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Cytotoxic effects of the Fusarium mycotoxins deoxynivalenol, T2-toxin, fumonisin B1 and zearalenone on intestinal porcine epithelial cells derived from the jejunum

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Abstract
In the northern hemisphere, Fusarium species are the most common moulds invading grain and maize plants (1). Trichothecenes, fumonisins and zearalenone have been identified as the major Fusarium mycotoxins occurring on a worldwide basis (2). In vitro cytotoxicity assays are useful to define basal toxicity and are necessary to define the concentration range for further in vitro experiments (3). The purpose of this study was to determine the cytotoxic effect of the four main Fusarium toxins deoxynivalenol (DON), T2-toxin (T-2), fumonisin B1 (FB1) and zearalenone (ZEA) on the intestinal porcine epithelial cell line derived from the jejunum (IPEC-J2) using flow cytometry. IPEC-J2 cells were seeded at a concentration of 5 x 105 cells/ml in 24-well plates in 1 ml of complete medium. After overnight culture, the medium was replaced by 1 ml medium containing various concentrations of the mycotoxins. After 72h of incubation, cells were trypsinized, washed with PBS and stained for 20 min at room temperature with a solution containing 20 µl propidium iodide (PI) and 20 µl Annexin-V-FITC (fluorescein isothiocyanate). Cells were analyzed on a FACSCanto flow cytometer with laser excitation at 488 nm. The flow cytometric technique allows the discrimination between viable (Annexin-V-/PI-), early apoptotic (Annexin-V+/PI-) and late apoptotic/necrotic cells (Annexin-V+/PI+). Results showed that T-2, DON, ZEA and FB1 are toxic to IPEC-J2 cells starting from 3.0 ng/ml, 1.0 µg/ml, 5.0 µg/ml and 7.5 µg/ml respectively. The percentage of apoptotic and necrotic cells shows a concentration-dependent relationship. The ratio between apoptotic and necrotic cells however varies depending on the type of toxin. It can be concluded that DON and ZEA cause mainly apoptosis, that T-2 causes mainly necrosis, while an equal percentage of apoptotic and necrotic cells can be seen after treatment with FB1. This suggests that different mechanisms are responsible for the in vitro toxic effect of mycotoxins on IPEC-J2 cells. References [1] Nelson PE, Dignani MC, Anaissie EJ (1994). Taxonomy, biology and clinical aspects of Fusarium species. Clinical Microbiology Reviews 7(4):479-504. [2] D'Mello JPF, Placinta CM, Macdonald AMC (1999). Fusarium mycotoxins: a review of global implications for animal health, welfare and productivity. Animal Feed Science and Technology 80(3-4):183-205. [3] Bouaziz C, Abid-Essefi S, Bouslimi A, El Golli E, Bacha H (2006). Cytotoxicity and related effects of T-2 Toxin on cultured Vero cells. Toxicon 48:343-352.

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Chicago
Goossens, Joline, Frank Pasmans, Virginie Vandenbroucke, Evelyne Meyer, Freddy Haesebrouck, Patrick De Backer, and Siska Croubels. 2011. “Cytotoxic Effects of the Fusarium Mycotoxins Deoxynivalenol, T2-toxin, Fumonisin B1 and Zearalenone on Intestinal Porcine Epithelial Cells Derived from the Jejunum.” In Mycotoxins, 4th International Symposium, Abstracts, 111–111.
APA
Goossens, Joline, Pasmans, F., Vandenbroucke, V., Meyer, E., Haesebrouck, F., De Backer, P., & Croubels, S. (2011). Cytotoxic effects of the Fusarium mycotoxins deoxynivalenol, T2-toxin, fumonisin B1 and zearalenone on intestinal porcine epithelial cells derived from the jejunum. Mycotoxins, 4th International symposium, Abstracts (pp. 111–111). Presented at the 4th International symposium on Mycotoxins (MYTOX 2011) : Challenges and perspectives.
Vancouver
1.
