoaffinity column. Results: The reduction of aflatoxin B1 at all treatments was determined using HPLC with a detection limit of 0.25 mg/ml and a quantification limit of 0.75 mg/ml. The mycotoxin recovery rate was between 89% and 94% for AFB1. The aflatoxin B1 calibration curve with a correlation coefficient of 0.95 was linear in the concentration range of 1 to 10 ng/ml. The highest and lowest average removal percentage of aflatoxin B1 was observed for 5 and 1 treatments (58.8 ± 0.018 and 13.86 ± 0.017) respectively, where a significant difference in removal percentage was observed among six treatment groups. Conclusion: The results indicated that beside L. rhamnosus strain GG, gastric and small intestine juices are suitable to eliminate or reduce aflatoxin B1, as well.
Agriculture. 2016; 2(1): 119-125.
2. Cheraghali A, Yazdanpanah H, Doraki N, Abouhossain G, Hassibi M, Ali-Abadi S. Incidence
of aflatoxins in Iran pistachio nuts. Food Chem Toxicol. 2007; 45(5): 812-816.
3. Abdulkadar A, Al-Ali A, Al-Jedah J. Aflatoxin contamination in edible nuts imported in Qatar.
Food Control. 2000; 11(2): 157-160.
4. Udomkun P, Wiredu AN, Nagle M, Müller J, Vanlauwe B, Bandyopadhyay R. Innovative
technologies to manage aflatoxins in foods and feeds and the profitability of application– A
review. Food Control. 2017; 76(1): 127-138.
5. Zain ME. Impact of mycotoxins on humans and animals. J Sau Chemi Soci. 2011; 15(2):
129-144.
6. Kumar KS, Sastry N, Polaki H, Mishra V. Colon cancer prevention through probiotics: an
overview. J Cancer Sci Ther. 2015; 7(2): 81-92.
7. Berg T. How to establish international limits for mycotoxins in food and feed? Food Control.
2003; 14(4): 219-224.
8. Institute of Standards and Industrial Research of Iran. Maximum validity Maycotoxins in
human food. ISIRI no 7133. ISIRI; 1992 [In Persian].
9. Park DL, Ayala CE, Guzman-Perez SE, Lopez-Garcia R, Trujillo S. Microbial toxins in foods:
algal, fungal, and bacterial: CRC Press, Boca Raton, FL; 2000.
10. El-Nezami H, Kankaanpaa P, Salminen S, Ahokas J. Ability of dairy strains of lactic acid
bacteria to bind a common food carcinogen, aflatoxin B1. Food Chem Toxicol. 1998; 36(4):
321-326.
11. Salim A-B, Zohair A, Hegazy AE-S, Said A. Effect of some strains of probiotic bacteria
against toxicity induced by aflatoxins in vivo. J Am Sci. 2011; 7(1): 1-12.
12. Kabak B, Ozbey F. Aflatoxin M1 in UHT milk consumed in Turkey and first assessment of its
bio-accessibility using an in vitro digestion model. Food Control. 2012; 28(2): 338-344.
13. Panwar R, Kumar N, Kashyap V, Ram C, Kapila R. Aflatoxin M 1 detoxification ability of
probiotic lactobacilli of Indian origin in In vitro digestion model. Probiotics Antimicrob
Proteins. 2018: 1: 1-10.
14. Martínez MP, González Pereyra ML, Fernandez Juri MG, Poloni V, Cavaglieri L. Probiotic
characteristics and aflatoxin B1 binding ability of Debaryomyces hansenii and Kazaschtania
exigua from rainbow trout environment. Aquacult Res. 2018; 49(4): 1588-1597.
15. Peltonen K, El-Nezami H, Haskard C, Ahokas J, Salminen S. Aflatoxin B1 binding by dairy
strains of lactic acid bacteria and bifidobacteria. Int Dairy J. 2001; 84(10): 2152-2156.
16. Saladino F, Posarelli E, Luz C, Luciano F, Rodriguez-Estrada M, Mañes J. Influence of
probiotic microorganisms on aflatoxins B1 and B2 bioaccessibility evaluated with a simulated
gastrointestinal digestion. J Food Compost Anal. 2018; 68: 128-132.
