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[Objectives] Heterogeneous nuclear ribonucleoproteins (hnRNP) represent a large family of RNA-binding proteins that contribute to multiple aspects of RNA metabolism. In Fusarium graminearum, FGSG_07478 was identified as the homologous protein of annotated Fusarium hnRNP G. The objective of this study was to analyze the function of FGSG_07478 in growth and development, stress, mycotoxins production and pathogenicity of F.graminearum. [Methods] Based on homologous recombination and polyethylene glycol (PEG) mediated protoplast transformation, the FGSG_07478 gene knockout mutant strain ΔFGSG_07478 was obtained to investigate its function in mycelial growth, asexual and sexual reproduction, and stress resistance of F.graminearum. Pathogenicity of the mutant ΔFGSG_07478 was assessed by infection assays using wheat coleoptiles. Deoxynivalenol (DON) produced by ΔFGSG_07478 was quantified using the ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method, and the relative expression levels of seven TRI involved in DON biosynthesis were determined by RT-qPCR. [Results] Compared with the wild type strain PH-1, the growth rate of ΔFGSG_07478 was reduced by 16.2%, and the spore production decreased by 43.4%. In mating cultures, the ΔFGSG_07478 mutant produced less perithecia, verifying the weakened sexual reproduction. External environmental stress assays showed that ΔFGSG_07478 mutant became sensitive to H2O2 and SDS, but more resistant to Congo red. Although no significant difference in pathogenicity, DON biosynthesis and expression levels of TRI4, TRI5, TRI6, TRI8, TRI10, TRI12 and TRI101 were significantly reduced in ΔFGSG_07478 mutant compared with the wild type. [Conclusions] FGSG_07478 played an important role in fungal growth and development, sexual reproduction, responses to various environmental stresses and mycotoxins production in F.graminearum. [ABSTRACT FROM AUTHOR]

Fusarium graminearum is the main pathogenic fungus causing wheat head blight. It not only causes severe crop yield reduction, but also produces fungal toxins such as deoxynivalenol (DON) and 15-acetyl deoxynivalenol (15-ADON), which are dangerous food pollutants to people and livestock. For the genetic functional study of Fusarium graminearum, it is often necessary to culture mycelium on LTB medium in the laboratory to investigate its regulatory effect on DON and 15-ADON synthesis. However, the detection of DON and 15-ADON in the mycelium is focused on extracellular, with less focus on the detection of intracellular. Simultaneous detection of intracellular and extracellular DON and 15-ADON can provide more in-depth information of fungal toxin production and secretion functions. In the study, a method based on gas chromatography-mass spectrometry (GC-MS) for simultaneous detection of DON and 15-ADON was developed. The solvents for extracting DON and 15-ADON can be completely separated and the calibration curves of each extract show good linear relationship within a certain range. The mass spectrometric characteristic scan shows high ionization efficiency for both DON and 15-ADON. The retention time of DON and 15-ADON in chromatogram are 6.98 and 7.68 min, respectively. Qualitative analysis of DON was performed using m/z 295, 235, and 193 under single ion monitor (SIM) mode, while m/z 392, 235, and 193 were used for 15-ADON. The highest intensity ion m/z 235 was selected for the quantification of DON, and m/z 193 for the quantification of 15-ADON. This method was used to detect the intracellular and extracellular levels of DON and 15-ADON in wild-type PH-1 and tri5 mutant with DON biosynthesis deficiency. The intracellular DON and 15-ADON levels in PH-1 are determined to be (149.13±9.15) μg/g and (1833.31±185.33) μg/g, extracellular levels are (5910.35±468.23) μg/g and (45222.12±2726.81) μg/g, respectively. The study identified a mutant strain with the deletion of the first enzyme involved in the synthesis pathway of DON and 15-ADON，both DON and 15-ADON are not detected in the extracellular and intracellular of the tri5 mutant. Additionally, this research confirms that after 7 days of cultivation in LTB medium, the wild-type strain exhibits higher levels of 15-ADON in both intracellular and extracellular compared to DON, with extracellular levels of DON and 15-ADON being higher than intracellular levels. This method can provide a methodological reference for the functional study of toxin synthesis in Fusarium graminearum.