Goossens J, Pasmans F, Vandenbroucke V, Meyer E, Haesebrouck F, De Backer P, et al. Cytotoxic effects of the Fusarium mycotoxins deoxynivalenol, T2-toxin, fumonisin B1 and zearalenone on intestinal porcine epithelial cells derived from the jejunum. Mycotoxins, 4th International symposium, Abstracts. 2011. p. 111–111.
MLA
Goossens, Joline, Frank Pasmans, Virginie Vandenbroucke, et al. “Cytotoxic Effects of the Fusarium Mycotoxins Deoxynivalenol, T2-toxin, Fumonisin B1 and Zearalenone on Intestinal Porcine Epithelial Cells Derived from the Jejunum.” Mycotoxins, 4th International Symposium, Abstracts. 2011. 111–111. Print.
@inproceedings{1279640,
  abstract     = {In the northern hemisphere, Fusarium species are the most common moulds invading grain and maize plants (1).  Trichothecenes, fumonisins and zearalenone have been identified as the major Fusarium mycotoxins occurring on a worldwide basis (2).
In vitro cytotoxicity assays are useful to define basal toxicity and are necessary to define the concentration range for further in vitro experiments (3). The purpose of this study was to determine the cytotoxic effect of the four main Fusarium toxins deoxynivalenol (DON), T2-toxin (T-2), fumonisin B1 (FB1) and zearalenone (ZEA) on the intestinal porcine epithelial cell line derived from the jejunum (IPEC-J2) using flow cytometry.
IPEC-J2 cells were seeded at a concentration of 5 x 105 cells/ml in 24-well plates in 1 ml of complete medium. After overnight culture, the medium was replaced by 1 ml medium containing various concentrations of the mycotoxins. After 72h of incubation, cells were trypsinized, washed with PBS and stained for 20 min at room temperature with a solution containing 20 {\textmu}l propidium iodide (PI) and 20 {\textmu}l Annexin-V-FITC (fluorescein isothiocyanate). Cells were analyzed on a FACSCanto flow cytometer with laser excitation at 488 nm.  
The flow cytometric technique allows the discrimination between viable (Annexin-V-/PI-), early apoptotic (Annexin-V+/PI-) and late apoptotic/necrotic cells (Annexin-V+/PI+). 
Results showed that T-2, DON, ZEA and FB1 are toxic to IPEC-J2 cells starting from 3.0 ng/ml, 1.0 {\textmu}g/ml, 5.0 {\textmu}g/ml and 7.5 {\textmu}g/ml respectively. The percentage of apoptotic and necrotic cells shows a concentration-dependent relationship. The ratio between apoptotic and necrotic cells however varies depending on the type of toxin. It can be concluded that DON and ZEA cause mainly apoptosis, that T-2 causes mainly necrosis, while an equal percentage of apoptotic and necrotic cells can be seen after treatment with FB1. This suggests that different mechanisms are responsible for the in vitro toxic effect of mycotoxins on IPEC-J2 cells.
References
[1] Nelson PE, Dignani MC, Anaissie EJ (1994). Taxonomy, biology and clinical aspects of Fusarium species. Clinical Microbiology Reviews 7(4):479-504.
[2] D'Mello JPF, Placinta CM, Macdonald AMC (1999). Fusarium mycotoxins: a review of global implications for animal health, welfare and productivity. Animal Feed Science and Technology 80(3-4):183-205.
[3] Bouaziz C, Abid-Essefi S, Bouslimi A, El Golli E, Bacha H (2006). Cytotoxicity and related effects of T-2 Toxin on cultured Vero cells. Toxicon 48:343-352.},
  author       = {Goossens, Joline and Pasmans, Frank and Vandenbroucke, Virginie and Meyer, Evelyne and Haesebrouck, Freddy and De Backer, Patrick and Croubels, Siska},
  booktitle    = {Mycotoxins, 4th International symposium, Abstracts},
  language     = {eng},
  location     = {Ghent, Belgium},
  pages        = {111--111},
  title        = {Cytotoxic effects of the Fusarium mycotoxins deoxynivalenol, T2-toxin, fumonisin B1 and zearalenone on intestinal porcine epithelial cells derived from the jejunum},
  year         = {2011},
}