Close

Not a member yet? Register now and get started.

lock and key

Sign in to your account.

Account Login

Forgot your password?

Lowering Effect of Selenium and Yogurt on Nuts Contaminated With Aflatoxins Induced Hepatotoxicity in Rats

Amnah M. A. Alsuhaibani

Nutrition and Food Sciences Dept, Home Economic College, Nourah Bint, Abdulrahman University, Riyadh, Saudi Arabia

Corresponding author Email: amalsuhaibani@pnu.edu.sa

ABSTRACT:

Nuts which contaminated with aflatoxins are potent to hepatotoxic and hepatocarcinogenic agents. Herein, we were assessed the ability of selenium and yogurt to ameliorate aflatoxin-contaminated nut-induced hepatotoxicity in experimental rats. Relative to the control group, the aflatoxin-contaminated nut-fed rats has been reduced body weight gain and feed efficiency ratio (FER), whereas those rats given selenium or yogurt, or both, and consumed 3% aflatoxin-contaminated nuts showed no significant decrease in body weight gain or decrease in FER. Food intake did not vary significantly between the groups. After 60 days, alanine and aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyltransferase activities were increased in the serum of rats fed aflatoxin-contaminated nuts, suggesting hepatic damage. The 3% aflatoxin-contaminated nut-fed group has been reduced total protein and serum, liver glutathione peroxidase and superoxide dismutase(GPX and SOD) enzymes but elevated creatinine, urea, uric acid, bilirubin and malondialdehyde (MDA) levels, as well as liver MDA, compared to the control group. Moreover, we were found that feeding of the rats by selenium, yogurt or both could be normalize of liver and antioxidant enzyme levels (GPX, SOD, and MDA), as well as total protein, albumin, globulin, and uric acid contents. Based on our findings, we were proposed that selenium and yogurt could reduce the side effects of hepatotoxicity in experimental rats that have consumed aflatoxin-contaminated nuts.

KEYWORDS:

Nuts; aflatoxins; antioxidants enzymes; selenium; yogurt; liver; rats



Copy the following to cite this article:

Alsuhaibani A. M. A. Lowering Effect of Selenium and Yogurt on Nuts Contaminated With Aflatoxins Induced Hepatotoxicity in Rats. Curr Res Nutr Food Sci 2018;6(1).


Copy the following to cite this URL:

Alsuhaibani A. M. A. Lowering Effect of Selenium and Yogurt on Nuts Contaminated With Aflatoxins Induced Hepatotoxicity in Rats. Curr Res Nutr Food Sci 2018;6(1). http://www.foodandnutritionjournal.org/?p=4867


Introduction

Nuts are rich in an unsaturated fatty acids and various bioactive compounds, which have been including high-quality vegetable protein, minerals, fiber, phytosterols, tocopherols and phenolic compounds. Almonds (Prunusamigdalis), hazelnuts (Corylusavellana), walnuts (Juglansregia), caju (Anacardiumoccidentale), and pistachios (Pistachiavera) are among popularly consumed edible tree nuts.1,2

Fungi have growing on nuts which were formed the mycotoxin and can be decrease the quality of nuts by reducing their nutritive quality.3 Both of the mycotoxins and aflatoxin have been formed by the molds Aspergillus flavus and Aspergillus parasiticus. Blue 1 (B1), B2, Green 1 (G1), and G2 are the most common aflatoxin strains that were found in foods, thus named because of their fluorescence properties and chromatography patterns4. Optimal aflatoxin production occurs at temperatures nearly to 30 °C.5

Mammals convert aflatoxins into the  M1 and M2 metabolites, which are carcinogenic.6,7Aflatoxin contamination usually occurs either by slowly acquiring aflatoxin over time in smaller quantities or by consuming large amounts at once. Aflatoxin contamination can lead to a variety of health problems, including cancers, mental and digestive problems, hemorrhages, and malabsorption.8

Aflatoxins reduce growth and suppress immune functions in animals. Aflatoxin induces hepatic and renal tumors in rodents, and has been implicated in esophageal cancer. Humans are exposed to mycotoxins through the consumption of contaminated foods, as well as by exposure to dust and air containing these toxins. Aflatoxins are hepatocarcinogenic, predominantly in conjunction with chronic hepatitis B virus infection, and form aflatoxicosis in episodic poisoning outbreaks.9,10

Here we have been evaluated the potential for yogurt and selenium to ameliorate aflatoxin-contaminated nut-induced hepatotoxicity in rats.

