Screening of Aflatoxin and Non-Aflatoxin Producing Strains of Aspergillus flavus in Maize

Main Article Content

Wajiha Seerat, 1Abida Akram, Nafeesa Qudsia Hanif, Ali Imran, Brian Gagosh Nayyar, Shaista Akhund, Sajid Hussain

Keywords

Aflatoxin, TLC, Toxigenic, Atoxigenic, Pothwar

Abstract

The present study aims to detect aflatoxin and non-aflatoxin producing ability of A. flavus isolates which were recovered from district of Punjab Province (Attock, Chakwal, Jhelum and Rawalpindi). The base of aflatoxin production was characterized using Thin Layer Chromatography and were successfully categorized into toxigenic (aflatoxin producing) and atoxigenic (non-aflatoxin producing) strains. Frequency of aflatoxins production of A. flavus isolates varied among the four districts of Punjab Province (Attock, Chakwal, Jhelum and Rawalpindi). Out of 212 A. flavus isolates, 200 isolates (94.34%) were found positive with aflatoxin B1 and B2 and these were classified as aflatoxin producing strains of A. flavus while 12 isolates (5.66%) did not show presence of any aflatoxin and these were classified as non-aflatoxin producing strains of A. flavus. However, AFG1 and AFG2 were not detected in any sample. The percent incidence of B1 was highest in Rawalpindi region (95.23%) whereas Attock (95.12%), Chakwal and Jhelum (93.18%) each. While AFB2 were recorded as the highest in Rawalpindi region (76.19%) followed by Chakwal and Jhelum (68.18%) and Attock (60.97%).

