Turmeric Powder Counteracts Oxidative Stress and Reduces AFB1 Content in the Liver of Broilers Exposed to the EU Maximum Levels of the Mycotoxin.

The most frequent adverse effects of AFB1 in chicken are low performance, the depression of the immune system, and a reduced quality of both eggs and meat, leading to economic losses. Since oxidative stress plays a major role in AFB1 toxicity, natural products are increasingly being used as an alternative to mineral binders to tackle AFB1 toxicosis in farm animals. In this study, an in vivo trial was performed by exposing broilers for 10 days to AFB1 at dietary concentrations approaching the maximum limits set by the EU (0.02 mg/kg feed) in the presence or absence of turmeric powder (TP) (included in the feed at 400 mg/kg). The aims were to evaluate (i) the effects of AFB1 on lipid peroxidation, antioxidant parameters, histology, and the expression of drug transporters and biotransformation enzymes in the liver; (ii) the hepatic accumulation of AFB1 and its main metabolites (assessed using an in-house-validated HPLC-FLD method); (iii) the possible modulation of the above parameters elicited by TP. Broilers exposed to AFB1 alone displayed a significant increase in lipid peroxidation in the liver, which was completely reverted by the concomitant administration of TP. Although no changes in glutathione levels and antioxidant enzyme activities were detected in any treatment group, AFB1 significantly upregulated and downregulated the mRNA expression of CYP2A6 and Nrf2, respectively. TP counteracted such negative effects and increased the hepatic gene expression of selected antioxidant enzymes (i.e., CAT and SOD2) and drug transporters (i.e., ABCG2), which were further enhanced in combination with AFB1. Moreover, both AFB1 and TP increased the mRNA levels of ABCC2 and ABCG2 in the duodenum. The latter changes might be implicated in the decrease in hepatic AFB1 to undetectable levels (

Occurrence and Determination of Alternaria Mycotoxins Alternariol, Alternariol Monomethyl Ether, and Tentoxin in Wheat Grains by QuEChERS Method.

The Alternaria mycotoxins such as alternariol (AOH), alternariol monomethyl ether (AME), and tentoxin (TEN) are mycotoxins, which can contaminate cereal-based raw materials. Today, wheat is one of the most important crops in temperate zones, and it is in increasing demand in the Western Balkans countries that are urbanizing and industrializing. This research aimed to investigate the occurrence and determine the concentration of Alternaria mycotoxins AOH, AME, and TEN in wheat samples from the Republic of Serbia and the Republic of Albania, harvested in the year 2020 in the period between 15 June and 15 July. A total of 80 wheat grain samples, 40 from each country, were analyzed by an QuEChERS (quick, easy, cheap, effective, rugged, and safe) method. From the obtained results, it can be seen that the mean concentration of AOH was 3.3 µg/kg and AME was 2.2 µg/kg in wheat samples from Serbia, while TEN from both Serbia and Albania was under the limit of quantification (

Simultaneous Removal of Mycotoxins by a New Feed Additive Containing a Tri-Octahedral Smectite Mixed with Lignocellulose.

Simultaneous removal of mycotoxins has been poorly addressed, and a limited number of studies have reported the efficacy of feed additives in sequestering a large spectrum of mycotoxins. In this study, a new mycotoxin-adsorbing agent was obtained by properly mixing a tri-octahedral smectite with a lignocellulose-based material. At a dosage of 1 mg mL-1, these materials simultaneously adsorbed frequently occurring mycotoxins and did not exert a cytotoxic effect on intestinal cells. Chyme samples obtained by a simulated GI digestion did not affect the viability of Caco-2TC7 cells as measured by the MTT test. In addition, the chyme of the lignocellulose showed a high content of polyphenols (210 mg mL-1 catechin equivalent) and good antioxidant activity. The properties of the individual constituents were maintained in the final composite, and were unaffected by their combination. When tested with a pool of seven mycotoxins at 1 µg mL-1 each and pH 5, the composite (5 mg mL-1) simultaneously sequestered AFB1 (95%), FB1 (99%), ZEA (93%), OTA (80%), T-2 (63%), and DON (22%). HT-2 adsorption did not occur. Mycotoxin adsorption increased exponentially as dosage increased, and occurred at physiological pH values. AFB1, ZEA and T-2 adsorption was not affected by pH in the range 3-9, whereas OTA and FB1 were adsorbed at pH values of 3-5. The adsorbed amount of AFB1, ZEA and T-2 was not released when pH rose from 3 to 7. FB1 and OTA desorption was less than 38%. Langmuir adsorption isotherms revealed high capacity and affinity for adsorption of the target mycotoxins. Results of this study are promising and show the potential of the new composite to remove mycotoxins in practical scenarios where several mycotoxins can co-occur.