Abstract Background Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly found in cereals and grains worldwide. The presence of this fungal secondary-metabolite raises public-health concerns at both the agriculture and food industry level. Recently, we have shown that DON has a negative impact on gut integrity, a feature also noticed for Campylobacter (C.) jejuni. We further demonstrated that DON increased the load of C. jejuni in the gut and inner organs. In contrast, feeding the less toxic DON metabolite deepoxy-deoxynivalenol (DOM-1) to broilers reduced the Campylobacter load in vivo. Consequently, it can be hypothesized that DON and DOM-1 have a direct effect on the growth profile of C. jejuni. The aim of the present study was to further resolve the nature of this interaction in vitro by co-incubation and RNA-sequencing. Results The co-incubation of C. jejuni with DON resulted in significantly higher bacterial growth rates from 30 h of incubation onwards. On the contrary, the co-incubation of C. jejuni with DOM-1 reduced the CFU counts, indicating that this DON metabolite might contribute to reduce the burden of C. jejuni in birds, altogether confirming in vivo data. Furthermore, the transcriptomic profile of C. jejuni following incubation with either DON or DOM-1 differed. Co-incubation of C. jejuni with DON significantly increased the expression of multiple genes which are critical for Campylobacter growth, particularly members of the Flagella gene family, frr (ribosome-recycling factor), PBP2 futA-like (Fe3+ periplasmic binding family) and PotA (ATP-binding subunit). Flagella are responsible for motility, biofilm formation and host colonization, which may explain the high Campylobacter load in the gut of DON-fed broiler chickens. On the contrary, DOM-1 downregulated the Flagella gene family and upregulated ribosomal proteins. Conclusion The results highlight the adaptive mechanisms involved in the transcriptional response of C. jejuni to DON and its metabolite DOM-1, based on the following effects: (a) ribosomal proteins; (b) flagellar proteins; (c) engagement of different metabolic pathways. The results provide insight into the response of an important intestinal microbial pathogen against DON and lead to a better understanding of the luminal or environmental acclimation mechanisms in chickens.

Deoxynivalenol (DON) is a mycotoxin that is produced by various species of Fusarium and is ubiquitous in food and feed. At low concentrations, it can cause metabolic disorders in animals and humans and, at high concentrations, it can lead to pathological changes in the body. The impact of DON on human/animal health and animal productivity has thus attracted a great deal of attention around the world. DON causes severe damage to the intestine, including compromised intestinal barrier, mucosal damage, weakened immune function, and alterations in gut microbiota composition. These effects exacerbate intestinal infections and inflammation in livestock and poultry, posing adverse effects on overall health. Furthermore, research into biological methods for DON detoxification is a crucial avenue for future studies. This includes the utilization of adsorption, enzymatic degradation, and other biological approaches to mitigate DON’s impact, offering new strategies for prevention and treatment of DON-induced diseases. Future research will focus on identifying highly efficient detoxifying microorganisms or enzymes to reduce DON levels in food and feed, thereby mitigating its risks to both animals and human health.

Fusarium graminearum is the main pathogenic fungus causing wheat head blight. It not only causes severe crop yield reduction, but also produces fungal toxins such as deoxynivalenol (DON) and 15-acetyl deoxynivalenol (15-ADON), which are dangerous food pollutants to people and livestock. For the genetic functional study of Fusarium graminearum, it is often necessary to culture mycelium on LTB medium in the laboratory to investigate its regulatory effect on DON and 15-ADON synthesis. However, the detection of DON and 15-ADON in the mycelium is focused on extracellular, with less focus on the detection of intracellular. Simultaneous detection of intracellular and extracellular DON and 15-ADON can provide more in-depth information of fungal toxin production and secretion functions. In the study, a method based on gas chromatography-mass spectrometry (GC-MS) for simultaneous detection of DON and 15-ADON was developed. The solvents for extracting DON and 15-ADON can be completely separated and the calibration curves of each extract show good linear relationship within a certain range. The mass spectrometric characteristic scan shows high ionization efficiency for both DON and 15-ADON. The retention time of DON and 15-ADON in chromatogram are 6.98 and 7.68 min, respectively. Qualitative analysis of DON was performed using m/z 295, 235, and 193 under single ion monitor (SIM) mode, while m/z 392, 235, and 193 were used for 15-ADON. The highest intensity ion m/z 235 was selected for the quantification of DON, and m/z 193 for the quantification of 15-ADON. This method was used to detect the intracellular and extracellular levels of DON and 15-ADON in wild-type PH-1 and tri5 mutant with DON biosynthesis deficiency. The intracellular DON and 15-ADON levels in PH-1 are determined to be (149.13±9.15) μg/g and (1 833.31±185.33) μg/g, extracellular levels are (5 910.35± 468.23) μg/g and (45 222.12±2 726.81) μg/g, respectively. The study identified a mutant strain with the deletion of the first enzyme involved in the synthesis pathway of DON and 15-ADON，both DON and 15-ADON are not detected in the extracellular and intracellular of the tri5 mutant. Additionally, this research confirms that after 7 days of cultivation in LTB medium, the wild-type strain exhibits higher levels of 15-ADON in both intracellular and extracellular compared to DON, with extracellular levels of DON and 15-ADON being higher than intracellular levels. This method can provide a methodological reference for the functional study of toxin synthesis in Fusarium graminearum. [ABSTRACT FROM AUTHOR]

