Detection of Aflatoxigenic Aspergillus Flavus in Maize Grains and Soils in Sulaimani Province using Molecular Approaches.

Authors

  • Khattab A. M. Shekhany Faculty of Science, School of Science, Biology Department, University of Sulaimani, Kurdistan Region, Iraq. Author
  • Shkar R. K. Rostam Faculty of Science, School of Science, Biology Department, University of Sulaimani, Kurdistan Region, Iraq. Author

DOI:

https://doi.org/10.17656/jzs.10557

Keywords:

Aspergillus flavus aflatoxin, Polymerase chain reaction (PCR) detection

Abstract

Aflatoxins are potent carcinogens produced by A. flavus as secondary metabolites. They have the ability to contaminate a large number of food, which ultimately affect the human population. Investigating the incidences of aflatoxigenic species of A. flavus in the local area samples were the aim of current study. Samples were collected randomly from soil and maize grains in the Sulaimani governorate. Morphological features and specific PCR-based protocol were used for identification of A. flavus isolates. This PCR protocol was based on the multi-copy internal transcribed region of the rDNA unit (ITS1-5.8S-ITS2 rDNA), which allowed discrimination from other closely related species. Molecular approach has been used for detection of aflatoxigenicity of the A. flavus isolates. In this study out of eighteen isolates only seven isolates were molecularly and morphologically identified as A. flavus. The aflatoxigenicity of these seven isolates were detected by molecularly method which successfully amplify two structural genes (aflD and aflO) out of three structural genes, and one regulatory gene (aflR). Furthermore, no specific amplicon for (aflP) was obtained in this study.

References

Othman, G. and Al-Delamiy, K. “Aflatoxin B1 Production by Aspergillus flavus In Different Media and Containers and The Antifungal Activity of Garlic and Black Cumin”, Research J. In Engineering and Applied Sciences, Vol. 1, No. 2, pp 117-121. (2012).

Abdulateef, S., Aljubori, M. and Abdulbaqi, N. “Genetic Diversity among Some Aspergillus flavus Isolates by Using Inter Simple Sequence Repeats (ISSR)”, Iraqi J. Science Vol.55, No. 3A, pp 986-993. (2014).

Teniente, L., Chapa-Oliver, A., Vazquez-Cruz, M., Torres-Pacheco, I. and Guevara-González, R. “Chapter:16 Aflatoxins Biochemistry and Molecular Biology - Biotechnological Approaches for Control in Crops”, In: Aflatoxins - Biochemistry and Molecular Biology edited by R. Guevara-González, pp.317-343.Rijeka and Croatia. (2011).

Hussein, H. (2001) “Toxicity, Metabolism, and Impact of Mycotoxins on Humans and Animals”, Toxicology, Vol. 167, No. 2, pp 101-134. (2011). DOI: https://doi.org/10.1016/S0300-483X(01)00471-1

Bennett, J. and Klich, M. “Mycotoxins”, Clin. Microbiol.Rev Vol.16, No.3, pp 497–516. (2003). DOI: https://doi.org/10.1128/CMR.16.3.497-516.2003

González-Salgado, A., González-Jaén, T., Vázquez, C. and Patiño, B. “Highly Sensitive PCR-based Detection Method Specific for Aspergillus flavus in Wheat Flour”, Food Additives & Contaminants Part A - Chemistry, Analysis, Control, Exposure & Risk Assessment, Vol. 25, No.6, pp 758 -764. (2008). DOI: https://doi.org/10.1080/02652030701765715

Ritter, A., Hoeltz, M. And Noll, I. (2011) “Toxigenic Potential of Aspergillus flavus Tested in Different Culture Conditions”,Ciênc. Tecnol. Aliment. Campinas Vol. 31, No.3, pp 623-628. (2008). DOI: https://doi.org/10.1590/S0101-20612011000300011

Shweta. S, Madhavan.., S, Paranidharan., V and R, V. "Detection of Aspergillus flavus in Maize Kernels by Conventional and Real-time PCR assays". Int. Food Rese. J., Vol. 20, No. 6, pp 3329-3335. (2013).

Nazir, K., Hassan, J., Durairaj, P. and Yun, H. "Isolation and Identification of Aspergillus flavus From Poultry Feed Samples Using Combined Traditional-Molecular Approach and Expression of Cyp64a1 At mRNA Level". Pak. J. Agri. Sci., Vol.51, No.2, pp 287-291. (2014).

