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Keywords = Ochratoxin A

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15 pages, 2791 KiB  
Article
1H NMR Spectroscopy Primitivo Red Wine Screening After Grape Pomace Repassage for Possible Toxin Contamination Removal
by Laura Del Coco, Chiara Roberta Girelli, Lucia Gambacorta, Michele Solfrizzo and Francesco Paolo Fanizzi
Foods 2025, 14(5), 734; https://doi.org/10.3390/foods14050734 - 21 Feb 2025
Viewed by 180
Abstract
Food safety and quality awareness have reached significant relevance as consumers are more interested in authentic foods and drinks with specific organoleptic values. Among foodstuffs, grape products can be contaminated by Ochratoxin A (OTA), a mycotoxin that can occur in red grape after [...] Read more.
Food safety and quality awareness have reached significant relevance as consumers are more interested in authentic foods and drinks with specific organoleptic values. Among foodstuffs, grape products can be contaminated by Ochratoxin A (OTA), a mycotoxin that can occur in red grape after infection with Aspergillus carbonarius. The high affinity of grape pomace with OTA makes its use advantageous as an adsorbing/decontaminating material whether the pomace is fresh, has undergone pressing, or has undergone a stabilizing process. The effects of different grape repassage treatments on wine metabolic profiles were studied by 1H NMR spectroscopy coupled with metabolomics. The relative quantification of discriminating metabolites for activated-carbon-treated samples revealed higher levels of ethyl acetate and succinate than for the grape-pomace-repassed wine samples. On the contrary, the latter exhibited a relatively high content of glycerol, lactate, tartaric, isobutanol, isopentanol, and polyphenols. Although a specific decrease in aromatic compounds such as gallic acid, tyrosine, and tyrosol was also observed compared with the controls, for the pomace-based processes, the activated carbon treatment led to a marked general impoverishment of the metabolomic profiles, with a reduction in organic acids and glycerol. The repassage of wine over the grape pomace did not significantly affect the quality attributes of the wine, offering an alternative natural adsorbing/decontaminating material for the removal of OTA. Full article
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<p><sup>1</sup>H NMR relative expansions of significant spectral regions (400 MHz, D<sub>2</sub>O) of a specimen of analyzed wine spectra, acquired with automated noesygpps1d.comp1 Bruker sequence.</p>
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<p>(<b>a</b>) PLS-DA t [1]/t [2] score plot (two components, R<sup>2</sup>X = 0.59, R<sup>2</sup>Y = 0.53, Q<sup>2</sup> = 0.20) for the whole wine sample data set, classified according to the performed OTA decontamination treatment: Primitivo grape pomace repassage (sky-blue circle), Aglianico grape pomace repassage (blue circle), activated carbon (red circle) and control samples (green circles). (<b>b</b>). Loading S-line plot for the PLS-DA model, colored according to the correlation scaled coefficient. The <span class="html-italic">x</span>-axis indicates binned signals (ppm) in the <sup>1</sup>H NMR spectrum.</p>
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<p>Pairwise OPLS-DA t [1]/t [2] score plots for untreated (controls) vs. (<b>a</b>) Primitivo (1 + 2 + 0; R<sup>2</sup>X = 0.82, R<sup>2</sup>Y = 0.98 and Q<sup>2</sup> = 0.83) and (<b>c</b>) Aglianico (1 + 2 + 0; R<sup>2</sup>X = 0.84, R<sup>2</sup>Y = 0.99 and Q<sup>2</sup> = 0.91) repassed wine samples. Sample symbols are colored according to the different OTA decontamination treatment: control, green circle; Aglianico, blue circles; and Primitivo, green circles. Metabolites responsible for the class separation can be observed in the model-related S-line plots, (<b>b</b>,<b>d</b>) colored according to the correlation scaled coefficient. The <span class="html-italic">x</span>-axis indicates binned signals (ppm) in the <sup>1</sup>H NMR spectrum.</p>
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<p>(<b>a</b>) Focused aromatic spectral region PLS-DA t [1]/t [2] score plot (five components, R<sup>2</sup>X = 0.92, R<sup>2</sup>Y = 0.85, Q<sup>2</sup> = 0.56) for the wine samples, classified according to the performed OTA decontamination treatment: Primitivo grape pomace repassage (sky-blue circle), Aglianico grape pomace repassage (blue circle), activated carbon (red circle) and control samples (green circles). (<b>b</b>) Metabolites responsible for the class separation could be observed in the model-related loading line plots, colored according to the correlation scaled coefficient. The <span class="html-italic">x</span>-axis indicates binned signals (ppm) in the <sup>1</sup>H NMR spectrum. Quality parameters (Correct Classification Rate), CCR; area under the Receiver Operating Characteristic (ROC) curve (AUC); intercepts of R<sup>2</sup> and Q<sup>2</sup> values on the <span class="html-italic">y</span>-axis; Fisher’s probability index) of the model are reported in <a href="#app1-foods-14-00734" class="html-app">Table S3</a>.</p>
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<p>Focused aromatic spectral region OPLS-DA t [1]/to [1] score plot, for the control vs. (<b>a</b>) activated carbon (1 + 1 + 0; R<sup>2</sup>X = 0.69, R<sup>2</sup>Y = 0.99, Q<sup>2</sup> = 0.96); (<b>b</b>) Primitivo pomace (1 + 1 + 0; R<sup>2</sup>X = 0.79, R<sup>2</sup>Y = 0.96, Q<sup>2</sup> = 0.90) and (<b>c</b>) Aglianico pomace (1 + 1 + 0; R<sup>2</sup>X = 0.78, R<sup>2</sup>Y = 0.94, Q<sup>2</sup> = 0.83) wine samples. Sample symbols are colored according to the different OTA decontamination treatment: control, green circle; Aglianico, blue circles; and Primitivo green circles. Metabolites responsible for the class separation can be observed in the OPLS-DA model-related loading S-line plots (<b>d</b>–<b>f</b>), colored according to the correlation scaled coefficient. The <span class="html-italic">x</span>-axis indicates binned signals (ppm) in the <sup>1</sup>H NMR spectrum.</p>
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16 pages, 1652 KiB  
Article
Removal Capacity and Mechanism of Modified Chitosan for Ochratoxin A Based on Rapid Magnetic Separation Technology
by Xueyan Xin, Mina Nan, Yang Bi, Huali Xue, Liang Lyu, Daiwei Jiang, Hongjuan Chen and Qifang Luo
Foods 2025, 14(4), 666; https://doi.org/10.3390/foods14040666 - 15 Feb 2025
Viewed by 429
Abstract
Ochratoxin A (OTA) exposure in food is very dangerous to human health. Therefore, the development of a fast and efficient technique for OTA removal has become an urgent research topic in the field of food safety. Nano Fe3O4 modified chitosan [...] Read more.