Background: Deoxynivalenol (DON) is a type B trichothecene mycotoxin that is commonly found in cereals and grains worldwide. The presence of this fungal secondary-metabolite raises public-health concerns at both the agriculture and food industry level. Recently, we have shown that DON has a negative impact on gut integrity, a feature also noticed for Campylobacter (C.) jejuni. We further demonstrated that DON increased the load of C. jejuni in the gut and inner organs. In contrast, feeding the less toxic DON metabolite deepoxy-deoxynivalenol (DOM-1) to broilers reduced the Campylobacter load in vivo. Consequently, it can be hypothesized that DON and DOM-1 have a direct effect on the growth profile of C. jejuni. The aim of the present study was to further resolve the nature of this interaction in vitro by co-incubation and RNA-sequencing. Results: The co-incubation of C. jejuni with DON resulted in significantly higher bacterial growth rates from 30 h of incubation onwards. On the contrary, the co-incubation of C. jejuni with DOM-1 reduced the CFU counts, indicating that this DON metabolite might contribute to reduce the burden of C. jejuni in birds, altogether confirming in vivo data. Furthermore, the transcriptomic profile of C. jejuni following incubation with either DON or DOM-1 differed. Co-incubation of C. jejuni with DON significantly increased the expression of multiple genes which are critical for Campylobacter growth, particularly members of the Flagella gene family, frr (ribosome-recycling factor), PBP2 futA-like (Fe3+ periplasmic binding family) and PotA (ATP-binding subunit). Flagella are responsible for motility, biofilm formation and host colonization, which may explain the high Campylobacter load in the gut of DON-fed broiler chickens. On the contrary, DOM-1 downregulated the Flagella gene family and upregulated ribosomal proteins. Conclusion: The results highlight the adaptive mechanisms involved in the transcriptional response of C. jejuni to DON and its metabolite DOM-1, based on the following effects: (a) ribosomal proteins; (b) flagellar proteins; (c) engagement of different metabolic pathways. The results provide insight into the response of an important intestinal microbial pathogen against DON and lead to a better understanding of the luminal or environmental acclimation mechanisms in chickens. [ABSTRACT FROM AUTHOR]

Abstract Polyamines, such as putrescine, spermidine, and spermine, are crucial for plant defense against both abiotic and biotic stresses. Putrescine is also known as a significant inducer of deoxynivalenol (DON) production in Fusarium graminearum, the primary causal agent of Fusarium head blight (FHB). However, the impact of other polyamines on DON production and whether modifying polyamine biosynthesis could improve wheat resistance to FHB are currently unknown. In this study, we demonstrate that key precursor components of putrescine synthesis, including arginine, ornithine, and agmatine, can induce DON production, albeit to a lesser extent than putrescine in trichothecene biosynthesis-inducing (TBI) culture under the same total nitrogen conditions. Intriguingly, spermidine and spermine, downstream products of putrescine in the polyamine biosynthesis pathway, do not induce DON production under the same conditions. Additionally, externally applying either spermidine or spermine to wheat heads significantly reduces the diseased spikelet number caused by F. graminearum. Furthermore, our results show that overexpression of the wheat spermidine synthase (SPDS) gene TaSPDS-7D1 significantly enhances the spermidine content and wheat resistance to FHB. In addition, the TaSPDS-7D1-overexpressing line OE3 exhibited a 1000-grain weight and plant height increase compared to the wild type. Our findings reveal that overexpression of the spermidine synthase gene can enhance wheat resistance to FHB without compromising wheat yield.

