Abstract
Enniatins, the most prevalent emerging mycotoxins, represent an emerging food safety issue, because of their common co-occurrence with other fusariotoxins such as trichothecenes co-produced by Fusarium spp on field grains and because of their extensive prevalence in grains. In this study, the intestinal toxicity of enniatin B1 (ENN) alone and mixed with the most toxic trichothecene T-2 toxin (T2) was characterized by using two biological models from pig, the most sensitive species: the intestinal cell line IPEC1 (in vitro exposure) and jejunal explants (ex vivo exposure). Dose-dependent decreases in cell proliferation in IPEC1 and in the histopathological scores of explants were observed for ENN at μM-levels and for T2 at nM-levels, with IC50 values for ENN of 15.8 and 29.7 μM, and for T2 of 9.3 and 15.1 nM in vitro and ex vivo, respectively. Interaction analysis by probabilistic and by determinist approaches showed a less than additive effect both in vitro and ex vivo, at IC50 values, with increasing antagonism with decreasing concentrations of toxins. The results obtained by the determinist median-effect dose analysis and by the nonlinear regression analysis were concordant. All the median-effect doses estimated for IPEC cells were included in the IC50 confidence intervals of the nonlinear regression fitting. Given the occurrence of enniatins, potential synergy following the co-occurrence of enniatins and the major fusariotoxins, especially trichothecene B deoxynivalenol should be investigated.
Similar content being viewed by others
References
Behm C, Degen GH, Föllmann W (2009) The Fusarium toxin enniatin B exerts no genotoxic activity, but pronounced cytotoxicity in vitro. Mol Nutr Food Res 53:423–430
Behm C, Föllmann W, Degen GH (2012) Cytotoxic Potency of Mycotoxins in Cultures of V79 Lung Fibroblast Cells. J Toxicol Environ Health Part A 75:1226–1231
Berenbaum MC (1985) The expected effect of a combination of agents: the general solution. J Theor Biol 114:413–431
Borgert CJ, Price B, Wells CS (2001) Evaluating chemical interaction studies for mixture risk assessment. Hum Ecol Risk Assess 7:259–306
Bosch U, Mirocha CJ, Abbas HK, di Menna M (1989) Toxicity and toxin production by Fusarium isolates from New Zealand. Mycopathologia 108:73–79
Bouhet S, Le Dorze E, Pérès SY, Fairbrother JM, Oswald IP (2006) Mycotoxin fumonisin B1 selectively down-regulates the basal IL-8 expression in pig intestine: in vivo and in vitro studies. Food Chem Toxicol 44:1768–1773
Chou TC (2006) Theoretical basis, experimental design, and computerized simulation of synergism and antagonism in drug combination studies. Pharmacol Rev 58:621–681
Chou TC, Talalay P (1984) Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 22:27–55
Doi K, Shinozuka J, Sehata S (2006) T-2 toxin and apoptosis. J Toxicol Pathol 19:15-27
EFSA (2011) Scientific Opinion on the risks for animal and public health related to the presence of T-2 and HT-2 toxin in food and feed. EFSA J 9:1–187
Föllmann W, Behm C, Degen GH (2009) The emerging Fusarium toxin enniatin B: in vitro studies on its genotoxic potential and cytotoxicity in V79 cells in relation to other mycotoxins. Mycotoxin Res 25:11–19
Gammelsrud A, Solhaug A, Dendelé B, Sandberg WJ, Ivanova L, Kocbach Bølling A, Lagadic-Gossmann D, Refsnes M, Becher R, Eriksen G, Holme JA (2012) Enniatin B-induced cell death and inflammatory responses in RAW 267.4 murine macrophages. Toxicol Appl Pharmacol 261:74–87
Gaümann E, Naef-Roth S, Ettlinger L (1950) Zur Gewinnung von enniatinen aus dem myzel verschiedener fusarien. Phytopathol Z 16:289–299
Hymery N, Sibiril Y, Parent-Massin D (2006) In vitro effects of trichothecenes on human dendritic cells. Toxicol In Vitro 20:899–909
Ivanova L, Skjerve E, Eriksen GS, Uhlig S (2006) Cytotoxicity of enniatins A, A1, B, B1, B2 and B3 from Fusarium avenaceum. Toxicon 47:868–876
Ivanova L, Uhlig S, Eriksen GS, Johannessen LE (2010) Enniatin B1 is a substrate of intestinal P-glycoprotein, multidrug resistance-associated protein 2 and breast cancer resistance protein. World Mycotoxin J 3:271–281
Ivanova L, Egge-Jacobsen WM, Solhaug A, Thoen E, Fæste CK (2012) Lysosomes as a possible target of enniatin B-induced toxicity in Caco-2 cells. Chem Res Toxicol 25:1662–1674
Jestoi M (2008) Emerging fusarium-mycotoxins fusaprofilin, beauvericin, enniatins, and moniliformin: a review. Crit Rev Food Sci Nutr 48:21–49
Kolf-Clauw M, Castellote J, Joly B, Bourges-Abella N, Raymond-Letron I, Pinton P et al (2009) Development of a pig jejunal explant culture for studying the gastrointestinal toxicity of the mycotoxin deoxynivalenol: histopathological analysis. Toxicol In Vitro 23:1580–1584
