{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,29]],"date-time":"2026-05-29T09:32:25Z","timestamp":1780047145570,"version":"3.53.1"},"reference-count":46,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2025,5,18]],"date-time":"2025-05-18T00:00:00Z","timestamp":1747526400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation (MCIU)","doi-asserted-by":"publisher","award":["PID2022-140722OB-I00"],"award-info":[{"award-number":["PID2022-140722OB-I00"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004837","name":"Spanish Ministry of Science and Innovation (MCIU)","doi-asserted-by":"publisher","award":["10.13039\/501100011033"],"award-info":[{"award-number":["10.13039\/501100011033"]}],"id":[{"id":"10.13039\/501100004837","id-type":"DOI","asserted-by":"publisher"}]},{"name":"European Regional Development Fund (FEDER)","award":["PID2022-140722OB-I00"],"award-info":[{"award-number":["PID2022-140722OB-I00"]}]},{"name":"European Regional Development Fund (FEDER)","award":["10.13039\/501100011033"],"award-info":[{"award-number":["10.13039\/501100011033"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) are two of the most potent mycotoxins, recognized for their severe toxicity. In recent years, the consumption of bioactive substances has proven to be a valuable ally in combating their harmful effects on human health. For this purpose, this study evaluates the protective effects of fermented whey (FW) and pumpkin (P), as functional ingredients in bread, on duodenum tissue against sub-chronic toxicity induced by AFB1 and OTA. Nine groups of male and female Wistar rats (n = 5 per sex\/group) were exposed to different combinations of AFB1, OTA, FW, and P for 28 days. The gene expression of apoptotic and antioxidant markers, including p53, Bax, Hmox1, NF-\u03baB, and occludin, was measured by quantitative real-time PCR (RT-qPCR). AFB1 + OTA exposure led to an increased expression of p53 and NF-\u03baB, with the downregulation of Bax and Hmox1. Occludin expression, which supports tight junction integrity, remained largely unaffected. Supplementation with FW and FW + P modulated gene expression favorably, offering protection against AFB1 and OTA toxicity. These bioactive components effectively mitigated oxidative stress and apoptosis in duodenal tissue. Notably, the results indicate that the protective effects of FW and P are not sex-dependent. These findings highlight the potential of FW and P as functional ingredients in combating the toxic effects of AFB1 and OTA in vivo.<\/jats:p>","DOI":"10.3390\/foods14101793","type":"journal-article","created":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T05:37:13Z","timestamp":1747633033000},"page":"1793","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Evaluation of Dietary Bioactive Agents Against Aflatoxin B1 and Ochratoxin A-Induced Duodenal Toxicity in Rats"],"prefix":"10.3390","volume":"14","author":[{"given":"Sarra","family":"Rafai","sequence":"first","affiliation":[{"name":"Biotech Agrifood, Faculty of Pharmacy and Food Sciences, Universitat de Val\u00e8ncia, Avda, Vicent Andr\u00e9s Estell\u00e9s s\/n, 46100 Burjassot, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4741-3967","authenticated-orcid":false,"given":"Alessandra","family":"Cimbalo","sequence":"additional","affiliation":[{"name":"Biotech Agrifood, Faculty of Pharmacy and Food Sciences, Universitat de Val\u00e8ncia, Avda, Vicent Andr\u00e9s Estell\u00e9s s\/n, 46100 Burjassot, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5777-6107","authenticated-orcid":false,"given":"Lara","family":"Manyes","sequence":"additional","affiliation":[{"name":"Biotech Agrifood, Faculty of Pharmacy and Food Sciences, Universitat de Val\u00e8ncia, Avda, Vicent Andr\u00e9s Estell\u00e9s s\/n, 46100 Burjassot, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Manyes, L., and Font, G. (2023). Mycotoxins: Toxicity, Occurrence, Risk Assessment and Prevention. Encyclopedia of Human Nutrition, Elsevier.","DOI":"10.1016\/B978-0-12-821848-8.00147-5"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Marchese, S., Polo, A., Ariano, A., Velotto, S., Costantini, S., and Severino, L. (2018). Aflatoxin B1 and M1: Biological Properties and Their Involvement in Cancer Development. Toxins, 10.","DOI":"10.3390\/toxins10060214"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"984","DOI":"10.1016\/j.fct.2007.10.005","article-title":"On the Occurrence of Aflatoxin M1 in Milk and Dairy Products","volume":"47","author":"Prandini","year":"2009","journal-title":"Food Chem. Toxicol."