FUNGUS ISOLATION, DETERMINATION OF AFLATOXIN AND OCHRATOXIN A FROM Corylus avellana (HAZELNUT) SAMPLES COLLECTED FROM LOCAL MARKETS IN AFYONKARAHİSAR PROVINCE

Öz

In our study, hazelnut samples taken randomly from 20 different markets in Afyonkarahisar were evaluated by High Performance Liquid Chromatography (HPLC) for their aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2), total aflatoxin (AF) and ochratoxin A (OTA) contents. In addition, in order to determine the fungal contamination in the samples, fungus was isolated and classified at the genus level. According to the results obtained, the fungus was isolated from hazelnut samples in all regions except two localities. 1.8070 ng/ml AFG1 was found to be the highest aflatoxin amount in the central 5th sampling area. When the data obtained was evaluated, the detected aflatoxin amounts did not exceed the Turkish Food Codex limits. Therefore, no high potential risk was detected in the hazelnut samples in the study. However, in order to make a potential risk assessment; even if aflatoxin and ochratoxin are present in small amounts, they should be evaluated for the possibility of being taken into the body with various foods and accumulating in the liver in the long term, and this study will the basis for future studies. Additionally, there has been no previous study on this subject in Afyonkarahisar.

Anahtar Kelimeler

• Aflatoxin
• Corylus avellana
• fungal contamination
• ochratoxin.

AFYONKARAHİSAR İLİNDE YEREL MARKETLERDEN TOPLANAN CORYLUS AVELLANA (FINDIK) ÖRNEKLERİNDEN FUNGUS İZOLASYONU, AFLATOKSİN VE OKRATOKSİN A MİKTARININ BELİRLENMESİ

Öz

Çalışmamızda Afyonkarahisar’da rastgele 20 farklı marketten alınan fındık örnekleri aflatoksin B1 (AFB1), aflatoksin B2 (AFB2), aflatoksin G1 (AFG1), aflatoksin G2 (AFG2, toplam aflatoksin (AF) ve okratoksin A (OTA) içerikleri bakımından Yüksek Performanslı Sıvı Kromotografisi (HPLC) ile incelenmiştir. Ayrıca örneklerdeki fungal kontaminasyonu belirlemek amacıyla, fungus izolasyonu yapılarak genus seviyesinde sınıflandırılmaları gerçekleştirilmiştir. Elde edilen sonuçlara göre iki lokalite dışında kalan tüm bölgelerdeki fındık örneklerinden fungus izole edilmiştir. Aflatoksin taramalarında merkez 5. örneklem alanında 1.8070 ng/ml AFG1 en yüksek aflatoksin miktarı olarak bulunmuştur. Elde edilen veriler değerlendirildiğinde tespit edilen aflatoksin miktarları Türk Gıda Kodeksi sınırlarını aşmamıştır. Dolayısıyla yapılan çalışma ile fındık örneklerinde yüksek potansiyel bir risk tespit edilmemiştir. Ancak potansiyel risk değerlendirmesinin yapılabilmesi için; az miktarlarda aflatoksin ve okratoksin varlığının çeşitli gıdalarla vücuda alınarak uzun vadede karaciğerde birikme ihtimali açısından da değerlendirilmesi gerekmekte olup, bu çalışma ileri çalışmalar için bir temel oluşturmaktadır. Ayrıca daha önce bu konuda Afyonkarahisar’da yapılmış herhangi bir çalışma bulunmamaktadır.

Anahtar Kelimeler

• Aflatoksin
• Corylus avellana
• fungal kontaminasyon
• okratoksin.