Materials and Methods

Nuts

Five kilograms of commonly consumed nuts (pistachio, caju, walnut, almond, and hazelnut) were obtained from a local market of Riyadh in the Kingdom of Saudi Arabia. These nuts are more susceptible to mold growth.

Reagents

All the materials used for this experiment were of analytical grade and used without further purifications. BioMeriuex Kits were purchased from Alkan Co. for Chemicals and Biodiagnostics. Selenium was obtained from Sigma-Aldrich.

Probiotic Bacteria

Lactobacillus delbrueckiisubspeciesbulgaricusCH-2 and Streptococcus thermophilus ST-36 were obtained from the HansenLab (University of Denmark).

Experimental animals

Fifty adult male Sprague-Dawley strain (albino) rats, weighs within 130–140g, were provided by the experimental animals of the center in Research Center in Prince Sultan Military Medical City, Riyadh. Rats were housed as groups in wire cages under laboratory conditions and, during a 1-week adaption period, were fed a standard diet. Food and water were provided adlibitum. Animal management ethical guidelines were followed throughout the study and permission was obtained from the concerned department.

Standard diet

The standard experimental diet was composed of corn starch(598 g kg-1), casein (200 g kg-1), soybean oil (100 g kg-1), vitamin mixture (10 g kg-1), salts mixture (40 g kg-1), cellulose (50 g kg-1), and choline chloride (2 g kg-1), according to Ref.11.

Preparation of ordinary yogurt

L.delbrueckii subspecies bulgaricus CH-2 was cultured in MRS broth at 37 °C for 24 h. S. thermophilus ST-36 was grown in M17 broth at 40 °C for 24 h. Whole milk was heated to boiling temperature to reduce its volume by approximately 20% after cooling;this milk was then heated to 90 °C for 5 min and then cooled to 42°C and inoculated with 1% of L.delbrueckii sub-species bulgaricus CH-2 and S. thermophilusST-36, then incubated at 40°C until coagulation (about 4 h).12

Nuts storage and quantification of aflatoxin content

Raw nuts were stored in glass dishes at 25 oC and 60% relative humidity for 6 months. Then, these nuts were crushed to estimate their aflatoxins content, as previously described.13,14 The total aflatoxins were 23.25, 23.66, 22.07, 26.02 and 28.6μgkg-1 in the pistachios, caju, walnuts, almonds, and hazelnuts, respectively. Mixed crushed nuts were added as 3%(w/w) to the standard diet, with consideration of the nutritional value of the nuts.

Treatment schedule

Rats were divided into five groups, 10 rats per group, as follows: (1)  normal control group fed the standard diet; (2) positive control group fed the standard diet plus aflatoxin-contaminated nuts to induce hepatotoxicity, as reported in previous work;9,10 (3) selenium group fed the standard diet plus aflatoxin-contaminated nuts plus selenium by stomach tube (3 mg kg-1 body weight); (4) yogurt group, fed the standard diet plus aflatoxin-contaminated nuts plus yogurt by stomach tube (160 mlkg-1 body weight); (5) yogurt plus selenium group, fed the standard diet plus aflatoxin-contaminated nuts plus selenium and yogurt by stomach tube (3 mg kg-1 and 160 mlkg-1 body weight, respectively).

The daily food intake and weekly body weight of the rats were recorded. The feed efficiency ratio (FER) was calculated using the method of Ref.15 After completion of the experimental period (60 days), rats were fasted overnight and sacrificed to obtain blood and liver samples for biochemical analyses.

Serum analyses

From these samples, we quantified the serum alanine and aspartate aminotransferase (ALT +AST), alkaline phosphatase (ALP), and gamma-glutamyltransferase (γGT) enzyme activities. Also, serum total protein,  globulin, albumin, creatinine, uric acid, urea, and bilirubin contents were enzymatically determined as mentioned in the method of Ref.16 The activity of glutathione peroxidase (GPX), malondialdehyde (MDA), and superoxide dismutase(SOD) enzymes were determined as previously described17–19.