Mycotoxin Removal by Lactobacillus spp. and Their Application in Animal Liquid Feed.

The removal of mycotoxins from contaminated feed using lactic acid bacteria (LAB) has been proposed as an inexpensive, safe, and promising mycotoxin decontamination strategy. In this study, viable and heat-inactivated L. acidophilus CIP 76.13T and L. delbrueckii subsp. bulgaricus CIP 101027T cells were investigated for their ability to remove aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEA), and deoxynivalenol (DON) from MRS medium and PBS buffer over a 24 h period at 37 °C. LAB decontamination activity was also assessed in a ZEA-contaminated liquid feed (LF). Residual mycotoxin concentrations were determined by UHPLC-FLD/DAD analysis. In PBS, viable L. acidophilus CIP 76.13T and L. delbrueckii subsp. bulgaricus CIP 101027T cells removed up to 57% and 30% of ZEA and DON, respectively, while AFB1 and OTA reductions were lower than 15%. In MRS, 28% and 33% of ZEA and AFB1 were removed, respectively; OTA and DON reductions were small (≤15%). Regardless of the medium, heat-inactivated cells produced significantly lower mycotoxin reductions than those obtained with viable cells. An adsorption mechanism was suggested to explain the reductions in AFB1 and OTA, while biodegradation could be responsible for the removal of ZEA and DON. Both viable LAB strains reduced ZEA by 23% in contaminated LF after 48 h of incubation. These findings suggest that LAB strains of L. acidophilus CIP 76.13T and L. delbrueckii subsp. bulgaricus CIP 101027T may be applied in the feed industry to reduce mycotoxin contamination.

Curcumin Supplementation Protects Broiler Chickens Against the Renal Oxidative Stress Induced by the Dietary Exposure to Low Levels of Aflatoxin B1.

Aflatoxin B1 (AFB1) causes hepatotoxicity, immunotoxicity, and kidney damage, and it is included in group I of human carcinogens. The European Commission has established maximum limits of AFB1 in feed, ranging from 5 to 20 µg/kg. Chicken is moderately sensitive to AFB1, which results in reduced growth performance and economic losses. Oxidative stress triggered by AFB1 plays a crucial role in kidney damage and the antioxidant activity of Curcumin (CURC) could help in preventing such adverse effect. Twenty-days-old broilers were treated for 10 days with AFB1 (0.02 mg/kg feed), alone or in combination with CURC (400 mg/kg feed), to explore the effects on the renal tissue. Animals exposed to AFB1 alone displayed alterations of the oxidative stress parameters compared with controls: serum antioxidant capacity, and enzymatic activity of kidney superoxide dismutase, catalase and glutathione peroxidase were decreased, while renal malondialdehyde levels and NADPH oxidase complex expression were increased. The administration of CURC attenuates all the oxidative stress parameters modified by AFB1 in the chicken kidney, opening new perspectives in the management of aflatoxicosis.

Development of a DNA-based biosensor for the fast and sensitive detection of ochratoxin A in urine.

In this paper, a novel DNA-based biosensor is proposed, which is based on paramagnetic microbeads carrying an ochratoxin A (OTA) capture aptamer. A sandwich-like detection complex is linked to the capture aptamer and is able to trigger, in presence of OTA, an isothermal rolling circle amplification (RCA) reaction. This latter generated autocatalytic units with a peroxidase activity (DNAzyme) that, in presence of a proper substrate, gave a blue-coloured product visible by the naked eye. The capture aptamer, blocked onto magnetic beads, allowed the specific capture of OTA in liquid samples. The modified detection aptamer, annealed to a circularized probe, was then used to detect the toxin capture event. Indeed, in the presence of OTA and an isothermal enzyme, the circular DNA was amplified, producing a single-stranded and tandem repeated long homologous copy of its sequence. In the DNA strand, a self-catalytic structure was formed with hemin as the catalytic core, inducing the development of blue colour in the presence of ABTS and hydrogen peroxide. The results showed that the biosensor has high sensitivity and selectivity for the detection of OTA, as low as 1.09 × 10-12 ng/mL. Moreover, the proposed biosensor was successfully used for the detection of OTA in naturally contaminated rat urine. Accuracy and repeatability data obtained in recovery experiments were satisfying, being recoveries >95% with relative standard deviations in the range 3.6-15%. For the first time, an aptasensor was successfully applied to detect OTA in biological fluids. It can be used for mycotoxin biomonitoring and assessment of individual exposure.