Rodrigues, P., Soares, C., Kozakiewicz, Z., Paterson, R., Lima, N. and Venâncio, A. “Identification and Characterization of Aspergillus flavus and Aflatoxins”, Communicating Current Research and Educational Topics and Trends in Applied Microbiology, A (Méndez-Vilas), pp 528-534. (2007).

Yu, J. "Current Understanding on Aflatoxin Biosynthesis and Future Perspective in Reducing Aflatoxin Contamination". Toxins, Vol.4, No.12, pp 1024-1057. (2012). DOI: https://doi.org/10.3390/toxins4111024

Shekhany, K. M. "Study on Toxicity and Detoxification of Zearalenone". PhD. University of Sulaimani. (2008).

Yazdani,D.,Abidin,Z.,A,M. and Kamaruzaman,S. "Evaluation of the Detection Techniques of Toxigenic Aspergillus Isolates", African J.Biotechnol., Vol. 9, No.45, pp 7654-7659. (2010).

Saadullah, A. "Identification and Determination of Aflatoxin G1 and Aflatoxigenic Aspergillus Isolates from Dried Vine Fruits in Duhok by LCMs-MS Technique", J. Basrah Researches, Vol. 39, No. 1, pp 66-75. (2013).

Nair, S., Bhagobaty, R., Nampoothiri, K., Kalaigandhi, V. and Menon, K. "Detection of Aflatoxin Production by Fungi in Spice Samples Using HPLC and Direct Visual Cultural Methods", Innovative Romanian Food Biotechnology, Vol. 14(march 2014), pp 1-12. (2014).

Hussain, A., Afzal, A., Irfan, M. and Malik, K. "Molecular Detection of Aflatoxin Producing Strains of Aspergillus flavus from Peanut (Arachis Hypogaea)", Turkish J. of Agriculture - Food Sci. and Techno., Vol. 3, No. 5, pp 335-341. (2015). DOI: https://doi.org/10.24925/turjaf.v3i5.335-341.123

Amadi, E. and Adeniyi, O. "Mycotoxin Production by Fungi Isolated from Stored Grains", African J. Biotechnology, Vol. 8m No.7, pp 1219-1221.

Gaddeyya, G., Niharika, P., Bharathi, P. and Kumar, P. "Isolation and Identification of Soil Mycoflora in Different Crop Fields at Salur Mandal", Advances in Applied Science Research, Vol.3, No.4, pp 2020-2026. (2012).

Rohilla, K. and Salar, K. "Isolation and Characterization of Various Fungal Strains from Agricultural Soil Contaminated with Pesticides", Research Journal of Recent Sciences, Vol.1, pp 297-303. (2012).

Jasuja, N., Saxena, R., Chandra, S. and Joshi, S. "Isolation and Identification of Microorganism from Polyhouse Ggriculture Soil of Rajasthan", African J.Microbiol. Res., Vol.7, No.41, pp 4886-4891. (2013). DOI: https://doi.org/10.5897/AJMR2012.2413

Abdullah, S., AL-Saad, I. and Essa, R. "Mycobiota and Natural Occurrence of Sterigmatocystin in Herbal Drugs ln lraq", Basrah J. Science, Vol. 20, No. 2, pp 1-8. (2002).

Bennett, J. "An Overview of the Genus Aspergillus. In: M. Machida and K. Gomi, ed., Aspergillus Molecular Biology and Genomics", 1st ed. Ibaraki, Japan: National Institute of Advanced Industrial Science and Technology. pp.1-17. (2010).

Houbraken, J., de Vries, R. and Samson, R. "Modern taxonomy of Biotechnologically Important Aspergillus and Penicillium species", Advances in Applied Microbiology, Vol. 86, pp 199-249. (2014). DOI: https://doi.org/10.1016/B978-0-12-800262-9.00004-4

Afzal, H., Shazad, S. and Qamar Un Nisa, S. "Morphological Identification of Aspergillus Species from the Soil of Larkana District (SINDH, PAKISTAN)". Asian J Agri Biol, Vol.1, No. 3, pp 105-117. (2013).

