Development of an UPLC-MS/MS Method for the Analysis of Mycotoxins in Rumen Fluid with and without Maize Silage Emphasizes the Importance of Using Matrix-Matched Calibration

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Ruminants are generally considered to be less susceptible to the effects of mycotoxins than monogastric animals as the rumen microbiota are capable of detoxifying some of these toxins. Despite this potential degradation, mycotoxin-associated subclinical health problems are seen in dairy cows. In this research, the disappearance of several mycotoxins was determined in an in vitro rumen model and the effect of realistic concentrations of those mycotoxins on fermentation was assessed by volatile fatty acid production. In addition, two hypotheses were tested: (1) a lower rumen pH leads to a decreased degradation of mycotoxins and (2) rumen fluid of lactating cows degrade mycotoxins better than rumen fluid of non-lactating cows. Maize silage was spiked with a mixture of deoxynivalenol (DON), nivalenol (NIV), enniatin B (ENN B), mycophenolic acid (MPA), roquefortine C (ROQ-C) and zearalenone (ZEN). Fresh rumen fluid of two lactating cows (L) and two non-lactating cows (N) was added to a buffer of normal pH (6.8) and low pH (5.8), leading to four combinations (L6.8, L5.8, N6.8, N5.8), which were added to the spiked maize substrate. In this study, mycotoxins had no effect on volatile fatty acid production. However, not all mycotoxins fully disappeared during incubation. ENN B and ROQ-C disappeared only partially, whereas MPA showed almost no disappearance. The disappearance of DON, NIV, and ENN B was hampered when pH was low, especially when the inoculum of non-lactating cows was used. For ZEN, a limited transformation of ZEN to α-ZEL and β-ZEL was observed, but only at pH 6.8. In conclusion, based on the type of mycotoxin and the ruminal conditions, mycotoxins can stay intact in the rumen.

Section: Rumen Fluid Sample Extraction and Mycotoxin Analysismentioning

confidence: 99%

Section: Rumen Fluid Sample Extraction and Mycotoxin Analysismentioning

confidence: 99%

Section: Rumen Fluid Sample Extraction and Mycotoxin Analysismentioning

confidence: 99%

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In Vitro Rumen Simulations Show a Reduced Disappearance of Deoxynivalenol, Nivalenol and Enniatin B at Conditions of Rumen Acidosis and Lower Microbial Activity

Debevere

Cools

Baere

et al. 2020

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“…Binder 2 adsorbed ENN B by 28%, although the difference between the ENN B concentration in the control and binder 2 treatment decreased over time, which could indicate an unstable and reversible adsorption of ENN B. Nivalenol concentrations were statistically higher with binder 2 compared to the control treatment, although the biological relevance of the small difference of 6.7% (p = 0.00587) is considered to be negligible as this difference also falls within the range of precision of the UPLC-MS/MS method (RSD for NIV: ±7.8%) [29]. No effect of this binder on the ROQ-C, MPA, DON, DOM-1, ZEN, and α-ZEL concentrations was noted compared to the control treatment.…”

Section: Binders In Feed Normal Rumen Ph (68)mentioning

confidence: 91%

Evaluation of the Efficacy of Mycotoxin Modifiers and Mycotoxin Binders by Using an In Vitro Rumen Model as a First Screening Tool

