Toxicological evaluation of a glycan preparation from an enzymatic hydrolysis of Saccharomyces cerevisiae.

The aim of this paper was to provide a comprehensive toxicological and safety evaluation of a yeast cell wall preparation (YCWP) for use as an animal feed ingredient. The following toxicological assessments were carried out: the mutagenic activity was tested using the Ames' Test in five Salmonella typhimurium strains; clastogenic activity was investigated using the mammalian micronucleus test in Swiss ICO OF1 (IOPS Caw) mice; genotoxic activity was assessed using the in vitro mammalian chromosomal aberration test in human lymphocytes; acute oral toxicity was tested by administration of a single dose of 2000 mg/kg BW. Eye and skin irritation were assessed in rabbits according to OECD guidelines; skin sensitivity was established in guinea pigs by means of the Buehler test, while acute dermal and inhalation studies in rats were further completed, also according to OECD guidelines. All conducted tests were considered valid under the experimental conditions. No significant mutagenic activity or genotoxic activity was observed, and it was concluded that the test article did not induce any clastogenic effect. YCWP was found to be mildly irritating to the eye, slightly irritating to the skin but was found to be non-sensitizing in the guinea pig. The acute oral, dermal and inhalation studies did not yield any evidence of gross toxicity or pharmacological effects.

Mycotoxin-contaminated diets and an adsorbent affect the performance of Nellore bulls finished in feedlots.

Mycotoxins are present in almost all feedstuffs used in animal nutrition but are often ignored in beef cattle systems, even though they can affect animal performance. The objective of this study was to evaluate the effects of mycotoxins and a mycotoxin adsorbent (ADS) on performance of Nellore cattle finished in a feedlot. One hundred Nellore cattle (430 ± 13 kg) were used in a randomized complete block design with a 2 × 2 factorial arrangement of treatments. The factors consisted of two diets with either natural contamination (NC) or exogenous contamination (EC) and the presence (1 g/kg of DM; ADS) or absence of a mycotoxin adsorbent. The NC and EC diets had the following contaminations, respectively: 0.00 and 10.0 µg/kg aflatoxins, 5114 and 5754 µg/kg fumonisins, 0.00 and 42.1 µg/kg trichothecenes B, 0.00 and 22.1 µg/kg trichothecenes A and 42.9 and 42.9 µg/kg fusaric acid. At the beginning of the experiment, all animals were weighed, and four randomly selected animals were slaughtered to evaluate the initial carcass weight. After 97 days of treatment, all animals were weighed and slaughtered. There was no interaction among factors for the DM intake (DMI; P = 0.92); however, there was a tendency for the EC diets to decrease the DMI by 650 g/day compared to animals fed NC diets (P = 0.09). There was a trend for interaction among factors (P = 0.08) for the average daily gain (ADG), where the greatest ADG was observed for cattle fed the NC diet (1.77 kg), and the lowest was observed for those fed the EC diet (1.51 kg). The NC + ADS and EC + ADS treatments presented intermediate values for ADG. The animals fed the NC diet had a greater final BW (596 kg) than animals fed the EC treatment (582 kg; P = 0.04). There was a tendency for interaction among factors for carcass gain (P = 0.08). Similarly to ADG, the highest carcass gain was observed for animals fed the NC diet (1.20 kg), and the lowest was observed for those fed the EC diet (1.05 kg). The NC + ADS and EC + ADS treatments presented intermediate values. The natural contamination groups had greater carcass gain than that of the EC groups, and the use of the ADS recovered part of the weight gain in animals fed the EC diet. In conclusion, mycotoxins at the levels evaluated affected the performance of beef cattle, and adsorbents may mitigate their impact.

Do mycotoxin contaminated diets and yeast cell wall adsorbent affect meat quality of Nellore bulls finished in feedlot? - A short communication.