Liver biochemical estimations

Livers of rats were rapidly removed and parts of them perfused with 50 to 100 ml of ice-cold 0.9% NaCl solution for estimation of the liver glutathione peroxidase (GPX), superoxide dismutase (SOD) and malondialdehyde (MDA) activities according to Refs.18,20,21

Statistical analysis

Data were subjected to ANOVA. Comparison of means was performed using Duncan’s multiple-range test with level of significance 0.05, complemented by Kruskal-Wallis correlation method to analyze correlations between parameters at significance levels of 0.05.

Results and Discussion

Herein, we found that rats fed aflatoxin-contaminated nuts have been decreased the body weight gain and FER, but that selenium or yogurt, or both, could ameliorate these effects. Food intake did not vary among the five test groups (Table 1). It has been well-documented nut contamination with aflatoxins (from A. flavusand A. parasiticus) is a major health concern, especially in hot and humid regions. Under conventional storage conditions, aflatoxin-producing molds are able to grow exponentially.23Aflatoxins have a low molecular weight therefore quickly absorbed in the gastrointestinal tract and appear as metabolites in theblood.24,25 Following exposure to aflatoxins, they can be detected covalently bound to DNA, thereby reducing protein synthesis; this effect can be persist for up to 5 days. Also, animals exposed to aflatoxin are less efficient at food use and dietary animals exposed to aflatoxin have a reduced growth rate and productivity. These effects are likely due to an increased degradation of lipids and proteins.26,27

Co-administration of selenium and yogurt has made improvement in the nutritional results. Indeed, Navarro-Alarcon and Cabrera-Vique have been detected the increasing in growth and development of the selenium supplementation.28 Fermented milk and yogurt are considered safe and nutritious. Yogurt is related to its component and proteolytic bacteria. Milk is a source of protein, calcium, and the B-group vitamins, as well as vitamin A, vitamin C, magnesium, and zinc.29 The consumption of fermented dairy products containing lactic acid bacteria, as well as the probiotic bacteria which was found in yogurt. These can be proposed to reduce the risk of liver cancer by binding to the mutagens produced by intestinal bacteria.30,31

Table 1: Mean values ± standard deviation (SD) of body weight gain, food intake, and feed efficiency ratio (FER)of the control group, rats fed aflatoxin-contaminated nuts, and those supplemented with selenium, yogurt or both selenium and yogurt.

Groups

Variables

 Normal  Control Rats fed aflatoxin-contaminated nuts (3%)
 Positive control Selenium yogurt

Yogurt with selenium

Body weight gain     (g) 130.20±12.51ab 100.21±8.99d 115.51±11.33bc 116.81±12.45bc 126.11±12.24bc
Food intake (g/w) 15.41±1.40a 14.50±1.69a 15.77±1.57a 15.86±1.37a 15.93±1.28a
FER 0.140±0.003a 0.114±0.002d 0.121±0.005c 0.121±0.004c 0.131±0.001b

Values with the different letters indicate significant difference (P<0.05) and vice versa.

Because of the liver’s role in the detoxification of environmental xenobiotics, consuming aflatoxins-contaminated nuts can cause liver injury and induce hepatotoxicity, which is agreement with our results. The group fed aflatoxins-contaminated nuts had increased serum ALT, AST, ALP, and γGT enzyme activities, compared to the control group. Supplemental selenium or yogurt, or both, could significantly lower these enzyme activities to control levels (Table 2). Multiple studies reported that aflatoxins, potent hepatotoxics, and hepatocarcinogenic mycotoxincan induce lipid peroxidation. These compounds have been also associated with various diseases, including aflatoxicosis and hepatocellular carcinoma.27,32 Here, we demonstrate that the consumption of aflatoxin-contaminated nuts by rats forced a marked elevation in liver enzyme activities, demonstrating hepatocellular damage, as previously reported.33Selenium tended to alleviate serum liver enzymes.34,35The improvement in the liver enzyme levels followed by the consumption of yogurt, that is likely related to the yogurt’s probiotic organisms. Therefore, directly affect multiple processes, including digestion and immune function.36,37

Table 2: Mean values ± standard deviation (SD)of serum ALT, AST, ALP and γ GT enzymes of the control group, rats fed aflatoxin-contaminated nuts, and those supplemented with selenium, yogurt or both selenium and yogurt.

