First global registration of an isolate producing aflatoxin B1 from the fungus (Rhizopus microsporus) from stored grains and nuts in Iraq
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Abstract
This study delineates the mycotoxin-producing capabilities of Rhizopus microsporus strains isolated from stored grains and nuts. Aflatoxins, particularly aflatoxin B1, are significant due to their deleterious effects on human and animal health and their economic impact on vital agricultural commodities. Employing polymerase chain reaction (PCR) for identification, one isolate was confirmed to produce aflatoxin B1 and was associated with R. microsporus. Thin-layer chromatography (TLC) and ammonia gas tests substantiated the isolate’s toxin-producing ability. High-performance liquid chromatography (HPLC) quantified aflatoxin B1 concentration, revealing a yield of 395.19 PPb. This study’s findings are unprecedented, marking the first known instance of such isolates producing toxins globally. DNA extraction from fungal hyphae, PCR analysis, nucleotide sequencing, and submission to the National Center for Biotechnology Information (NCBI) database further corroborated the isolate’s classification as R. microsporus. The isolate was formally registered in the Gene Bank with serial number OP205376.1, representing a notable contribution to Iraqi and international scientific communities.
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Keywords
Aflatoxin B1, HPLC (High-performance liquid chromatography), Mycotoxin, NCBI, Rhizopus microspores
References
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Raper, K. B. & Fennell, D. I. (1965). The genus Aspergillus. The Genus Aspergillus.
Saito, M., & Machida, S. (1999). A rapid identification method for aflatoxin-producing strains of Aspergillus flavus and A. parasiticus by ammonia vapor. Mycoscience, 40(2), 205–208.
Savita, & Sharma, A. (2019). Fungi as biological control agents. Biofertilizers for Sustainable Agriculture and Environment, 395–411.
Sobolev, V. S. & Dorner, J. W. (2002). Cleanup procedure for determination of aflatoxins in major agricultural commodities by liquid chromatography. Journal of AOAC International, 85(3), 642–645.
Thakur Deepshikha & Teja, S. (2024). Mycotoxins and Toxic Fungus in Food: Prevention and Sustainable Management Techniques. In M. Thakur (Ed.), Sustainable Food Systems (Volume II): SFS: Novel Sustainable Green Technologies, Circular Strategies, Food Safety & Diversity (pp. 343–363). Springer Nature Switzerland. https://doi.org/10.1007/978-3-031-46046-3_17
Wani, A. K., Akhtar, N., Sher, F., Navarrete, A. A, & Américo-Pinheiro, J. H. P. (2022). Microbial adaptation to different environmental conditions: molecular perspective of evolved genetic and cellular systems. Archives of Microbiology, 204(2), 144. https://doi.org/10.1007/s00203-022-02757-5
Zheng, R., Chen, G., Huang, H. & Liu, X. (2007). A monograph of Rhizopus. SYDOWIA-HORN-, 59(2), 273.
Agrios, G. N. (2005). Plant pathology. Elsevier.
Al-Jumaili, Sami Abdel Reda (1996). Integrated resistance against infection with the fungus Aspergillus flavus and contamination with aflatoxin B1 in field pistachioyield. Doctoral thesis. College of Agriculture - University of Baghdad. (n.d.).
Alameri, M. M., Kong, A. S. Y., Aljaafari, M. N., Ali, H. Al, Eid, K., Sallagi, M. Al, Cheng, W. H., Abushelaibi, A., Lim, S. H. E., Loh, J. Y. & Lai, K. S. (2023). Aflatoxin Contamination: An Overview on Health Issues, Detection and Management Strategies. In Toxins (Vol. 15, Issue 4). MDPI. https://doi.org/10.3390/toxins15040246
Aziz, M. F., & Caetano-Anollés, G. (2021). Evolution of networks of protein domain organization. Scientific Reports, 11(1). https://doi.org/10.1038/s41598-021-90498-8
Beyene, A. M., Du, X., Schrunk, D., Ensley, S. & Rumbeiha, W. K. (2019). High-performance liquid chromatography and Enzyme-Linked Immunosorbent Assay techniques for detection and quantification of aflatoxin B1 in feed samples: A comparative study. BMC Research Notes, 12(1). https://doi.org/10.1186/s13104-019-4538-z
Bhat, R. V. & Vasanthi, S. (2003). Food safety in food security and food trade: Mycotoxin food safety risk in developing countries. International Food Policy Research Institute, 10(3), 119-156.
Birol, D. & Gunyar, O. A. (2021). Investigation of presence of endofungal bacteria in Rhizopus spp. ısolated from the different food samples. Archives of Microbiology, 203(5), 2269–2277. https://doi.org/10.1007/s00203-021-02251-4
Garnier, L., Valence, F., Pawtowski, A., Auhustsinava-Galerne, L., Frotté, N., Baroncelli, R., Deniel, F., Coton E. & Mounier, J. (2017). Diversity of spoilage fungi associated with various French dairy products. International Journal of Food Microbiology, 241, 191–197.
Gizachew, D., Szonyi, B., Tegegne, A., Hanson, J. & Grace, D. (2016). Aflatoxin contamination of milk and dairy feeds in the Greater Addis Ababa milk shed, Ethiopia. Food Control, 59, 773–779.