17. Damayanti E, Istiqomah L, Saragih JE, Purwoko T. Characterization of lactic acid bacteria as
poultry probiotic candidates with aflatoxin B1 binding activities. Earth Environ Sci. 2017; 101:
120-130.
18. Liu N, Wang J, Deng Q, Gu K, Wang J. Detoxification of aflatoxin B1 by lactic acid bacteria
and hydrated sodium calcium aluminosilicate in broiler chickens. Livest Sci. 2018; 208: 28-32.
19. Hojsa nova A dović ajews a H iša olače . Lactobacillus GG in the
prevention of gastrointestinal and respiratory tract infections in children who attend day care
centers: a randomized, double-blind, placebo-controlled trial. Clin Nutr. 2010; 29(3): 312-316.
20. Assaf JC, Atoui A, Khoury AE, Chokr A, Louka N. A comparative study of procedures for
binding of aflatoxin M1 to Lactobacillus rhamnosus GG. Braz J Microbiol. 2018; 49(1):
120-127.
21. Martin A, Cubillos-Ruiz A, Von Groll A, Del Portillo P, Portaels F, Palomino JC. Nitrate
reductase assay for the rapid detection of pyrazinamide resistance in Mycobacterium
tuberculosis using nicotinamide. J Antimicrob Chemother. 2007; 61(1): 123-127.
22. Haddadin M. Effect of olive leaf extracts on the growth and metabolism of two probiotic
bacteria of intestinal origin. Pakistan J Nutr. 2010; 9(8): 787-793.
23. De Palencia PF, López P, Corbí AL, Peláez C, Requena T. Probiotic strains: survival under
simulated gastrointestinal conditions, in vitro adhesion to Caco-2 cells and effect on cytokine
secretion. Eur Food Res Technol. 2008; 227(5): 1475-1484.
24. Neeff D, Ledoux D, Rottinghaus G, Bermudez A, Dakovic A, Murarolli R, Oliveira C. In
vitro and in vivo efficacy of a hydrated sodium calcium aluminosilicate to bind and reduce
aflatoxin residues in tissues of broiler chicks fed aflatoxin B1. Poult Sci. 2013; 92(1): 131-137.
25. Bognanno M, La Fauci L, Ritieni A, Tafuri A, De Lorenzo A, Micari P, Di Renzo L,
Ciappellano S, Sarullo, V. Survey of the occurrence of Aflatoxin M1 in ovine milk by HPLC
and its confirmation by MS. Mol Nutr Food Res. 2006; 50(3): 300-305.
26. Association of official analytical chemists. Official methods of association of official analytical chemists (17th Ed.). Gaithersburg, MD: AOAC; 2000.
27. Wei R, Qiu F, Kong W, Wei J, Yang M, Luo Z. Co-occurrence of aflatoxin B1, B2, G1, G2
and ochrotoxin A in Glycyrrhiza uralensis analyzed by HPLC-MS/MS. Food Control. 2013; 32
(1): 216-221.
28. Peltonen KD, El‐Nezami HS, Salminen SJ, Ahokas JT. Binding of aflatoxin B1 by probiotic
bacteria. J Sci Food Agric. 2000; 80(13): 1942-1945.
29. Nurul Adilah Z, Liew WP, Mohd Redzwan S, Amin I. Effect of high protein diet and
probiotic Lactobacillus casei shirota supplementation in aflatoxin B1-induced rats. Bio Med
Res Int. 2018; 2018.
30. Ismail A, Levin RE, Riaz M, Akhtar S, Gong YY, de Oliveira CA. Effect of different
microbial concentrations on binding of aflatoxin M1 and stability testing. Food Control.
2017; 73: 492-496.
31. Hernandez-Mendoza A, Garcia H, Steele J. Screening of Lactobacillus casei strains for their
ability to bind aflatoxin B1. Food Chem Toxicol. 2009; 47(6): 1064-1068.