Groups

Variables

  Normal  Control Rats fed aflatoxin-contaminated nuts (3%)
 Positive control Selenium yogurt

Yogurt with selenium

AST(µ ml-1) 45.87±5.41bc 75.755±8.13a 54.21±6.41b 47.74±3.45bc 46.53±6.21bc
ALT(µ ml-1) 39.55±5.22c 61.95±9.67a 42.11±5.60bc 45.83±5.75bc 48.25±6.52b
ALP(µ ml-1) 40. 55±3.99c 68.88±9.50a 48. 88±4. 66b 46.37±4.85bc 43. 41±7.15bc
γGT(µ ml-1) 6.11±0.55bc 9.63±1. 65a 6.87±1.07bc 7. 53±1. 15b 7. 76±1. 12b

Values with the different letters indicate significant difference (P<0.05) and vice versa.

Compared to the control group, the consumption of aflatoxin-contaminated nuts has been reduced the total protein and elevated the creatinine, urea, uric acid, and bilirubin levels. Supplemental selenium decreased globulin and increased creatinine, urea, and bilirubin levels, whereas yogurt increased urea compared to the control group. Compared to the group fed aflatoxin-contaminated nuts, selenium or yogurt, or both, significantly increased serum total protein and decreased creatinine, urea, uric acid, and bilirubin levels (Table 3). Uric acid is the metabolic end product of purine metabolism. The observed decreasing level of uric acid in the treated groups is likely a result of an increased utilization of uric acid, thereby inhibiting the generation of free radicals.27Selenium can reduce nephrotoxicityby attenuating oxidative-stress-associated kidney injury through the reduction of oxygen free radicals and lipid peroxidation in rats treated with gentamicin. Selenium is a cofactor of several enzymes that participate in the regulation of enzymatic antioxidant defenses.38,39 Here we show that improvement of this renal parameter is related to the bioactive peptides that are generated during the production of fermented dairy products, which have antioxidative and growth promoting properties.40–42

Table 3: Mean values ± standard deviation (SD) of serum total protein, albumin, globulin, creatinine, urea, uric acid and bilirubin of the control group, rats fed aflatoxin-contaminated nuts, and those supplemented with selenium, yogurt or both selenium and yogurt.

Groups

Variables

  Normal  Control Rats fed aflatoxin-contaminated nuts (3%)
 Positive control Selenium yogurt

Yogurt with selenium

Total protein (gdl-1) 6.99±0.80a 5.66±0.52b 6.41±0.55a 6.75±0.63a 6.25±0.90a
Albumin(gdl-1) 3.63±0.49a 2.70±0.49ab 2. 81±0.26ab 3.16±0.40a 3.02±0.34a
Globulin(gdl-1) 3.36±0.52a 2.96±0.23ab 2.15±0.24b 3.24±0.33a 3.23±0.25a
Creatinine(mgdl-1) 0.66±0.21c 1.91±0.43a 0.88±0.10b 0.73±0.19bc 0.69±0.13c
Urea(µ mg-1) 35.65±3.51c 65. 50±6.81a 48.87±4. 16b 49. 69±4.51b 41.40±5. 91bc
Uric  acid(mgdl-1) 3.99±0.39c 6.09±0.80a 4.59±0.34bc 4.62±0.54bc 4.01±0.44bc
Bilirubin(mgdl-1) 0.65±0.14cd 1.76±0.53a 1.09±0.32b 0.88±0.25c 0.79±0.21c

Values with the different  letters indicate significant difference (P<0.05) and vice versa