Ninety-six Nellore bulls (430 ±â€¯13 kg and 24 months) were assigned to a completely randomized block design (2 × 2 factorial arrangement of treatments) to evaluate meat quality. Dietary treatments consisted of natural or exogenous contamination with mycotoxins (Factor 1), with or without yeast cell wall adsorbent (10 g/animal/d; Factor 2). The diets were provided during 97 d. The meat chemical composition was unaffected (P ≥ .37) by the factors and the averages of variables were 74.2% moisture, 22.7% protein, 1.04% ether extract, and 2.10% ash. The L*, a*, b*, E*, C* (P ≥ .11), cooking loss (P ≥ .24) and Warner-Bratzler shear force (P ≥ .17) were also similar among factors. In conclusion, low mycotoxin contamination and yeast cell wall based adsorbent do not affect meat quality of Nellore bulls finished in feedlot.

Comparison of the sequestering properties of yeast cell wall extract and hydrated sodium calcium aluminosilicate in three in vitro models accounting for the animal physiological bioavailability of zearalenone.

The sequestration/inactivation of the oestrogenic mycotoxin zearalenone (ZEA) by two adsorbents--yeast cell wall extract (YCW) and hydrated sodium calcium aluminosilicate (HSCAS)--was studied in three laboratory models: (1) an in vitro model was adapted from referenced methods to test for the sequestrant sorption capabilities under buffer conditions at two pH values using liquid chromatography coupled to a fluorescence detector for toxin quantification; (2) a second in vitro model was used to evaluate the sequestrant sorption stability according to pH variations and using ³H-labelled ZEA at low toxin concentration; and (3) an original, ex vivo Ussing chamber model was developed to further understand the transfer of ZEA through intestinal tissue and the impact of each sequestrant on the mycotoxin bioavailability of ³H-labelled ZEA. YCW was a more efficient ZEA adsorbent than HSCAS in all three models, except under very acidic conditions (pH 2.5 or 3.0). The Ussing chamber model offered a novel, ex vivo, alternative method for understanding the effect of sequestrant on the bioavailability of ZEA. The results showed that compared with HSCAS, YCW was more efficient in sequestering ZEA and that it reduced the accumulation of ZEA in the intestinal tissue by 40% (p < 0.001).

Selenium metabolomics in yeast using complementary reversed-phase/hydrophilic ion interaction (HILIC) liquid chromatography-electrospray hybrid quadrupole trap/Orbitrap mass spectrometry.

A high efficiency chromatographic separation on a porous graphitic carbon stationary phase was developed for a large-scale separation of selenium metabolites in Se-rich yeast prior to their identification by electrospray hybrid quadrupole trap/Orbitrap mass spectrometry (Orbitrap MS(n)). The reversed-phase (RP) separation mode offered distinctly higher separation efficiency than the hydrophilic ion interaction (HILIC) mode. The latter was nevertheless complementary and useful to validate the detection of several compounds. The method allowed the detection of 64 metabolites including 30 SeSe or SeS conjugates (3 triple S/Se/S ones) and 14 selenoethers. 21 previously unreported metabolites were detected on the basis of the selenium isotopic pattern usually matched with the sub-ppm mass accuracy. 9 of these metabolites were subsequently identified using the multi-stage high mass accuracy (<5ppm) mass spectrometry. The identified metabolites (and their groups) were quantified on-line by ICP-MS fitted with a frequency-matching generator allowing a quasi-uniform response over the large (20-90%) acetonitrile mobile phase concentration range. The morphology of HPLC-ICP-MS chromatograms was remarkably similar to that of HPLC multi-ion extracted ESI-MS chromatograms. The detection limits obtained by ICP MS and ESI MS were 1 and 2ppb, respectively.

Effectiveness of modified yeast cell wall extracts to reduce aflatoxin B1 absorption in dairy ewes.