Endometrial cancer is one of the most common cancer types among women. Many factors can contribute to the development of this disease, including environmental factors and, thus, eating habits. Our study aims to determine the levels of various mycotoxins and their metabolites in the blood serum and endometrial tissue samples of participants with previously proven endometrial cancer and to find possible contributions to cancer development. In the cohort clinical trial, 52 participants aged between 44 and 86 were studied. The participants were divided into two groups: patients or matched controls. All patients had previously histologically diagnosed endometrial cancer. The cancer patients were divided into low-grade endometrioid and low- plus high-grade endometrioid groups. Controls had no history of endometrial malignancy or premalignancy. Blood serum and endometrial tissue samples were obtained from all study patients. We compared the concentrations of total Aflatoxins (Afs), Deoxynivalenol (DON), Ochratoxin-A (OTA), T2-toxin and HT2 toxin (T2/HT2 toxin), Zearalenone (ZEN), alpha-Zearalenol (α-ZOL), and Fumonisin B1 (FB1) in the serum and endometrium between the different study groups. As a result, we can see a significant correlation between the higher levels of Afs and zearalenone and the presence of endometrial cancer. In the case of Afs, DON, OTA, T2/HT2 toxins, ZEN, and alpha-ZOL, we measured higher endometrial concentrations than in serum. Considering the effect of mycotoxins and eating habits on cancer development, our results might lead to further research exploring the relationship between certain mycotoxins and endometrium cancer. [ABSTRACT FROM AUTHOR]

Fusarium crown rot (FCR) is an important and devastating disease of wheat (Triticum aestivum) caused by the fungus Fusarium pseudograminearum and related pathogens. Using two distinct susceptible cultivars, we investigated the isolation frequencies of F. pseudograminearum and quantified its biomass accumulation and the levels of the associated toxins deoxynivalenol (DON) and DON-3-glucoside (D3G) in inoculated field-grown wheat plants. We detected F. pseudograminearum in stem, peduncle, rachis, and husk tissues, but not in grains, whereas DON and D3G accumulated in stem, rachis, husk, and grain tissues. Disease severity was positively correlated with the frequency of pathogen isolation, F. pseudograminearum biomass, and mycotoxin levels. The amount of F. pseudograminearum biomass and mycotoxin contents in asymptomatic tissue of diseased plants were associated with the distance of the tissue from the diseased internode and the disease severity of the plant. Thus, apparently healthy tissue may harbor F. pseudograminearum and contain associated mycotoxins. This research helps clarify the relationship between F. pseudograminearum occurrence, F. pseudograminearum biomass, and mycotoxin accumulation in tissues of susceptible wheat cultivars with or without disease symptoms, providing information that can lead to more effective control measures. [ABSTRACT FROM AUTHOR]

The control and assessment of the level of mycotoxins in food is a very important desideratum for the food industry and, in particular, for products of plant origin. Control of food products represents a priority to get safe the population. Products intended for consumers must comply with the legislation. Mycotoxins are compounds with high stability, with very few ways and possibilities of elimination from the body. In order to eliminate the risk, a rigorous control of the feed is necessary. The purpose of this study was to evaluate the content of zearalenone (ZEA) and deoxynivalenol (DON) in cereals, bread and bread with potatoes. As a result of the determinations, it was found that the ZEA and the DON are well below the values provided by the legislation, due to the proper application of the food safety systems. In the case of ZEA and DON, the highest amount was observed in the case of whole grains, and the smallest amount was observed in homemade bread with potatoes [ABSTRACT FROM AUTHOR]

