Mycotoxin Contamination Levels Research Articles

Enhancing bread quality and extending shelf life using dried sourdough

This study investigated the physicochemical properties, microbiological analysis, antifungal activity, and mycotoxin analysis of bread. Eight breads were prepared, including control Bread (CB), control Bread with calcium propionate (CPB), commercial sourdough breads (CCB), control bread made with oven-dried sourdough (SB), and bread baked with oven-dried sourdough fermented by Pediococcus pentosaceus TI6 (SB-TI6). These breads were contaminated with Aspergillus flavus and Penicillium verrucosum, and their shelf life was monitored over 7 days. The results indicated that incorporating sourdough powder into bread significantly extends the shelf life and reduces fungal contamination. This reduction resulted in lower levels of mycotoxins, with the use of dry sourdough fermented by P. pentosaceus TI6 leading to the lowest levels of aflatoxins. In the case of SB-TI6, minor levels of aflatoxin B1 (AFB1), B2 (AFB2), and G1(AFG1) were detected, with only 0.003 mg of AFG2/Kg of the product. Conversely, CB loaves exhibited the highest mycotoxin contamination levels, reaching values of 330 mg of AFB1/Kg. The findings suggested that incorporating dry sourdough in breadmaking served as a viable alternative to synthetic preservatives, effectively reducing mycotoxin levels and enhancing bread safety and shelf life.

PSV-20 Mycotoxin contamination trends in United States corn grain: 2019-2023

Abstract Mycotoxins are secondary fungal metabolites which may affect animal health, reproduction, and performance. These naturally occurring toxic metabolites are found in a variety of feedstuffs worldwide. Classic signs of mycotoxicosis in livestock can include reduced feed intake, diarrhea, and vomiting. Consideration of clinical signs alone will underestimate the true impact of mycotoxins on animals as inflammation, decreased nutrient digestibility, impaired gut integrity, and altered immune responses have also been associated with mycotoxin exposure. The objective of this study was to compare mycotoxin contamination levels of the 2023 corn crop with previous year trends. Samples were analyzed utilizing the liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for six major mycotoxin groups: aflatoxins (Afla), type A trichothecenes (A-Trich), type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN), and ochratoxin A (OTA). Statistical analyses were performed with JMP Pro 16 software using one-way ANOVA and means were separated using Tukey’s HSD with significance reported at P ≤ 0.05. A limited number of 2023 samples were screened by time of abstract submission (n = 37); therefore, the contamination profile of this crop year will likely change as the sample pool expands and includes more samples originating from the Midwest. Contamination levels in corn for the tested toxin groups have remained consistent over the survey period. To date, 100% of the samples evaluated contained at least one mycotoxin, compared with 91% that was observed in 2022. Aflatoxin occurrence was numerically greater in 2023 corn compared with previous harvest years (27 vs. 11, 7, 5, and 3%, 2022 through 2019, respectively) while Afla mean contamination levels were similar across the study period (P > 0.05). The occurrence of FUM in 2023 corn was also numerically the greatest (97 vs. 79, 64, 64, and 78%, 2022 through 2019, respectively) and mean FUM contamination levels did not differ across the study period (P > 0.05). Occurrence of A-Trich was numerically increased in 2023 vs. 2022 (22 vs. 3%) but mean contamination level of positive samples for A-Trich was significantly less (P < 0.05) in 2023 vs. 2019 (21 vs. 330 parts per billion) and did not differ from other years in the study. The occurrence of B-Trich was numerically less in 2023 vs. 2022 (35 vs. 68%) while B-Trich mean contamination levels were similar across the study period (P > 0.05). Preliminary results of the 2023 US corn grain survey indicate mycotoxin occurrence is numerically increased while mean contamination levels are similar for most mycotoxin groups other than A-Trich. Continued surveillance is advised to assess the risk to livestock as new crop corn is fed out over the coming months.

Occurrence of mycotoxins in swine feed from South Korea.

