Document Type : Original Paper


1 Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Islamic Republic of Iran

2 Nour Danesh Institute of Higher Education, Mimeh, Isfahan, Islamic Republic of Iran

3 Department of Medical Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Islamic Republic of Iran

4 Department of Mycology, Pasture Institute of Iran, Tehran, Islamic Republic of Iran


The present study was designed to examine the effects of Lactobacillus plantaraum and L. paracasei on the growth and aflatoxin-producing ability of Aspergillus parasiticus. In these experiments, direct and indirect interactions of two different probiotic lactobacilli with A. parasiticus was examined.A co-culture system which supported both fungal growth and probiotic bacteria showed the inhibitory actions of the lactobacilli on A. parasiticus growth. Disk Diffusion Assay was developed to examine the anti-fungal action of the probiotic cell free extract.  The probiotic extracts were also applied in a micro-dilution assay to evaluate the anti-fungal and anti-aflatoxin properties of the probiotic extracts. The results showed that direct interaction of the bacteria with A. parasiticus in a spot-culture method (two-layer semi-solid/solid culture) caused a clear zone of inhibition of A. parasiticus growth. Fungal growth inhibition was found to be dependent on the concentration of the probiotic extracts added to A. parasiticus culture (micro dilution assay). The inhibitory effects of the probiotic extracts was further confirmed using Disc Diffusion Assay by showing that discs loaded with 50 or 100 µl of extracts resulted in a significant (4-5 folds) inhibition in A. parasiticus growth. The extract prepared from the probiotics inhibited the fungal growth and at higher levels. Accordingly, the probiotic extract (50% v/v) could significantly suppress aflatoxin levels in mycelia and fungal culture media.  In conclusion, probiotic bacteria and their cell-free extracts produced under optimized condition exhibit anti-fungal and anti-mycotoxin effects provided that these bacteria undergo screening and selection prior totheir application. 


Main Subjects

  1. Fulle R. A., Review:  Probiotics in man and animals. J. Appl. Bacteriology 66:365-378 (1989).
  2. Shoaib A., Dachang W. and Y. Xin Y. Determining the role of a probiotic in the restoration of intestinal microbial balance by molecular and cultural techniques. Genet. Mol. Res. 14 (1): 1526 –1537 (2015).
  3. Ng S.C., Hart A.L., Kamm M.A, Stagg A.J. and Knight S.C. Mechanisms of action of probiotics: Recent advances. Inflam. Bowel Dis. 15: 300-310 (2009).
  4. Wacoo, A.P., Mukisa, I.M., Meeme, R., Byakika, S., Wendiro D. Sybesma, W. and Kort, R. Probiotic enrichment and reduction of aflatoxins in a traditional African maize-based fermented food. Nutrients 11, 265 (2019).
  5. El-Nezami H., Kankaanpaa P., Salminen S. and Ahokas J. Ability of dairy strains of lactic acid bacteria to bind a common food carcinogen, aflatoxin B1. Food Chem. Toxico. 36: 321-326 (1998). 
  6. Haskard C., Binnion C. and Ahokas J. Factors affecting the sequestration of aflatoxin by Lactobacillus rhamnosus strain GG. Chem. Biol. Interact. 128: 39–49 (2000).
  7. Gratz S., Täubel M., Juvonen R.O., Viluksela M, Turner P.C., Mykkänen H. and El-Nezami H. Lactobacillus rhamnosus strai GG modulates intestinal absorption, fecal excretion, and toxicity of aflatoxin B (1) in rats. Appl. Environ. Microbiol. 72(11):7398-7400 (2006).
  8. Bagherzadeh-Kasmani F., Karimi-Torshizi, M.A. and Allameh A., Shariatmadari F. A novel aflatoxin-binding Bacillus probiotic: Performance, serum biochemistry, and immunological parameters in Japanese quail.  Poult. Sci. 91:1846-1853 (2012).
  9. Khanian M., Karimi-Torshizi M.A. and Allameh A. Alleviation of aflatoxin-related oxidative damage to liver and improvement of growth performance in broiler chickens consumed Lactobacillus plantaraum 299v for entire growth period. Toxicon 158:57–62 (2019).
  10. El-Nezami H.S., Polychronaki N.N., Ma J., Zhu H., Ling W., Salminen E.K., Juvonen R.O, Salminen S.J, Poussa T. and Mykkänen HM. Probiotic supplementation reduces a biomarker for increased risk of liver cancer in young men from Southern China. Am. J. Clin. Nutr. 83(5):1199-203 (2006).
  11. Slizewska K., Nowak A., Libudzisz Z. and Blasiak J. Probiotic preparation reduces the faecal water genotoxicity in chickens fed with aflatoxin B1 contaminated fodder. Res. Vet. Sci 89: 391-395 (2010).
  12. Rawal S., Kim J.E. and Coulombe R. Jr. Aflatoxin B1 in poultry: toxicology, metabolism and prevention. Res. Vet. Sci., 89(3):325-31. (2010).
  13. Takagi R., Tsujikawa Y., Nomoto R. and Osawa R. Comparison of the growth of Lactobacillus delbrueckii, L. paracasei and L. plantarum on Inulin in co-culture systems.  Biosci. Microbiota Food Health 33.139 (2014).
  14. Saadatzadeh A., Fazeli M.R., Jamalifar H. and Dinarvand R. Probiotic Properties of lyophilized cell free extract of lactobacillus casei. Jundishapur J. Natural Pharmaceut. Products 8 (3): 131-137 (2013).
  15. Allameh, A., Razzaghi M., Shams M., Rezaee M.B. and  Jaimand K.  Effects of neem leaf extract on production of aflatoxins and activities of fatty acid synthetase, isocitrate dehydrogenase and glutathione S-transferase Aspergillus parasiticus. Mycopathologia 154;79-84 (2002).
  16. Namazi  M.,   Allameh  A.,  Aminshahidi  M.,  Nohee  A.  and  Malekzadeh  F.  Inhibitory  effects  of  ammonia  solution  on  growth  and  aflatoxins  production  by Aspergillus parasiticus NRRL-2999. Acta Poloniae Toxicologica, 10: 65-72 (2002).

