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In vitro rumen simulations show a lower disappearance of deoxynivalenol, nivalenol, zearalenone and enniatin B at conditions of rumen acidosis and at dry conditions

Sandra Debevere (UGent) , Siegrid De Baere (UGent) , Geert Haesaert (UGent) , Michael Rychlik, Siska Croubels (UGent) and Veerle Fievez (UGent)
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Abstract
Ruminants are generally considered less susceptible to the effects of mycotoxins than monogastric animals as their rumen microbiota are able to degrade and deactivate some of these toxic molecules. Despite this potential degradation, mycotoxin-associated subclinical health problems have been reported in high productive dairy cows, reflected by vague and non-specific symptoms and periodic decrease in milk production. The significantly increased milk yield per cow per lactation during the last decades is not only associated with a higher incidence of metabolic disorders such as subacute rumen acidosis (SARA), but also with a higher proportion of highly fermentable maize silage in the ration, the latter being more vulnerable to contamination with multiple mycotoxins than grassland products. In this research, the effect of lower rumen pH and lactation stage on degradation of mycotoxins that are often present in maize silage (DON: deoxynivalenol, NIV: nivalenol, ENN B: enniatin B, MPA: mycophenolic acid, ROQ-C: roquefortine C and ZEN: zearalenone) was investigated by means of in vitro rumen simulations. Maize silage was spiked with a mixture of DON (12 μg/g), NIV (60 μg/g), ENN B (1 μg/g), MPA (6 μg/g), ROQ-C (2 μg/g) and ZEN (3 μg/g). Fresh rumen fluid of 2 lactating cows (L) and 2 dry cows (D) was added to a MES (2-(N-morpholino)ethanesulfonic acid) buffer with normal pH (6.8) and low pH (5.8), leading to 4 different combinations (L6.8, L5.8, D6.8, D5.8) that were subsequently added to the spiked maize substrate under CO2 conditions. After 1.5h, 3h, 6h, 24h and 48h of incubation, samples were taken. Volatile fatty acids (VFA) were measured with gas chromatography with flame ionization detector (GC-FID) and mycotoxin concentrations with an in-house developed and validated ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS) multimethod. The incubations were performed in triplicate. Mycotoxins had no significant effect on VFA production after 48h of incubation. After 24h of incubation, a complete detoxification of DON occurred at normal rumen pH, in contrast with the low pH, where 14% of DON was still present for L5.8 and even 91% for D5.8. No NIV could be detected anymore after 24h of incubation at a normal pH, in contrast with the low pH, where 38% was still present for L5.8 and even 100% for D5.8. For ZEN, after 48h of incubation, a partial transformation of ZEN to α-ZEL (13-22%) and β-ZEL (4-5%) was observed, but did not occur at the low pH. After 48h of incubation, disappearance of ENN B could be seen at normal pH (L: 71%, D: 43%), but not at low pH. For MPA and ROQ-C, neither pH nor lactation stage had an effect on the disappearance of the mycotoxin in the rumen. In conclusion, low ruminal pH (e.g. SARA conditions) and lactation stage can have a lower ruminal degradation of certain mycotoxins such as DON, NIV, ZEN and ENN B. Hence, these mycotoxins could reach the small intestine intact and exert their toxic effects.
Keywords
Mycotoxins, maize silage, SARA, lactation, rumen fluid, UPLC-MS/MS, multi-method

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Chicago
Debevere, Sandra, Siegrid De Baere, Geert Haesaert, Michael Rychlik, Siska Croubels, and Veerle Fievez. 2019. “In Vitro Rumen Simulations Show a Lower Disappearance of Deoxynivalenol, Nivalenol, Zearalenone and Enniatin B at Conditions of Rumen Acidosis and at Dry Conditions.” In Mycotoxin Workshop, 41st, Abstracts, 61–61.
APA
Debevere, S., De Baere, S., Haesaert, G., Rychlik, M., Croubels, S., & Fievez, V. (2019). In vitro rumen simulations show a lower disappearance of deoxynivalenol, nivalenol, zearalenone and enniatin B at conditions of rumen acidosis and at dry conditions. Mycotoxin workshop, 41st, Abstracts (pp. 61–61). Presented at the 41st Mycotoxin workshop.
Vancouver
1.