The Effectiveness of Durian Peel as a Multi-Mycotoxin Adsorbent.

Durian peel (DP) is an agricultural waste that is widely used in dyes and for organic and inorganic pollutant adsorption. In this study, durian peel was acid-treated to enhance its mycotoxin adsorption efficacy. The acid-treated durian peel (ATDP) was assessed for simultaneous adsorption of aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), and fumonisin B1 (FB1). The structure of the ATDP was also characterized by SEM-EDS, FT-IR, a zetasizer, and a surface-area analyzer. The results indicated that ATDP exhibited the highest mycotoxin adsorption towards AFB1 (98.4%), ZEA (98.4%), and OTA (97.3%), followed by FB1 (86.1%) and DON (2.0%). The pH significantly affected OTA and FB1 adsorption, whereas AFB1 and ZEA adsorption was not affected. Toxin adsorption by ATDP was dose-dependent and increased exponentially as the ATDP dosage increased. The maximum adsorption capacity (Qmax), determined at pH 3 and pH 7, was 40.7 and 41.6 mmol kg-1 for AFB1, 15.4 and 17.3 mmol kg-1 for ZEA, 46.6 and 0.6 mmol kg-1 for OTA, and 28.9 and 0.1 mmol kg-1 for FB1, respectively. Interestingly, ATDP reduced the bioaccessibility of these mycotoxins after gastrointestinal digestion using an in vitro, validated, static model. The ATDP showed a more porous structure, with a larger surface area and a surface charge modification. These structural changes following acid treatment may explain the higher efficacy of ATDP in adsorbing mycotoxins. Hence, ATDP can be considered as a promising waste material for mycotoxin biosorption.

Decontamination of Mycotoxin-Contaminated Feedstuffs and Compound Feed.

Mycotoxins are known worldwide as fungus-produced toxins that adulterate a wide heterogeneity of raw feed ingredients and final products. Consumption of mycotoxins-contaminated feed causes a plethora of harmful responses from acute toxicity to many persistent health disorders with lethal outcomes; such as mycotoxicosis when ingested by animals. Therefore, the main task for feed producers is to minimize the concentration of mycotoxin by applying different strategies aimed at minimizing the risk of mycotoxin effects on animals and human health. Once mycotoxins enter the production chain it is hard to eliminate or inactivate them. This paper examines the most recent findings on different processes and strategies for the reduction of toxicity of mycotoxins in animals. The review gives detailed information about the decontamination approaches to mitigate mycotoxin contamination of feedstuffs and compound feed, which could be implemented in practice.

Antimicrobial Activity of Yeast Cell Wall Products Against Clostridium perfringens.

Yeast cell wall (YCW) products are used worldwide as alternatives to antibiotics growth promoters for health and performances improvement in livestock. The success of yeast and YCW products as feed additives in farm animals' nutrition relies on their capacity to bind enteropathogenic bacteria and on their immunomodulatory activity. In vivo studies report their anti-infectious activity on Gram-positive pathogens like clostridia. However, the in vitro antimicrobial activity of YCW products seems to be limited to some Gram-negative enteropathogens, and literature lacks in vitro evidences for antimicrobial effect of YCW products against Clostridium perfringens. This study aims to measure the antimicrobial activity of YCW products on C. perfringens. Five different YCW products were assayed for their capacity to inhibit the growth of C. perfringens, by analyzing the growth kinetics of the pathogen. All YCW products inhibited the growth of the pathogen, by reducing the growth rate and the maximum growth value and extending the lag phase duration. The effect on the growth parameters was product and dosage dependent. The most effective YCW (namely YCW2), at the minimum effective concentration of 1.25 mg/mL, increased the lag phase duration by 3.6 h, reduced the maximum growth rate by >50%, and reduced the final cell count by 102 colony-forming unit per milliliter in 24 h, with respect to the control. YCW products did not show a strain-dependent impact on C. perfringens growth when tested on different strains of the bacterium.