To update recent information on contamination levels of mycotoxins in South Korea. A total of 208 samples sourced from the feeds of swine farms were collected. The contamination levels of mycotoxins, which are aflatoxin (Afla), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN), fumonisin (FUM), and T-2 toxin, were investigated by enzyme-linked immunosorbent assays (ELISAs). The detection levels of the total samples were 78.91% for DON, 75.24% for Afla, 47.02% for ZEN, 68.31% for FUM, and 5.94% for OTA and T-2, which were not detected at all. Most of the analyzed mycotoxins showed significant high occurrences in 47.02%-78.91% of the swine feed samples. 11 of the 152 alfa-positive samples exceeded the maximum residue limit (MRL) of Afla proposed by the Korean regulation. In the analysis of mycotoxin detection levels by growth stage, ZEN was found in the nursery stage at a remarkably high concentration level (126.46 ± 63.76 ppb), exceeding the MRL of ZEN for piglets proposed by the European Commission. This mycotoxin was also found in the samples from the gestation barn (89.04 ± 46.05 ppb) and the farrowing house (105.58 ± 94.12) at a high concentration level. Afla was found in the nursery stage at a high concentration (8.00 ± 2.22 ppb), approaching the MRL (10 ppb) of Afla proposed by the Korean regulation. These results indicate that many swine farms in South Korea are still exposed to mycotoxin risk, and special attention and surveillance are necessary for these mycotoxin risks in swine farms.

Mycotoxin detection in selected medicinal plants using chromatographic techniques.

Mycotoxins are toxic mycological products that when consumed, absorbed or inhaled cause sickness or even the death of humans. Therefore, the present study aimed to evaluate the contamination levels of mycotoxins (aflatoxins, AFB1 , AFB2 , AFG1 , AFG2 , and ochratoxin A, OTA) in selected medicinal herbs and shrubs using thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). A total of 15 samples of medicinal herbs and shrubs were selected. Among them, four samples were aflatoxin contaminated while two samples were ochratoxin A contaminated. The highest level of aflatoxin was detected in Justicia adhathoda (4,704.94 ppb) through HPLC (153.4ppb) and through TLC, while the lowest level of aflatoxin was detected in Pegnum harmala (205.1ppb) through HPLC. Similarly, the highest level of OTA was detected in Dodonia viscosa (0.53 ppb) through HPLC (0.5ppb) and through TLC, while the lowest level was detected in J. adhathoda (O.11 ppb) through HPLC (0.4ppb) and through TLC. The OTA concentration was very low, being negligible and below permissible limits. The present study concludes that there is a potential risk for the consumption of herbal decoctions. Therefore, regular monitoring and proper management of mycotoxins, including aflatoxins and OTA, in herbal medicines are needed to ensure the safety of herbal drugs to protect consumers.

An Occurrence Study of Mycotoxins in Plant-Based Beverages Using Liquid Chromatography-Mass Spectrometry.

Mycotoxins are toxic mold metabolites that can adversely affect human and animal health. More than 400 mycotoxins have been identified so far. Cereals and nuts are the predominant mycotoxin-contaminated foodstuffs. Plant-based drinks produced from cereals, nuts, and legumes have grown in popularity. The mycotoxins accumulated in these crops may transfer to these beverages. A liquid chromatography-tandem mass spectrometry method was developed and optimized for the assessment of 22 mycotoxins in commercially available plant-based drinks in Latvia and Lithuania. A total of 64% of the seventy-two analyzed beverages were positive for one to sixteen mycotoxins, with deoxynivalenol, beauvericin, and enniatins A, B, B1, T-2, and HT-2 toxins detected most frequently. The European Commission has not yet set guidelines for the maximum mycotoxin concentrations in plant-based beverages, nor has the European Food Safety Authority conducted a risk assessment. Therefore, acute exposure studies were provided for the Latvian population based on the assumed replacement of dairy milk with plant-based beverages to ascertain the safety of plant-based milk substitutes. Based on the observed levels of mycotoxin prevalence and contamination levels and assumed exposure, it can be concluded that tested plant-based beverages may be relatively safe. However, exposure to emerging mycotoxins should be considered.