Ochratoxin A (OTA) exposure in food is very dangerous to human health. Therefore, the development of a fast and efficient technique for OTA removal has become an urgent research topic in the field of food safety. Nano Fe3O4 modified chitosan nanocomposite (nano-Fe3O4@CTS) was synthesized as a rapidly separable and safe adsorbent and was used to adsorb OTA in wine. FT-IR, XRD, and VSM characterization methods indicated that chitosan was successfully modified by Fe3O4 and exhibited good magnetism. The adsorption and kinetics isotherms between OTA and nano-Fe3O4@CTS were studied by the Langmuir equation and the pseudo-second order kinetics equation. The mechanism of OTA adsorption on nano-Fe3O4@CTS nanoparticles was the combined effect of physical adsorption and chemisorption. The negative ΔH°, ΔG° and ΔS° values proved that the adsorption was a spontaneous and exothermic process. Nano-Fe3O4@CTS with a high maximum adsorption capacity of 5018.07 ng/g at 25 °C can rapidly separate the matrix immobilized OTA from wine, and to a certain extent retains some of the wine quality after OTA removal. Full article
(This article belongs to the Section Food Engineering and Technology)
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<p>Schematic illustrations of the synthesis strategy used to prepare nano-Fe<sub>3</sub>O<sub>4</sub>@CTS.</p>
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<p>(<b>A</b>) XRD of CTS, Fe<sub>3</sub>O<sub>4</sub> and nano-Fe<sub>3</sub>O<sub>4</sub>@CTS; (<b>B</b>) VSM of Fe<sub>3</sub>O<sub>4</sub> and nano-Fe<sub>3</sub>O<sub>4</sub>@CTS; (<b>C</b>) FT-IR of CTS, Fe<sub>3</sub>O<sub>4</sub> and nano-Fe<sub>3</sub>O<sub>4</sub>@CTS; (<b>D</b>) Particle size distribution of nano-Fe<sub>3</sub>O<sub>4</sub>@CTS.</p>
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<p>(<b>A</b>) Effect of pH, (<b>B</b>) Effects of adsorbent dosage, (<b>C</b>) Effect of adsorption temperature, (<b>D</b>) Effect of adsorption time. The error bar is expressed by standard deviation.</p>
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<p>Removal schematic of OTA with nano-Fe<sub>3</sub>O<sub>4</sub>@CTS.</p>
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18 pages, 4261 KiB  
Article
Investigating the Correlations Between Weather Factors and Mycotoxin Contamination in Corn: Evidence from Long-Term Data
by Alexander Platzer, Younos Cherkaoui, Barbara Novak and Gerd Schatzmayr
Toxins 2025, 17(2), 77; https://doi.org/10.3390/toxins17020077 - 8 Feb 2025
Viewed by 454
Abstract
Mycotoxins are secondary metabolites produced by certain fungi, posing significant health risks to humans and animals through contaminated food and feed. These fungi, and consequently the mycotoxins which they produce, are strongly influenced by weather, and this shifts over time due to climate [...] Read more.
Mycotoxins are secondary metabolites produced by certain fungi, posing significant health risks to humans and animals through contaminated food and feed. These fungi, and consequently the mycotoxins which they produce, are strongly influenced by weather, and this shifts over time due to climate change, leading to more frequent and severe events, such as heat waves, storms, and heavy rainfall. This study investigates how long-term weather trends and climatic factors impacted mycotoxin levels in corn samples over a 17-year period (2006–2022) across 12 countries, with a focus on 136 specific weather features. Among all potential relationships, we found Aspergillus toxins and fumonisins to be positively correlated with temperature, while deoxynivalenol and zearalenone are negatively correlated. Additionally, the dew point, particularly its 90th percentile value, is positively correlated with Aspergillus mycotoxins. We also identified significant patterns associated with wind direction. Collectively, these findings offer a comprehensive overview of mycotoxin–weather correlations, which may also be projected into future scenarios. Full article
(This article belongs to the Collection Impact of Climate Change on Fungal Population and Mycotoxins)
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<p>Weather trends from 2006 to 2022 (17 years). Boxplots per year show the values of different weather features for 12 countries—so exactly 12 values per year. Golden points and dashed lines indicate median values. The transparent orange line shows the weak trend (ordinary least squares). The following countries were included: Argentina, Austria, China, France, Germany, Hungary, Italy, Poland, South Africa, Spain, Turkey, and the United States. The weather features, represented on the y-axis, are specified in the title line of each plot. (<b>A</b>–<b>C</b>) represent the median, minimum, and maximum temperatures in the time frame of 60 days before harvest to harvest (m60_to0). (<b>D</b>) shows the averages of the 90th percentiles of the dew point for location nodes per country across two different periods. (<b>E</b>,<b>F</b>) show the ratios of time the wind comes from the north and south. (<b>D</b>–<b>F</b>) exhibit significant trends over these 17 years: the year-to-year absolute difference decreases. The string following the abbreviated weather feature name indicates the period, e.g., “m20_to10” denotes the time from 20 days before harvest to 10 days after harvest.</p>
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<p>Scatter plots of temperature and mycotoxins. (<b>A</b>–<b>D</b>) Median temperature correlates with different mycotoxins: <span class="html-italic">Aspergillus</span> toxins (aflatoxins, ochratoxin A), fumonisins, deoxynivalenol, and zearalenone. Positive correlations in (<b>A</b>,<b>B</b>) and negative correlations in (<b>C</b>,<b>D</b>) are clearly visible. The correlations of other temperature weather features for these 4 mycotoxins (groups) are going in the same direction, and there is no significant opposite—see text and <a href="#app1-toxins-17-00077" class="html-app">Table S3</a> for more information. The upper legend on the right pertains to (<b>A</b>–<b>D</b>), and the lower legend pertains to (<b>E</b>,<b>F</b>). The sizes of the (semi-) circles are scaled equally in all 6 diagrams. (<b>E</b>,<b>F</b>) The T-2 toxin abundance is positively correlated with minimum temperature and negatively correlated with maximum temperature. The lower legend on the right pertains to (<b>E</b>,<b>F</b>). The scale for sample sizes is the same for all diagrams; the scale is linear for size, but 0 corresponds to a certain size, not to invisibility. The minimal sample size is 5 by filtering. The colors for each point in the diagram depict its country on the left and the year on the right. The weather data correspond to the period from 60 days before harvest until harvest. The FDR-corrected <span class="html-italic">p</span>-values of Spearman correlation coefficients for (<b>A</b>–<b>F</b>) are as follows: 9.01 × 10<sup>−11</sup>, 5.98 × 10<sup>−9</sup>, 7.45 × 10<sup>−6</sup>, 0.004, 0.024, and 0.01.</p>
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<p>Relationship between dew point and <span class="html-italic">Aspergillus</span> toxins normalized. “Dewpoint90Percentile” represents the 90th percentile value of dew points in each country during the period from 60 days before harvest until harvest. (<b>A</b>) Scatter plot. The minimal sample size is 5 by filtering. The colors for each point in the diagram depict its country on the left side and the year on the right side. (<b>B</b>,<b>C</b>) China and Spain as line charts, as these two countries particularly drive the correlation. However, the relationship between dew points and <span class="html-italic">Aspergillus</span> toxins is not significant for any individual country, primarily due to sample size. The Spearman correlation is significant only when considering all countries together (FDR-corrected <span class="html-italic">p</span>-value: 6.87 × 10<sup>−9</sup>).</p>
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<p>Correlations between mycotoxin concentrations and weather features. The FDR-significant, “most covering” correlations are shown from all pairwise calculations; only statistically significant results are depicted. The labels “long”, “medium”, and “short” indicate time intervals, as defined in <a href="#sec4dot2dot1-toxins-17-00077" class="html-sec">Section 4.2.1</a>. For a complete list of correlations, refer to <a href="#app1-toxins-17-00077" class="html-app">Table S3</a>.</p>
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<p>Scatter plots of wind direction and mycotoxins (<b>A</b>–<b>D</b>). “WinddirectionNorthRatio” represents the proportion of time the wind originates from the north in each country during the period spanning 20 days before harvest to 10 days after harvest, while “WinddirectionSouthRatio” reflects the proportion of time the wind originates from the south. Each point is colored by country (left side) and year (right side). The minimal sample size was set to five after filtering. FDR-corrected <span class="html-italic">p</span>-values are 0.011, 2.8 × 10<sup>−5</sup>, 0.001, and 0.07. The last correlation is not significant when considering only Europe—shown here for consistency—and an alpha level of 0.05, but is significant across all countries combined (FDR-corrected <span class="html-italic">p</span> = 0.0005).</p>
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<p>Correlations between weather features. One representative feature is taken for each strongly correlated weather feature cluster. All pairwise correlations were calculated. The time period is from 20 days before harvest to 10 days after harvest. Only significant correlations are shown (Bonferroni-corrected). The label colors are used for grouping by type: one color for temperature-related features, another color for precipitation, etc. The labels WinddirectionSouthRatio and WinddirectionNorthRatio are more prominent because these are most discussed in the main text.</p>
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13 pages, 1396 KiB  
Article
Detection of Ochratoxin A in Tissues of Wild Boars (Sus scrofa) from Southern Italy
by Sara Damiano, Consiglia Longobardi, Lucia De Marchi, Nadia Piscopo, Valentina Meucci, Alessio Lenzi and Roberto Ciarcia
Toxins 2025, 17(2), 74; https://doi.org/10.3390/toxins17020074 - 6 Feb 2025
Viewed by 392
Abstract
Ochratoxin A (OTA) is a secondary metabolite produced by fungi of the genera Aspergillus and Penicillium, known to contaminate various food substrates. Its toxic effects include direct nephrotoxicity, as well as observed teratogenic, immunogenic, and carcinogenic effects. Climate change may contribute to [...] Read more.