This study investigated the effect of a modified yeast cell wall extract preparation (YCW) on the excretion of aflatoxin B1 (AFB1) and aflatoxin M1 (AFM1) in feces, urine, and milk of dairy ewes fed an aflatoxin-contaminated diet. Sixteen ewes in mid-lactation were assigned to 4 treatment groups: control, AF (60 µg of AFB1/kg of feed), YCW (2 g/kg of feed), and AF+YCW. The trial consisted of a short-term (3-d) exposure period followed by a long-term (21-d) exposure period. At the end of each exposure period, milk, urine, and feces were collected over 72 h. The treatments did not affect feed intake, milk production, milk composition, or body weight. The presence of AFM1 was detected in all matrices, whereas AFB1 was only present in feces. Daily excretion was higher following long-term exposure and reached 26.9 µg of AFB1/d in feces, 37.2 µg of AFM1/d in feces, and 10.7 µg of AFM1/d in urine. Supplementation with YCW was effective in increasing aflatoxin excretion in feces in the long-term exposure (up to 156% increase). The effect was accompanied by a trend of decreasing urinary excretion of AFM1. In contrast, the addition of YCW to the contaminated diet did not affect the transfer of aflatoxins from feed to milk under the present experimental conditions with low-producing ewes. The transfer rates of AFM1 in milk ranged from 0.24 to 0.54%. In conclusion, feed supplementation with YCW reduced the absorption of AFB1 and increased the elimination of AFB1 and AFM1 in ewe feces. Yeast cell wall extract could be used to protect ruminants from chronic exposure to aflatoxins present in feeds.

Adsorption of Zearalenone by beta-D-glucans in the Saccharomyces cerevisiae cell wall.

Cell walls of yeasts and bacteria are able to complex with mycotoxins and limit their bioavailability in the digestive tract when these yeasts and bacteria are given as feed additives to animals. To identify the component(s) of the yeast cell wall and the chemical interaction(s) involved in complex formation with zearalenone, four strains of Saccharomyces cerevisiae differing in their cell wall glucan and mannan content were tested. Laboratory strains wt292, fks1, and mnn9 were compared with industrial S. cerevisiae strain sc1026. The complex-forming capacity of the yeast cell walls was determined in vitro by modelling the plots of amount of toxin bound versus amount of toxin added using Hill's model. A cooperative relationship between toxin and adsorbent was shown, and a correlation between the amount of beta-D-glucans in cell walls and complex-forming efficacy was revealed (R2 = 0.889). Cell walls of strains wt292 and mnn9, which have higher levels of beta-D-glucans, were able to complex larger amounts of zearalenone, with higher association constants and higher affinity rates than those of the fks1 and sc1026 strains. The high chitin content in strains mnn9 and fks1 increased the alkali insolubility of beta-D-glucans from isolated cell walls and decreased the flexibility of these cell walls, which restricted access of zearalenone to the chemical sites of the beta-D-glucans involved in complex formation. The strains with high chitin content thus had a lower complex-forming capacity than expected based on their beta-D-glucans content. Cooperativity and the three-dimensional structure of beta-D-glucans indicate that weak noncovalent bonds are involved in the complex-forming mechanisms associated with zearalenone. The chemical interactions between beta-D-glucans and zearalenone are therefore more of an adsorption type than a binding type.

Influence of pH on complexing of model beta-d-glucans with zearalenone.

Previous studies have shown that isolated beta-(1,3 and 1,6)-D-glucans and related alkali-extracted fractions from the cell wall of Saccharomyces cerevisiae are able to complex with zearalenone in vitro (affinity up to 50%) and thus may reduce the bioavailability of toxins in the digestive tract. The complexation mechanisms involve cooperative interaction between the two chemical entities that can be computed by Hill's model. Various linear or branched soluble or insoluble beta-D-glucans were evaluated to elucidate their roles in the adsorption mechanisms under three pH conditions (3.0, 6.0, and 8.0) found in the digestive tract. A constant quantity of each beta-D-glucans (1 mg/ml) was mixed at 39 degrees C with increasing amounts of zearalenone (2 to 100 microg/ml), and the amount of bound toxin was measured. Acidic and neutral conditions gave the highest affinity rates (64 to 77%) by beta-(1,3)-D-glucans, whereas alkaline conditions decreased adsorption except when beta-(1,6)-D-glucan side chains were branched on beta-(1,3)-D-glucans. Alkaline conditions appear to impede the active three dimensional conformation of beta-D-glucans and favor single helix and/or random coil structures. Study of the equilibrium between beta-D-glucan-bound and free toxins revealed that two types of chemical interactions occur during toxin complexation with beta-D-glucans, identified as weak chemical linkages such as hydrogen and van der Waals bonds.