Al-Wathiqi, F., Ahmad, S. and Khan, Z. "Molecular Identification and Antifungal Susceptibility Profile of Aspergillus flavus Isolates Recovered from Clinical Specimens in. Kuwait", BMC Infect Dis, Vol. 13, No.126, pp1-9. (2013). DOI: https://doi.org/10.1186/1471-2334-13-126

Almoammar, H., Bahkali, A. and Abd-Elsalam, K. "A polyphasic Method for the Identification of Aflatoxigenic Aspergillus Species Isolated from Camel Feeds", Australian J.Crop Science, Vol. 7, No. 11, pp 1707-1713. (2013).

Criseo, G., Bagnara, A. and Bisignano, G. "Differentiation of Aflatoxin-producing and non-producing Strains of Aspergillus flavus group". Letters in Appl. Microbiol., Vol. 33, No.4, pp 291-295. (2001). DOI: https://doi.org/10.1046/j.1472-765X.2001.00998.x

Degola, F., Berni, E., Dall'Asta, C., Spotti, E., Marchelli, R., Ferrero, I. and Restivo, F. "A multiplex RT-PCR Approach to Detect Aflatoxigenic Strains of Aspergillus flavus". J Appl Microbiol, Vol. 103, No. 2, pp 409-417. (2007). DOI: https://doi.org/10.1111/j.1365-2672.2006.03256.x

Abdel-Hadi, A., Carter, D. and Magan, N. "Discrimination Between Aflatoxigenic and non-aflatoxigenic Aspergillus Section Flavi Strains from Egyptian Peanuts Using Molecular and Analytical Techniques". World Mycotoxin J., Vol.4, No.1, pp 69-77. (2011). DOI: https://doi.org/10.3920/WMJ2010.1223

Hashim, A., Al-Kazaz, A. and Abdulmalek, H. "PCR Detection of Aspergillus flavus Isolates for Aflatoxin B1 Producer", Journal of Biotechnology Research Center, Vol.7, No.3, pp 81-89. (2013). DOI: https://doi.org/10.24126/jobrc.2013.7.3.286

Hashemi, E., Pourbakhsh, S., Shahsavandi, S., Mohammadi, S., Shooshtari, S. and Jahanshiri, A. Application of PCR on Detection of Aflatoxinogenic Fungi. Jahanshiri, Vol. 62, No.2, pp 95-100. (2007).

Scherm, B., Palomba, M., Serra, D., Marcello, A. and Migheli, Q. "Detection of Transcripts of the Aflatoxin genes aflD, aflO, and aflP by Reverse Transcription–polymerase Chain Reaction Allows Differentiation of Aflatoxin-producing and non-producing Isolates of Aspergillus flavus and Aspergillus parasiticus". Int. J. Food Microbiol.,Vol. 98, No.2, pp 201-210. (2005). DOI: https://doi.org/10.1016/j.ijfoodmicro.2004.06.004

Scheidegger, K. and Payne, G. "Unlocking the Secrets Behind Secondary Metabolism: A Review of Aspergillus flavus from Pathogenicity to Functional Genomics". In: H. Abbas, ed., Aflatoxin and Food Safety, 1st ed. Boca Raton London and New York: Taylor & Francis Group, pp.138-155. (2005). DOI: https://doi.org/10.1201/9781420028171.ch8

Das, A., Bhattacharya, S., Palaniswamy, M. and Angayarkanni, J. "Molecular identification of Aspergillus flavus GHBF09 Involved in Aflatoxin B1 Production in Rice Straw". Scholars Research Library, Vol. 4, No.6, pp 102-110. (2013).

Gallo, A., Stea, G., Battilani, P., Logrico, A. and Perroni, G. "Molecular Characterization of an Aspergillus flavus Population Isolated from Maize During the First Outbreak of Aflatoxin Contamination in Italy". Phytopathologia Mediterranea, Vol. 51, No.1, pp 198−206. (2012).