32. Kaak B, Var I. Binding of aflatoxin M1 by Lactobacillus and Bifidobacterium strains.
Milchwissenschaft. 2004; 59(5-6): 301-303.
33. Marrez DA, Shahy EM, El-Sayed HS, Sultan YY. Detoxification of aflatoxin B1 in milk
using lactic acid bacteria. J Biol Sci. 2018; 18(3): 144-151.
34. El Khoury A, Atoui A, Yaghi J. Analysis of aflatoxin M1 in milk and yogurt and AFM1
reduction by lactic acid bacteria used in Lebanese industry. Food Control. 2011; 22(10):
1695-1699.
35. Serrano-Niño J, Cavazos-Garduño A, Hernandez-Mendoza A, Applegate B, Ferruzzi M, San
Martin-González M. Assessment of probiotic strains ability to reduce the bio-accessibility of
aflatoxin M1 in artificially contaminated milk using an in vitro digestive model. Food
Control. 2013; 31(1): 202-207.
36. Oozeer R, Leplingard A, Mater DD, Mogenet A, Michelin R, Seksek I, Marteau P, Dore J,
Bresson J, Corthier G. Survival of Lactobacillus casei in the human digestive tract after
consumption of fermented milk. J Appl Environ Microbiol. 2006; 72(8): 5615-5627.
37. Kheadr EE. Impact of acid and oxgall on antibiotic susceptibility of probiotic lactobacilli.
Afr J Agric Res. 2006; 1(5): 172-181.
38. Madureira A, Pereira C, Truszkowska K, Gomes A, Pintado M, Malcata F. Survival of
probiotic bacteria in a whey cheese vector submitted to environmental conditions prevailing
in the gastrointestinal tract. Int Dairy J. 2005; 15(6-9): 921-927.
39. Pacheco KC, del Toro GV, Martínez FR, Duran-Paramo E. Viability of Lactobacillus
delbrueckii under human gastrointestinal conditions simulated in vitro. Am J Agric Biol Sci.
2010; 5: 37-42.
40. Shetty PH, Hald B, Jespersen L. Surface binding of aflatoxin B1 by Saccharomyces
cerevisiae strains with potential decontaminating abilities in indigenous fermented foods. Int
J Food Microbiol. 2007; 113(1): 41-46.
41. Haskard C, Binnion C, Ahokas J. Factors affecting the sequestration of aflatoxin by
Lactobacillus rhamnosus strain GG. Chem Biol Interact. 2000; 128(1): 39-49.
_||_
Agriculture. 2016; 2(1): 119-125.
2. Cheraghali A, Yazdanpanah H, Doraki N, Abouhossain G, Hassibi M, Ali-Abadi S. Incidence
of aflatoxins in Iran pistachio nuts. Food Chem Toxicol. 2007; 45(5): 812-816.
3. Abdulkadar A, Al-Ali A, Al-Jedah J. Aflatoxin contamination in edible nuts imported in Qatar.
Food Control. 2000; 11(2): 157-160.
4. Udomkun P, Wiredu AN, Nagle M, Müller J, Vanlauwe B, Bandyopadhyay R. Innovative
technologies to manage aflatoxins in foods and feeds and the profitability of application– A
review. Food Control. 2017; 76(1): 127-138.
5. Zain ME. Impact of mycotoxins on humans and animals. J Sau Chemi Soci. 2011; 15(2):
129-144.
6. Kumar KS, Sastry N, Polaki H, Mishra V. Colon cancer prevention through probiotics: an
overview. J Cancer Sci Ther. 2015; 7(2): 81-92.
7. Berg T. How to establish international limits for mycotoxins in food and feed? Food Control.
2003; 14(4): 219-224.
8. Institute of Standards and Industrial Research of Iran. Maximum validity Maycotoxins in
human food. ISIRI no 7133. ISIRI; 1992 [In Persian].
9. Park DL, Ayala CE, Guzman-Perez SE, Lopez-Garcia R, Trujillo S. Microbial toxins in foods:
algal, fungal, and bacterial: CRC Press, Boca Raton, FL; 2000.