Equilibrium Isotherm Approach to Measure the Capability of Yeast Cell Wall to Adsorb Clostridium perfringens.

Yeast cell wall (YCW) products are currently used as substitutes to antibiotic growth promoters, to improve animal performances, and to reduce the incidence of infectious diseases in livestock. They are claimed to bind enteropathogens, thus interfering with their colonization in the intestinal mucosa. Although the anti-infectious activity of YCW products on Gram-positive pathogens like Clostridium perfringens has been reported in vivo, in vitro evidences on the adsorption of C. perfringens by YCW fractions are not yet available. Preliminary results showed that purified YCW products exert antimicrobial activity toward C. perfringens. Using the adsorption isotherm approach, we measured the ability of YCW products in adsorbing C. perfringens, thus reducing its viability. Dosages of YCW products >1 mg/mL adsorbed 4 Log colony-forming unit (CFU)/mL of C. perfringens in buffered solution. The maximum adsorption of the bacterium was reached in 3 h, whereas only one product of four YCW products retained the adsorption up to 6 h. The analysis of equilibrium isotherms and adsorption kinetics revealed that all products adsorb C. perfringens in a dose- and time-dependent manner, with high affinity and capacity, sequestering up to 4 Log CFU/mg of product. The determination of adsorption parameters allows to differentiate among adsorbents and select the most efficient product. This approach discriminated among YCW products more efficiently than the antimicrobial assay. In conclusion, this study suggests that the ability of YCW products in reducing C. perfringens viability can be the result of an adsorption mechanism.

Comparative efficacy of agricultural by-products in sequestering mycotoxins.

BACKGROUND: Biosorption using agricultural by-products has been proven as a low-cost and safe way to sequester mycotoxins. Few agricultural by-products have been studied for their efficacy in adsorbing simultaneously a large range of mycotoxins. The present work compared the ability of 51 agricultural by-products to adsorb mycotoxins from liquid mediums simulating physiological pH values, and it studied the mechanism for mycotoxin adsorption by isotherm adsorption experiments. RESULTS: Grape pomaces, artichoke wastes, and almond hulls were selected as promising biosorbents for mycotoxins, being quite effective towards aflatoxin B1 (AFB1 ), zearalenone (ZEA), and ochratoxin A (OTA). Their adsorption was not affected by medium pH, and the adsorbed fraction was not released when pH rose from acid to neutral values. Fumonisin B1 (FB1 ) was adsorbed to a lesser extent, and deoxynivalenol adsorption was not recorded. For the selected biosorbents, maximum adsorption capacity calculated by the best fitting model (Freundlich, Langmuir, or Sips equation) ranged from 1.2 to 2.9 µg mg-1 for AFB1 , 1.3 to 2.7 µg mg-1 for ZEA, 0.03 from 2.9 µg mg-1 for OTA, and 0.01-1.1 µg mg-1 for FB1 . CONCLUSION: This study confirms that some agricultural by-products can find technological applications as feed/food additives for mycotoxin reduction. © 2018 Society of Chemical Industry.

Antimicrobial properties of rosin acids-loaded nanoparticles against antibiotic-sensitive and antibiotic-resistant foodborne pathogens.

Rosin acids (RA) from coniferous trees are used in folk medicine for healing various skin infections. Despite the antimicrobial potential of RA, their poor solubility in aqueous media may limit their use. In this work RA-loaded polyethylene glycol-poly(lactic-co-glycolic acid) nanoparticles (RA-NPs) with enhanced antimicrobial properties against foodborne bacterial pathogens were produced. RA-NPs were prepared by solvent displacement technique and characterized for relevant colloidal features by dynamic light scattering, laser Doppler anemometry and transmission electron microscopy. Association of RA to NPs occurred with high yields (86% w/w). RA and RA-NPs (∼130 nm) were strongly active against antibiotic-sensitive Gram + pathogens, i.e. Clostridium perfringens, Listeria monocytogenes and antibiotic-resistant Staphylococcus aureus. However, both failed in inhibiting the growth of Gram - pathogens (Campylobacter jejuni, Campylobacter coli, Escherichia coli and Salmonella enterica). Association to NPs enhanced the antimicrobial activity of RA. MIC, IC50, IC90, and MBC values of RA-NPs were ten-times lower than RA. RA-NPs did not change the intrinsic toxicity potential of RA. This is the first study on the enhancement of the antimicrobial activity of RA when associated to nanocarriers. This approach may be an effective strategy to produce aqueous-based RA solutions with enhanced antimicrobial activity against antibiotic-sensitive and antibiotic-resistant Gram + pathogens.