Kaynakça

1. Adetunji, M.C., Alika, O.P., Awa, N.P., Atanda, O.O., Mwanza, M. (2018). Microbiological quality and risk assessment for aflatoxins in groundnuts androasted cashew nuts meant for human consumption. Journal of Toxicology, http://dx.doi.org/10.1155/2018/1308748.
2. Afolabi, C.G., Ezekiel, C.N., Ogunbiyi, A.E., Oluwadairo, O.J., Sulyok, M. (2020). Fungi and mycotoxins in cowpea (Vigna unguiculata L) on Nigerian markets. Food Additives & Contaminants: Part B, 13(1):52-58.
3. Akçin, Y., Bostan, S.Z. (2019). Farklı Sulama Programlarının ‘Tombul’ Fındık Çeşidinde Depolama Süresince Su Aktivitesine Etkisi. Uluslararası Tarım ve Yaban Hayatı Bilimleri Dergisi, 5(2):308- 313.
4. Anonymous (2021). Minimizing risks posed by mycotoxins utilizing the HACCP concept. https://www.fao.org/3/x2100t/x2100t08.htm (Erişim Tarihi: 05.11.2021)
5. Arroyo-Manzanares, N., Huertas-Pérez, J., Gámiz-Gracia, L., García-Campaña, A.M. (2013). A new approach in sample treatment combined with UHPLC-MS/MS for the determination of multiclass mycotoxins in edible nuts and seeds. Talanta, 115:61-67.
6. Asghar, M.A., Ahmed, A., Iqbal, J. (2016). Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan. Food Additives & Contaminants: Part B, 9(1):51-58. http://dx.doi.org/10.1080/19393210.2015.1127293.
7. Baquião, A.C., Lopes, E.L., Corrêa, B. (2016). Molecular and mycotoxigenic biodiversity of Aspergillus flavus isolated from Brazil nuts. Food Research International, 89:266-271.
8. Barnett, H.L., Hunter, B.B. (1998). Illustrated genera of imperfect fungi, 4th Ed. USA. Prentice-Hall, Inc. https://www.academia.edu/35499449/ Illustrated_genera_of_imperfect_fungi_fourth_ edition. Barnett_y_Hunter._pdf.pdf, 19.03.2020.

