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Porcine intestinal epithelial barrier disruption by the Fusarium mycotoxins deoxynivalenol and T-2 toxin promotes transepithelial passage of doxycycline and paromomycin

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Abstract
Background: The gastrointestinal tract is the first target for the potentially harmful effects of mycotoxins after intake of mycotoxin contaminated food or feed. With deoxynivalenol (DON), T-2 toxin (T-2), fumonisin B1 (FB1) and zearalenone (ZEA) being important Fusarium toxins in the northern hemisphere, this study aimed to investigate in vitro the toxic effect of these mycotoxins on intestinal porcine epithelial cells derived from the jejunum (IPEC-J2 cells). Viability of IPEC-J2 cells as well as the proportion of apoptotic and necrotic IPEC-J2 cells was determined by flow cytometry after 72 h of exposure to the toxins. Correlatively, the integrity of the intestinal epithelial cell monolayer was studied using Transwell (R) inserts, in which the trans-epithelial electrical resistance (TEER) and passage of the antibiotics doxycycline and paromomycin were used as endpoints. Results: We demonstrated that the percentage of Annexin-V-FITC and PI negative (viable) cells, Annexin-V-FITC positive and PI negative (apoptotic) cells and Annexin-V-FITC and PI positive (necrotic) IPEC-J2 cells showed a mycotoxin concentration-dependent relationship with T-2 toxin being the most toxic. Moreover, the ratio between Annexin-V-FITC positive and PI negative cells and Annexin-V-FITC and PI positive cells varied depending on the type of toxin. More Annexin-V-FITC and PI positive cells could be found after treatment with T-2 toxin, while more Annexin-V-FITC positive and PI negative cells were found after exposure to DON. Consistent with the cytotoxicity results, both DON and T-2 decreased TEER and increased cellular permeability to doxycycline and paromomycin in a time-and concentration-dependent manner. Conclusions: It was concluded that Fusarium mycotoxins may severely disturb the intestinal epithelial barrier and promote passage of antibiotics.
Keywords
Mycotoxins, Cytotoxicity, Flow cytometry, Permeability, Intestinal cells, RIBOTOXIC STRESS-RESPONSE, IN-VITRO, FUMONISIN B-1, CACO-2 CELLS, P-GLYCOPROTEIN, PROTEIN-KINASE, LINE CACO-2, CYTOTOXICITY, ZEARALENONE, METABOLISM

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Chicago
Goossens, Joline, Frank Pasmans, Elin Verbrugghe, Virginie Vandenbroucke, Siegrid De Baere, Evelyne Meyer, Freddy Haesebrouck, Patrick De Backer, and Siska Croubels. 2012. “Porcine Intestinal Epithelial Barrier Disruption by the Fusarium Mycotoxins Deoxynivalenol and T-2 Toxin Promotes Transepithelial Passage of Doxycycline and Paromomycin.” Bmc Veterinary Research 8.
APA
Goossens, Joline, Pasmans, F., Verbrugghe, E., Vandenbroucke, V., De Baere, S., Meyer, E., Haesebrouck, F., et al. (2012). Porcine intestinal epithelial barrier disruption by the Fusarium mycotoxins deoxynivalenol and T-2 toxin promotes transepithelial passage of doxycycline and paromomycin. BMC VETERINARY RESEARCH, 8.
Vancouver
1.
Goossens J, Pasmans F, Verbrugghe E, Vandenbroucke V, De Baere S, Meyer E, et al. Porcine intestinal epithelial barrier disruption by the Fusarium mycotoxins deoxynivalenol and T-2 toxin promotes transepithelial passage of doxycycline and paromomycin. BMC VETERINARY RESEARCH. 2012;8.
MLA
Goossens, Joline, Frank Pasmans, Elin Verbrugghe, et al. “Porcine Intestinal Epithelial Barrier Disruption by the Fusarium Mycotoxins Deoxynivalenol and T-2 Toxin Promotes Transepithelial Passage of Doxycycline and Paromomycin.” BMC VETERINARY RESEARCH 8 (2012): n. pag. Print.
@article{3156970,
  abstract     = {Background: The gastrointestinal tract is the first target for the potentially harmful effects of mycotoxins after intake of mycotoxin contaminated food or feed. With deoxynivalenol (DON), T-2 toxin (T-2), fumonisin B1 (FB1) and zearalenone (ZEA) being important Fusarium toxins in the northern hemisphere, this study aimed to investigate in vitro the toxic effect of these mycotoxins on intestinal porcine epithelial cells derived from the jejunum (IPEC-J2 cells). Viability of IPEC-J2 cells as well as the proportion of apoptotic and necrotic IPEC-J2 cells was determined by flow cytometry after 72 h of exposure to the toxins. Correlatively, the integrity of the intestinal epithelial cell monolayer was studied using Transwell (R) inserts, in which the trans-epithelial electrical resistance (TEER) and passage of the antibiotics doxycycline and paromomycin were used as endpoints. 
Results: We demonstrated that the percentage of Annexin-V-FITC and PI negative (viable) cells, Annexin-V-FITC positive and PI negative (apoptotic) cells and Annexin-V-FITC and PI positive (necrotic) IPEC-J2 cells showed a mycotoxin concentration-dependent relationship with T-2 toxin being the most toxic. Moreover, the ratio between Annexin-V-FITC positive and PI negative cells and Annexin-V-FITC and PI positive cells varied depending on the type of toxin. More Annexin-V-FITC and PI positive cells could be found after treatment with T-2 toxin, while more Annexin-V-FITC positive and PI negative cells were found after exposure to DON. Consistent with the cytotoxicity results, both DON and T-2 decreased TEER and increased cellular permeability to doxycycline and paromomycin in a time-and concentration-dependent manner. 
Conclusions: It was concluded that Fusarium mycotoxins may severely disturb the intestinal epithelial barrier and promote passage of antibiotics.},
  articleno    = {245},
  author       = {Goossens, Joline and Pasmans, Frank and Verbrugghe, Elin and Vandenbroucke, Virginie and De Baere, Siegrid and Meyer, Evelyne and Haesebrouck, Freddy and De Backer, Patrick and Croubels, Siska},
  issn         = {1746-6148},
  journal      = {BMC VETERINARY RESEARCH},
  keyword      = {Mycotoxins,Cytotoxicity,Flow cytometry,Permeability,Intestinal cells,RIBOTOXIC STRESS-RESPONSE,IN-VITRO,FUMONISIN B-1,CACO-2 CELLS,P-GLYCOPROTEIN,PROTEIN-KINASE,LINE CACO-2,CYTOTOXICITY,ZEARALENONE,METABOLISM},
  language     = {eng},
  pages        = {10},
  title        = {Porcine intestinal epithelial barrier disruption by the Fusarium mycotoxins deoxynivalenol and T-2 toxin promotes transepithelial passage of doxycycline and paromomycin},
  url          = {http://dx.doi.org/10.1186/1746-6148-8-245},
  volume       = {8},
  year         = {2012},
}

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