Eubiotics for Food Security at Farm Level: Yeast Cell Wall Products and Their Antimicrobial Potential Against Pathogenic Bacteria.

The population increase in the last century was the first cause of the industrialization of animal productions, together with the necessity to satisfy the high food demand and the lack of space and land for the husbandry practices. As a consequence, the farmers moved from extensive to intensive agricultural systems and introduced new practices, such as the administration of antimicrobial drugs. Antibiotics were then used as growth promoters and for disease prevention. The uncontrolled and continuous use of antibiotics contributed to the spread of antibiotic resistance in animals, and this had adverse impacts on human health. This emergence led the European Union, in 2003, to ban the marketing and use of antibiotics as growth promoters, and for prophylaxis purposes from January 2006. This ban caused problems in farms, due to the decrease in animal performances (weight gain, feed conversion ratio, reproduction, etc.), and the rise in the incidence of certain diseases, such as those induced by Clostridium perfringens, Salmonella, Escherichia coli, and Listeria monocytogenes. The economic losses due to the ban increased the interest in researching alternative strategies for the prophylaxis of infectious diseases and for health and growth promotion, such as feed additives. Yeast-based materials, such as cell wall extract, represent promising alternatives to antibiotics, on the base of their prebiotic activity and their claimed capacity to bind enteropathogenic bacteria. Several authors reported examples of the effectiveness of yeast cell wall products in adsorbing bacteria, but there is a lack of knowledge on the mechanisms involved in this interaction. The purpose of this review is to provide an overview of the current approaches used for the control of pathogenic bacteria in feed, with a particular focus on the use of yeast-derived materials proposed to control zoonoses at farm level, and on their effect on animal health.

Grape Pomace, an Agricultural Byproduct Reducing Mycotoxin Absorption: In Vivo Assessment in Pig Using Urinary Biomarkers.

The efficacy of four agricultural byproducts (ABPs) and two commercial binders (CBs) to reduce the gastrointestinal absorption of a mixture of mycotoxins was tested in piglets using urinary mycotoxin biomarkers as indicator of the absorbed mycotoxins. Twenty-eight piglets were administered a bolus contaminated with the mycotoxin mixture containing or not ABP or CB. Twenty-four hour urine was collected and analyzed for mycotoxin biomarkers by using a multiantibody immunoaffinity-based LC-MS/MS method. Each bolus contained 769 µg of fumonisin B1 (FB1), 275 µg of deoxynivalenol (DON), 29 µg of zearalenone (ZEN), 6.5 µg of aflatoxin B1 (AFB1) and 6.6 µg of ochratoxin A (OTA) corresponding to 2.2, 0.8, 0.08, 0.02, and 0.02 µg/g in the daily diet, respectively. The percentage of ABP in each bolus was 50%, whereas for the two CBs the percentages were 5.2 and 17%, corresponding to 2.8, 0.3, and 0.9% in the daily diet, respectively. The reduction of mycotoxin absorption was up to 69 and 54% for ABPs and CBs, respectively. White grape pomace of Malvasia was the most effective material as it reduced significantly (p < 0.05) urinary mycotoxin biomarker of AFB1 (67%) and ZEN (69%), whereas reductions statistically not significant were observed for FB1 (57%), DON (40%), and OTA (27%). This study demonstrates that grape pomace reduces the gastrointestinal absorption of mycotoxins. This agricultural byproduct can be considered an alternative to commercial products and used in the feed industries as an effective, cheap, and natural binder for multiple mycotoxins.

Assessment of multi-mycotoxin adsorption efficacy of grape pomace.