Also, the aflatoxin-contaminated nut-fed group had decreased serum and liver GPX and SOD enzyme activities and increased serum and liver MDA, as compared to the control group. Selenium or yogurt, or both, was able to normalize serum and liver GPX, SOD, and MDA levels (Table 4). In the current study, consumption of aflatoxin-contaminated nuts induced oxidative damage through the generation of reactive oxygen species. This was accompanied by an increase in the expression of liver enzymes and decreased biological activities of some liver antioxidant enzymes. Glutathione is exhausted by glutathione-related enzymes to detoxify the peroxides formed from increased lipid peroxidation under oxidative stress. The increase in hepatic MDA level might be because aflatoxins are metabolized by the cellular cytochrome P450 enzyme system to form the reactive intermediate, aflatoxin-8,9-epoxide, which in-turn reacts with macromolecules, such as lipids and DNA. This leads to cellular injury and lipid peroxidation.43Also, MDA was elevated in the treatment groups compared to the control group, most possibly reflecting an adaptive reaction towards free radical damage in the liver.44

Table 4: Mean values ± SD of serum and liver GPX, SOD and MDA of the control group, rats fedaflatoxin-contaminated nuts, and those supplemented with selenium, yogurt or both selenium and yogurt.

Groups

Variables

Normal  Control Rats fed aflatoxin-contaminated nuts (3%)
 Positive control Selenium yogurt

Yogurt with selenium

GPX(mmol L blood-1) 8.27±1.47a 4.11±0.64b 7.51±0.54a 7.50±0.67a 7.70±0.68a
SOD(mmol L blood-1) 35.20±4.51a 22.95±2.93c 32.51±3.29a 31.31±3.91ab 33.86±3.59a
MDA(mmol L blood-1) 4.02±0.45bc 7.80±1.34a 5.07±0.67b 5.14±0.83b 5.15±0.54b
GPX(µ mg liver-1) 65.01±6.24a 31.85±4. 71c 59.91±5.84ab 58.91±6.32ab 61.14±6.39a
SOD(µ mg liver-1) 44.91±5.16a 25. 89±2. 16b 45.01±4. 12a 42. 87±4.27a 45. 41±3.16a
MDA(mmolg liver-1) 13.97±2.53bc 27.91±2.99a 15.51±2. 91b 14. 73±2.41b 15. 17±3. 16b

Values with the different letters indicate significant difference (P<0.05) and vice versa

Selenium’s ability to enhance GPX activity might be because of increased selenium bioavailability, thereby preventing the formation of free radicals and protecting both integrity and functions of tissues, thus protecting the liver from peroxidation of  lipid and changes in glutathione and antioxidant enzyme activities.45,46The administration of the lactic acid bacteria in yogurt results in increased antioxidative enzyme activity and modulated circulatory oxidative stress, thereby protecting the cells against damage.31,42,47

Conclusion

Selenium and yogurt reduce the hepatotoxicity in rats caused by consuming gaflatoxin-contaminated nuts. Further studies are now needed to better understand the in vivo mechanism(s) by which selenium and yogurt reduce aflatoxin toxicity.

Acknowledgement

I would like to record my profound gratitude to Princess Nourah bint Abdulrahman University for its moral support in accomplishing this research paper.

Competing interest

I declare that there is no conflict of interest regarding the publication of this paper which titled “Lowering effect of selenium and yoghurt on nuts contaminated with aflatoxins induced hepatotoxicity in rats.”

Funding sources

I confirm that The funding of this research was on my personal account .