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References

Achakzai, A. K. K., & Bazai, Z. A. (2015). Occurrence and health hazard status of aflatoxin in human food and animal feed of wheat from Pakistan: A review paper. Pure and Applied Biology, 4(4), 611.
Aikore, M. O., Ortega-Beltran, A., Eruvbetine, D., Atehnkeng, J., Falade, T., Cotty, P. J., & Bandyopadhyay, R. (2019). Performance of broilers fed with maize colonized by either toxigenic or atoxigenic strains of Aspergillus flavus with and without an aflatoxin-sequestering agent. Toxins, 11(10), 565.
Anthony, M. H., Francis, D. M., Patrick, N., Berka, G. T. A., & Haruna, O. G. (2012). Aflatoxin contamination in foods and feeds: A special focus on Africa. Trends in Vital Food and Control Engineering, 187.
Atehnkeng, J., Ojiambo, P. S., Donner, M., Ikotun, T., Sikora, R. A., Cotty, P. J., & Bandyopadhyay, R. (2008). Distribution and toxigenicity of Aspergillus species isolated from maize kernels from three agro-ecological zones in Nigeria. International Journal of Food Microbiology, 122(1-2), 74-84.
Bhatnagar, D., Ehrlich, K. C., & Cleveland, T. E. (2003). Molecular genetic analysis and regulation of aflatoxin biosynthesis. Applied Microbiology and Biotechnology, 61(2), 83-93.
Brown, R. L., Zhi-Yuan, C., Menkir, A., & Cleveland, T. E. (2003). Using biotechnology to enhance host resistance to aflatoxin contamination of corn. African Journal of Biotechnology, 2(12), 557-562.
Castellari, C., Marcos Valle, F., Mutti, J., Cardoso, L., & Bartosik, R. (2010). Toxigenic fungi in corn (maize) stored in hermetic plastic bags. Julius-Kuhn-Archiv, (425), 501-504.
Choochuay, S., Phakam, J., Jala, P., Maneeboon, T., & Tansakul, N. (2018). Determination of aflatoxin B1 in feedstuffs without clean-up step by high-performance liquid chromatography. International Journal of Analytical Chemistry.
Donner, M., Atehnkeng, J., Sikora, R. A., Bandyopadhyay, R., & Cotty, P. J. (2010). Molecular characterization of atoxigenic strains for biological control of aflatoxins in Nigeria. Food Additives and Contaminants, 27(5), 576-590.
Food and Agriculture Organization (FAO). 2018. Integrated management of the fall armyworm on maize. A Guide for Farmer Field Schools in Africa.
Fuchs, B., Sub, R., Teuber, K., Eibisch, M., & Schiller, J. (2011). Lipid analysis by thin-layer chromatography - a review of the current state. Journal of Chromatography A, 1218(19), 2754-2774.
Garcia-Cela, E., Kiaitsi, E., Sulyok, M., Krska, R., Medina, A., Petit Damico, I., & Magan, N. (2019). Influence of storage environment on maize grain: CO2 production, dry matter losses and aflatoxins contamination. Food Additives and Contaminants: Part A, 36(1), 175-185.
Giorni, P., Battilani, P., & Magan, N. (2008). Effect of solute and matric potential on in vitro growth and sporulation of strains from a new population of Aspergillus flavus isolated in Italy. Fungal Ecology, 1(2-3), 102-106.
Guan, D., Li, P., Zhang, Q., Zhang, W., Zhang, D., & Jiang, J. (2011). An ultra sensitive monoclonal antibody-based competitive enzyme immunoassay for aflatoxin M1 in milk and infant milk products. Food Chemistry, 125(4), 1359-1364.
Horn, B. W. (2007). Biodiversity of Aspergillus section Flavi in the United States: a review. Food Additives and Contaminants, 24(10), 1088-1101.
Hua, S. S. T., Parfitt, D. E., Sarreal, S. B. L., & Sidhu, G. (2019). Dual culture of atoxigenic and toxigenic strains of Aspergillus flavus to gain insight into repression of aflatoxin biosynthesis and fungal interaction. Mycotoxin Research, 35(4), 381-389.
Iqbal, N., Hussain, S., Ahmed, Z., Yang, F., Wang, X., Liu, W., ... & Liu, J. (2019). Comparative analysis of maize–soybean strip intercropping systems: a review. Plant Production Science, 22(2), 131-142.
Iram, W., Anjum, T., Iqbal, M., Ghaffar, A., & Abbas, M. (2016). Structural elucidation and toxicity assessment of degraded products of aflatoxin B1 and B2 by aqueous extracts of Trachyspermum ammi. Frontiers in Microbiology, 7, 346.
Kensler, T. W., Roebuck, B. D., Wogan, G. N., & Groopman, J. D. (2011). Aflatoxin: a 50-year odyssey of mechanistic and translational toxicology. Toxicological Sciences, 120(suppl_1), S28-S48.
Khatoon, S., Hanif, N. Q., Tahira, I., Sultana, N., Sultana, K., & Ayub, N. (2012). Natural occurrence of aflatoxins, zearalenone and trichothecenes in maize grown in Pakistan. Pakistan Journal of Botany, 44(1), 231-236.
Khlangwiset, P., & Wu, F. (2010). Costs and efficacy of public health interventions to reduce aflatoxin-induced human disease. Food Additives and Contaminants, 27(7), 998-1014.
Klich, M. A. (2007). Aspergillus flavus: the major producer of aflatoxin. Molecular Plant Pathology, 8(6), 713-722.
Lagogianni, C. S., & Tsitsigiannis, D. I. (2019). Effective biopesticides and biostimulants to reduce aflatoxins in maize fields. Frontiers in Microbiology, 10, 2645.
Lanubile, A., Maschietto, V., Borrelli, V. M., Stagnati, L., Logrieco, A. F., & Marocco, A. (2017). Molecular basis of resistance to Fusarium ear rot in maize. Frontiers in Plant Science, 8, 1774.
Lanyasunya, T. P., Wamae, L. W., Musa, H. H., Olowofeso, O., & Lokwaleput, I. K. (2005). The risk of mycotoxins contamination of dairy feed and milk on smallholder dairy farms in Kenya. Pakistan Journal of Nutrition, 4(3), 162-169.
Lewis, L., Onsongo, M., Njapau, H., Schurz-Rogers, H., Luber, G., Kieszak, S., ... & Kenya Aflatoxicosis Investigation Group. (2005). Aflatoxin contamination of commercial maize products during an outbreak of acute aflatoxicosis in eastern and central Kenya. Environmental Health Perspectives, 113(12), 1763-1767. doi:10.1289/ehp.7998.
Mahmoud, M. A., Ali, H. M., El-Aziz, A. R. M., Al-Othman, M. R., & Al-Wadai, A. S. (2014). Molecular characterization of aflatoxigenic and non-aflatoxigenic Aspergillus flavus isolates collected from corn grains. Genetics and Molecular Research, 13, 9352-9370.