Guan, Y., Chen, J., Nepovimova, E., Long, M., Wu, W. & Kuca, K. (2021). Aflatoxin detoxification using microorganisms and enzymes. Toxins, 13(1), 46.
Gupta, A. & Kumar, R. (2020). Management of seed-borne diseases: an integrated approach. Seed-Borne Diseases of Agricultural Crops: Detection, Diagnosis & Management, 717–745.
Horky, P., Skalickova, S., Baholet, D. & Skladanka, J. (2018). Nanoparticles as a solution for eliminating the risk of mycotoxins. Nanomaterials, 8(9), 727.
Hymery, N., Vasseur, V., Coton, M., Mounier, J., Jany, J., Barbier, G. & Coton, E. (2014). Filamentous fungi and mycotoxins in cheese: A review. Comprehensive Reviews in Food Science and Food Safety, 13(4), 437–456.
Kumar, A., Pathak, H., Bhadauria, S., & Sudan, J. (2021). Aflatoxin contamination in food crops: causes, detection, and management: a review. In Food Production, Processing and Nutrition (Vol. 3, Issue 1). BioMed Central Ltd. https://doi.org/10.1186/s43014-021-00064-y
Ma, L.-J., Ibrahim, A. S., Skory, C., Grabherr, M. G., Burger, G., Butler, M., Elias, M., Idnurm, A., Lang, B. F. & Sone, T. (2009). Genomic analysis of the basal lineage fungus Rhizopus oryzae reveals a whole-genome duplication. PLoS Genetics, 5(7), e1000549.
Majeed, S., De Boevre, M., De Saeger, S., Rauf, W., Tawab, A., Rahman, M. & Iqbal, M. (2018). Multiple mycotoxins in rice: Occurrence and health risk assessment in children and adults of Punjab, Pakistan. Toxins, 10(2), 77.
Moghadam, M. M., Rezaee, M., Mohammadi, S. H., Zamanizadeh, A. H., Moradi, H. R. & Fani, M. (2020). The Potential of Aflatoxin Production in the Aspergillus Section Flavi Isolates of Pistachio in Iran. J Nutr Fast Health, 8(4), 254–263. https://doi.org/10.22038/jnfh
Mustafa, M. A., Mabhaudhi, T. & Massawe, F. (2021). Building a resilient and sustainable food system in a changing world–a case for climate-smart and nutrient dense crops. Global Food Security, 28, 100477.
Oadi, M. (2020). Mycotoxin: Global risk and silent killer in our food. Int. J. Agric. Sci. Vet. Med, 8(1), 26–32.
Okasha, H., Song, B. & Song, Z. (2024). Hidden Hazards Revealed: Mycotoxins and Their Masked Forms in Poultry. In Toxins (Vol. 16, Issue 3). Multidisciplinary Digital Publishing Institute (MDPI). https://doi.org/10.3390/toxins16030137
Op De Beeck, M., Lievens, B., Busschaert, P., Declerck, S., Vangronsveld, J. & Colpaert, J. V. (2014). Comparison and validation of some ITS primer pairs useful for fungal metabarcoding studies. PLoS ONE, 9(6). https://doi.org/10.1371/journal.pone.0097629
Pitt, J. I. & Hocking, A. D. (2009). Fungi and food spoilage (Vol. 519). Springer.
Raper, K. B. & Fennell, D. I. (1965). The genus Aspergillus. The Genus Aspergillus.
Saito, M., & Machida, S. (1999). A rapid identification method for aflatoxin-producing strains of Aspergillus flavus and A. parasiticus by ammonia vapor. Mycoscience, 40(2), 205–208.
Savita, & Sharma, A. (2019). Fungi as biological control agents. Biofertilizers for Sustainable Agriculture and Environment, 395–411.
Sobolev, V. S. & Dorner, J. W. (2002). Cleanup procedure for determination of aflatoxins in major agricultural commodities by liquid chromatography. Journal of AOAC International, 85(3), 642–645.
Thakur Deepshikha & Teja, S. (2024). Mycotoxins and Toxic Fungus in Food: Prevention and Sustainable Management Techniques. In M. Thakur (Ed.), Sustainable Food Systems (Volume II): SFS: Novel Sustainable Green Technologies, Circular Strategies, Food Safety & Diversity (pp. 343–363). Springer Nature Switzerland. https://doi.org/10.1007/978-3-031-46046-3_17
Wani, A. K., Akhtar, N., Sher, F., Navarrete, A. A, & Américo-Pinheiro, J. H. P. (2022). Microbial adaptation to different environmental conditions: molecular perspective of evolved genetic and cellular systems. Archives of Microbiology, 204(2), 144. https://doi.org/10.1007/s00203-022-02757-5
Zheng, R., Chen, G., Huang, H. & Liu, X. (2007). A monograph of Rhizopus. SYDOWIA-HORN-, 59(2), 273.
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How to Cite
First global registration of an isolate producing aflatoxin B1 from the fungus (Rhizopus microsporus) from stored grains and nuts in Iraq. (2024). Journal of Applied and Natural Science, 16(3), 1222-1232. https://doi.org/10.31018/jans.v16i3.5652