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Fouad, A.M., Ruan, D., El-Senousey, H.K., Chen, W., Jiang, S., and Zheng, C. (2019). Harmful Effects and Control Strategies of Aflatoxin B1 Produced by Aspergillus Flavus and Aspergillus Parasiticus Strains on Poultry: Review. Toxins, 11.","DOI":"10.3390\/toxins11030176"},{"key":"ref_5","first-page":"1","article-title":"Mycotoxicity and Embryonic Development: I- Aflatoxin B1 Reduces Quality and Birth Rate during Mice Embryonic Development","volume":"6","author":"Ali","year":"2023","journal-title":"SVU-Int. J. Vet. Sci."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.trsl.2018.09.001","article-title":"Epigenetic Alterations Caused by Aflatoxin B1: A Public Health Risk in the Induction of Hepatocellular Carcinoma","volume":"204","author":"Ferreira","year":"2019","journal-title":"Transl. Res."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Yang, X., Li, X., Gao, Y., Wang, J., and Zheng, N. (2023). Integrated Metabolomics and Lipidomics Analysis Reveals Lipid Metabolic Disorder in NCM460 Cells Caused by Aflatoxin B1 and Aflatoxin M1 Alone and in Combination. Toxins, 15.","DOI":"10.3390\/toxins15040255"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"461","DOI":"10.3390\/toxins2040461","article-title":"Ochratoxin A: General Overview and Actual Molecular Status","volume":"2","author":"Atoui","year":"2010","journal-title":"Toxins"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"107230","DOI":"10.1016\/j.foodcont.2020.107230","article-title":"Ochratoxin\u2014Characteristics, Influences and Challenges of Global Research","volume":"114","author":"Braun","year":"2020","journal-title":"Food Control"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Longobardi, C., Damiano, S., Andretta, E., Prisco, F., Russo, V., Pagnini, F., Florio, S., and Ciarcia, R. (2021). Curcumin Modulates Nitrosative Stress, Inflammation, and DNA Damage and Protects against Ochratoxin A-Induced Hepatotoxicity and Nephrotoxicity in Rats. Antioxidants, 10.","DOI":"10.3390\/antiox10081239"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1007\/s00204-019-02434-5","article-title":"Nephrotoxicity Instead of Immunotoxicity of OTA Is Induced through DNMT1-Dependent Activation of JAK2\/STAT3 Signaling Pathway by Targeting SOCS3","volume":"93","author":"Gan","year":"2019","journal-title":"Arch. Toxicol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"106","DOI":"10.1016\/j.envpol.2018.06.062","article-title":"OTA Induces Intestinal Epithelial Barrier Dysfunction and Tight Junction Disruption in IPEC-J2 Cells through ROS\/Ca2+-Mediated MLCK Activation","volume":"242","author":"Wang","year":"2018","journal-title":"Environ. Pollut."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"320","DOI":"10.1016\/j.fct.2018.01.002","article-title":"Ochratoxin A: Toxicity, Oxidative Stress and Metabolism","volume":"112","author":"Tao","year":"2018","journal-title":"Food Chem. Toxicol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.toxicon.2020.07.004","article-title":"Genotoxic Effects of Mycotoxins","volume":"185","author":"Yilmaz","year":"2020","journal-title":"Toxicon"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"230","DOI":"10.47836\/ifrj.28.2.02","article-title":"Mycotoxins in Foods, from the Field to the Plate: A Review","volume":"28","year":"2021","journal-title":"Int. Food Res. J."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.1021\/acsfoodscitech.1c00220","article-title":"Food Mycotoxins: Dietary Interventions Implicated in the Prevention of Mycotoxicosis","volume":"1","author":"Sharma","year":"2021","journal-title":"ACS Food Sci. Technol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"11250","DOI":"10.1039\/D1FO02059E","article-title":"Bioaccessibility and Bioavailability of Bioactive Compounds from Yellow Mustard Flour and Milk Whey Fermented with Lactic Acid Bacteria","volume":"12","author":"Manyes","year":"2021","journal-title":"Food Funct."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1111\/1471-0307.12660","article-title":"A Natural Strategy to Improve the Shelf Life of the Loaf Bread against Toxigenic Fungi: The Employment of Fermented Whey Powder","volume":"73","author":"Luz","year":"2020","journal-title":"Int. J. Dairy. Tech."