Grape pomace (pulp and skins) was investigated as a new biosorbent for removing mycotoxins from liquid media. In vitro adsorption experiments showed that the pomace obtained from Primitivo grapes is able to sequester rapidly and simultaneously different mycotoxins. Aflatoxin B1 (AFB1) was the most adsorbed mycotoxin followed by zearalenone (ZEA), ochratoxin A (OTA), and fumonisin B1 (FB1), whereas the adsorption of deoxynivalenol (DON) was negligible. AFB1 and ZEA adsorptions were not affected by changing pH values in the pH 3-8 range, whereas OTA and FB1 adsorptions were significantly affected by pH. Equilibrium adsorption isotherms obtained at different temperatures (5-70 °C) and pH values (3 and 7) were modeled and evaluated using the Freundlich, Langmuir, Sips, and Hill models. The goodness of the fits and the parameters involved in the adsorption mechanism were calculated by the nonlinear regression analysis method. The best-fitting models to describe AFB1, ZEA, and OTA adsorption by grape pomace were the Sips, Langmuir, and Freundlich models, respectively. The Langmuir and Sips models were the best models for FB1 adsorption at pH 7 and 3, respectively. The theoretical maximum adsorption capacities (mmol/kg dried pomace) calculated at pH 7 and 3 decreased in the following order: AFB1 (15.0 and 15.1) > ZEA (8.6 and 8.3) > OTA (6.3-6.9) > FB1 (2.2 and 0.4). Single- and multi-mycotoxin adsorption isotherms showed that toxin adsorption is not affected by the simultaneous presence of different mycotoxins in the liquid medium. The profiles of adsorption isotherms obtained at different temperatures and pH and the thermodynamic parameters (ΔG°, ΔH°, ΔS°) suggest that mycotoxin adsorption is an exothermic and spontaneous process, which involves physisorption weak associations. Hydrophobic interactions may be associated with AFB1 and ZEA adsorption, whereas polar noncovalent interactions may be associated with OTA and FB1 adsorption. In conclusion, this study suggests that biosorption of mycotoxins onto grape pomace may be a reasonably low-cost decontamination method.

Removal of ochratoxin A from contaminated red wines by repassage over grape pomaces.

Ochratoxin A contamination of red wines might be quite severe in certain high-risk regions and vintages, thus requiring corrective measures to fulfill acceptable standards for human consumption. This work proposes an innovative and environmentally friendly corrective measure to reduce ochratoxin A levels by repassage of contaminated musts or wines over grape pomaces having no or little ochratoxin A contamination. Grape pomaces have a high affinity for ochratoxin A and have been shown to remove ochratoxin A from must and wine during vinification. Time course experiments showed that ochratoxin A adsorption by pomaces is a rapid process, reaching equilibrium in less than 10 h, and is not affected by the tested toxin concentrations. Repassage of wine from Primitivo grapes spiked with 2-10 microg/kg ochratoxin A over pomaces obtained from the same grapes removed up to 65% ochratoxin A within 24 h. Similar results (50-65% ochratoxin A reduction) were obtained with Primitivo or Negroamaro wines repassed over pomaces from different grape varieties including white grapes (Malvasia, Greco di Tufo) and red grapes (Sangiovese, Aglianico). Grape pomaces maintained a good efficacy in removing ochratoxin A after being reused four times. Unlike other enological fining agents, the use of grape pomaces to adsorb ochratoxin A from red wines of the same grape variety (Primitivo) did not affect relevant wine quality parameters, including color intensity and health-promoting phenolic content (trans-resveratrol, quercetin, total polyphenols). These quality parameters were instead positively or negatively affected when contaminated wines were repassed over grape pomaces from other grape varieties, the effect being related to the intrinsic characteristics of the pomace variety. The proposed decontamination procedure can be applied in a modern winery provided that contaminated grapes are identified early and processed separately from uncontaminated grapes.

Assessment of the multi-mycotoxin-binding efficacy of a carbon/aluminosilicate-based product in an in vitro gastrointestinal model.

A laboratory model, set to simulate the in vivo conditions of the porcine gastrointestinal tract, was used to study the small intestinal absorption of several mycotoxins and the effectiveness of Standard Q/FIS (a carbon/aluminosilicate-based product) in reducing mycotoxin absorption when added to multitoxin-contaminated diets. Mycotoxins were quickly absorbed in the proximal part of the small intestine at levels of 105 and 89% for fumonisins B1 and B2, respectively, 87% for ochratoxin A, 74% for deoxynivalenol, 44% for aflatoxin B1, and 25% for zearalenone. Addition of Standard Q/FIS to the diet (up to 2%, w/w) significantly reduced mycotoxin absorption, in a dose-dependent manner, up to 88% for aflatoxin B1, 44% for zearalenone, and 29% for the fumonisins and ochratoxin. Standard Q/FIS was ineffective in reducing deoxynivalenol uptake. These findings suggest that Standard Q/FIS can be used as a multitoxin adsorbent material to prevent the individual and combined adverse effects of mycotoxins in animals.