    Shams-Ghahfarokhi M., Kalantari S., Razzaghi-Abyaneh M. Terrestrial bacteria from agricultural soils: versatile weapons against aflatoxigenic fungi. Razzaghi-Abyaneh M. ed(s). In: Aflatoxins recent advances and future prospects. InTech.. pp: 22-39 (2013).
  1. Barry A.L., Jones R.N., Thornsberry C. Cefuroxime, cefamandole, cefoxitin, and cephalothin in vitro susceptibility tests: reassessment of the "class representative" concept, confirmation of disc interpretive criteria, and proposed quality control guidelines. Am. J. Clin. Pathol. 80 (2):182–189.53 (1983).
  2. Azeem, N.; Nawaz M., Anjum A.A., Saeed S., Sana S., Mustafa A. and Yousuf M.R. Activity and anti-aflatoxigenic effect of indigenously characterized probiotic Lactobacilli against Aspergillus flavus- A common poultry feed contaminant. Animals 9: 166- (2019).
  3. El-Gendy S.M. and Marth E.H. Growth and aflatoxin production by Aspergillus parasiticus in the presence of Lactobacillus casei. J. Food Prot. 44: 211-212 (1981).
  4. Muñoz, R., Arena M.E., Silva, J. and González S.N. Inhibition of mycotoxin-producing Aspergillus nomius VSC 23 by lactic acid bacteria and Saccharomyces cerevisiae. Braz. J. Microbio. 41: 1019-1026 (2010).
  5. Huang L., Duan C., Zhao Y., Gao L., Niu C, Xu J. and Li S. Reduction of aflatoxin B1 toxicity by bacillus plantaraum C88: A potential probiotic strain isolated from Chinese traditional fermented food ªTofu.  PLOS One, 12 (1): (2017).
  6. Allameh   A.,   Saxena   M.,   and   Raj   H.G. Lack   of   influence   of   butylated hydroxytoluene on modification of lung microsome-mediated aflatoxin B1-DNA binding. Role of pulmonary glutathione S-transferase. Cancer Let. 43: 125-131 (1988).