Conversion of mycotoxin-contaminated maize by black soldier fly larvae into feed and fertilizer

Abstract Globally, large amounts of various crops such as cereals, oilseeds, nuts and spices are contaminated with mycotoxins during pre-harvest, postharvest handling, processing and/or storage. Mycotoxin contamination results into economic and health issues, and valorisation options of contaminated crops are urgently needed. The aim of this research was to evaluate whether quality feed and fertilizer can be safely produced from naturally mycotoxin contaminated crops using black soldier fly larvae (BSFL, Hermetia illucens L.) under realistic field conditions in East Africa. Naturally mycotoxin contaminated maize (corn; Zea mays L.) was used as a model due its prevalence as food and feed and utilized by BSFL together with local agri-food by-products at a research facility in Rwanda. To assess the influence of the initial maize mycotoxin contamination and maize inclusion, larval diets with three mycotoxin contamination levels and two maize inclusion levels were tested. BSFL were tolerant against the high mycotoxin concentrations (e.g. 99.4 μg aflatoxin B1 kg dry mass-1) as the presence of mycotoxins in the substrate did not affect BSFL mass at harvest. Product safety was assessed by quantifying the presence of 38 common and emerging mycotoxins and metabolites in the maize, substrates and BSFL products (e.g. larvae and frass). The results show that it is possible to produce feed and fertilizer with BSFL considered safe within the European Union and East African legal limits with maize contaminated with mycotoxins typical for East Africa. Thereby, this research works towards the safe recycling of nutrients from mycotoxin contaminated maize within the food system in East Africa and beyond.

Plasma treatment: An alternative and sustainable green approach for decontamination of mycotoxin in dried food products

In the realm of food safety, dried foods have traditionally been perceived as having a lower risk profile. However, recent reports have highlighted concerning levels of mycotoxin contamination in dried food products, such as dried fruits, vegetables, and spices, exceeding permissible limits. This contamination not only compromises shelf life but also poses a significant threat to consumers, while depleting the bioactive components of these products. The food industry grapples with considerable challenges arising from the substantial risk of mycotoxin contamination in dried foods. To meet the demands of a disc; erning food market, there is an imperative need for sustainable mycotoxin-degrading technologies. These technologies must be non-thermal and chemical-free, minimizing thermal damage to treated food to preserve essential nutrients, ultimately enhancing food quality and safety. Developing non-thermal decontamination technologies for dried foods holds enormous economic and public health significance.This review centers its focus on the decontamination of dried foods, proposing a green approach through the utilization of Cold Plasma Technology (CPT). We delve into the working principles and applications of CPT in dried foods, highlight the influential factors affecting its efficacy, and acknowledge the limitations of this innovative technique. Furthermore, we pinpoint and explore emerging research opportunities in the decontamination of dried foods, particularly in conjunction with other complementary methods. CPT, an evolving and environmentally friendly technology, exhibits promising capabilities in controlling pathogens in dried food products and extending their shelf life. Critical determinants encompass the parameters of cold plasma, operational conditions, food-related variables, and microbiological factors, which collectively influence the efficacy of CPT. We advocate for future research dedicated to assessing the impact of CPT on various mycotoxins, food composition, and its interaction with food-related factors. In this vein, it is imperative to emphasize the scaling up of this technology for practical commercial applications, addressing the growing need for enhanced food safety in dried products.

PSVI-20 Trends in Mycotoxin Contamination of the 2022 United States Corn Crop

Abstract Mycotoxins are harmful secondary fungal metabolites detrimental to animal health, even at reduced quantities. Focus on clinical signs such as decreased feed intake and vomiting overlook impacts of mycotoxicosis, including increased disease incidence and severity, immune modulation, inflammation, and gastrointestinal disorders. This study was conducted to compare mycotoxin contamination of initial samples of the 2022 corn crop with previous year trends. Samples were analyzed utilizing the liquid chromatography with tandem mass spectrometry (LC-MS/MS) method for six major mycotoxin groups: aflatoxins (Afla), type A trichothecenes (A-Trich), type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN), and ochratoxin A (OTA). The 2022 crop year data were compared with the prior four harvest years using GLIMMIX procedure of SAS with harvest year as fixed effect and sample as the experimental unit. A limited number of samples is available thus far (n = 44) as harvest is still progressing, therefore the risk profile of this crop year is likely to change as the sample pool expands. Contamination levels in corn for the tested toxin groups have remained consistent over the survey period. To date, 97% of the samples evaluated contained at least one mycotoxin, compared with 90% that was observed in 2021. The percentage of Afla contaminated samples increased (P < 0.05) in 2022 corn compared with the 2020 and 2021 harvest years (42 vs. 5 and 7%). The percentage of FUM contaminated samples in 2022 corn is greater (P < 0.05) compared with 2020 and 2021 (88 vs. 64 and 64%). The preliminary results of the 2022 US corn crop survey indicate mycotoxin occurrence and contamination levels are similar or greater (Afla and FUM) to the prior crop year and will continue to challenge animal health and performance as these ingredients are fed out.