Debevere S, De Baere S, Haesaert G, Rychlik M, Croubels S, Fievez V. In vitro rumen simulations show a lower disappearance of deoxynivalenol, nivalenol, zearalenone and enniatin B at conditions of rumen acidosis and at dry conditions. Mycotoxin workshop, 41st, Abstracts. 2019. p. 61–61.
MLA
Debevere, Sandra et al. “In Vitro Rumen Simulations Show a Lower Disappearance of Deoxynivalenol, Nivalenol, Zearalenone and Enniatin B at Conditions of Rumen Acidosis and at Dry Conditions.” Mycotoxin Workshop, 41st, Abstracts. 2019. 61–61. Print.
@inproceedings{8615949,
  abstract     = {Ruminants are generally considered less susceptible to the effects of mycotoxins than monogastric animals as their rumen microbiota are able to degrade and deactivate some of these toxic molecules. Despite this potential degradation, mycotoxin-associated subclinical health problems have been reported in high productive dairy cows, reflected by vague and non-specific symptoms and periodic decrease in milk production. The significantly increased milk yield per cow per lactation during the last decades is not only associated with a higher incidence of metabolic disorders such as subacute rumen acidosis (SARA), but also with a higher proportion of highly fermentable maize silage in the ration, the latter being more vulnerable to contamination with multiple mycotoxins than grassland products. In this research, the effect of lower rumen pH and lactation stage on degradation of mycotoxins that are often present in maize silage (DON: deoxynivalenol, NIV: nivalenol, ENN B: enniatin B, MPA: mycophenolic acid, ROQ-C: roquefortine C and ZEN: zearalenone) was investigated by means of in vitro rumen simulations.
Maize silage was spiked with a mixture of DON (12 μg/g), NIV (60 μg/g), ENN B (1 μg/g), MPA (6 μg/g), ROQ-C (2 μg/g) and ZEN (3 μg/g). Fresh rumen fluid of 2 lactating cows (L) and 2 dry cows (D) was added to a MES (2-(N-morpholino)ethanesulfonic acid) buffer with normal pH (6.8) and low pH (5.8), leading to 4 different combinations (L6.8, L5.8, D6.8, D5.8) that were subsequently added to the spiked maize substrate under CO2 conditions. After 1.5h, 3h, 6h, 24h and 48h of incubation, samples were taken. Volatile fatty acids (VFA) were measured with gas chromatography with flame ionization detector (GC-FID) and mycotoxin concentrations with an in-house developed and validated ultra-performance liquid-chromatography tandem mass spectrometry (UPLC-MS/MS) multimethod. The incubations were performed in triplicate.
Mycotoxins had no significant effect on VFA production after 48h of incubation. After 24h of incubation, a complete detoxification of DON occurred at normal rumen pH, in contrast with the low pH, where 14% of DON was still present for L5.8 and even 91% for D5.8. No NIV could be detected anymore after 24h of incubation at a normal pH, in contrast with the low pH, where 38% was still present for L5.8 and even 100% for D5.8. For ZEN, after 48h of incubation, a partial transformation of ZEN to α-ZEL (13-22%) and β-ZEL (4-5%) was observed, but did not occur at the low pH. After 48h of incubation, disappearance of ENN B could be seen at normal pH (L: 71%, D: 43%), but not at low pH. For MPA and ROQ-C, neither pH nor lactation stage had an effect on the disappearance of the mycotoxin in the rumen.
In conclusion, low ruminal pH (e.g. SARA conditions) and lactation stage can have a lower ruminal degradation of certain mycotoxins such as DON, NIV, ZEN and ENN B. Hence, these mycotoxins could reach the small intestine intact and exert their toxic effects.},
  author       = {Debevere, Sandra and De Baere, Siegrid and Haesaert, Geert and Rychlik, Michael and Croubels, Siska and Fievez, Veerle},
  booktitle    = {Mycotoxin workshop, 41st, Abstracts},
  keywords     = {Mycotoxins,maize silage,SARA,lactation,rumen fluid,UPLC-MS/MS,multi-method},
  language     = {eng},
  location     = {Lisbon, Portugal},
  pages        = {61--61},
  title        = {In vitro rumen simulations show a lower disappearance of deoxynivalenol, nivalenol, zearalenone and enniatin B at conditions of rumen acidosis and at dry conditions},
  year         = {2019},
}