Ochratoxin A (OTA) is a secondary metabolite produced by fungi of the genera Aspergillus and Penicillium, known to contaminate various food substrates. Its toxic effects include direct nephrotoxicity, as well as observed teratogenic, immunogenic, and carcinogenic effects. Climate change may contribute to increased humidity and temperature, favouring fungal growth and, consequently, OTA spreading. Recent increases in wild boar populations, along with their omnivorous nature and their varied diet, define them as environmental bioindicators for contaminants like mycotoxins. This study aimed to assess the concentrations of OTA in kidney, liver, and muscle tissue samples from 74 wild boars that were hunted in different areas of Avellino, Campania region, between 2021 and 2022. Tissue samples underwent extraction, purification, and analysis using high-performance liquid chromatography (HPLC) coupled with a fluorescence detector. Results revealed OTA presence in 35.1% of tested wild boars. The highest OTA concentration was observed in the kidney and liver, with less in the muscle, indicating the presence of this mycotoxin in the wild boars and their surrounding environment. Consequently, there is a need to formulate rules for edible wildlife products. These findings emphasize the significant risk of OTA contamination in wild boar tissues, suggesting their potential as reliable environmental markers for mycotoxin prevalence and as a toxicological concern for human health. Full article
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<p>HPLC-FLD chromatograms of: (<b>A</b>) a liver sample naturally contaminated with OTA; (<b>B</b>) a kidney sample naturally contaminated with OTA; and (<b>C</b>) a muscle sample naturally contaminated with OTA, and (<b>D</b>) OTA and OTB standard solutions (at 10 ng/mL).</p>
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<p>HPLC-FLD chromatograms of: (<b>A</b>) a liver sample naturally contaminated with OTA; (<b>B</b>) a kidney sample naturally contaminated with OTA; and (<b>C</b>) a muscle sample naturally contaminated with OTA, and (<b>D</b>) OTA and OTB standard solutions (at 10 ng/mL).</p>
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<p>Median ± range of OTA concentrations (μg/kg) in liver, muscle, and kidneys of wild boars.</p>
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<p>Linear scatterplots depicting the Spearman correlation analysis of OTA concentrations between kidney and muscle (<b>A</b>), kidney and liver (<b>B</b>), and muscle and liver (<b>C</b>) tissues.</p>
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<p>Cartographic representation of the province of Avellino (Campania region, southern Italy) delineating hunting zones (in blue) designated for wild boar (<span class="html-italic">Sus scrofa</span>) sampling. The image was created with Epi Info version 7.2.</p>
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15 pages, 1631 KiB  
Article
Impact of Bioactive Ingredients on the Fecal Excretion of Aflatoxin B1 and Ochratoxin A in Wistar Rats
by Pilar Vila-Donat, Dora Sánchez, Lara Manyes and Alessandra Cimbalo
Molecules 2025, 30(3), 647; https://doi.org/10.3390/molecules30030647 - 1 Feb 2025
Viewed by 468
Abstract
This study evaluates the effects of fermented whey (FW) and pumpkin (P) on the excretion of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in rats using immunoaffinity column cleanup and high-performance liquid chromatography–fluorescence detection (IAC-LC-FLD). The method achieved detection limits of 0.1 µg/kg [...] Read more.
This study evaluates the effects of fermented whey (FW) and pumpkin (P) on the excretion of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in rats using immunoaffinity column cleanup and high-performance liquid chromatography–fluorescence detection (IAC-LC-FLD). The method achieved detection limits of 0.1 µg/kg for AFB1 and 0.3 µg/kg for OTA, with recovery rates ranging from 72–92% for AFB1 and 88–98% for OTA. A fecal analysis of 100 rats showed peak AFB1 concentrations of 418 µg/kg and OTA of 1729 µg/kg. In the toxin-exposed groups, OTA levels were higher than AFB1, with males in the OTA-only group showing significantly higher OTA (1729 ± 712 µg/kg) than females (933 ± 512 µg/kg). In the AFB1-only group, the fecal levels were 52 ± 61 µg/kg in males and 91 ± 77 µg/kg in females. The AFB1 + FW group showed notable AFB1 concentrations (211 ± 51 µg/kg in males, 230 ± 36 µg/kg in females). The FW + P combination further influenced excretion, with higher AFB1 and OTA levels. These findings suggest that FW and P modulate mycotoxin excretion and may play a role in mycotoxin detoxification, providing insight into dietary strategies to reduce mycotoxin exposure and its harmful effects. Full article
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<p>Effects of fermented whey (FW) and pumpkin (P) on fecal aflatoxin B1 (AFB1) levels in male and female Wistar rats. (*) indicates statistically significant differences (<span class="html-italic">p</span> ≤ 0.05) in AFB1 fecal levels between experimental groups. (**) indicates statistically significant differences (<span class="html-italic">p</span> ≤ 0.01) in AFB1 fecal levels between experimental groups.</p>
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<p>Effects of fermented whey (FW) and pumpkin (P) on ochratoxin A (OTA) fecal levels in male and female Wistar rats. (*) indicates statistically significant differences (<span class="html-italic">p</span> ≤ 0.05) in OTA fecal levels between experimental groups.</p>
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<p>In vivo study scheme.</p>
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<p>Extraction method of aflatoxin B1 (AFB1) and ochratoxin A (OTA) in feces using AflaOchra immunoaffinity columns (IACs) and analysis via liquid chromatography with fluorescence detection (LC-FLD).</p>
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17 pages, 480 KiB  
Article
Monitoring of Ochratoxin A Occurrence and Dietary Intake in Tarhana, a Fermented Cereal-Based Product
by Esra Akkaya, Meryem Akhan, Burcu Cakmak Sancar, Hamparsun Hampikyan, Ayse Seray Engin, Omer Cetin, Enver Baris Bingol and Hilal Colak
Foods 2025, 14(3), 443; https://doi.org/10.3390/foods14030443 - 29 Jan 2025
Viewed by 468
Abstract
The aim of this study was to determine the mold and ochratoxin A (OTA) contamination of tarhana, a traditional product widely consumed in Turkish cuisine. For this purpose, a total of 350 tarhana samples (homemade and industrially produced) were randomly collected from retail [...] Read more.
