Project Title

Effects of Essential Oils as Antifungal agents against Aspergillus flavus and Aspergillus parasiticus and detection of Aflatoxin B1

Presenters

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Faculty Sponsor Name

Dr. Premila Achar

Abstract (300 words maximum)

Effects of Essential Oils as Antifungal agents against Aspergillus flavus and Aspergillus parasiticus and detection of Aflatoxin B1

Zolome, Y; khorsandi, S.; P.N. Achar1 and H. Msimanga2

1Dept. of Molecular and Cellular Biology, Kennesaw State University, Kennesaw, GA

2Dept. of Chemistry and Biochemistry, Kennesaw State University, Kennesaw, GA

Aspergillus flavus and Aspergillus parasiticus are the most common aflatoxin B1 (AFB1) producing fungi in peanuts. Aflatoxin contaminated peanuts continue to be a serious problem in Georgia. AFB1 is the most toxic and dangerous carcinogen to humans and animals. There has been considerable interest in the use of essential oils (EOs) with antimicrobial activities against Aspergillus spp in peanuts. EOs is known for their GRAS (Generally Recognized As Safe) status. Previous work in our lab screened several essential oils against this mold in peanut. Our current study was to test the Minimum Inhibition Concentration (MIC) and Minimal Fungicidal Concentration (MFC) of selected EOs, Clove and Cinnamon. MIC and MFC were determined by the plate diffusion procedure (Hadaceck and Greger, 2000). Filter paper disc was impregnated with EOs at various concentrations: 500ppm, 1000ppm, 2000ppm, and 2500ppm, respectively. Treated disc was placed at the center of each agar plate containing mycelia and spores. Experiments were repeated for synergistic activity of the two oils (Pinto et al. 2009). All experimental set up were incubated at 28±2°C for 24, 72, and 96h and replicated thrice. AFB1 in treated samples were tested by biochemical methods. Aflatoxin production was promoted on specific media, Rose Bengal and Czapek dox agar. After incubation period, we observed that Clove oils showed the maximum MIC and MFC effect over Cinnamon at 2500ppm against both Aspergillus flavus and Aspergillus parasiticus.We detected presence of AFB1 in fungal extract by Thin Layer Chromatography. Quantification of AFB1 is in progress using GC-MS and HPLC.

Key word: A. flavus, A. parasiticus, Essential oils, Aflatoxin and Peanuts

Acknowledgment

Authors would like to thank the Dept. of Molecular and Cellular Biology, Kennesaw State University for financial support.

References

Pinto, E., Vale-Silva, L., Cavaleiro, C. and Salgueiro, L 2009. Antifungal activity of the clove essential oil from Syzygium aromaticum (Eugenia caryophyllus) on Candida, Aspergillus and dermatophyte species. J. Med. Microbial., DOI: 10.1099/jmm.0.010538-0.

Viudas-Martos, M., Ruiz-Navajas, Y., Fernandez-Lopez, J. and Perez-Alvarez, J.A. 2006. Antifungal activities of thyme, clove and oregano essential oils. J. Food Safety, 27:91-101.

Frank Hadacek and Harald Greger 2000. Testing of Antifungal Natural Products: Methodologies, Comparability of Results and Assay Choice Phytochem. Anal 11, 137-147.

Project Type

Poster

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Effects of Essential Oils as Antifungal agents against Aspergillus flavus and Aspergillus parasiticus and detection of Aflatoxin B1

Effects of Essential Oils as Antifungal agents against Aspergillus flavus and Aspergillus parasiticus and detection of Aflatoxin B1

Zolome, Y; khorsandi, S.; P.N. Achar1 and H. Msimanga2

1Dept. of Molecular and Cellular Biology, Kennesaw State University, Kennesaw, GA

2Dept. of Chemistry and Biochemistry, Kennesaw State University, Kennesaw, GA

Aspergillus flavus and Aspergillus parasiticus are the most common aflatoxin B1 (AFB1) producing fungi in peanuts. Aflatoxin contaminated peanuts continue to be a serious problem in Georgia. AFB1 is the most toxic and dangerous carcinogen to humans and animals. There has been considerable interest in the use of essential oils (EOs) with antimicrobial activities against Aspergillus spp in peanuts. EOs is known for their GRAS (Generally Recognized As Safe) status. Previous work in our lab screened several essential oils against this mold in peanut. Our current study was to test the Minimum Inhibition Concentration (MIC) and Minimal Fungicidal Concentration (MFC) of selected EOs, Clove and Cinnamon. MIC and MFC were determined by the plate diffusion procedure (Hadaceck and Greger, 2000). Filter paper disc was impregnated with EOs at various concentrations: 500ppm, 1000ppm, 2000ppm, and 2500ppm, respectively. Treated disc was placed at the center of each agar plate containing mycelia and spores. Experiments were repeated for synergistic activity of the two oils (Pinto et al. 2009). All experimental set up were incubated at 28±2°C for 24, 72, and 96h and replicated thrice. AFB1 in treated samples were tested by biochemical methods. Aflatoxin production was promoted on specific media, Rose Bengal and Czapek dox agar. After incubation period, we observed that Clove oils showed the maximum MIC and MFC effect over Cinnamon at 2500ppm against both Aspergillus flavus and Aspergillus parasiticus.We detected presence of AFB1 in fungal extract by Thin Layer Chromatography. Quantification of AFB1 is in progress using GC-MS and HPLC.

Key word: A. flavus, A. parasiticus, Essential oils, Aflatoxin and Peanuts

Acknowledgment

Authors would like to thank the Dept. of Molecular and Cellular Biology, Kennesaw State University for financial support.

References

Pinto, E., Vale-Silva, L., Cavaleiro, C. and Salgueiro, L 2009. Antifungal activity of the clove essential oil from Syzygium aromaticum (Eugenia caryophyllus) on Candida, Aspergillus and dermatophyte species. J. Med. Microbial., DOI: 10.1099/jmm.0.010538-0.

Viudas-Martos, M., Ruiz-Navajas, Y., Fernandez-Lopez, J. and Perez-Alvarez, J.A. 2006. Antifungal activities of thyme, clove and oregano essential oils. J. Food Safety, 27:91-101.

Frank Hadacek and Harald Greger 2000. Testing of Antifungal Natural Products: Methodologies, Comparability of Results and Assay Choice Phytochem. Anal 11, 137-147.