Polyamines, such as putrescine, spermidine, and spermine, are crucial for plant defense against both abiotic and biotic stresses. Putrescine is also known as a significant inducer of deoxynivalenol (DON) production in Fusarium graminearum, the primary causal agent of Fusarium head blight (FHB). However, the impact of other polyamines on DON production and whether modifying polyamine biosynthesis could improve wheat resistance to FHB are currently unknown. In this study, we demonstrate that key precursor components of putrescine synthesis, including arginine, ornithine, and agmatine, can induce DON production, albeit to a lesser extent than putrescine in trichothecene biosynthesis-inducing (TBI) culture under the same total nitrogen conditions. Intriguingly, spermidine and spermine, downstream products of putrescine in the polyamine biosynthesis pathway, do not induce DON production under the same conditions. Additionally, externally applying either spermidine or spermine to wheat heads significantly reduces the diseased spikelet number caused by F. graminearum. Furthermore, our results show that overexpression of the wheat spermidine synthase (SPDS) gene TaSPDS-7D1 significantly enhances the spermidine content and wheat resistance to FHB. In addition, the TaSPDS-7D1-overexpressing line OE3 exhibited a 1000-grain weight and plant height increase compared to the wild type. Our findings reveal that overexpression of the spermidine synthase gene can enhance wheat resistance to FHB without compromising wheat yield. [ABSTRACT FROM AUTHOR]

This study randomly selected a total of 1 160 feed raw materials and feed samples including corn, corn by-products (corn distillers dried grains with solubles, corn germ meal, corn gluten meal, corn bran), wheat, wheat bran, miscellaneous meal (cottonseed meal, palm meal, rapeseed meal, peanut meal, sunflower seed meal) and feed (creep feed, nursery feed, piglet feed, growing feed, sow feed, broiler feed, meat duck feed, laying hen feed and laying duck feed) from different provinces and cities, and determined aflatoxin (AFB1), zearalenone (ZEN) and deoxynivalenol (DON) levels using the immune affinity column-high performance liquid chromatography (HPLC) method to understand the Chinese mycotoxins contamination of feed raw materials and feed. The results indicated: the detection rate of AFB1 in corn, corn by-products, wheat, wheat bran, oil seed meals and feed was 98.22%, 100.00%,100.00%, 98.84% and 94.24%, respectively, while the exceeding standard rate was 12.81%, 5.00%, 8.07%, 20.93% and 11.82%, respectively. The detection rate of ZEN in corn, corn by-products, wheat, wheat bran, oil seed meals and feed was 93.24%, 100.00%, 100.00%, 95.35% and 100.00%, respectively, while the exceeding standard rate was 9.25%, 22.50%, 14.35%, 5.81% and 18.18%, respectively. The detection rate of DON in corn, corn by-products, wheat, wheat bran, oil seed meals and feed was 97.15%, 100.00%, 100.00%, 88.23% and 100.00%, respectively, while the exceeding standard rate was 6.05%, 17.50%, 17.49%, 0 and 28.18%, respectively. In conclusion, the survey results indicated corn by-products and feed were seriously polluted by ZEN and DON, which is a key focus for the prevention and control of toxin pollution. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON), primarily generated by Fusarium species, often exists in agricultural products. It can be transformed to 3-epi-deoxynivalenol (3-epi-DON), with a relatively low toxicity, via two steps. DDH in Pelagibacterium halotolerans ANSP101 was proved to convert DON to 3-keto-deoxynivalenol (3-keto-DON). In the present research, AKR4, a NADPH-dependent aldo/keto reductase from P. halotolerans ANSP101, was identified to be capable of converting 3-keto-DON into 3-epi-DON. Our results demonstrated that AKR4 is clearly a NADPH-dependent enzyme, for its utilization of NADPH is higher than that of NADH. AKR4 functions at a range of pH 5–10 and temperatures of 20–60 °C. AKR4 is able to degrade 89% of 3-keto-DON in 90 min at pH 7 and 50 °C with NADPH as the cofactor. The discovery of AKR4, serving as an enzyme involved in the final step in DON degradation, might provide an option for the final detoxification of DON in food and feed. [ABSTRACT FROM AUTHOR]