The aim of this study was to determine the mold and ochratoxin A (OTA) contamination of tarhana, a traditional product widely consumed in Turkish cuisine. For this purpose, a total of 350 tarhana samples (homemade and industrially produced) were randomly collected from retail stores, markets, and bazaars in different regions of Türkiye and analyzed by means of LC-MS/MS for the occurrence of OTA. According to the results, OTA was detected in 36 of 150 (24%) industrially produced tarhana samples, with a concentration range of 0.12–2.34 µg/kg, while 118 of 200 (59%) homemade tarhana samples contained OTA, with the range from 0.16 to 4.15 µg/kg. Only 8 of 350 (4%) homemade tarhana samples were found to be above the maximum permissible limit (3.0 µg/kg) for OTA. The mold contamination was found to be higher in homemade tarhana (3.756 log CFU/g) than in the industrially produced samples (2.742 log CFU/g). The estimated weekly intake values of OTA with tarhana consumption were well below the provisional tolerable weekly intake values for both industrially produced and homemade tarhana samples, even when consumed every day of the week, indicating that dietary intake of OTA through tarhana consumption does not pose a health risk. In conclusion, optimizing the fermentation and drying conditions applied during tarhana production and ensuring proper hygiene conditions can help to reduce the risk of OTA contamination. Moreover, monitoring and testing the OTA levels in tarhana on a regular basis can also ensure the food safety of this product. Full article
(This article belongs to the Section Food Toxicology)
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<p>The detection rate of OTA in tarhana samples (<span class="html-italic">n</span> = 350). * ND: not detected (detection limit: 0.10 μg/kg). <sup>a</sup>: The maximum permissible limit of OTA set by the Turkish Food Codex and European Commission.</p>
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22 pages, 2303 KiB  
Article
Real-Time Classification of Ochratoxin a Contamination in Grapes Using AI-Enhanced IoT
by Mohamed Riad Sebti, Zohra Dakhia, Sonia Carabetta, Rosa Di Sanzo, Mariateresa Russo and Massimo Merenda
Sensors 2025, 25(3), 784; https://doi.org/10.3390/s25030784 - 28 Jan 2025
Viewed by 446
Abstract
Ochratoxin A (OTA) contamination presents significant risks in viticulture, affecting the safety and quality of wine and grape-derived products. This study introduces a groundbreaking method for early detection and management of OTA, leveraging environmental data such as temperature and humidity. A function derived [...] Read more.
Ochratoxin A (OTA) contamination presents significant risks in viticulture, affecting the safety and quality of wine and grape-derived products. This study introduces a groundbreaking method for early detection and management of OTA, leveraging environmental data such as temperature and humidity. A function derived from chemical analysis was developed to estimate OTA concentrations and used to label a synthetic dataset, establishing safe thresholds. Two AI models were trained: one for the detecting of OTA presence and the other for classifying the concentration range. These models were deployed on a M5Stick C+, a microcontroller designed for real-time data processing. The inference process is optimized for rapid response, requiring minimal time to deliver results. Additionally, the low power consumption of the M5Stick C+ ensures that the device can operate throughout the harvest period on a single charge. The system is able to transmit inference data via MQTT for real-time analysis. This comprehensive approach offers a scalable, cost-effective, on-site solution that is autonomous, eliminating the need for domain experts and extensive resources. The robustness of the system was demonstrated through its consistent performance across multiple test sets, providing an effective enhancement to food safety in grape and wine production. The study also details the system architecture, describes the function used for data labeling, outlines the training and deployment processes of the models, and finally, assesses the testing of the overall system. Full article
(This article belongs to the Special Issue Sensor-Based Crop and Soil Monitoring in Precise Agriculture)
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<p>Overview of the proposed system’s architecture, detailing three main sections: (1) Model training and deployment; (2) On-device inference and data processing; (3) Data transmission and storage.</p>
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<p>AI model flowchart showing the workflow from training to deployment for OTA prediction. The model is trained and optimized in (2) on a synthetic dataset in (1), converted from .h5 to .cc in (3), and deployed on an M5C+ device in (4). The M5C+ is setup with sensors in (5) for real-time data collection in (6) and OTA concentration prediction in (7).</p>
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<p>Evaluation function over time for OTA.</p>
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<p>M5stick C plus [<a href="#B36-sensors-25-00784" class="html-bibr">36</a>].</p>
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<p>PCD model architecture.</p>
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<p>A test set (from real-world) distribution.</p>
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<p>Test set samples distribution: ADD and PCD.</p>
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<p>Confusion matrixes.</p>
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<p>Classification results using M5C+. The screen shows the predicted class and the inference time in milliseconds.</p>
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<p>Classification and measurement results using M5C+ for ADD.</p>
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<p>Classification and measurement results using M5C+ for PCD.</p>
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<p>Commercial harvesting box with integration of the M5C+.</p>
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16 pages, 2463 KiB  
Article
Enzymatic Degradation of Ochratoxin A: The Role of Ultra-Pure Water
by Joana Santos, Constança Oliveira, Filipe Teixeira, Armando Venâncio and Carla Silva
Foods 2025, 14(3), 397; https://doi.org/10.3390/foods14030397 - 25 Jan 2025
Viewed by 526
Abstract
Ochratoxin A (OTA) is a toxic mycotoxin, making its removal from food essential for public health. This study examines OTA degradation by porcine pancreatic lipase (PPL) in ultra-pure water versus buffer systems through in vitro assays and molecular modeling. The results show that [...] Read more.
Ochratoxin A (OTA) is a toxic mycotoxin, making its removal from food essential for public health. This study examines OTA degradation by porcine pancreatic lipase (PPL) in ultra-pure water versus buffer systems through in vitro assays and molecular modeling. The results show that PPL fully degrades OTA in ultra-pure water within 7 h at 44 °C, whereas only partial degradation occurs in phosphate buffer. After 4 h, PPL in water degrades 91% of OTA, compared to only 12% in buffer. The enzyme’s half-life is longer in water (~4 h 4 min) than in phosphate buffer (~2 h 30 min), suggesting better stability in water. Other buffers, including acetate, citrate, and borate, confirmed higher degradation efficiency in low-conductivity, acidic environments similar to ultra-pure water. Additionally, using the model compound p-nitrophenyl octanoate (p-NPO), it was found that p-NPO degrades faster in buffer, likely due to a salting-out effect. Molecular modeling and circular dichroism analysis indicate that PPL’s secondary structure in water promotes an ideal conformation for OTA binding. This study suggests ultra-pure water as a greener, sustainable option for reducing mycotoxins in food, with broad industrial applications. Full article
(This article belongs to the Special Issue Recent Advances of Enzymes in Food Industry)
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<p>Evaluation of the optimal conditions for OTA hydrolysis by PPL in two different media. The reaction was monitored at different (<b>a</b>) temperatures (21, 30, 37, 44, 50, 57, and 60 °C) and (<b>b</b>) incubation times (15 and 30 min; 1, 2, 4, 6, 7, and 10 h). The optimal temperature for OTA hydrolysis was 44 °C in water and buffer. The incubation time required for the complete hydrolysis of OTA was 7 h in pure water (44 °C).</p>
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<p>PPL enzymatic activity in pure water and in phosphate buffer (100 mM, pH 7.5) evaluated as OTA degradation for 4 h of incubation at 44 °C.</p>
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<p>The hydrolysis of OTA by PPL in different buffers and ultra-pure water at 44 °C is represented by bars. The conductivity in mS/cm of each solution under study is represented by a black dot above each bar.</p>
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<p>The hydrolytic behavior of PPL in water and phosphate buffer (pH 7.5, 100 mM) across different substrates was investigated.</p>
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<p>Cartoon representation of the last recorded conformation of PPL in water (<b>a</b>) and PBS (<b>b</b>), the cavities of the active centers are highlighted for chain A (magenta surface) and chain C (green surface). Both images are cropped depth-wise to aid visualization of the active center cavities.</p>
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<p>Radial-pair distribution function of the oxygen atom of the water molecules (Ow) with respect to the average position of the active centers of PPL’s chains A and C in water and PBS (<b>a</b>). RDF of Ow with respect to the average position of the histidine residues in PPL in water and PBS (<b>b</b>).</p>
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27 pages, 31119 KiB  
Article
Hazards in Products from Northern Mediterranean Countries Reported in the Rapid Alert System for Food and Feed (RASFF) in 1997–2021 in the Context of Sustainability
by Marcin Pigłowski, Alberto Nogales and Maria Śmiechowska
Sustainability 2025, 17(3), 889; https://doi.org/10.3390/su17030889 - 22 Jan 2025
Viewed by 875
Abstract
The European Green Deal attaches great importance to sustainability, including food security, which is also linked to food safety. This is particularly relevant in such a sensitive region as the Mediterranean. The goal of this study was to investigate Rapid Alert System for [...] Read more.