10. El-Nezami H, Kankaanpaa P, Salminen S, Ahokas J. Ability of dairy strains of lactic acid
bacteria to bind a common food carcinogen, aflatoxin B1. Food Chem Toxicol. 1998; 36(4):
321-326.
11. Salim A-B, Zohair A, Hegazy AE-S, Said A. Effect of some strains of probiotic bacteria
against toxicity induced by aflatoxins in vivo. J Am Sci. 2011; 7(1): 1-12.
12. Kabak B, Ozbey F. Aflatoxin M1 in UHT milk consumed in Turkey and first assessment of its
bio-accessibility using an in vitro digestion model. Food Control. 2012; 28(2): 338-344.
13. Panwar R, Kumar N, Kashyap V, Ram C, Kapila R. Aflatoxin M 1 detoxification ability of
probiotic lactobacilli of Indian origin in In vitro digestion model. Probiotics Antimicrob
Proteins. 2018: 1: 1-10.
14. Martínez MP, González Pereyra ML, Fernandez Juri MG, Poloni V, Cavaglieri L. Probiotic
characteristics and aflatoxin B1 binding ability of Debaryomyces hansenii and Kazaschtania
exigua from rainbow trout environment. Aquacult Res. 2018; 49(4): 1588-1597.
15. Peltonen K, El-Nezami H, Haskard C, Ahokas J, Salminen S. Aflatoxin B1 binding by dairy
strains of lactic acid bacteria and bifidobacteria. Int Dairy J. 2001; 84(10): 2152-2156.
16. Saladino F, Posarelli E, Luz C, Luciano F, Rodriguez-Estrada M, Mañes J. Influence of
probiotic microorganisms on aflatoxins B1 and B2 bioaccessibility evaluated with a simulated
gastrointestinal digestion. J Food Compost Anal. 2018; 68: 128-132.
17. Damayanti E, Istiqomah L, Saragih JE, Purwoko T. Characterization of lactic acid bacteria as
poultry probiotic candidates with aflatoxin B1 binding activities. Earth Environ Sci. 2017; 101:
120-130.
18. Liu N, Wang J, Deng Q, Gu K, Wang J. Detoxification of aflatoxin B1 by lactic acid bacteria
and hydrated sodium calcium aluminosilicate in broiler chickens. Livest Sci. 2018; 208: 28-32.
19. Hojsa nova A dović ajews a H iša olače . Lactobacillus GG in the
prevention of gastrointestinal and respiratory tract infections in children who attend day care
centers: a randomized, double-blind, placebo-controlled trial. Clin Nutr. 2010; 29(3): 312-316.
20. Assaf JC, Atoui A, Khoury AE, Chokr A, Louka N. A comparative study of procedures for
binding of aflatoxin M1 to Lactobacillus rhamnosus GG. Braz J Microbiol. 2018; 49(1):
120-127.
21. Martin A, Cubillos-Ruiz A, Von Groll A, Del Portillo P, Portaels F, Palomino JC. Nitrate
reductase assay for the rapid detection of pyrazinamide resistance in Mycobacterium
tuberculosis using nicotinamide. J Antimicrob Chemother. 2007; 61(1): 123-127.
22. Haddadin M. Effect of olive leaf extracts on the growth and metabolism of two probiotic
bacteria of intestinal origin. Pakistan J Nutr. 2010; 9(8): 787-793.
23. De Palencia PF, López P, Corbí AL, Peláez C, Requena T. Probiotic strains: survival under
simulated gastrointestinal conditions, in vitro adhesion to Caco-2 cells and effect on cytokine
secretion. Eur Food Res Technol. 2008; 227(5): 1475-1484.
24. Neeff D, Ledoux D, Rottinghaus G, Bermudez A, Dakovic A, Murarolli R, Oliveira C. In
vitro and in vivo efficacy of a hydrated sodium calcium aluminosilicate to bind and reduce
aflatoxin residues in tissues of broiler chicks fed aflatoxin B1. Poult Sci. 2013; 92(1): 131-137.