Simple Summary: Barley is the fourth largest cereal crop in the world. Fusarium head blight (FHB) is one of the diseases in barley producing mycotoxins, such as deoxynivalenol (DON), that could affect grain quality as well as human and animal health worldwide. Due to limited reliable biomarkers to identify and develop FHB-resistant cultivars in barley, we investigated the composition of phenolic compounds in ten barley cultivars under clean and FHB-infected conditions. We analyzed free and bound forms of phenolic compounds and identified differences among tested cultivars. Analysis of mycotoxin DON content showed that resistant cultivars produced less compared to susceptible cultivars. In addition, the resistant cultivars showed higher amounts of major phenolic compounds compared to the known susceptible cultivar. The results of this study suggest that phenolic compounds in barley could have a role as potential biomarkers to identify and develop FHB-resistant barley cultivars. Barley (Hordeum vulgare L.) is the fourth largest cereal crop in the world. One of the most devastating diseases in barley worldwide is Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe. Several mycotoxins are produced by FHB infection, and deoxynivalenol (DON) is one of them responsible for the deterioration of grain quality. The current limited number of reliable molecular markers makes the development of FHB-resistant cultivars rather difficult and laborious. Moreover, there is a limited number of designed specific biomarkers that could distinguish the FHB resistance and mycotoxin accumulation in barley cultivars. This study investigated the phenolic compounds of ten different Canadian barley cultivars, grown in artificially FHB-infected and non-infected field trials. The enzyme-linked immunosorbent assay (ELISA) was used to assess the presence of DON in the harvested infected grains of each tested variety. High-performance liquid chromatography (HPLC) analysis was performed using both infected and non-infected samples. We identified differences among cultivars tested in non-infected samples through quantitative analysis of free and bound phenolic compounds. The resistant cultivars showed higher amounts of major bound phenolic compounds compared to the susceptible check CDC Bold. Additionally, the FHB-infected cultivars produced significantly higher amounts of sinapic acid (SIN) () and catechin (CAT) in the soluble free form of phenolics in barley compared to the non-infected subjects. This study suggests that phenolic compounds in barley could allow barley breeders to precisely identify and develop FHB-resistant barley germplasm and cultivars. [ABSTRACT FROM AUTHOR]

Endometrial cancer is one of the most common cancer types among women. Many factors can contribute to the development of this disease, including environmental factors and, thus, eating habits. Our study aims to determine the levels of various mycotoxins and their metabolites in the blood serum and endometrial tissue samples of participants with previously proven endometrial cancer and to find possible contributions to cancer development. In the cohort clinical trial, 52 participants aged between 44 and 86 were studied. The participants were divided into two groups: patients or matched controls. All patients had previously histologically diagnosed endometrial cancer. The cancer patients were divided into low-grade endometrioid and low- plus high-grade endometrioid groups. Controls had no history of endometrial malignancy or premalignancy. Blood serum and endometrial tissue samples were obtained from all study patients. We compared the concentrations of total Aflatoxins (Afs), Deoxynivalenol (DON), Ochratoxin-A (OTA), T2-toxin and HT2 toxin (T2/HT2 toxin), Zearalenone (ZEN), alpha-Zearalenol (α-ZOL), and Fumonisin B1 (FB1) in the serum and endometrium between the different study groups. As a result, we can see a significant correlation between the higher levels of Afs and zearalenone and the presence of endometrial cancer. In the case of Afs, DON, OTA, T2/HT2 toxins, ZEN, and alpha-ZOL, we measured higher endometrial concentrations than in serum. Considering the effect of mycotoxins and eating habits on cancer development, our results might lead to further research exploring the relationship between certain mycotoxins and endometrium cancer.