The European Green Deal attaches great importance to sustainability, including food security, which is also linked to food safety. This is particularly relevant in such a sensitive region as the Mediterranean. The goal of this study was to investigate Rapid Alert System for Food and Feed (RASFF) notifications of hazards reported in 1997–2021 (a 25-year period) in products from northern Mediterranean countries considering products and other variables. A two-way joining cluster analysis was used. The most notable hazards in the latter years of the reported period were as follows: ochratoxin A and pesticide residues in fruits and vegetables imported from Turkey and ethylene oxide in various products, as well as Salmonella in chicken, Listeria in cheese, Escherichia coli in cheese and mussels from France, mercury in swordfish from Spain, and Anisakis in seafood from France and Morocco. The increasing number of notifications of ochratoxin A and pathogenic micro-organisms in recent years may be caused by climate change. This also results in the need to use more pesticides and the appearance of related hazards, i.e., residues of such compounds in food products. It is, therefore, vitally important that border posts and control authorities in particular European Union countries are vigilant. Full article
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<p>The number of notifications in the RASFF reported on products from Mediterranean countries per year over the period 1997–2021.</p>
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<p>The findings of cluster analysis for hazard categories and countries of origin on products from northern Mediterranean countries reported in the RASFF in 1997–2021. Explanations for hazard categories: biological contaminants—biological contaminants (other); chemical contaminants—chemical contaminants (other); food additives…—food additives and flavourings; genetically modified—genetically modified food or feed; labelling—labelling absent/incomplete/incorrect; microbial contaminants (other)— microbial contaminants (other); packaging—packaging defective/incorrect; veterinary products—residues of veterinary medicinal products; TSEs—transmissible spongiform encephalopathies. Explanations for countries of origin: B&amp;H—Bosnia and Herzegovina.</p>
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<p>The findings of cluster analysis for hazard categories and product categories on products from northern Mediterranean countries reported in the RASFF in 1997–2021. Explanations for hazard categories: biological contaminants—biological contaminants (other); chemical contaminants—chemical contaminants (other); food additives…—food additives and flavourings; genetically modified—genetically modified food or feed; labelling—labelling absent/incomplete/incorrect; microbial contaminants (other)— microbial contaminants (other); packaging—packaging defective/incorrect; veterinary products—residues of veterinary medicinal products; TSEs—transmissible spongiform encephalopathies. Explanations for product categories: bivalve molluscs—bivalve molluscs and products thereof; cephalopods—cephalopods and products thereof; cereals and bakery—cereals and bakery products; cocoa, coffee and tea—cocoa and cocoa preparations, coffee, and tea; crustaceans—crustaceans and products thereof; dietetic foods…—dietetic foods, food supplements, and fortified foods; eggs—eggs and egg products; fish—fish and fish products; food additives…—food additives and flavourings; meat—meat and meat products (other than poultry); milk—milk and milk products; molluscs—molluscs and products thereof; nuts and seeds—nuts, nut products, and seeds; poultry meat—poultry meat and poultry meat products; soups, broths…—soups, broths, sauces, and condiments; water—water for human consumption (other); crustaceans (wild-caught)—wild-caught crustaceans and products thereof; fish (wild caught)—wild-caught fish and products thereof (other than crustaceans and molluscs).</p>
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<p>Imports of fruits and vegetables from Turkey to the European Union in 1999–2021.</p>
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<p>The network visualization carried out in VOSviewer taking into account the keywords indicated by authors of scientific works containing the words “Mediterranean*” and “food”.</p>
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<p>The overlay visualization carried out in VOSviewer taking into account the keywords indicated by authors of scientific works containing the words “Mediterranean*” and “food”.</p>
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20 pages, 813 KiB  
Review
Mycotoxins in Cheese: Assessing Risks, Fungal Contaminants, and Control Strategies for Food Safety
by Camila Aranda, Rodrigo Rodriguez, Martín A. Fernández-Baldo and Paola Durán
Foods 2025, 14(3), 351; https://doi.org/10.3390/foods14030351 - 22 Jan 2025
Viewed by 819
Abstract
According to the scientific information reviewed, cheese is highly susceptible to contamination by mycotoxin-producing fungi, primarily species from the genera Aspergillus (A. niger, A. flavus) and Penicillium (P. commune, P. solitum, P. palitans, and P. crustosum [...] Read more.
According to the scientific information reviewed, cheese is highly susceptible to contamination by mycotoxin-producing fungi, primarily species from the genera Aspergillus (A. niger, A. flavus) and Penicillium (P. commune, P. solitum, P. palitans, and P. crustosum). Studies on various types of cheese made from cow’s milk report an average concentration of Aflatoxin M1 (AFM1) at 13,000 ng kg−1, which is alarming since the regulatory limits for AFM1 in cheese range from 250 to 500 ng kg−1. For instance, limits set by Codex Alimentarius, the European Commission (EC), Turkey, and Iran are 250 ng kg−1. In the Netherlands, the limit is 200 ng kg−1, and in Italy, it is 450 ng kg−1. However, the concentration of mycotoxins frequently exceeds these regulatory limits, including critical mycotoxins such as ochratoxin A, citrinin, and cyclopiazonic acid, which pose significant global health concerns. Therefore, this study aims to review the mycobiota responsible for producing key mycotoxins in cheese and to assess the influence of physicochemical factors on fungal growth and mycotoxin production. By incorporating control strategies such as hygiene practices, pasteurization, and the use of preservatives, this study seeks to improve methodologies in the cheese production chain and mitigate contamination by fungi and mycotoxins. Full article
(This article belongs to the Section Food Microbiology)
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<p>The key physicochemical parameters that affect fungal growth and mycotoxin production in cheese. These include temperature, water activity (a<sub>w</sub>), pH, NaCl content, moisture, carbon and nitrogen sources, C/N ratio, and redox potential (E°). The interaction of these factors determines fungal spore germination, colony growth, and toxin synthesis, highlighting their critical roles in cheese contamination dynamics.</p>
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11 pages, 1896 KiB  
Communication
Nobiletin and Eriodictyol Suppress Release of IL-1β, CXCL8, IL-6, and MMP-9 from LPS, SARS-CoV-2 Spike Protein, and Ochratoxin A-Stimulated Human Microglia
by Irene Tsilioni, Duraisamy Kempuraj and Theoharis C. Theoharides
Int. J. Mol. Sci. 2025, 26(2), 636; https://doi.org/10.3390/ijms26020636 - 14 Jan 2025
Viewed by 1396
Abstract
Neuroinflammation is involved in various neurological and neurodegenerative disorders in which the activation of microglia is one of the key factors. In this study, we examined the anti-inflammatory effects of the flavonoids nobiletin (5,6,7,8,3′,4′-hexamethoxyflavone) and eriodictyol (3′,4′,5,7-tetraxydroxyflavanone) on human microglia cell line activation [...] Read more.
