Investigation the Effects of Lactobacillus acidophilus and Lactobacillus casei on aflR Gene expression in Aspergillus parasiticus by Real Time-PCR
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Abstract
Background: The effect of probiotic bacteria (Lactobacillus acidophilus and L. casei) as safe organisms was examined on fungal growth and aflatoxin gene regulation in Aspergillus parasiticus.
Methods: The fungus was cultured in presence of two different concentrations of L. acidophilus and L. casei in MRS broth medium. Mycelia dry weight is indicated as criteria to evaluate fungal growth. Besides, investigation of aflR gene expression by Real Time PCR was performed for analysis of gene regulatory effects in aflatoxin biosynthetic pathway.
Results: Both Lactobacillus strongly inhibited fungal growth in the concentrations of 1. 5 102, 1. 5 103 . Expression analysis of aflatoxin genes pathway by real time PCR showed inhibitory effect of L. acidophilus and L. casei on expression of aflR gene. The gene expression revealed to be reduced at the approximate rates of 99. 7% and 98% respectively by L. acidopholus and L. casei in concentrations of 1. 5 102 and more.
Conclusion: L. acidophilus and L. casei may be used successfully as suitable candidates in controlling of A. parasiticus growth on food and feed as well as reducing of aflatoxin contamination.
Jamali M, Ebrahimi MA, Karimipour M, Shams-Ghahfarokhi M, et al. (2012). An insight into the distribution, genetic di-versity and my¬cotoxin production of As-pergillus section Flavi in soils of pistachio orchards. Folia Microbiol (Praha), 57(1):27-36.
Cotty PJ (1988). Aflatoxin and sclerotial produc¬tion by Aspergillus flavus: influence of pH. Phyto¬pathology, 78:1250-1253.
Boutrif E (1998). Prevention of Aflatoxin in Pis¬tachios. Food Nutr Agri, 21, FAO.
Trail F, Mahanti N, Linz J (1995). Molecular biol¬ogy of aflatoxin biosynthesis. Microbi-ology, 141 (Pt 4): 755–765.
Yuan GF, Liu CS, Chen CC (1995). Differ-entia¬tion of Aspergillus parasiticus from Aspergillus sojae by random amplification of polymorphic DNA. Appl Environ Mi-crobiol, 61(6): 2384–2387.
Woloshuk CP, Prieto R (1998). Genetic or-ganiza¬tion and function of the aflatoxin B1 biosyn¬thetic genes. FEMS Microbiol Lett, 160(2): 169–176.
Chang PK, Cary JW, Bhatnagar D, Cleve-land TE, Bevvett JE, Line JE, Woloshuk CP, Payne GA (1993). Cloning of the As-pergillus parasiticus apa-2 gene associated with the regulation of afla¬toxin biosyn-thesis. Appl Environ Microbiol, 59(10): 3273–3279.
Payne GA, Nystrom GJ, Bhatnagar D, Cleveland TE, Woloshuk CP (1993). Cloning of the afl-2 gene involved in af-latoxin biosynthesis from Aspergillus flavus. Appl Environ Microbiol, 59(1): 156–162.
Woloshuk CP, Foutz KR, Brewer JF, Bhatnagar D, Cleveland TE and Payne GA (1994). Mo¬lecular characterization of aflR, a regulatory lo¬cus for aflatoxin bio-synthesis. Appl Environ Mi¬crobiol, 60(7): 2408–2414.
Abdel-Hadi A, Carter D, Magan N (2011). Dis¬crimination between aflatoxigenic and non-af¬latoxigenic strains of Aspergillus section Flavi group contaminating Egyp-tian peanuts using molecular and analyti-cal techniques. World My¬cotoxin J, Vol 4(1): 69-77.
Sheng Wang B, Zhao Q, Wang Y. (2011). The un¬certainty of assessing aflatoxin B1-producing ability using aflR gene in Asper-gillus species. Afr J Microbio Res, 5(31):5603-5606.
Jahanshiri Z, Shams-Ghahfarokhi M, Al-lameh A, Razzaghi-Abyaneh M (2012). Effect of curcu¬min on Aspergillus parasiticus growth and expres¬sion of major genes involved in the early and late stages of af-latoxin biosynthesis. Iran J Public Health, 41(6):72-79.
Muñoz R, Arena ME, Silva J, González SN. (2010). Inhibition of mycotoxin-producing As¬pergillus nomius VSC 23 by lactic acid bacteria and Saccharomyces cerevisiae. Braz J Microbiol, 41(4): 1019-1026.
Oliveira RBP, Oliveira AL, Glória MBA (2008). Screening of lactic acid bacteria from vacuum packaged beef for antimi-crobial activity. Braz J Microbiol, 39(2): 368-374.
Rezaie S, Ban J, Mildner M, Poitschek Ch, Brna Ch, Tschachler E (2000). Character-ization of cDNA clone, encoding a70 kDa heat shock protein from the dermathophyte pathogen Tri¬chophyton rubrum. Gene, 241(1):27-33.
Chang I, Kim J (2007). Inhibition of Afla-toxin Production of Aspergillus flavus by Lactobacillus casei. Mycobiology, 35(2):76-81.
Oluwafemi F, Kumar M, Bandyopadhyay R, Ogunbanwo T, Ayanwande K (2010). Bio-de¬toxification of aflatoxin B1 in arti-ficially con¬taminated maize grains using lactic acid bacte¬ria. Toxin Reviews, 29(3-4):115–122.
Yoshinari T, Akiyama T, Nakamura K, Kondo T, Takahashi Y, Muraoka Y, Nonomura Y, Na¬gasawa H, Sakuda S (2007). Dioctatin A is a strong inhibitor of aflatoxin production by As¬pergillus para-siticus. Microbiology, 153(Pt 8):2774-80.
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| Issue | Vol 45 No 6 (2016) | |
| Section | Original Article(s) | |
| Keywords | ||
| Aflatoxin Aspergillus parasiticus Lactobacilus acidophilus Lactobacilus casei | ||
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