This study investigated if dietary composition influences the effects of Deoxynivalenol (DON) on the health and performance of rainbow trout (Oncorhynchus mykiss). Four experimental diets (2 × 2 factorial design) were formulated which differed in 1) the diet composition; fishmeal (FM) versus soybean meal-based (SBM) and 2) the DON content of wheat; clean versus naturally contaminated wheat. Triplicate groups of n = 30 fish were assigned to each diet: (1) CON-FM; DON= 0 µg/kg feed; (2) DON-FM; DON= 1200 µg/kg feed; (3) CON-SBM; DON= 46 µg/kg feed; (4) DON-SBM; DON= 1300 µg/kg feed. The 8 week experiment was divided into two feeding periods: after 6 weeks of restrictive feeding, fish were fed ad libitum for 2 weeks. Influences on performance were evaluated by determining growth, protein and energy gain metrics, and on health parameters through the determination of histopathological changes in the liver and gastrointestinal tract (GIT). Restrictive feeding showed negative effects of DON and dietary composition on performance but did not show an interaction between DON and diet composition. Similarly, subsequent ad libitum feeding showed effects of DON and/or diet composition on growth, feed efficiency and body biometrics, but no interaction effects. These data confirmed the challenging nature of the SBM-based diet and previously noted negative effects of DON on performance. Only the histopathological assessment of mucosal fold width, enterocyte width and goblet cell density indicated significant interaction effects between DON and diet composition in the midgut. Yet, the differences were generally small and interaction effects were restricted to the midgut and not to the hindgut which is usually challenged by SBM. The combined information on growth performance and health suggests that DON challenges rainbow trout regardless of being fed a FM- or SBM-based diet, allowing more flexible formulations in the aquafeeds.

Fusarium head blight (FHB), caused by Fusarium spp., is a destructive disease of cereal grains. Apart from grain yield loss, a major quality concern is contamination with Fusarium-produced mycotoxins, specifically deoxynivalenol (DON). Mycotoxins accumulate in the grain, making it unfit for consumption by humans and animals. Breeding cultivars with high disease resistance and low mycotoxin contamination is a priority for wheat breeders. However, DON measurement in breeding programs is expensive and time consuming due to the lack of efficient quantification methods. In this study, we established a simple fast chromatography-tandem mass spectrometry method, which employed a one-step acetonitrile extraction protocol with a short guard column to reduce complexity, cost, and analysis time. To ensure robustness and reproducibility, the method was validated according to the U.S. Food and Drug Administration Guidance for Bioanalytical Method Validation. Furthermore, the method was applied for determination of DON in 102 wheat grain samples. Obtained results highly correlated with the conventional immunological method for all tested samples. With its ease of use, rapid sample analysis, and high sensitivity and accuracy, the method could be integrated into current FHB breeding programs to increase breeding efficiency and accelerate screening progress to identify germplasm with increased resistance to DON accumulation. [Figure: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

The COP9 signalosome (Csn) complex is an evolutionarily conserved complex that regulates various important cellular processes. However, the function of the Csn complex in pathogenic fungi remains elusive. Here, the distribution of Csn subunits in the fungal kingdom was surveyed, and their biological functions were systematically characterized in the fungal pathogen Fusarium graminearum, which is among the top 10 plant fungal pathogens. The results obtained from bioinformatic analyses suggested that the F. graminearum Csn complex consisted of seven subunits (Csn1–Csn7) and that Csn5 was the most conserved subunit across the fungi kingdom. Yeast two-hybrid assays demonstrated that the seven Csn subunits formed a complex in F. graminearum. The Csn complex was localized to both the nucleus and cytoplasm and necessary for hyphal growth, asexual and sexual development and stress response. Transcriptome profiling revealed that the Csn complex regulated the transcription abundance of TRI genes necessary for mycotoxin deoxynivalenol (DON) biosynthesis, subsequently regulating DON production to control fungal virulence. Collectively, the roles of the Csn complex in F. graminearum were comprehensively analyzed, providing new insights into the functions of the Csn complex in fungal virulence and suggesting that the complex may be a potential target for combating fungal diseases [ABSTRACT FROM AUTHOR]