Neuroinflammation is involved in various neurological and neurodegenerative disorders in which the activation of microglia is one of the key factors. In this study, we examined the anti-inflammatory effects of the flavonoids nobiletin (5,6,7,8,3′,4′-hexamethoxyflavone) and eriodictyol (3′,4′,5,7-tetraxydroxyflavanone) on human microglia cell line activation stimulated by either lipopolysaccharide (LPS), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) full-length Spike protein (FL-Spike), or the mycotoxin ochratoxin A (OTA). Human microglia were preincubated with the flavonoids (10, 50, and 100 µM) for 2 h, following which, they were stimulated for 24 h. The inflammatory mediators interleukin-1 beta (IL-1β), chemokine (C-X-C motif) ligand 8 (CXCL8), IL-6, and matrix metalloproteinase-9 (MMP-9) were quantified in the cell culture supernatant by enzyme-linked immunosorbent assay (ELISA). Both nobiletin and eriodictyol significantly inhibited the LPS, FL-Spike, and OTA-stimulated release of IL-1β, CXCL8, IL-6, and MMP-9 at 50 and 100 µM, while, in most cases, nobiletin was also effective at 10 µM, with the most pronounced reductions at 100 µM. These findings suggest that both nobiletin and eriodictyol are potent inhibitors of the pathogen-stimulated microglial release of inflammatory mediators, highlighting their potential for therapeutic application in neuroinflammatory diseases, such as long COVID. Full article
(This article belongs to the Special Issue Natural Bioactives and Inflammation, 2nd Edition)
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<p>Inhibition of the release of inflammatory mediators from human microglia stimulated by LPS. Human microglia (0.5 × 10<sup>5</sup> cells/1 mL/well in 24-well plates) were first preincubated with nobiletin (&gt;98% purity; 10, 50, and 100 μM), eriodictyol (10, 50, and 100 μM), luteolin (50 μM), and methoxyluteolin (50 μM) for 2 h and then incubated with LPS (10 ng/mL) for 24 h. Control cells were treated with an equal volume of culture medium. Then, the cell culture supernatant fluids were collected and assayed for IL-1β (<b>A</b>), CXCL8 (<b>B</b>), IL-6 (<b>C</b>), and MMP-9 (<b>D</b>) by commercial ELISA kits. All assays were performed in triplicate. LPS was compared to the control; each of the other conditions was compared to LPS (n = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>Inhibition of the release of inflammatory mediators from human microglia stimulated by SARS-CoV-2 Spike protein. Human microglia (0.5 × 10<sup>5</sup> cells/1 mL /well in 24-well plates) were preincubated with nobiletin (10, 50, and 100 μM), eriodictyol (10, 50, and 100 μM), luteolin (50 μM), and methoxy luteolin (50 μM) for 2 h and then incubated with SARS-CoV-2 Spike protein (10 ng/mL) for 24 h. Control cells were treated with an equal volume of culture medium. Following this, cell culture supernatant fluids were collected and assayed for IL-1β (<b>A</b>), CXCL8 (<b>B</b>), IL-6 (<b>C</b>), and MMP-9 (<b>D</b>) by commercial ELISA kits. FL Spike was compared to the control; each of the other conditions was compared to FL Spike (n = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001).</p>
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<p>Inhibition of the release of inflammatory mediators from human microglia stimulated by OTA. Human microglia (0.5 × 10<sup>5</sup> cells/500 μL/well in 24-well plates) were preincubated with nobiletin (10, 50, and 100 μM), luteolin (50 μM), and methoxy luteolin (50 μM) for 2 h and then incubated with OTA (10 nM) for 24 h (n = 3). Control cells were treated with an equal volume of culture medium. Following this, cell culture supernatant fluids were collected and assayed for IL-1β (<b>A</b>), CXCL8 (<b>B</b>), IL-6 (<b>C</b>), and MMP-9 (<b>D</b>) by commercial ELISA kits. OTA was compared to the control; each of the other conditions was compared to OTA (n = 3, * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001).</p>
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18 pages, 4143 KiB  
Article
Proteomic Analysis of the Murine Liver Response to Oral Exposure to Aflatoxin B1 and Ochratoxin A: The Protective Role to Bioactive Compounds
by Silvia Trombetti, Alessandra Cimbalo, Michela Grosso, Pilar Vila-Donat, Jordi Mañes and Lara Manyes
Toxins 2025, 17(1), 29; https://doi.org/10.3390/toxins17010029 - 9 Jan 2025
Viewed by 679
Abstract
Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) are considered the most important mycotoxins in terms of food safety. The aim of this study was to evaluate the hepatotoxicity of AFB1 and OTA exposure in Wistar rats and to assess the beneficial effect of [...] Read more.
Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) are considered the most important mycotoxins in terms of food safety. The aim of this study was to evaluate the hepatotoxicity of AFB1 and OTA exposure in Wistar rats and to assess the beneficial effect of fermented whey (FW) and pumpkin (P) as functional ingredients through a proteomic approach. For the experimental procedures, rats were fed AFB1 and OTA individually or in combination, with the addition of FW or a FW-P mixture during 28 days. For proteomics analysis, peptides were separated using a LC-MS/MS-QTOF system and differentially expressed proteins (DEPs) were statistically filtered (p < 0.05) distinguishing males from females. Gene ontology visualization allowed the identification of proteins involved in important biological processes such as the response to xenobiotic stimuli and liver development. Likewise, KEGG pathway analysis reported the metabolic routes as the most affected, followed by carbon metabolism and biosynthesis of amino acids. Overall, the results highlighted a strong downregulation of DEPs in the presence of AFB1 and OTA individually but not with the mixture of both, suggesting a synergistic effect. However, FW and P have helped in the mitigation of processes triggered by mycotoxins. Full article
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<p>Venn diagram representation of common DEPs for male (<b>A</b>) and female (<b>B</b>) rats exposed to mycotoxins versus the control. <span class="html-italic">p</span> &lt; 0.05 were significantly different from the control.</p>
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<p>Venn diagram representation of common DEPs for male (<b>A</b>) and female (<b>B</b>) rats exposed to FW and mycotoxins versus the corresponding mycotoxin. <span class="html-italic">p</span> &lt; 0.05 were significantly different from mycotoxins group.</p>
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<p>Venn diagram representation of common DEPs for male (<b>A</b>) and female (<b>B</b>) rats exposed to FW + P and mycotoxins versus the corresponding mycotoxin. <span class="html-italic">p</span> &lt; 0.05 were significantly different from mycotoxins group.</p>
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<p>Gene ontology (GO) functional annotation of differentially expressed proteins for biological processes and molecular functions of male (<b>A</b>,<b>C</b>) and female (<b>B</b>,<b>D</b>) rats exposed to FW + AFB1, FW + OTA, and FW + AFB1 + OTA compared with respective mycotoxins without functional ingredient.</p>
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<p>Gene ontology (GO) functional annotation of differentially expressed proteins for biological processes and molecular functions of male (<b>A</b>,<b>C</b>) and female (<b>B</b>,<b>D</b>) rats exposed to FW + P + AFB1, FW + P + OTA, and FW + P + AFB1 + OTA compared with respective mycotoxins without functional ingredients.</p>
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<p>Heatmap representation on the expression of DEPs involved in the main biological processes after AFB1, OTA, and the combination (AFB1 + OTA) exposure in presence of FW or FW + P in male (<b>A</b>) and female (<b>B</b>) rats compared to control. The red-to-green gradient represents the logarithmic fold change value for upregulation (Log2FC &gt; 0) and downregulation (Log2FC &lt; 0), respectively. Black box is log2FC = 0. <span class="html-italic">p</span> &lt; 0.05 significantly different from the mycotoxin groups.</p>
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<p>KEGG pathway visualization of significant signaling pathways in rats exposed to mycotoxins in combination with fermented whey (FW) (<b>A</b>) or fermented whey + pumpkin (FW + P) (<b>B</b>) feed related to the number of proteins involved compared with the exposure without functional ingredients.</p>
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<p>Heatmap representation of the expression of DEPs involved in the main signaling pathways after AFB1, OTA, and the combination (AFB1 + OTA) exposure in presence of FW or FW + P in male (<b>A</b>) and female (<b>B</b>) rat livers compared with the expression after exposure to mycotoxins without functional ingredients. The red-to-green gradient represents the logarithmic fold change value for upregulation (LogFC &gt; 0) and downregulation (LogFC &lt; 0), respectively. Black box is log2FC = 0. <span class="html-italic">p</span> &lt; 0.05 significantly different from the mycotoxin’s groups.</p>
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<p>KEGG pathway visualization showing key molecular events involved in the development of hepatocellular carcinoma (HCC). The diagram highlights the critical signaling pathways, including those related to cell cycle regulation, apoptosis, and metabolic alterations, which contribute to the initiation and progression of liver cancer. Red stars indicate DEPs found in this study after AFB1 and OTA exposure and bioactive ingredients.</p>
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19 pages, 3481 KiB  
Article
Holocellulose from a Winemaking By-Product to Develop a Biopolymeric System for Bacterial Immobilization: Adsorption of Ochratoxin A in Wine Model Solutions (Box–Behnken Design)
by Verónica Carrasco-Sánchez, V. Felipe Laurie, Marcelo Muñoz-Vera and Ricardo Ignacio Castro
Toxins 2025, 17(1), 26; https://doi.org/10.3390/toxins17010026 - 6 Jan 2025
Viewed by 766
Abstract
Significant agro-industrial waste is produced during the winemaking process, including grape stalks, which are a rich source of the valuable biopolymer holocellulose that can be utilized for biotechnological processes. The purpose of this study was to delignify grape stalks in order to extract [...] Read more.