9. Basaran, P., Özcan, M. (2008). Occurrence of aflatoxins in various nuts commercialized in Turkey. https://onlinelibrary.wiley.com/ doi/pdf/10.1111/j.1745-4565.2008.00143.
10. Belli, N., Ramos, A.J., Coronas, I., Sanchis, V., Marin, S. (2005). Aspergillus carbonarius growth and ochratoxin A production on a synthetic grape medium in relation to environmental factors. Journal of Applied Microbiology, 98:839-844.
11. Blomhoff, R., Carlsen, M.H., Andersen, L.F., Jacobs, D.R. (2006). Health benefitsof nuts: potential role of antioxidants. British Journal of Nutrition, 96(2):52-60. http://dx.doi.org/ 10.1017/BJN20061864.
12. Borchers, A., Teuber, S.S., Keen, C.L., Gershwin, M.E. (2010). Food safety. Clinical Reviews in Allergy and Immunology, 9:95-141.
13. Butler, E.E., Mann, M.P. (1959). Use of cellophane tape for mounting and photographing phytopathogenic fungi. Phytopathology, 49:231-232.
14. Cary, J.W., Ehrlich, K.C. (2006). Aflatoxigenicity in Aspergillus:molecular genetics, phylogenetic relationships and evolutionary implications. Mycopathologia, 162:167-177.
15. Cheraghali, A.M., Yazdanpanah, H., Doraki, N., Abouhossain, G., Hassibi, M., Aliabad, S., Aliakbarpoor, M., Amirahmadi, M., Askarian, A., Fallah, N., Hashemi, T., Jalali, M., Kalantari, N., Khodadadi, E., Maddah, B., Mohit, R., Mohseny, M., Phaghihy, Z., Rahmani, A., Setoodeh, L., Soleimany, E., Zamanian, F. (2007). Incidence of aflatoxins in Iran pistachio nuts. Food and Chemical Toxicology, 45:812-816.
16. Chulze, S.N., Magnoli, C.E., Dalcero, A.M. (2006). Occurrence of ochratoxin A in wine and ochratoxigenic mycoflora in grapes and dried vine fruits in South America. International Journal of Food Microbiology, 111:5-9.
17. Commission Regulation (EC) No. 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs consolidated version 01 Sept (2012). European Community, Brussels. http://eur-ex.europa.eu/LexUriServ/ LexUriServ.do?uri =CONSLEG:2006R1881:20120901:EN:PDF. Accessed 31 December 2012.
18. Commission Regulation. 2012. (EC) No. 1881/2006 of 19 December 2006 setting maximum levels for certain contaminants in foodstuffs consolidated version 01 Sept (2012). European Community, Brussels.http:// eurlex.europa.eu/LexUriServ/LexUriServ.do?uri= CONSLEG:2006R1881:20120901:EN:PDF. Accessed 31 December 2012.
19. Cunha, S.C., Sá, S.V.M., Fernandes, J.O. (2018). Multiple mycotoxin analysis in nut products: Occurrence and risk characterization. Food and Chemical Toxicology, 114:260-269.
20. Deabes, M. (2010). Fungi and aflatoxins contamination in nuts imported to Saudi Arabia. National Research Center, Saudi Arabia. P310-008 Abstracts/Toxicol Letter, 196S, S37–S351 doi:10.1016/j.toxlet.2010.03.1096.
21. Ekinci, R., Otag, M., Kadakal, C. (2014). Patulin & ergosterol: New quality parameters together with aflatoxins in hazelnuts. Food Chemistry, 150:17-21.
22. Elzupir, A.O., Alamer, A.S. (2015). Quantitative cancer risk of aflatoxin in peanutbutter and vegetable oils: Sudan case study. Toxin Reviews, 33:202-205.
23. Elzupir, A.O. (2018). Seasonal variation and health implications due to aflatoxins in nuts sold in Riyadh region. Revue Francaise d'Allergologie, 59(1):15-21.
24. Emilio, R. (2010). Health benefits of nut consumption. Nutrients, 2:652-682.
25. Fernane, F., Cano-Sancho, G., Sanchis, V., Marín, S., Ramos, A.J. (2010). Aflatoxins and ochratoxin A in pistachios sampled in Spain: occurrence and presence of mycotoxigenic fungi. Food Additives and Contaminants: Part B Surveillance, 3(3):185-192.
26. Ghali, R., Hmaissia-Khlifa, K., Ghorbel, H., Maaroufi, K., Hedili, A. (2009). HPLC determination of ochratoxin A in high consumption Tunisian foods. Food Control, 20:716-720.
27. Gonzalez, L., Juan, C., Soriano, J.M., Molto, J.C., Manes, J. (2006). Occurrence and daily intake of ochratoxin A of organic and nonorganic rice and rice products. International Journal of Food Microbiology, 107:223-227.
28. Grajewski, J., Blajet-Kosicka, A., Twaruzek, M., Kosicki, R. (2012). Occurrence of mycotoxins in polish animal feed in years 2006–2009. Journal of Animal Physiology and Animal Nutrition, 96:870-877.
29. Gürses, M. (2006). Mycoflora and aflatoxin content of hazelnuts, walnuts, peanuts, almonds and roasted chickpeas (leblebi) sold in Turkey. International Journal of Food Properties, 9:395-399.
30. Hammami, W., Fiori, S., Al Thani, R., Ali Kali, N., Balmas, V., Migheli, Q., Jaoua, S. (2014). Fungal and aflatoxin contamination of marketed spices. Food Control, 37:177-181.
31. Hepsag, F., Golge, O., Kabak, B. (2014). Quantitation of aflatoxins in pistachios and groundnuts using HPLC-FLD method. Food Control, 38:75-81.
32. Hepsağ, F., Hayoğlu., İ. (2022). Çukurova ve Doğu Akdeniz Bölgesi’nde satışa sunulan kırmızı pul biber ve kuru incirler’de aflatoksin B1 ve toplam aflatoksin (B1,B2,G1,G2) içeriğinin yüksek performans sıvı kromatografi yöntemi ile belirlenmesi. Osmaniye Korkut Ata Üniversitesi Fen Bilimleri Enstitüsü Dergisi, 5(3): 1393-1406.