Ecological characterisation of the mycotoxin Alternariol in the green mass of spring wheat and white mustard under the conditions of the Moscow region

The article presents the results of the study of the influence of agro-ecological factors on the content of mycotoxins in the green mass of white mustard (Sinapis alba) and the spring wheat variety Lyubava (Triticum aestivum L.). The study was conducted at the experimental field «Yuzhnoe» of the Ecological Station of the Russian State Agrarian University – Moscow Timiryazev Agricultural Academy using chemical and microbiological methods. Agro-ecological factors were evaluated by the following parameters: humus content, soil acidity, phosphorus, potassium and lead content. The immunoenzyme method was used to determine the content of mycotoxins in the vegetative mass of white mustard and spring wheat (according to development stages). As a result, the highest levels of mycotoxin contamination were found in the mid-ripe stage of the crops, exceeding the average level of contamination by 8‑11 times in wheat and 4‑5 times in white mustard. The highest inverse correlation was found between the Alternariol content in the studied crops and the soil temperature of the upper layer of the humus horizon of 0‑5 cm. This fact justifies a more detailed study of the effect of this environmental factor.

Mycotoxin reduction and metabolite profiles of ogi produced using traditional fermentation methods

Mycotoxins are widely present in maize, a favourite staple food in sub-Saharan Africa. Food processing methods, like fermentation, have been suggested as potential ways to reduce mycotoxin contamination levels in the grain and, as a result, limit the exposure of crop consumers to the harmful effects of the toxins. The influence of four traditional fermentation processes [cold (with changed steeping liquor (CSL) and unchanged steeping liquor (USL), Fon and Goun procedures] on the mycotoxin reduction and metabolites profile of ogi, a fermented maize product, was studied. Ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) and gas chromatography linked to high resolution time-of-flight mass spectrometry (GC-HR-TOF-MS) were respectively employed for the mycotoxin and metabolite profiles analyses of the samples. Among the nine mycotoxins detected in the raw maize samples, aflatoxin B1 (AFB1) and fumonisin B1 (FB1) + fumonisin B2 (FB2) concentrations were found to exceed the European Union (EU) maximum limit. Both cold (containing USL and CSL) and Goun fermentation techniques were able to lower the AFB1 concentration below this threshold. The metabolomics result revealed that ogi produced using the cold (USL) and Fon fermentation processes had the highest number of most of the detected important compounds, whereas the Goun fermentation process produced the fewest compounds in total. There was no statistically significant difference in the ability of the specified natural fermentation processes to lower FB1, FB2, FB3, deoxynivalenol (DON), sterigmatocystin (STERIG), and zearalenone concentrations in maize (ZEN). In addition, the results demonstrated that the four natural fermentation processes evaluated had varying effects.

Epiphytic microflora and mycotoxin content in meadows-Is plant biodiversity affecting fungal contamination?