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Fiorbelli, E., Lapris, M., Errico, M., Della Badia, A., Riahi, I., Rocchetti, G., and Gallo, A. (2024). Mycotoxin Challenge in Dairy Cows: Assessment of the Efficacy of an Anti-Mycotoxin Agent by Adopting an In Vitro Rumen Simulation Method. Toxins, 16.","DOI":"10.3390\/toxins16110490"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Zavistanaviciute, P., Ruzauskas, M., Antanaitis, R., Televicius, M., Lele, V., Santini, A., and Bartkiene, E. (2023). Antimicrobial and Mycotoxin Reducing Properties of Lactic Acid Bacteria and Their Influence on Blood and Feces Parameters of Newborn Calves. Animals, 13.","DOI":"10.3390\/ani13213345"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Bergantin, C., Maietti, A., Tedeschi, P., Font, G., Manyes, L., and Marchetti, N. (2018). HPLC-UV\/Vis-APCI-MS\/MS Determination of Major Carotenoids and Their Bioaccessibility from \u201cDelica\u201d (Cucurbita Maxima) and \u201cViolina\u201d (Cucurbita Moschata) Pumpkins as Food Traceability Markers. Molecules, 23.","DOI":"10.3390\/molecules23112791"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"2200902","DOI":"10.1002\/mnfr.202200902","article-title":"The Protective Effect of Pumpkin and Fermented Whey Mixture against AFB1 and OTA Immune Toxicity In Vitro. A Transcriptomic Approach","volume":"67","author":"Frangiamone","year":"2023","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Vila-Donat, P., S\u00e1nchez, D., Cimbalo, A., Ma\u00f1es, J., and Manyes, L. (2024). Effect of Bioactive Ingredients on Urinary Excretion of Aflatoxin B1 and Ochratoxin A in Rats, as Measured by Liquid Chromatography with Fluorescence Detection. Toxins, 16.","DOI":"10.3390\/toxins16080363"},{"key":"ref_24","unstructured":"World Health Organization, and Food and Agriculture Organization of the United Nations and Joint FAO\/WHO Expert Committee on Food Additives (2022). Evaluation of Certain Food Additives and Contaminants: Ninety-First Report of the Joint FAO\/WHO Expert Committee on Food Additives, World Health Organization. [1st ed.]."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Rotimi, O.A., Rotimi, S.O., Goodrich, J.M., Adelani, I.B., Agbonihale, E., and Talabi, G. (2019). Time-Course Effects of Acute Aflatoxin B1 Exposure on Hepatic Mitochondrial Lipids and Oxidative Stress in Rats. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.00467"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1016\/j.fct.2017.11.003","article-title":"Sex Differences in Ochratoxin a Toxicity in F344 Rats after 7 and 21 Days of Daily Oral Administration","volume":"111","author":"Pastor","year":"2018","journal-title":"Food Chem. Toxicol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1857","DOI":"10.1002\/tox.23306","article-title":"Toxicogenicity and Mechanistic Pathways of Aflatoxin B1 Induced Renal Injury","volume":"36","author":"Dlamini","year":"2021","journal-title":"Environ. Toxicol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"107085","DOI":"10.1016\/j.toxicon.2023.107085","article-title":"Toxicogenic Fungal Profile, Ochratoxin A Exposure and Cancer Risk Characterization through Maize (Zea Mays) Consumed by Different Age Populations in the Volta Region of Ghana","volume":"226","author":"Kortei","year":"2023","journal-title":"Toxicon"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Sobral, M.M.C., Gon\u00e7alves, T., Martins, Z.E., B\u00e4uerl, C., Cort\u00e9s-Mac\u00edas, E., Collado, M.C., and Ferreira, I.M.P.L.V.O. (2022). Mycotoxin Interactions along the Gastrointestinal Tract: In Vitro Semi-Dynamic Digestion and Static Colonic Fermentation of a Contaminated Meal. Toxins, 14.","DOI":"10.3390\/toxins14010028"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"121639","DOI":"10.1016\/j.jhazmat.2019.121639","article-title":"Neurotoxic Effects of Aflatoxin B1 on Human Astrocytes in Vitro and on Glial Cell Development in Zebrafish in Vivo","volume":"386","author":"Park","year":"2020","journal-title":"J. Hazard. Mater."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Xiao, S., Wu, Y., Gao, S., Zhou, M., Liu, Z., Xiong, Q., Jiang, L., Yuan, G., Li, L., and Yang, L. (2023). Deciphering the Hazardous Effects of AFB1 and T-2 Toxins: Unveiling Toxicity and Oxidative Stress Mechanisms in PK15 Cells and Mouse Kidneys. Toxins, 15.","DOI":"10.3390\/toxins15080503"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"138909","DOI":"10.1016\/j.foodchem.2024.138909","article-title":"In Vitro and in Vivo Assessment of AFB1 and OTA Toxic Effects and the Beneficial Role of Bioactive Compounds. A Systematic Review","volume":"447","author":"Frangiamone","year":"2024","journal-title":"Food Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1038\/cdd.2017.169","article-title":"How Does P53 Induce Apoptosis and How Does This Relate to P53-Mediated Tumour Suppression?","volume":"25","author":"Aubrey","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1038\/cdd.2017.179","article-title":"MOMP, Cell Suicide as a BCL-2 Family Business","volume":"25","author":"Kalkavan","year":"2018","journal-title":"Cell Death Differ."