The overall objective of the MycoPig-project was to elucidate the impact of trichothecene- and other mycotoxins on pig health in Norway. This has been achieved by combining work packages comprising of different related and unrelated tasks.
WP1: We have evaluated the microbe Coriobacteriaceum DSM 11798 (known as the active ingredient in Biomin® BBSH 797) as a trichothecene-reducing feed additive in piglets. The piglets were fed with pelletized oats-based feed containing up to ca. 5000 µg/kg of DON, with or without the microbe over a period of six weeks. During the first seven days of exposure, pigs in the highest dose group showed 5-8% reduction in feed intake and 7-10% reduction in weight gain compared to pigs in the control and low-dose groups. These differences were, however, leveled out at study end. Supplementation of the experimental feed with DSM 11798 had no effect on the observed DON-related clinical effects, nor on the plasma concentrations of DON and its metabolites. Thus, neither uptake of DON nor DON-related toxicity in growing pigs could be alleviated by DSM 11798 under the chosen experimental conditions.
WP2: Additionally, we have studied the conjugation of DON with certain types of sulfur-containing molecules (thiols) and investigated if such conjugates occur in nature. The reaction between DON and different thiols, including biologically significant compounds, such as L-cysteine and L-glutathione, was investigated in order to get insight into the kinetics of the reaction and to characterize the reaction products chemically as well as their biological effects. The reaction was shown to occur at two different sites in the molecule, and adducts of DON with 2-mercaptoethanol (as model molecule), L-cysteine and L-glutathione were purified and characterized. The presence of some adducts was shown in naturally contaminated samples from Norway, though at low concentrations.
WP3: We have studied the relationship between different levels of the major trichothecene mycotoxin 4-deoxynivalenol (DON) in high-yielding sows. The sows (three weeks before labor) in a commercial setting received either ca. 1300 μg/kg of DON or a control diet. The focus of the trial was to test the effect of DON on different parameters of reproduction and the transfer of the toxin to the piglets.
WP4: Samples were collected at selected pig farms in Southeastern Norway and analysed for microfungi and mycotoxins. Samples of air, bedding and feed were collected at seven pig farms. Fungal metabolites were chemically analysed using a screening method covering several hundred compounds, while fungi were were generally determined on genus level. The results show that animals (and farmers) are exposed to a complex mixture of fungal metabolites and mycotoxins from many major toxigenic fungi, e.g Penicillium, Fusarium and Aspergillus.
The MycoPig project was a cooperation between the Norwegian Veterinary Institute, the Norwegain Univeristy of Life Sciences, The National Institute of Occupational Health and the University of Natural Resources and Life Sciences in Vienna, Austria. The project was financed by the Norwegian Research Council (Grant 225332) with additional support by Felleskjøpet Fôrutvikling, Lantmännen, Biomin GmbH and Animalia.
Two PhD students were trained:
- Amin Sayyari (2019). Deoxynivalenol in pigs: Effects, toxicokinetics, vertical transmission and microbial detoxification. Dissertation for the degree of Ph.D at Faculty of Veterinary Medicine, Norwegian University of Life Sciences, Oslo.
- Ana Stanic (2017). Preparation of thiol conjugates of the mycotoxin deoxynivalenol and their occurrence in nature. Dissertation for the degree of Ph.D at Faculty of Mathematics and Sciences, University of Oslo.
Publications
- Sayyari, A., Uhlig, S., Fæste, C.K., Framstad, T., Sivertsen T. Transfer of deoxynivalenol (DON) through placenta, colostrum and milk from sows to their offspring during late gestation and lactation. Toxins 2018, 10, 517–537.
- Sayyari, A., Framstad, T., Krogenæs, A.K., Sivertsen, T. Effects of feeding naturally contaminated deoxynivalenol diets to sows during late gestation and lactation in a high-yield specific pathogen-free herd. Porcine Health Management 2018, 4, 26–36.
- Fæste CK, Ivanova L, Sayyari A, Hansen U, Sivertsen T, Uhlig S. Prediction of deoxynivalenol toxicokinetics in humans by in vitro-to-in vivo extrapolation and allometric scaling of in vivo animal data. Arch Toxicol 2018, 92:2195-2216.
- Sayyari A, Fæste CK, Hansen U, Uhlig S, Framstad T, Schatzmayr D, Sivertsen T. Effects and biotransformation of the mycotoxin 4-deoxynivalenol (DON) in growing pigs fed naturally-contaminated grain pelleted with and without the addition of Coriobacteriaceum DSM 11798. J. Food Add Contam A 2018, 1-16.
- Uhlig, S., Stanic, A., Hussain, F., Miles, C.O. Selectivity of commercial immunoaffinity columns for modified forms of the mycotoxin 4-deoxynivalenol (DON), Journal of Chromatography B 2017, 1061–1062, 322–326.
- Stanic, A.; Uhlig, S.; Solhaug, A.; Rise, F.; Wilkins, A.L.; Miles, C.O. Preparation and characterization of cysteine adducts of deoxynivalenol. Journal of Agricultural and Food Chemistry 2016, 64, 4777−4785.
- Stanic, A.; Uhlig, S.; Sandvik, M.; Rise, F.; Wilkins, A.L.; Miles, C.O. Characterization of deoxynivalenol–glutathione conjugates using nuclear magnetic resonance spectroscopy and liquid chromatography–high resolution mass spectrometry. Journal of Agricultural and Food Chemistry 2016, 64, 6903-6910.
- Stanic, A.; Uhlig, S.; Solhaug, A.; Rise, F.; Wilkins, A.; Miles, C.O. Nucleophilic addition of thiols to deoxynivalenol, Journal of Agricultural and Food Chemistry 2015, 63, 7556-7566.
Lectures and poster presentations at conferences, workshops etc.
- Uhlig, S. Insight into the thiol chemistry of 4-deoxynivalenol (DON). Mycotoxin Summer Academy, 6.7.2018, Tulln, Austria.
- Sayyari A, Framstad T, Hansen U, Fæste CK, Uhlig S, Sivertsen T. Effectiveness of a microbial feed additive in preventing deoxynivalenol-related toxicity in growing pigs. ESPHM 3.-5.5.2017, Prague, Czech Republic.
- Ljøkjel, K. Mycotoxins in Norwegian pig farming – a project introduction and some preliminary results, Intercoop Workshop Swine, The Netherlands, October 2015.
- Stanic, A. Nucleophilic addition of thiols to the epoxide ring and double bond of trichothecenes, Gordon Research Conference on Mycotoxins and Phycotoxins, Stonehill College, Easton, MA, USA, June 2015.
- Stanic, A., Uhlig, S., Rise, F., Wilkins, A. L., Miles, C. O. Reaction of deoxynivalenol with model and biologically relevant thiols, Nordic Baltic Fusarium Seminar, Helsinki, Finland, November 2014.
- Ivanova, L., Hansen, U., Uhlig, S., Fæste, C. K. Detoxification kinetics of deoxynivalenol (DON) in nonruminants, Nordic Baltic Fusarium Seminar, Helsinki, Finland, November 2014.
- Fæste CK, Uhlig S, Ivanova L. In vitro kinetics as a tool to predict in vivo exposure. Nordic Baltic Fusarium Seminar 2014, 18.-19.11.2014, Helsinki, Finland.
- Stanic, A. Understanding the reaction of deoxynivalenol with model and biologically relevant nucleophiles, International Mycotoxin Conference 2014, Beijing, China, May 2014.
- Uhlig, S. Kan grisen bli glad i mykotoksiner? Veterinærinstituttets mandagsseminar, November 2013.