Ecosystem services are an important aspect of grasslands utilization; however, they are often contradictory to their main purpose, which is a production of good quality and safe feed. In this study, we evaluated the difference between grass monocultures and species-rich mixtures in terms of epiphytic microflora and mycotoxin contamination levels. We hypothesized that higher species diversity would lead to higher microbial counts, which could lead to higher mycotoxin contamination risk. Differences in epiphytic fungal, yeast and total amount of microorganisms (CFU g -1) depending on the species diversity in the field has been evaluated by cultivation method. Concentration of deoxynivalenol (DON), zearalenone (ZEN) and aflatoxin B1 (AFB1) was measured by ELISA. Results are suggesting that higher total amount of microorganisms were found in monocultures, however, fungal and yeast counts were higher in species-rich mixtures. Higher species diversity of grasses was related to higher total microbial count (TMC) and yeast colonization of phyllosphere. Our results suggest higher risk of fungal phyllosphere colonization of species-rich mixtures with higher biodiversity and therefore higher risk of mycotoxin contamination of such feed.

Comparative utility of hermetic and conventional grain storage bags for smallholder farmers: a meta-analysis.

Postharvest management is critical to attaining household food, nutrition, and income security. Hermetic grain storage bags offer an effective pesticide-free way to protect stored grain against fungal and insect infestation. We evaluated articles indexed in the Web of Science that included experiments comparing the storage efficacy of conventional and hermetic storage bags based on grain germination rate, insect infestation, physical damage, mycotoxin contamination, and changes in weight and moisture content. Compared with grain stored in hermetic bags, grain stored in conventional bags lost 3.6-fold more seed viability, contained 42-fold more insects, had 11-fold more physical damage, and lost 23-fold more grain weight, while grain moisture levels were similar for both hermetic and conventional storage bags. Mycotoxin contamination levels were not as frequently assessed. Levels could be low in grain stored in both types of bags, or levels could be low in hermetic bags and significantly higher in conventional bags. The improved properties of grain stored in hermetic bags can increase food security and household income by providing safe storage options for maintaining seed germinability, and for consumption and/or sale when food supplies are high, or when prices are low. Hermetic bags are economically feasible for use by subsistence farmers in Sub-Saharan Africa for grain for household consumption and for carrying-over seed for planting in the next season. Additional studies are needed to verify the mycotoxin contamination results and to determine if there are differences in functional food characteristics, e.g. flavor and cooking properties, that have not been as comprehensively studied. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Major Fusarium species and mycotoxins associated with freshly harvested maize grain in Uruguay.

Fusarium species are common fungal pathogens of maize. Fusarium graminearum and Fusarium verticillioides, among others, can cause maize ear rot, and they are also mycotoxin producers. The aims of this work were to determine the frequency and diversity of Fusarium species in Uruguayan maize kernels, evaluate the toxigenic potential of the isolates, determine toxin contamination levels on freshly harvested grain, and assess the sensitivity of main Fusarium species against fungicides. Fusarium verticillioides was the most frequent species isolated, followed by Fusarium graminearum sensu stricto. Of F. verticillioides isolates studied for fumonisin production, 72% produced fumonisin B1 and 32% fumonisin B2. Considering in vitro toxin production by F. graminearum sensu stricto isolates, deoxynivalenol was the main toxin produced, followed by zearalenone and nivalenol. Fumonisins were the most frequently found toxins on freshly harvested maize samples (98% in 2018 and 86% in 2019), and also, fumonisin B1 was the toxin with highest concentration in both years studied (4860µg/kg in 2018 and 1453µg/kg in 2019). Deoxynivalenol and zearalenone were also found as contaminants. Metconazole and epoxiconazole were the most effective fungicides tested on F. verticillioides isolates. Fusarium graminearum sensu stricto isolates also were more sensitive to metconazole compared to other fungicides; nevertheless, epoxiconazole was less efficient in controlling this species. This is the first study that reports Fusarium species and mycotoxin contamination levels associated with maize grain in Uruguay. Its detection is the main step to develop management strategies in order to minimize fungal infection in maize crops.