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e13338","DOI":"10.1111\/1541-4337.13338","article-title":"Toxicodynamic of Combined Mycotoxins: MicroRNAs and Acute-phase Proteins as Diagnostic Biomarkers","volume":"23","author":"Murtaza","year":"2024","journal-title":"Comp. Rev. Food Sci. Food Safe"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Cimbalo, A., Alonso-Garrido, M., Font, G., Frangiamone, M., and Manyes, L. (2021). Transcriptional Changes after Enniatins A, A1, B and B1 Ingestion in Rat Stomach, Liver, Kidney and Lower Intestine. Foods, 10.","DOI":"10.3390\/foods10071630"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Pfeffer, C., and Singh, A. (2018). Apoptosis: A Target for Anticancer Therapy. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19020448"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Serasanambati, M., and Chilakapati, S.R. (2016). Function of Nuclear Factor Kappa B (NF-kB) in Human Diseases-A Review. S. Indian J. Biol. Sci., 2.","DOI":"10.22205\/sijbs\/2016\/v2\/i4\/103443"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"618","DOI":"10.1002\/mco2.104","article-title":"NF-\u03baB Signaling in Inflammation and Cancer","volume":"2","author":"Zhang","year":"2021","journal-title":"MedComm"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"842","DOI":"10.5187\/jast.2022.e49","article-title":"Gene Expression Profiling after Ochratoxin A Treatment in Small Intestinal Epithelial Cells from Pigs","volume":"64","author":"Yoon","year":"2022","journal-title":"J. Anim. Sci. Technol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"19376","DOI":"10.18632\/oncotarget.14270","article-title":"Ochratoxin A Induces ER Stress and Apoptosis in Mesangial Cells via a NADPH Oxidase-Derived Reactive Oxygen Species-Mediated Calpain Activation Pathway","volume":"8","author":"Sheu","year":"2017","journal-title":"Oncotarget"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Wu, J., He, C., Bu, J., Luo, Y., Yang, S., Ye, C., Yu, S., He, B., Yin, Y., and Yang, X. (2020). Betaine Attenuates LPS-Induced Downregulation of Occludin and Claudin-1 and Restores Intestinal Barrier Function. BMC Vet. Res., 16.","DOI":"10.1186\/s12917-020-02298-3"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Escriv\u00e1, L., Agahi, F., Vila-Donat, P., Ma\u00f1es, J., Meca, G., and Manyes, L. (2021). Bioaccessibility Study of Aflatoxin B1 and Ochratoxin A in Bread Enriched with Fermented Milk Whey and\/or Pumpkin. Toxins, 14.","DOI":"10.3390\/toxins14010006"},{"key":"ref_44","first-page":"173","article-title":"The Role of Pumpkin Pulp Extract Carotenoids against Mycotoxin Damage in the Blood Brain Barrier In Vitro","volume":"72","author":"Font","year":"2021","journal-title":"Arch. Ind. Hyg. Toxicol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"103039","DOI":"10.1016\/j.algal.2023.103039","article-title":"Biological Detoxification of Mycotoxins: Emphasizing the Role of Algae","volume":"71","author":"Yadavalli","year":"2023","journal-title":"Algal Res."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Pauletto, M., Giantin, M., Tolosi, R., Bassan, I., Barbarossa, A., Zaghini, A., and Dacasto, M. (2021). Discovering the Protective Effects of Resveratrol on Aflatoxin B1-Induced Toxicity: A Whole Transcriptomic Study in a Bovine Hepatocyte Cell Line. Antioxidants, 10.","DOI":"10.3390\/antiox10081225"}],"container-title":["Foods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-8158\/14\/10\/1793\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:34:46Z","timestamp":1760031286000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-8158\/14\/10\/1793"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,18]]},"references-count":46,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2025,5]]}},"alternative-id":["foods14101793"],"URL":"https:\/\/doi.org\/10.3390\/foods14101793","relation":{},"ISSN":["2304-8158"],"issn-type":[{"value":"2304-8158","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,18]]}}}