Debevere

Schatzmayr²,

Reisinger³

et al. 2020

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Ruminal microbiota of cattle are not able to detoxify all mycotoxins. In addition, detoxification can be hampered by adverse ruminal conditions (e.g., low ruminal pH). Hence, in the cattle husbandry, mycotoxin binders and modifiers could be used to prevent animal exposure to mycotoxins. In this study, an in vitro rumen model, including feed matrix, was established as first screening tool to test the efficacy of five products claiming to detoxify mycotoxins. The detoxifiers had different modes of action: (a) binding (three products); (b) enzymatic detoxification of zearalenone (ZEN; one product, ZenA); and (c) bacterial transformation of trichothecenes (one product, BBSH 797). For the mycotoxin binders, the binding to the mycotoxins enniatin B (ENN B), roquefortine C (ROQ-C), mycophenolic acid (MPA), deoxynivalenol (DON), nivalenol (NIV), and zearalenone (ZEN) were tested at a dose recommended by the manufacturers. The in vitro model demonstrated that all binders adsorbed ENN B to a certain extent, while only one of the binders also partially adsorbed ROQ-C. The binders did not change the concentrations of the other mycotoxins in the ruminal fluid. The enzyme ZenA detoxified ZEN very quickly and prevented the formation of the more toxic metabolite α-zearalenol (α-ZEL), both at normal (6.8) and low ruminal pH (5.8). The addition of BBSH 797 enhanced detoxification of DON and NIV, both at normal and low ruminal pH. The in vitro rumen model demonstrated that the addition of ZenA seems to be a very promising strategy to prevent estrogenic effects of ZEN contaminated feed, and BBSH 797 is efficient in the detoxification of trichothecenes.

“…In addition to foodstuff and feed, the determination of mycotoxins in biological fluids is also one of the topics addressed in this Special Issue. Debevere et al develop and fully-validate a UHPLC-MS/MS method for the quantitative determination in the rumen fluid of some of the most relevant mycotoxins found in maize silage [16]. Extraction recovery and matrix effects were determined in the rumen fluid with and without maize silage.…”

mentioning

confidence: 99%

Application of LC–MS/MS in the Mycotoxins Studies

Gámiz‐Gracia

Garcı́a-Campaña

Arroyo‐Manzanares

2020

Mycotoxins are secondary metabolites produced by fungi of different species (mainly Aspergillus, Fusarium, and Penicillium) with toxic effects for humans and animals that can contaminate food and feed. Notifications on the Rapid Alert System for Food and Feed (RASFF) concerning mycotoxins are becoming frequent, being among the "top 10" hazards reported on food products, mainly cereals and nuts [1]. The European Union (EU), among others, has established maximum permitted or recommended levels for several mycotoxins, including aflatoxins (AF) B1, B2, G1, G2 and M1, ochratoxin A (OTA), patulin (PAT), deoxynivalenol (DON), zearalenone (ZEN), fumonisins (FB1 and FB2), HT-2 and T-2 toxins, or citrinin (CIT) in different food commodities [2]. However, there are other mycotoxins not included in regulations, the so-called "emerging mycotoxins", whose toxicity is still not clear, that could also pose a risk to humans and animals. This group includes Alternaria toxins, sterigmatocystin (STC), phomopsins, and Fusarium toxins as enniatins (ENN) or beauvericin (BEA), among others [3]. Moreover, the so-called "modified or masked mycotoxins" (produced as a consequence of a detoxification strategy of the host plant of the fungus or during food processing) could be equally or even more toxic than the original mycotoxin, and should also be taken into account [4].All these facts make it necessary to develop analytical methods for the accurate determination of mycotoxins in different food matrices and feeds. In this sense, liquid chromatography tandem mass spectrometry (LC-MS/MS) is a powerful tool for the unique identification and quantification of analytes. Moreover, the use of high-resolution mass spectrometry (HRMS) allows the identification of novel mycotoxins and targeted/untargeted approaches for their study.This Special Issue is dedicated to recent applications of LC-MS/MS in mycotoxin studies, including the development and validation of new analytical methods for their identification and quantification in different food matrices and feed, occurrence studies and biomonitoring of mycotoxins, and their metabolites in biological fluids.Although most of the papers propose methods for the simultaneous determination of several mycotoxins, Bertuzzi et al. focus their work on the improvement of determination of moniliformin (MON) by adding lanthanide ions to the mobile phase during chromatographic separation [5]. MON is weakly retained in reversed-phase chromatography, making the separation difficult. The addition of La 3+ , Tb 3+ or Eu 3+ to the mobile phase is investigated, improving peak shape and MON separation, probably due to the formation of coordination complexes lanthanide-MON. After extraction and purification, the method is applied to the determination of MON in maize and wheat samples.The potential of LC-MS/MS for multi-mycotoxin determination in different food matrices is shown in different contributions of this Special Issue. For instance, Guo et al. propose a modified QuEChERS (Quick, Easy, Cheap, Efficient, Rugged, an...