Prevalence and stakeholders’ perception of mycotoxins in the Ghanaian poultry feed value chain

The poultry sector in Ghana has the potential to contribute significantly to domestic food and nutrition security. There is limited information regarding the knowledge of feed industry stakeholders on the presence of mycotoxins in poultry feed. This work was aimed at assessing the prevalence of aflatoxin and fumonisin in poultry feed, and mycotoxins awareness among stakeholders of the poultry feed value chain in Ghana. Three hundred and fifty (350) respondents consisting of 255 commercial poultry farmers, 85 feed processors and 16 feed vendors were involved in the study. AgraStrip® Quantitative test from RomerLab, USA was used to quantify total fumonisin and aflatoxin levels in the prepared feed samples. The study revealed that commercial feed producers, feed mills operators and on-farm feed processors are the three main sources of feed and ingredients for commercial poultry farmers in Ghana. The study identified a high level of mycotoxin contamination of feed and ingredients. Feed samples from the commercial feed processors recorded significantly lower aflatoxin levels (21.63+7.05 ppb, p<0.008). However, the threshold of 15 ppb for the Ghana Standards Authority, FAO/WHO maximum permissible limit of 30 ppb and EU regulatory limit of 20 ppb for poultry feeds were exceeded by 70% of all the samples. Fumonisin levels were less than the 30mg/kg FDA guidance levels for maize and maize by-products intended for consumption by breeding poultry. From the study, a majority of the farmers (85%), vendors (87.5%) and all of the feed processors have their own feed storage facilities or warehouses. Storage problems identified by these stakeholders included pest infestation and mold development, which usually occur after a few weeks of storage to the end of the storage period. Sanitation was poor along the feed chain, particularly during feed formulation and storage. Awareness of mycotoxins and their effect on animal and human health was low among the respondents. Some of those who have heard about aflatoxins were not aware of what it is and its effect on the health of humans and animals. This threatens the management of mycotoxins in the poultry feed chain. Key words: aflatoxin, fumonisin, mycotoxin contamination, mycotoxin awareness, sanitation, poultry feed, feed processors

Centrifugation-Assisted Solid-Phase Extraction Coupled with UPLC-MS/MS for the Determination of Mycotoxins in ARECAE Semen and Its Processed Products.

Mycotoxins can occur naturally in a variety of agriculture products, including cereals, feeds, and Chinese herbal medicines (TCMs), via pre- and post-harvest contamination and are regulated worldwide. However, risk mitigation by monitoring for multiple mycotoxins remains a challenge using existing methods due to their complex matrices. A multi-toxin method for 22 mycotoxins (aflatoxin B1, B2, G1, G2, M1, M2; ochratoxin A, B, C; Fumonisin B1, B2, B3; 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol, diace-toxyscirpenol, HT-2, T-2, deepoxy-deoxynivalenol, deoxynivalenol, neosolaniol, zearalenone, and sterigmatocystin) using centrifugation-assisted solid-phase extraction (SPE) clean-up prior to ultra-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis for Arecae Semen and its processed products was developed and validated. Several experimental parameters affecting the extraction and clean-up efficiency were systematically optimized. The results indicated good linearity in the range of 0.1-1000 μg/kg (r2 > 0.99), low limits of detection (ranging from 0.04 μg/kg to 1.5 μg/kg), acceptable precisions, and satisfactory recoveries for the selected mycotoxins. The validated method was then applied to investigate mycotoxin contamination levels in Areca catechu and its processed products. The mycotoxins frequently contaminating Areca catechu were aflatoxins (AFs), and the average contamination level and number of co-occurring mycotoxins in the Arecae Semen slices (Binlangpian) were higher than those in commercially whole Arecae Semen and Arecae Semen Tostum (Jiaobinlang). Sterigmatocystin was detected in 5 out of 30 Arecae Semen slices. None of the investigated mycotoxins were detected in Arecae pericarpium (Dafupi). The results demonstrated that centrifugation-assisted SPE coupled with UHPLC-MS/MS can be a useful tool for the analysis of multiple mycotoxins in Areca catechu and its processed products.