Significant agro-industrial waste is produced during the winemaking process, including grape stalks, which are a rich source of the valuable biopolymer holocellulose that can be utilized for biotechnological processes. The purpose of this study was to delignify grape stalks in order to extract holocellulose. Then Lactobacillus plantarum (LP) was immobilized in the interstitial spaces of holocellulose and then coated with natural polymers (chitosan, Ch; and alginate, Al) to create the Holo-LP/Ch/Al complex. A physicochemical analysis of the system revealed strong bacterial immobilization and stability. The efficiency of the complex in adsorbing ochratoxin A (OTA) from wine model solutions was assessed using a Box–Behnken design under various pH, time, and concentration conditions. The results showed that at pH 3.0, 75.39 min, and a complex concentration of 43.82 mg mL−1, the best OTA removal (53.68%) took place. Because of its physicochemical interactions, the complex showed improved OTA adsorption in acidic environments. This study demonstrates the potential of biopolymeric systems based on holocellulose for reducing mycotoxin contamination in beverages and stabilizing bacterial cells. These results offer a viable way to increase food safety and value winemaking by-products. Full article
(This article belongs to the Special Issue Mitigation and Detoxification Strategies of Mycotoxins)
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<p>Development of a biopolymeric system for bacterial immobilization. (<b>A</b>) Lignin elimination from stalk grape; (<b>B</b>) Holo-LP complex formation; (<b>C</b>) Holo-LP/Ch/Al Complex Formation.</p>
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<p>Analysis of control and holocellulose: (<b>A</b>) TG curve; (<b>B</b>) DTG curves; (<b>C</b>) deconvolution curves of control; (<b>D</b>) deconvolution curves of holocellulose.</p>
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<p>(<b>A</b>) FTIR spectra, holocellulose (red line), and control (blue line); (<b>B</b>) range between 3800 at 2700 cm<sup>−1</sup>; (<b>C</b>) range between 1200 at 800 cm<sup>−1</sup>; (<b>D</b>) range between 1800 at 1300 cm<sup>−1</sup>.</p>
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<p>Scanning electronic microscopy (SEM); (<b>A</b>,<b>B</b>) waste from grape stalk of <span class="html-italic">Vitis vinifera</span> L. (control); (<b>C</b>,<b>D</b>) changes in waste due to mechanical; (<b>E</b>,<b>F</b>) Holocellulose; (<b>G</b>,<b>H</b>) representation of the formation of holocellulose.</p>
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<p>TGA of <span class="html-italic">Holo-LP/Ch/Al</span> complex and controls: (<b>A</b>) TG curve; (<b>B</b>) DTG curves relative loss mass.</p>
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<p>FT-IR spectra of different samples.</p>
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<p>(<b>A</b>) Pareto chart standardized effects; (<b>B</b>–<b>D</b>) three-dimensional response surface plot for the analytical response vs. the effect of A (pH), B (time; min.), and C (Concentrations; mg mL<sup>−1</sup>).</p>
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26 pages, 7553 KiB  
Article
Chemical Composition, In Vivo, and In Silico Molecular Docking Studies of the Effect of Syzygium aromaticum (Clove) Essential Oil on Ochratoxin A-Induced Acute Neurotoxicity
by Mostapha Brahmi, Djallal Eddine H. Adli, Imane Kaoudj, Faisal K. Alkholifi, Wafaa Arabi, Soumia Sohbi, Kaddour Ziani, Khaled Kahloula, Miloud Slimani and Sherouk Hussein Sweilam
Plants 2025, 14(1), 130; https://doi.org/10.3390/plants14010130 - 4 Jan 2025
Viewed by 817
Abstract
The aim of our research was to understand the impact of ochratoxin A (OTA) exposure on various physiological and behavioral aspects in adult Wistar rats, and to evaluate the efficacy of a Syzygium aromaticum essential oil (EOC) treatment in restoring the damage caused [...] Read more.
The aim of our research was to understand the impact of ochratoxin A (OTA) exposure on various physiological and behavioral aspects in adult Wistar rats, and to evaluate the efficacy of a Syzygium aromaticum essential oil (EOC) treatment in restoring the damage caused by this toxin. The essential oils were extracted by hydrodistillation, a yield of 12.70% was obtained for EOC, and the GC-MS characterization of this essential oil revealed that its principal major components are eugenol (80.95%), eugenyl acetate (10.48%), β-caryophyllene (7.21%), and α-humulene (0.87%). Acute OTA intoxication was induced by an intraperitoneal (IP) injection of 289 µg/kg/b.w. every 48 h for 12 doses, resulting in significant reductions in the body and brain weights of exposed rats when compared with controls. The neurobehavioral analysis using several behavioral testing techniques, such as the forced swimming, the dark/light test, the Morris water maze, and the open field test, clearly revealed that OTA exposure causes neurobehavioral disorders, including decreased locomotor activity, a reduced willingness to explore the environment, reflecting a state of stress, anxiety and depression, as well as impaired memory and learning. In addition, OTA intoxication has been associated with metabolic disturbances such as hyperglycemia and hypercortisolemia. However, treatment with EOC mitigated these adverse effects by improving body and brain weights and restoring neurobehavioral function. The in silico analysis revealed significant affinities between clove oils and two tested esterase enzymes (ACh and BuChE) that were more than or similar to the four neurotransmitters “dopamine, serotonin, norepinephrine, and glutamic acid” and the co-crystallized ligands NAG, MES, and GZ5. These results highlight the therapeutic potential of EOC in combating the toxic effects of OTA and pave the way for future research into the mechanisms of action and therapeutic applications of natural compounds in the prevention and treatment of poison-induced diseases. Full article
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<p>Forced swimming immobility times of control, OTA-intoxicated, and intoxicated and EOC-treated young animals (rats). The data are expressed as means ±SEMs (*** <span class="html-italic">p</span> &lt; 0.001, * <span class="html-italic">p</span> &lt; 0.05). Eight rats were included in each group.</p>
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<p>A comparison of the various parameters of the open field test in control, OTA-intoxicated, OTA-intoxicated and EOC-treated rats. The data are expressed as means ± SEMs; OTA vs. C (*** <span class="html-italic">p</span> &lt; 0.001); OTA-EOC vs. OTA-EOC (*** <span class="html-italic">p</span> &lt; 0.001). Eight rats were included in each group.</p>
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<p>Time spent in the dark/light compartments by the four groups. The data are expressed as means ± SEMs; OTA vs. C (* <span class="html-italic">p</span> &lt; 0.05), and OTA vs. OTA-EOC (*: <span class="html-italic">p</span> &lt; 0.05). Eight rats were included in each group.</p>
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<p>Morris water maze test: latency during the learning phase (over 4 days) of the control (C), treated (EOC), intoxicated (OTA) and intoxicated and treated (OTA-EOC) rats. The data are expressed as means ± SEMs (*** <span class="html-italic">p</span> &lt; 0.001). Eight rats were included in each group.</p>
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<p>Time spent in the northwest quadrant (NW) during probe testing by control (C), treated (EOC), intoxicated (OTA), and intoxicated and treated (OTA-EOC) rats. The data are expressed as means ± SEMs (*** <span class="html-italic">p</span> &lt; 0.001). Eight rats were included in each group.</p>