References

  1. Kris-Etherton, P. M. et al. Nuts and their bioactive constituents: effects on serum lipids and other factors that affect disease risk. Am. J. Clin. Nutr.70, 504S–511S (1999).
  2. Aune, D. et al. Nut consumption and risk of cardiovascular disease, cancer, all-cause and cause-specific mortality: a systematic review and dose-response meta-analysis of prospective studies. BMC Med. 1–14 (2016). doi:10.1186/s12916-016-0730-3
  3. Singh, P. K. & Shukla, A. N. Survey of mycoflora counts, aflatoxin production and induced biochemical changes in walnut kernels. J. Stored Prod. Res.44, 169–172 (2008).
  4. Wogan, G. N., Hecht, S. S., Felton, J.S., Conney, A. H. & Loeb, L. A. Environmental and chemical carcinogenesis. Seminars in Cancer Biology14, 473–486 (2004).
  5. OBrian, G. R. et al. The effect of elevated temperature on gene transcription and aflatoxin biosynthesis. Mycologia99, 232–239 (2007).
  6. Miliţă, N. M., Mihăescu, G. & Chifiriuc, C. [Aflatoxins–health risk factors]. Bacteriol Virusol Parazitol Epidemiol55, 19–24 (2010).
  7. Sadeghi, E., Almasi, A., Mohammdi, M. & Bohlouli, S. The evaluation of Aflatoxin M1 level in collected raw milk for pasteurized dairy factories of Kermanshah in 2010-2011. Zahedan J Res Med Sci15, 26–29 (2013).
  8. Wild, C. P. & Montesano, R. A model of interaction: Aflatoxins and hepatitis viruses in liver cancer aetiology and prevention. Cancer Letters286, 22–28 (2009).
  9. Saleemullah, Iqbal, A., Khalil, I. A. & Shah, H. Aflatoxin contents of stored and artificially inoculated cereals and nuts. Food Chem.98, 699–703 (2006).
  10. Wild, C. P. & Gong, Y. Y. Mycotoxins and human disease: A largely ignored global health issue. Carcinogenesis31, 71–82 (2009).
  11. Reeves, Nielsen, F. H. & Fahey, G. C. AIN-93 Purified Diets for Laboratory Rodents: Final Report of the American Institute of Nutrition Ad Hoc Writing Committee on the Reformulation of the AIN-76A Rodent Diet. J. Nutr.123, 1939–1951 (1993).
  12. Tamime, A. & Robinson, R. Yogurt: Science and Technology. (Woodhead Publishers, 1999).
  13. Anonymous. Enzyme immunoassay for the quantitative analysis of aflatoxins. (2002).
  14. Senyuva, H. Z. et al. Immunoaffinity column cleanup with liquid chromatography using post-column bromination for determination of aflatoxins in hazelnut paste: Interlaboratory study.J. AOAC Int.88, 526–535 (2005).
  15. Chapman, D., Gastilla, R. & Campbell, T. Evaluation of protein in food: A method for the determination of protein efficiency ratio. Can. J. Biochem.Physio.1, 679–686 (1950).
  16. Henry, J. B. Clinical diagnosis and management by laboratory methods. (W.B. Saunders, 2001).
  17. Habig, W. H., Pabst, M. J. & Jakoby, W. B. Glutathione S transferases. The first enzymatic step in mercapturic acid formation. J. Biol. Chem.249, 7130–7139 (1974).
  18. Uchiyama, M. & Mihara, M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal. Biochem.86, 271–278 (1978).
  19. Kakkar, P., Das, B. & Viswanathan, P. N. A modified spectrophotometric assay of superoxide dismutase. Indian J. Biochem. Biophys.21, 130–132 (1984).
  20. Beauchamp, C. & Fridovich, I. Superoxide dismutase: Improved assays and an assay applicable to acrylamide gels. Anal. Biochem.44, 276–287 (1971).
  21. Weiss, C., Maker, H. S. & Lehrer, G. M. Sensitive fluorometric assays for glutathione peroxidase and reductase. Anal. Biochem.106, 512–516 (1980).
  22. Berry, G. Statistical guide-lines and statistical guidance. Medical Journal of Australia146, 408–409 (1987).
  23. Hell, K., Mutegi, C. & Fandohan, P. Aflatoxin control and prevention strategies in maize for Sub-Saharan Africa. in Publication of 10th International Working Conference on Stored Product Protection 534–540 (2010). doi:10.5073/jka.2010.425.388
  24. Yiannikouris, A. & Jouany, J. P. Les mycotoxines dans les aliments des ruminants, leur devenir et leurs effets chez l’animal. Prod. Anim.15, 3–16 (2002).