Manzoor, M., Farooq, Z., Iqbal, S., Mukhtar, H., & Nawaz, M. (2018). Quantification of aflatoxins in maize samples collected from various parts of the Punjab, Pakistan. JAPS: Journal of Animal and Plant Sciences, 28(6).
Muhammad, D., Chand, N., Khan, S., Sultan, A., & Mushtaq, M. (2012). Hepatoprotective Role of Milk Thistle (Silybum marianum) in Meat Type Chicken Fed Aflatoxin B 1 Contaminated Feed. Pakistan Veterinary Journal, 32(3), 443-446.
Orsi, R. B., Corrêa, B., Possi, C. R., Schammass, E. A., Nogueira, J. R., Dias, S. M., & Malozzi, M. A. (2000). Mycoflora and occurrence of fumonisins in freshly harvested and stored hybrid maize. Journal of Stored Products Research, 36(1), 75-87.
Ouakhssase, A., Chahid, A., Choubbane, H., Aitmazirt, A., & Addi, E. A. (2019). Optimization and validation of a liquid chromatography/tandem mass spectrometry (LC-MS/MS) method for the determination of aflatoxins in maize. Heliyon, 5(5), e01565.
Probst, C., Bandyopadhyay, R., & Cotty, P. J. (2014). Diversity of aflatoxin-producing fungi and their impact on food safety in sub-Saharan Africa. International Journal of Food Microbiology, 174, 113-122.
Probst, C., Bandyopadhyay, R., & Cotty, P. J. (2014). Diversity of aflatoxin-producing fungi and their impact on food safety in sub-Saharan Africa. International Journal of Food Microbiology, 174, 113-122.
Qazi, J. I., & Fayyaz, Z. (2006). Aflatoxin contaminated foods and health risk perspective for Pakistani population. Mycopathologia, 4(2), 27-34.
Rahimi, S., Sohrabi, N., Ebrahimi, M. A., Tebyanian, M., Taghyzadeh, M., & Rahimi, S. A. (2016). Studying the effect of aflatoxin genes Aflp and Aflq on Aspergillus flavus and Aspergillus parasiticus in the cattle feed used in industrial animal husbandries. Acta Medica Austriaca, 32, 2091.
Razzaghi-Abyaneh, M., Shams-Ghahfarokhi, M., Yoshinari, T., Rezaee, M. B., Jaimand, K., Nagasawa, H., & Sakuda, S. (2008). Inhibitory effects of Satureja hortensis L. essential oil on growth and aflatoxin production by Aspergillus parasiticus. International Journal of Food Microbiology, 123(3), 228-233.
Reddy, E. S., Sudhakar, C., & Reddy, N. E. (2011). Aflatoxin contamination in groundnut induced by Aspergillus flavus type fungi: a critical review. International Journal of Applied Biology and Pharmaceutical Technology, 2(2), 180-192.
Riba, A., Mokrane, S., Mathieu, F., Lebrihi, A., & Sabaou, N. (2008). Mycoflora and ochratoxin A producing strains of Aspergillus in Algerian wheat. International Journal of Food Microbiology, 122(1-2), 85-92.
Richard, J. L. (2007). Some major mycotoxins and their mycotoxicoses—An overview. International Journal of Food Microbiology, 119(1-2), 3-10.
Richard, J. L., Payne, G. A., Desjardins, A. E., Maragos, C., Norred, W. P., & Pestka, J. J. (2003). Mycotoxins: risks in plant, animal and human systems. CAST Task Force Report, 139, 101-103.
Rocha, L. O., Nakai, V. K., Braghini, R., Reis, T. A., Kobashigawa, E., & Corrêa, B. (2009). Mycoflora and co-occurrence of fumonisins and aflatoxins in freshly harvested corn in different regions of Brazil. International Journal of Molecular Sciences, 10(11), 5090-5103.
Ruiqian, L., Qian, Y., Thanaboripat, D., & Thansukon, P. (2004). Biocontrol of Aspergillus flavus and aflatoxin production. Current Applied Science and Technology, 4(1), 238-250.
Sabahat, A., Bhatti, I. A., Asi, M. R., Bhatti, H. N., & Sheikh, M. A. (2010). Occurrence of aflatoxins in maize grains from central areas of Punjab, Pakistan. International Journal of Agriculture and Biology, 12(4), 571-575.
Scherm, B., Palomba, M., Serra, D., Marcello, A., & Migheli, Q. (2005). 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. International Journal of Food Microbiology, 98(2), 201-210.
Seerat, W., Akram, A., Qureshi, R., Yaseen, G., Mukhtar, T., & Hanif, N. Q. (2022). Light and scanning electron microscopic characterization of aflatoxins producing Aspergillus flavus in the maize crop. Microscopy Research and Technique, 85(8), 2894-2903.
Shah, H. U., Simpson, T. J., Alam, S., Khattak, K. F., & Perveen, S. (2010). Mould incidence and mycotoxin contamination in maize kernels from Swat Valley, North West Frontier Province of Pakistan. Food and Chemical Toxicology, 48(4), 1111-1116.
Sherma, J. (2000). Thin-layer chromatography in food and agricultural analysis. Journal of Chromatography A, 880(1-2), 129-147.
Teja, R. M., Vijay Krishna Kumar, K., Srilakshmi, P., Sudini, H., Kishore Varma, P., & Koteswara Rao, S. R. (2017). Prevalence of Aspergillus flavus Infection and Aflatoxin Contamination of Groundnut in Telangana and Andhra Pradesh. International Journal of Pure and Applied Bioscience, 5(5), 1603-1614.
Upadhaya, S. D., Park, M. A., & Ha, J. K. (2010). Mycotoxins and their biotransformation in the rumen: a review. Asian-Australasian Journal of Animal Sciences, 23(9), 1250-1260.
Velmourougane, K., Bhat, R., Gopinandhan, T. N., & Panneerselvam, P. (2011). Management of Aspergillus ochraceus and Ochratoxin-A contamination in coffee during on-farm processing through commercial yeast inoculation. Biological Control, 57(3), 215-221.
Waliyar, F., Reddy, S. V., Subramaniam, K., Reddy, T. Y., Devi, K. R., Craufurd, P. Q., & Wheeler, T. R. (2003). Importance of mycotoxins in food and feed in India. Aspects of Applied Biology, 68, 147-154.
Yu, J. H., & Keller, N. (2005). Regulation of secondary metabolism in filamentous fungi. Annual Review of Phytopathology, 43, 437-458.
Zhou, R., Liu, M., Liang, X., Su, M., & Li, R. (2019). Clinical features of aflatoxin B1-exposed patients with liver cancer and the molecular mechanism of aflatoxin B1 on liver cancer cells. Environmental Toxicology and Pharmacology, 71, 103225.