The combined effect of deoxynivalenol and Fumonisin B1 on small intestinal inflammation mediated by pyroptosis in vivo and in vitro

Currently, due to the actual contamination levels of multiple mycotoxins, the limits for a single mycotoxin may be no longer applicable. Deoxynivalenol (DON) and Fumonisin B1 (FB1) had high positive rate in grain and feed worldwide. The intestine is the first target of mycotoxins. NLRP3 plays a crucial role in the gut’s defense against external stimuli, which contributes vitally to pyroptosis activation. However, whether pyroptosis is engaged in the regulation of intestinal toxicity induced by DON and FB1 remains unclear. In this study, we explored the combined toxicity of DON and FB1 on the intestine and its underlying mechanisms in vivo and in vitro. Our data demonstrated gavage with DON and FB1 led to intestinal damage and promoted the secretion of pro-inflammatory cytokines (IL-1β, IL-18, IL-6) in mice, especially in the group exposed to both mycotoxins. Meanwhile, the expressions of pyroptosis related genes (NLRP3, ASC, caspase-1, GSDMD) were significantly increased after mycotoxins exposure. Same as in vivo, DON and FB1 promoted pyroptosis and cellular inflammatory response in IPEC-J2 cells, especially in the group exposed to both mycotoxins. In addition, the pretreatment with MCC950 and VX765, inhibitors for NLRP3 and caspase-1, abolished the expression of GSDMD and the release of pro-inflammatory factors (IL-1β, IL-18) induced by DON and FB1 exposure in IPEC-J2 cells. Our data demonstrated that the combination of DON and FB1 exhibited a synergistic or additive effect in facilitating intestinal inflammation via pyroptosis. Our finding may contribute to improve mycotoxin limit standards in feed.

351 Mycotoxin Trends in US Total Mixed Rations and Corn Silage

Abstract Mycotoxins are secondary fungal metabolites identified in a variety of feed ingredients and may affect animal health and productivity. This study investigated occurrence and contamination levels of mycotoxins in 2022 US total mixed ration (TMR) and 2021 corn silage samples. Corn silage samples harvested in 2021 and submitted through March 2022 and TMR samples submitted between January through March 2022 were screened via the LC-MS/MS technique for the presence of six major mycotoxin groups: aflatoxins, type A trichothecenes, type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN) and ochratoxin A. Detailed information for each sample was provided including, state of origin and whether or not clinical signs of mycotoxicosis was observed on-farm. Parameters of the main toxins found for corn silage and TMR samples were analyzed for effect of clinical concern (yes/no) and harvest year on contamination levels using LSMeans statement with Tukey's adjustment in the GLIMMIX procedure (SAS 9.4, Cary, NC). Main effects of mean contamination by harvest year are presented in Table 1, since no interactions were significant (P > 0.05). To date, 20 TMR samples from seven states have been submitted for analysis in 2022. Preliminary results show detectable amounts of B-Trich in 95% of TMR samples, with ZEN and FUM detected less frequently in 55 and 10%, respectively. Mean contamination concentrations (parts per billion, ppb) are presented on dry matter basis and were greater in 2022 compared with 2021 for B-Trich and ZEN (P < 0.05) and similar for FUM (P = 0.022). Preliminary results for 72 corn silage samples show detectable quantities of B-Trich in 90% of samples with ZEN and FUM detected less frequently in 67 and 24%, respectively. Mean contamination levels are greater than the last two years and approaching those observed in the challenging 2018 crop for B-Trich, FUM, and ZEN.

60 Occurrence of Mycotoxins in US Corn Gluten Products and Wheat Middlings

Abstract Secondary fungal metabolites which may affect animal health and productivity are known as mycotoxins. These naturally occurring metabolites are often detected in a variety of feed ingredients. Mycotoxin occurrence and contamination levels in corn gluten products and wheat middlings or “midds” were monitored in this study. Samples submitted from the US were screened via LC-MS/MS for six major mycotoxin groups: aflatoxins, type A trichothecenes, type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN), and ochratoxin A (OTA). The presence of clinical health or performance concerns on-farm was reported for each sample. Results from samples without clinical concerns that were above the limit of detection for each respective mycotoxin group were included in the study. Out of 14 corn gluten product samples reported as 2021 crop year, 100% were positive for both B-Trich and FUM, while 71% were positive for some amount of ZEN. These mycotoxin groups were also the most frequently detected in 2020. Mean contamination levels (parts per billion, ppb) for 2021 were 1,314 ppb, 443 ppb, and 132 ppb, respectively. Of the 24 samples of 2021 wheat midds, B-Trich were detected in 92% while 54% were positive for OTA with mean contamination levels of 665 ppb and 3 ppb, respectively. Ergot alkaloids were not screened for in the current year, however, previous analyses have identified these metabolites frequently occur in wheat midds samples. Although represented by limited data sets to date, analysis of 2021 corn gluten products and wheat midds indicate mycotoxins frequently occur in these by-product feeds. High feed costs may drive increased feeding of alternative feeds, including these and other by-products, in addition to established routine usage in pelleted feeds and other diets. Therefore, continued screening is suggested to assess mycotoxin exposure in livestock diets.