33. Hontanaya, C., Meca, G., Luciano, F.B., Manes, J., Font, G. (2015). Inhibition of aflatoxin B1, B2, G1 and G2 production by Aspergillus parasiticus in nuts using yellow and oriental mustard flours. Food Control, 47:154-160.
34. Imperato, R., Campone, L., Piccinelli, A.L., Veneziano, A., Rastrelli, L. (2011). Survey of aflatoxins and ochratoxin a contamination in food products imported in Italy. Food Control, 22:1905-1910.
35. International Agency for Research on Cancer, IARC. 1993. Monographs on the evaluation of carcinogenic risks to humans. 56:257-263.
36. İnanç, A.L. (2024). Dökme ve ambalajlı baharatlık kırmızı biberlerin aflatoksin kontaminasyonunun belirlenmesi, Kahramanmaraş Sütçü İmam Üniversitesi Tarım ve Doğa Dergisi, 27(3):695-703.
37. Kafouris, D., Christofidou, M., Christodoulou, M., Christou, E., Ioannou-Kakouri, E. (2017). A validated UPLC-MS/MS multi-mycotoxin method for nuts and cereals: results of the official control in Cyprus within the EU requirements. Food and Agricultural Immunology, 28:90-108.
38. Karaosmanoğlu, H. (2022). The effect of late harvest on the biometric and color characteristics and aflatoxin level of Tombul hazelnuts. Harran Journal of Agricultural and Food Science, 26(4):549-559.
39. Karaosmanoğlu, H. (2023). Farklı Ambalaj Materyallerinin Depolanan Fındıkların Geometrik ve Renk Özellikleriyle Aflatoksin Oluşumuna Etkisi. Anadolu Tarım Bilimleri Dergisi, 38(2):331-352.
40. Moss, M.O. (1998). Recent studies of mycotoxins. Journal of Applied Microbiology, 84:62-76.
41. Nguyen, K.T.N., Ryu, D. (2014). Concentration of ochratoxin A in breakfast cereals and snacks consumed in the United States. Food Control, 40:140-144.
42. Özay, G., Seyhan, F., Pembeci, C., Saklar, S., Yılmaz, A. (2008). Factors influencing fungal and aflatoxin levels in Turkish hazelnuts (Corylus avellana L.) during growth, harvest, drying and storage: A 3-year study. Food Additives and Contaminants, 25(2):209-218.
43. Pittet, A. (1998). Natural occurrence of mycotoxins in foods and feeds e an updated review. Revue de Medecine Veterinaire, 149:479-492.
44. Prelle, A., Spadaro, D., Garibaldi, A., Gullino, M. (2012). Aflatoxin monitoring in Italian hazelnut products by LCeMS. Food Additives and Contaminants Part B, 5:279-285.
45. Saffari, E., Madani, M., Karbasizade, V., Shakib, P. (2021). Detection of Fungal and Bacterial Contamination of Hazelnut and Determination of Aflatoxin B by HPLC Method in Isfahan, Iran. Current Medical Mycology, 7(4):1-5.
46. Samimi, P., Aslani, R., Molaee‑Aghaee, E., Sadighara, P., Shariatifar, N., Khaniki, G.J., Ozcakmak, S., Reshadat, Z. (2024). Determination and Risk Assessment of Aflatoxin B1 in the Kernel of Imported Raw Hazelnuts from Eastern Azerbaijan Province of Iran. Scientific Reports, 14:6864.
47. Sedefoğlu, C. (2013). Antep fıstıklarında okratoksin A ve aflatoksin varlığının incelenmesi. İstanbul Teknik Üniversitesi, Fen Bilimleri Enstitüsü, Yüksek Lisans Tezi, İstanbul.
48. Stroka, J., Ankle, E., Jorissen, U., Gilbert, J. (2000). Immunoaffinity column clean up with liquid chromatography using post-column bromination for determination of aflatoxins in peanut butter, pistachio paste, fig paste, and paprika powder: collaborative study. The Journal of AOAC International, 83:320-340.
49. Şen, L., Nas, S. (2010). Fındık ve antep fıstığının mikotoksin problemi. Gıda Teknolojileri Elektronik Dergisi, 5(1):49-56.
50. Tolosa, J., Font, G., Mañes, J., Ferrer, E. (2013). Nuts and dried fruits: Natural occurrence of emerging Fusarium mycotoxins. Food Control, 33:215-220.
51. Toptaş, Ö., Erköse, Genç, G. (2023). Yaygın mikotoksinler: aflatoksinler, okratoksin A, fumonisinler, deoksinivalenol ve zearalenon. Journal of Ankara Health Sciences,12(1): 87-98.
52. Turan, A., İslam, A. (2016). Çakıldak Fındık Çeşidinde Kurutma Ortamları ve Muhafaza Süresine Bağlı Olarak Meydana Gelen Değişimler. Ordu Üniversitesi Bilim ve Teknoloji Dergisi, 6(2):272-285.
53. Varga, E., Glauner, T., Berthiller, F., Krska, R., Schuhmacher, R., Sulyok, M. (2013). Development and validation of a (semi-)quantitative UHPLC-MS/MS method for the determination of 191 mycotoxins and other fungal metabolites in almonds, hazelnuts, peanuts and pistachios. Analytical and Bioanalytical Chemistry Research, 405:5087–5104 DOI 10.1007/s00216-013-6831-3.
54. Wu, F., Groopman, J.D., Pestka, J.J. (2014). Public health impacts of foodborne mycotoxins. Annual Review of Food Science and Technology, 351-372.
55. Var, I., Tekin, A. (2023. Piyasada tüketime sunulan ayçiçek ve mısırözü yağlarında aflatoksin varlığının araştırılması. Gıda, 48(6):1304-1317.
56. Yang, J. (2009). Brazil nuts and associated health benefits: a review. LWT - Food Science and Technology, 42:1573-1580.
57. Yu-jiao, W., Ji-yun, N., Zhen, Y., Zhi-xia, L., Yang, C., Farooq, S. (2018). Multi-mycotoxin exposure and risk assessments for Chinese consumption of nuts and dried fruits. Journal of Integrative Agriculture, 17(7):1676-1690.