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<p>Latency during the visible platform trial of control (C), treated (EOC), intoxicated (OTA), and intoxicated and treated (OTA-EOC) rats. The data are expressed as means ± SEMs (*** <span class="html-italic">p</span> &lt; 0.001). Eight rats were included in each group.</p>
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<p>Blood glucose levels in the control, OTA-intoxicated, OTA/EOC-treated, and EOC-treated groups. The data are expressed as means ± SEMs; OTA vs. C (*: <span class="html-italic">p</span> &lt; 0.05) and OTA vs. OTA-EOC (*: <span class="html-italic">p</span> &lt; 0.05). Eight rats were included in each group.</p>
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<p>Blood cortisol levels in the control, OTA-intoxicated, OTA/EOC-treated, and EOC-treated groups. The data are expressed as means ± SEMs; OTA-EOC (*** <span class="html-italic">p</span> &lt; 0.001). Eight rats were included in each group.</p>
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<p>Histological sections of the rat brain. (<b>A</b>) Photomicrograph of a brain lesion in the hippocampus; pyramidal neurons are slightly eosinophilic (Black arrow) and a perivascular bulge is marked (Blue arrow). H&amp;E staining, 100×. (<b>B</b>) Photomicrograph of a cerebral lesion, neurodegenerative changes, and gliosis. H&amp;E staining, 100×. (<b>C</b>) Photomicrograph of a brain lesion with eosinophilic Purkinje neurons (Black arrow), marked congestion and hemorrhages in the brain and meninges (Blue arrow). H&amp;E staining, 100×. (<b>D</b>) Photonic microscopic view of tissue from the cerebellar cortex stained with hematoxylin and eosin G (×40). Rats treated with EOC and intoxicated with OTA appeared to have a normal architecture. (<b>E</b>) Photonic microscopic view of tissue from the cerebellar cortex of a control rat, stained with hematoxylin and eosin G (×40).</p>
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<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) dopamine; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligand “NAG” in the binding site of the human acetylcholinesterase receptor (PDB ID: 4EY5). Dimensions X, Y, and Z are 19.2037, 16.0972, and 19.7357, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
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<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) dopamine; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligand “NAG” in the binding site of the human acetylcholinesterase receptor (PDB ID: 4EY5). Dimensions X, Y, and Z are 19.2037, 16.0972, and 19.7357, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
Full article ">Figure 10 Cont.
<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) dopamine; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligand “NAG” in the binding site of the human acetylcholinesterase receptor (PDB ID: 4EY5). Dimensions X, Y, and Z are 19.2037, 16.0972, and 19.7357, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
Full article ">Figure 10 Cont.
<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) dopamine; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligand “NAG” in the binding site of the human acetylcholinesterase receptor (PDB ID: 4EY5). Dimensions X, Y, and Z are 19.2037, 16.0972, and 19.7357, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
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<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) glutamic acid; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligands “NAG, MES, and GZ5” at the docking site of the human butyrylcholinesterase receptor (PDB ID: 6I0C)/The dimensions X, Y, and Z are 18.0237, 19.1638, and 17.9918, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
Full article ">Figure 11 Cont.
<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) glutamic acid; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligands “NAG, MES, and GZ5” at the docking site of the human butyrylcholinesterase receptor (PDB ID: 6I0C)/The dimensions X, Y, and Z are 18.0237, 19.1638, and 17.9918, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
Full article ">Figure 11 Cont.
<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) glutamic acid; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligands “NAG, MES, and GZ5” at the docking site of the human butyrylcholinesterase receptor (PDB ID: 6I0C)/The dimensions X, Y, and Z are 18.0237, 19.1638, and 17.9918, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
Full article ">Figure 11 Cont.
<p>Two-dimensional and three-dimensional molecular interactions of the most prevalent essential compounds: (<b>A</b>) eugenol acetate; (<b>B</b>) eugenol identified in <span class="html-italic">Syzygium aromaticum</span> with the neurotransmitter “controls” (<b>C</b>) serotonin, (<b>D</b>) norepinephrine, and (<b>E</b>) glutamic acid; and the neurotoxic compound (<b>F</b>) ochratoxin A with the co-crystallized ligands “NAG, MES, and GZ5” at the docking site of the human butyrylcholinesterase receptor (PDB ID: 6I0C)/The dimensions X, Y, and Z are 18.0237, 19.1638, and 17.9918, respectively, the root mean square deviation (RMSD) &lt; 2, and components in 3D are highlighted in yellow.</p>
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Article
Mycotoxicological Assessment of Broiler Compound Feed: A Multi-Year Analysis of Five Mycotoxins in a Romanian Feed Mill
by Dragoș Mihai Lăpușneanu, Silvia-Ioana Petrescu, Cristina-Gabriela Radu-Rusu, Mădălina Matei and Ioan Mircea Pop
Agriculture 2025, 15(1), 84; https://doi.org/10.3390/agriculture15010084 - 2 Jan 2025
Viewed by 619
Abstract
Mycotoxins are secondary metabolites of filamentous fungi that cause massive agricultural losses worldwide and constitute a significant health problem for humans and animals. The aim of this five-year study was to investigate the contamination of compound feed for broiler chickens at all stages [...] Read more.
Mycotoxins are secondary metabolites of filamentous fungi that cause massive agricultural losses worldwide and constitute a significant health problem for humans and animals. The aim of this five-year study was to investigate the contamination of compound feed for broiler chickens at all stages (starter, grower and finisher) from a feed mill in Romania with mycotoxins such as total aflatoxins (AFT), deoxynivalenol (DON), fumonisins (FUMs), ochratoxin A (OTA) and zearalenone (ZEN). AFT was detected in 49.3–72.2% of the samples with concentrations ranging from 0.01 to 5.2 µg/kg. DON was detected in 77.6–98.9% of the samples, with maximum concentrations ranging from 330 to 1740 µg/kg. FUM contamination ranged from 42.7% to 87.2%, with maximum levels between 460 and 1400 µg/kg. OTA was present in 44.2–87.9% of the samples, with maximum concentrations reaching 21.4 µg/kg. ZEN was consistently elevated at all feeding stages, being detected in 86.5–97.4% of the samples, with maximum levels of 89.4 µg/kg. Mycotoxin co-occurrence was common in the samples, with the most common combination of four mycotoxins occurring in 38.51% of the samples. Samples were collected from storage bunkers, homogenised and analysed in certified laboratories, with sampling procedures varying according to batch size to ensure representativeness. Investigation of the transfer of mycotoxins into animal products and the combined effects of mycotoxins on animal health, including potential synergistic or antagonistic interactions, is particularly relevant. This study emphasises the essential role of comprehensive and continuous monitoring of mycotoxins in protecting animal health and food safety. Full article
(This article belongs to the Special Issue Farming Factors’ Influence on Animal Productions)
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<p>Co-occurrence of mycotoxins in broiler feed over the five-year study period.</p>
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