Kanbe, T., Yamaki, K. and Kikuchi, A. "Identification of the Pathogenic Aspergillus Species by Nested PCR Using a Mixture of Specific Primers to DNA Topoisomerase II Gene". Microbiol. and Immunol., Vol.46, No.12, pp 841-848. (2002). DOI: https://doi.org/10.1111/j.1348-0421.2002.tb02771.x

El Khoury, A., Atoui, A., Rizk, T., Lteif, R., Kallassy, M. and Lebrihi, A. "Differentiation Between Aspergillus flavus and Aspergillus parasiticus from Pure Culture and Aflatoxin-Contaminated Grapes Using PCR-RFLP Analysis of aflR-aflJ Intergenic Spacer". J.Food Science, Vol. 76, No.4, pp M247-M253. (2011). DOI: https://doi.org/10.1111/j.1750-3841.2011.02153.x

Henry, T., Iwen, P. and Hinrichs, S. "Identification of Aspergillus Species Using Internal Transcribed Spacer Regions 1 and 2". J. of Clinical Microbiol., Vol. 38, No.4, pp 1510–1515. (2000). DOI: https://doi.org/10.1128/JCM.38.4.1510-1515.2000

Geiser, D., Dorner, J., Horn, B. and Taylor, J. "The Phylogenetics of Mycotoxin and Sclerotium Production in Aspergillus flavus and Aspergillus oryzae". Fungal Genetics and Biology, Vol. 31, No. 3, pp 169-179. (2000). DOI: https://doi.org/10.1006/fgbi.2000.1215

Geisen, R. "Multiplex Polymerase Chain Reaction for the Detection of Potential Aflatoxin and Sterigmatocystin Producing Fungi". Systematic and Appl. Microbiol., Vol. 19, No.3, pp 388-392. (1996). DOI: https://doi.org/10.1016/S0723-2020(96)80067-1

Criseo, G., Racco, C. and Romeo, O. "High Genetic Variability in non-aflatoxigenic A. flavus Strains by Using Quadruplex PCR-based assay". Int. J. Food Microbiol, Vol. 125, No 3, pp 341-343. (2008). DOI: https://doi.org/10.1016/j.ijfoodmicro.2008.04.020

Rahimi, P., Sharifnabi, B. and Bahar, M. "Detection of Aflatoxin in Aspergillus Species Isolated from Pistachio in Iran". J.Phytopathology, Vol. 156, No.1, pp 15-20. (2008). DOI: https://doi.org/10.1111/j.1439-0434.2007.01312.x

Navya, M., Hariprasad, P., Naveen, J., Chandranayaka, S. and Niranjana, R. "Natural Occurrence of Aflatoxin, Aflatoxigenic and non-aflatoxigenic Aspergillus flavus in Groundnut Seeds Across India". African J. Biotechnol., Vol. 12, No.19, pp 2587-2597. (2013).

Latha, R. Manonmani, H. and, E. "Multiplex PCR Assay for the Detection of Aflatoxigenic and non-Aflatoxigenic Aspergilli". Research J. Microbiology, Vol.3, No.3, pp 136-142. (2008). DOI: https://doi.org/10.3923/jm.2008.136.142

Mayer, Z., Farber, P. and Geisen, R. "Monitoring the Production of Aflatoxin B1 in Wheat by Measuring the Concentration of nor-1 mRNA". Appl. and Environ. Microbiol. Vol. 69, No. 2, pp 1154-1158. (2003). DOI: https://doi.org/10.1128/AEM.69.2.1154-1158.2003

Yu, J., Bhatnagar, D. and Ehrlich, K. "Aflatoxin Biosynthesis". Rev. Iberoam Micol, Vol.19, pp 191-200. (2002).

Yu, J., Chang, P., Ehrlich, K., Cary, J., Bhatnagar, D., Cleveland, T., Payne, G., Linz, J., Woloshuk, C. and Bennett, J. "Clustered Pathway Genes in Aflatoxin Biosynthesis". Appl. and Environm. Microbiol Vol.70m No.3, pp 1253-1262. (2004). DOI: https://doi.org/10.1128/AEM.70.3.1253-1262.2004

Houshyarfard, M., Rouhani, H., Falahati-Rastegar, M., Malekzadeh-Shafaroudi, S., Mehdikhani-Moghaddam, E. and Chang, P. "Gene Deletion Patterns in non-aflatoxigenic Strains of Aspergillus flavus". Mycologia Iranica, Vol.1m No.2, pp 87 – 97. (2014).

Published

2016-12-20

How to Cite

Detection of Aflatoxigenic Aspergillus Flavus in Maize Grains and Soils in Sulaimani Province using Molecular Approaches. (2016). Journal of Zankoy Sulaimani - Part A, 18(4), 25-36. https://doi.org/10.17656/jzs.10557

Most read articles by the same author(s)

1 2 3 4 5 6 7 8 9 10 > >>