Li G, Shinozuka J, Uetsuka K, Nakayama H, Doi K (1997) T-2 toxin-induced apoptosis in intestinal crypt epithelial cells of mice. Exp Toxicol Pathol. 49:447-50
Meca G, Mañes J, Font G, Ruiz MJ (2012) Study of the potential toxicity of enniatins A, A1, B, B1 by evaluation of duodenal and colonic bioavailability applying an in vitro method by Caco-2 cells. Toxicon 59:1–11
Pinton P, Nougayrède JP, Del Rio JC, Moreno C, Marin DE, Ferrier L, Bracarense AP, Kolf-Clauw M, Oswald IP (2009) The food contaminant deoxynivalenol, decreases intestinal barrier permeability and reduces claudin expression. Toxicol Appl Pharmacol 237:41–48
Pinton P, Guzylack-Piriou L, Kolf-Clauw M, Oswald IP (2012a) Effects of some fungal toxins, the trichothecenes, on the intestine. Curr Immunol Rev 8:193–208
Pinton P, Tsybulskyy D, Lucioli J, Laffitte J, Callu P, Lyazhri F, Grosjean F, Bracarense AP, Kolf-Clauw M, Oswald IP (2012b) Toxicity of deoxynivalenol and its acetylated derivatives on the intestine: differential effects on morphology, barrier function, tight junctions proteins and MAPKinases. J Toxicol Sci 130:180–190
Rodrigues I, Naehrer K (2012) A three-year survey on the worldwide occurrence of mycotoxins in feedstuffs and feed. Toxins 4:663–675
Ruiz MJ, Franzova P, Juan-Garcia A, Font G (2011a) Toxicological interactions between the mycotoxins beauvericin, deoxynivalenol and t-2 toxin in cho-k1 cells in vitro. Toxicon 58:315–326
Ruiz MJ, Macakova P, Juan-Garcia A, Font G (2011b) Cytotoxic effects of mycotoxin combinations in mammalian kidney cells. Food Chem Toxicol 49:2718–2724
Santini A, Meca G, Uhlig S, Ritieni A (2012) Fusaproliferin, beauvericin and enniatins: occurrence in food – a review. World Mycotoxin J 5:71–81
Serrano AB, Font G, Mañes J, Ferrer E (2013) Emerging Fusarium mycotoxins in organic and conventional pasta collected in Spain. Food Chem Toxicol 51:259–266
Streit E, Schwab C, Sulyok M, Naehrer K, Krska R, Schatzmayr G (2013) Multi-Mycotoxin Screening Reveals the Occurrence of 139 Different Secondary Metabolites in Feed and Feed Ingredients. Toxins 5:504–523
Tallarida RJ (2006) An overview of drug combination analysis with isobolograms. J Pharmacol Experiment Therapeutics 319:1–7
Tedjiotsop Feudjio F, Dornetshuber R, Lemmens M, Hoffmann O, Lemmens-Gruber R, Berger W (2010) Beauvericin and enniatin: emerging toxins and/or remedies? World Mycotoxin J 3:415–430
Uhlig S, Torp M, Heier BT (2006) Beauvericin and enniatins A, A1, B and B1 in Norwegian grain: a survey. Food Chem 94:193–201
Van der Fels-Klerx HJ, Stratakou I (2010) T-2 toxin and HT-2 toxin in grain and grain-based commodities in Europe: occurrence, factors affecting occurrence, co-occurrence and toxicological effects. World Mycotoxin J 3:349–367
Van Der Fels-Klerx HJ, Klemsdal S, Hietaniemi V, Lindblad M, Ioannou-Kakouri E, Van Asselt ED (2012) Mycotoxin contamination of cereal grain commodities in relation to climate in North West Europe. Food Addit Contam A 29:1581–1592
Wätjen W, Debbab A, Hohlfeld A, Chovolou Y, Kampkötter A, Edrada RA, Ebel R, Hakiki A, Mosaddak M, Totzke F, Kubbutat MH (2009) Enniatins A1, B and B1 from an endophytic strain of Fusarium tricinctum induce apoptotic cell death in H4IIE hepatoma cells accompanied by inhibition of ERK phosphorylation. Mol Nutr Food Res 53(431):440
Yang GH, Jarvis BB, Chung YJ, Pestka JJ (2000) Apoptosis induction by satratoxins and other trichothecene mycotoxins: relationship to ERK, p38 MAPK, and SAPK/JNK activation. Toxicol Appl Pharmacol 164:149–160
Acknowledgments
MS, JL and JR-G were supported by post-doctoral or doctoral fellowships from CAPES-COFECUB (project 593/08) and IAK by a doctoral fellowship from the Government of the Republic of Benin. This study was supported by the ANR-CESA project DON&Co. The authors are grateful to Pr. TC Chou, Memorial Sloan-Kettering Cancer Center, New York City for allowing special access to his software, to AM Cossalter and P Pinton for technical assistance with the animal experiments and to John Woodley for language editing.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kolf-Clauw, M., Sassahara, M., Lucioli, J. et al. The emerging mycotoxin, enniatin B1, down-modulates the gastrointestinal toxicity of T-2 toxin in vitro on intestinal epithelial cells and ex vivo on intestinal explants. Arch Toxicol 87, 2233–2241 (2013). https://doi.org/10.1007/s00204-013-1067-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00204-013-1067-8