61 Mycotoxin Trends in US Corn Grain and Corn Distiller’s Dried Grains with Solubles

Abstract Mycotoxins are secondary fungal metabolites which may affect animal health and performance and are frequently detected in various feedstuffs. Mycotoxin occurrence and contamination levels in US corn grain and corn distiller’s dried grains with solubles (DDGS) samples were monitored in this study. Samples were analyzed via LC-MS/MS for six major mycotoxin groups: aflatoxins, type A trichothecenes, type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN), and ochratoxin A. The presence of clinical health or performance concerns on-farm was reported for each sample. Results from samples without clinical concerns that were above the limit of detection for each respective mycotoxin group were analyzed using the GLIMMIX procedure (SAS 9.4, Cary, NC) to test the effect of harvest year on mean contamination level. Results are presented in Table 1. Of the 219 samples of 2021 corn grain screened in the current data set, 69% were positive for some level of B-Trich, an increase from 58% positive in 2020. Fumonisins (61%) and ZEN (27%) were the next most frequently detected mycotoxins in 2021 samples. Although numerically lower, mean B-Trich level (parts per billion, ppb) was similar to 2020 crop while contamination with FUM was elevated (P = 0.0001) compared to the prior year. Mean ZEN levels are numerically greater, but similar to 2020 samples. Out of 35 corn DDGS samples, occurrence of B-Trich, FUM, and ZEN remained high at 100, 100, and 94%, respectively. Mean B-Trich and FUM concentrations were similar year-to-year across the study period for DDGS while current mean ZEN level was similar to 2020, but lower than 2019 (P < 0.001). Preliminary results of the 2021 US corn grain and DDGS survey suggest increased mycotoxin occurrence with comparable mean contamination levels to those observed in 2020. Continued surveillance is suggested to assess contamination in common livestock feed ingredients.

Exposure assessment of aflatoxins and zearalenone in edible vegetable oils in Shandong, China: health risks posed by mycotoxin immunotoxicity and reproductive toxicity in children.

Human exposure to aflatoxins (AFs) and zearalenone (ZEA) has not been sufficiently investigated. Here, we analyzed the exposure level and health risks posed by AFs (B1, B2, G1, G2) and ZEA through cooking oil consumption in Shandong, China. The individual daily consumption of cooking oil was calculated through 2745 questionnaires during 2017-2019. The average contamination levels of mycotoxins were estimated by examining 60 cooking oil samples. For the peanut oil, AFs ranged from <0.2 to 274 μg/kg, with a positive rate of 66.6% (20/30). Average levels of 36.62 μg/kg AFB1 and 44.43 μg/kg total AFs were found. Over-the-limit level (20 μg/kg) of AFB1 was detected in 8/30 samples. Estimated daily intake (EDI) and margin of exposure (MOE) for age-stratified population groups showed that children are facing highest adverse health risk with AFB1 (MOE 5.88-6.39). The liver cancer incidences attributable to AFB1 exposure are non-negligible as 0.896, 0.825, and 0.767 cases per 100,000 for 6-14 age group, 15-17 age group, and adult labor-intensive workers. Over-the-limit level (60 μg/kg) ZEA contamination was detected in 25/30 corn oil samples with a 50th percentile value of 97.95 μg/kg. Our health risk assessment suggested significant health risks of enterohepatic (inflammation and cancer), reproductive, and endocrine systems posed by AFs and ZEA. However, the health risk of immunotoxicity is unclear because currently animal study data are not available for the immunotoxicity induced after long-term exposure. In general, the health risks posed by mycotoxins are non-negligible and long-term mycotoxin surveillance is necessary.