Evaluation of the intestinal absorption of deoxynivalenol and nivalenol by an in vitro gastrointestinal model, and the binding efficacy of activated carbon and other adsorbent materials.

In vitro screening of 14 adsorbent materials, including some commercial products used to detoxify Fusarium-mycotoxins, were tested in the pH range of 3-8 for deoxynivalenol (DON)- and nivalenol (NIV)-binding ability. Only activated carbon showed to be effective with binding capacities of 35.1 micromol and 8.8 micromol DON and NIV/g adsorbent, respectively, calculated from the adsorption isotherms. A dynamic laboratory model simulating the gastrointestinal (GI) tract of healthy pigs (TIM system) was used to evaluate the small-intestinal absorption of DON and NIV and the efficacy of activated carbon in reducing the relevant absorption. The in vitro intestinal absorptions of DON and NIV were 51% and 21%, respectively, as referred to 170 microg DON and 230 microg NIV ingested through contaminated (spiked) wheat. Most absorption occurred in the jejunal compartment for both mycotoxins. The inclusion of activated carbon produced a significant reduction in the intestinal mycotoxin absorption. At 2% inclusion level the absorption with respect to the intake was lowered from 51% to 28% for DON and from 21% to 12% for NIV. The binding activity of activated carbon for these trichothecenes was lower than that observed for zearalenone, a mycotoxin frequently co-occurring with them in naturally contaminated cereals.

Effects of muffin processing on fumonisins from 14C-labeled toxins produced in cultured corn kernels.

The persistence of fumonisins during cooking is known to be affected by several factors, including thermal degradation and the presence of various ingredients in corn-based food recipes that can react with the toxin. A method for the production of corn kernels containing 14C-fumonisins was developed. The corn kernels were colonized by Fusarium verticillioides MRC 826 and supplemented with 1,2-14C-sodium acetate. The specific activity of 14C-FB1 produced made the study of its fate in cornmeal muffins possible. The double-extraction acetonitrile-water-methanol/immunoaffinity column/o-phthaldialdehyde high-performance liquid chromatography (HPLC) method was used to determine FB1 levels in cornmeal muffins. Reductions in FB1 levels in muffins spiked with 14C-labeled and unlabeled FB1 (43 and 48%, respectively) were similar, indicating that the extraction method was efficient and consistent with previous reports. However, with the labeled corn kernel material, recovery levels based on the 14C counts for the eluate from an immunoaffinity column were much higher (90%). This finding indicates that some fumonisin-related compounds other than FB1 that were present in the cornmeal were recognized by the antibodies but not by the HPLC method.

Assessing the zearalenone-binding activity of adsorbent materials during passage through a dynamic in vitro gastrointestinal model.

A novel approach is presented herein to study the intestinal absorption of mycotoxins by using a laboratory model that mimics the metabolic processes of the gastrointestinal (GI) tract of healthy pigs. This model was used to evaluate the small-intestinal absorption of zearalenone from contaminated wheat (4.1 mg/kg) and the effectiveness of activated carbon and cholestyramine at four inclusion levels (0.25, 0.5, 1 and 2%) in reducing toxin absorption. Approximately 32% of ZEA intake (247 microg) was released from the food matrix during 6 h of digestion and was rapidly absorbed at intestinal level. A significant reduction of intestinal absorption of ZEA was found after inclusion of activated carbon or cholestyramine, even at the lowest dose of adsorbents, with a more pronounced effect exhibited by activated carbon. In particular, when 2% of activated carbon or cholestyramine was added to the meal the ZEA intestinal absorption was lowered from 32% of ZEA intake to 5 and 16%, respectively. The sequestering effect of both adsorbents took place already during the first 2 h of digestion and persisted during the rest of the experiment. The GI-model is a rapid and physiologically relevant method to test the efficacy of adsorbent materials in binding mycotoxins and can be used to pre-screen mycotoxin/adsorbent combinations as an alternative to animal experiments.