  25. Moschini, R. C., Sisterna, M. N. & Carmona, M. A. Modelling of wheat black point incidence based on meteorological variables in the southern Argentinean Pampas region. Aust. J. Agric. Res.57, 1151–1156 (2006).
  26. Shane, S. in veterinary, and agricultural significance (eds. Eaton, D. & Groopman, J.) 513–27 (Academic Press, 1993).
  27. Devendran, G. &Balasubramanian, U. Biochemical and histopathological analysis of aflatoxin-induced toxicity in liver and kidney of rat. PelagiaRes. Libr. Asian J. Plant Sci. Res.1, 61–69 (2011).
  28. Navarro-Alarcon, M. & Cabrera-Vique, C. Selenium in food and the human body: A review. Science of the Total Environment400, 115–141 (2008).
  29. Miller, G. D., Jarvis, J. K. & McBean, L. D. in 1 (CRC, 2007).
  30. Fondén, R., Saarela, M., Mättö, J. & Matilla-Sandholm, T. in Functional Dairy Products1, 244–262 (2003).
  31. Kumar, M. et al. Anticarcinogenic effect of probiotic fermented milk and chlorophyllin on aflatoxin-B1-induced liver carcinogenesis in rats. Br. J. Nutr.107, 1006–1016 (2012).
  32. Plaa, G. L. & Hewitt, W. R. in Principles and Methods of Toxicology (ed. Hayes, A.) 401–41 (Raven Press, 1989).
  33. Adibpour, N., Soleimanian-Zad, S., Sarabi-Jamab, M. & Tajalli, F. Effect of storage time and concentration of aflatoxin m1on toxin binding capacity of L. acidophilus in fermented milk product. J. Agric. Sci. Technol.18, 1209–1220 (2016).
  34. El-Demerdash, F. M. Antioxidant effect of vitamin E and selenium on lipid peroxidation, enzyme activities and biochemical parameters in rats exposed to aluminium. J. Trace Elem. Med. Biol.18, 113–121 (2004).
  35. Soudani, N., Ben Amara, I., Sefi, M., Boudawara, T. & Zeghal, N. Effects of selenium on chromium (VI)-induced hepatotoxicity in adult rats. Exp. Toxicol. Pathol.63, 541–548 (2011).
  36. Bengmark, S. & Martindale, R. Prebiotics and Synbiotics in Clinical Medicine. Nutr. Clin. Pract.20, 244–261 (2005).
  37. Deabes, M. M., Darwish, H. R., Abdel-Aziz, K. B., Farag, I. M. & Nada, S, A. Protective effects of lactobacillus rhamnosus GG on aflatoxins induced toxicities in male albino mice. J Environ. Anal. Toxicol2, 132 (2012).
  38. Ghorbani, A., Omidvar, B. &Parsi, A. Protective effect of selenium on cisplatin induced nephrotoxicity: A double-blind controlled randomized clinical trial. J. Nephropathol.2, 129 (2013).
  39. Iglesias, P., Selgas, R., Romero, S. &Díez, J.J. Selenium and kidney disease. Journal of Nephrology26, 266–272 (2013).
  40. Rival, S. G., Boeriu, C. G. & Wichers, H. J.Caseins and casein hydrolysates. 2. Antioxidative properties and relevance to lipoxygenase inhibition. J. Agric. Food Chem.49, 295–302 (2001).
  41. Parodi, P. W.A role for milk proteins and their peptides in cancer prevention. Curr. Pharm. Des.13, 813–828 (2007).
  42. Mohammadi, R., Sohrabvandi, S. & MohammadMortazavian, A. The starter culture characteristics of probiotic microorganisms in fermented milks. Engineering in Life Sciences12, 399–409 (2012).
  43. Umarani, M., Shanthi, P. & Sachdanandam, P. Protective effect of Kalpaamruthaa in combating the oxidative stress posed by aflatoxin B1-induced hepatocellular carcinoma with special reference to flavonoid structure-activity relationship. Liver Int.28, 200–213 (2008).
  44. Jihen, E. H., Imed, M., Fatima, H. & Abdelhamid, K. Protective effects of selenium (Se) and zinc (Zn) on cadmium (Cd) toxicity in the liver of the rat: Effects on the oxidative stress. Ecotoxicol. Environ. Saf.72, 1559–1564 (2009).
  45. Li, X., Hill, K. E., Burk, R. F. & May, J. M. Selenium spares ascorbate and alpha-tocopherol in cultured liver cell lines under oxidant stress. FEBS Lett.508, 489–492 (2001).
  46. Ognjanović, B. I. et al. Effect of chronic cadmium exposure on antioxidant defense system in some tissues of rats: protective effect of selenium. Physiol. Res.57, 403–411 (2008).
  47. Kumar, M. et al. Effect of probiotic fermented milk and chlorophyllin on gene expressions and genotoxicity during AFB₁-induced hepatocellular carcinoma. Gene490, 54–9 (2011).

Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License.