25. Bognanno M, La Fauci L, Ritieni A, Tafuri A, De Lorenzo A, Micari P, Di Renzo L,
Ciappellano S, Sarullo, V. Survey of the occurrence of Aflatoxin M1 in ovine milk by HPLC
and its confirmation by MS. Mol Nutr Food Res. 2006; 50(3): 300-305.
26. Association of official analytical chemists. Official methods of association of official analytical chemists (17th Ed.). Gaithersburg, MD: AOAC; 2000.
27. Wei R, Qiu F, Kong W, Wei J, Yang M, Luo Z. Co-occurrence of aflatoxin B1, B2, G1, G2
and ochrotoxin A in Glycyrrhiza uralensis analyzed by HPLC-MS/MS. Food Control. 2013; 32
(1): 216-221.
28. Peltonen KD, El‐Nezami HS, Salminen SJ, Ahokas JT. Binding of aflatoxin B1 by probiotic
bacteria. J Sci Food Agric. 2000; 80(13): 1942-1945.
29. Nurul Adilah Z, Liew WP, Mohd Redzwan S, Amin I. Effect of high protein diet and
probiotic Lactobacillus casei shirota supplementation in aflatoxin B1-induced rats. Bio Med
Res Int. 2018; 2018.
30. Ismail A, Levin RE, Riaz M, Akhtar S, Gong YY, de Oliveira CA. Effect of different
microbial concentrations on binding of aflatoxin M1 and stability testing. Food Control.
2017; 73: 492-496.
31. Hernandez-Mendoza A, Garcia H, Steele J. Screening of Lactobacillus casei strains for their
ability to bind aflatoxin B1. Food Chem Toxicol. 2009; 47(6): 1064-1068.
32. Kaak B, Var I. Binding of aflatoxin M1 by Lactobacillus and Bifidobacterium strains.
Milchwissenschaft. 2004; 59(5-6): 301-303.
33. Marrez DA, Shahy EM, El-Sayed HS, Sultan YY. Detoxification of aflatoxin B1 in milk
using lactic acid bacteria. J Biol Sci. 2018; 18(3): 144-151.
34. El Khoury A, Atoui A, Yaghi J. Analysis of aflatoxin M1 in milk and yogurt and AFM1
reduction by lactic acid bacteria used in Lebanese industry. Food Control. 2011; 22(10):
1695-1699.
35. Serrano-Niño J, Cavazos-Garduño A, Hernandez-Mendoza A, Applegate B, Ferruzzi M, San
Martin-González M. Assessment of probiotic strains ability to reduce the bio-accessibility of
aflatoxin M1 in artificially contaminated milk using an in vitro digestive model. Food
Control. 2013; 31(1): 202-207.
36. Oozeer R, Leplingard A, Mater DD, Mogenet A, Michelin R, Seksek I, Marteau P, Dore J,
Bresson J, Corthier G. Survival of Lactobacillus casei in the human digestive tract after
consumption of fermented milk. J Appl Environ Microbiol. 2006; 72(8): 5615-5627.
37. Kheadr EE. Impact of acid and oxgall on antibiotic susceptibility of probiotic lactobacilli.
Afr J Agric Res. 2006; 1(5): 172-181.
38. Madureira A, Pereira C, Truszkowska K, Gomes A, Pintado M, Malcata F. Survival of
probiotic bacteria in a whey cheese vector submitted to environmental conditions prevailing
in the gastrointestinal tract. Int Dairy J. 2005; 15(6-9): 921-927.
39. Pacheco KC, del Toro GV, Martínez FR, Duran-Paramo E. Viability of Lactobacillus
delbrueckii under human gastrointestinal conditions simulated in vitro. Am J Agric Biol Sci.
2010; 5: 37-42.
40. Shetty PH, Hald B, Jespersen L. Surface binding of aflatoxin B1 by Saccharomyces
cerevisiae strains with potential decontaminating abilities in indigenous fermented foods. Int
J Food Microbiol. 2007; 113(1): 41-46.
41. Haskard C, Binnion C, Ahokas J. Factors affecting the sequestration of aflatoxin by
Lactobacillus rhamnosus strain GG. Chem Biol Interact. 2000; 128(1): 39-49.