Fusarium toxins are one of the most common contaminants in poultry diets. The co-occurrence of fumonisins (FUM) and deoxynivalenol (DON), even at a subclinical dose, negatively affects the growth performance, intestinal integrity and induce subclinical necrotic enteritis in broiler chickens. Loss of gut integrity can be expected to alter the intestinal microbiota’s composition. The objective of this study was to identify the effects of combined FUM and DON on the cecal microbiome profile and predicted metabolic functions and a short chain fatty acid profile in broilers challenged with Clostridium perfringens. A total of 240 1 day-old chicks were randomly assigned to two treatments: a control diet and the control diet with 3 mg/kg FUM + 4 mg/kg DON each with eight replications. All the birds were received cocci vaccine at d0. All birds in both treatment groups were challenged with C. perfringens 1 × 108 CFU via feed on d 19 and 20 to achieve 5% mortality. On d 35, the FUM and DON contaminated diet numerically (P = 0.06) decreased the body weight gain (BWG) by 84 g compared to the control group. The bacterial compositions of the cecal contents were analyzed by sequencing the V3–V4 region of the 16S rRNA gene. Overall, microbial richness and diversity increased (P < 0.02) during the studied period (d 21–35). Cecal contents of birds in the FUM + DON group had greater (P < 0.05) microbial evenness and diversity (Shannon index) compared to the control group. FUM + DON exposure decreased (P = 0.001) the relative abundance of Proteobacteria in the cecal content, compared to the control group. The combined FUM + DON significantly increased the relative abundance of the Defluviitaleaceae and Lachnospiraceae families (P < 0.05) but decreased the abundances of the Moraxellaceae and Streptococcaceae (P < 0.05) compared to the control group birds. At the genus level, FUM + DON exposure decreased (P < 0.05) Acinetobacter and Pseudomonas abundance and had a tendency (P = 0.08) to decrease Thermincola abundance compared to the control group. In the ileum, no NE-specific microscopic abnormalities were found; however, the tip of the ileal villi were compromised. The present findings showed that dietary FUM and DON contamination, even at subclinical levels, altered cecal microbial composition, dysregulated intestinal functions, and impaired the gut immune response, potentially predisposing the birds to necrotic enteritis. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) is a disease of small grains caused by the fungus Fusarium graminearum. In this study, we explored the use of hyperspectral imaging (HSI) to evaluate the damage caused by FHB in wheat kernels. We evaluated the use of HSI for disease classification and correlated the damage with the mycotoxin deoxynivalenol (DON) content. Computational analyses were carried out to determine which machine learning methods had the best accuracy to classify different levels of damage in wheat kernel samples. The classes of samples were based on the DON content obtained from Gas Chromatography–Mass Spectrometry (GC-MS). We found that G-Boost, an ensemble method, showed the best performance with 97% accuracy in classifying wheat kernels into different severity levels. Mask R-CNN, an instance segmentation method, was used to segment the wheat kernels from HSI data. The regions of interest (ROIs) obtained from Mask R-CNN achieved a high mAP of 0.97. The results from Mask R-CNN, when combined with the classification method, were able to correlate HSI data with the DON concentration in small grains with an R2 of 0.75. Our results show the potential of HSI to quantify DON in wheat kernels in commercial settings such as elevators or mills. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON), primarily generated by Fusarium species, often exists in agricultural products. It can be transformed to 3-epi-deoxynivalenol (3-epi-DON), with a relatively low toxicity, via two steps. DDH in Pelagibacterium halotolerans ANSP101 was proved to convert DON to 3-keto-deoxynivalenol (3-keto-DON). In the present research, AKR4, a NADPH-dependent aldo/keto reductase from P. halotolerans ANSP101, was identified to be capable of converting 3-keto-DON into 3-epi-DON. Our results demonstrated that AKR4 is clearly a NADPH-dependent enzyme, for its utilization of NADPH is higher than that of NADH. AKR4 functions at a range of pH 5–10 and temperatures of 20–60 °C. AKR4 is able to degrade 89% of 3-keto-DON in 90 min at pH 7 and 50 °C with NADPH as the cofactor. The discovery of AKR4, serving as an enzyme involved in the final step in DON degradation, might provide an option for the final detoxification of DON in food and feed.

Fusarium toxins are one of the most common contaminants in poultry diets. The co-occurrence of fumonisins (FUM) and deoxynivalenol (DON), even at a subclinical dose, negatively affects the growth performance, intestinal integrity and induce subclinical necrotic enteritis in broiler chickens. Loss of gut integrity can be expected to alter the intestinal microbiota’s composition. The objective of this study was to identify the effects of combined FUM and DON on the cecal microbiome profile and predicted metabolic functions and a short chain fatty acid profile in broilers challenged with Clostridium perfringens. A total of 240 1 day-old chicks were randomly assigned to two treatments: a control diet and the control diet with 3 mg/kg FUM + 4 mg/kg DON each with eight replications. All the birds were received cocci vaccine at d0. All birds in both treatment groups were challenged with C. perfringens 1 × 108 CFU via feed on d 19 and 20 to achieve 5% mortality. On d 35, the FUM and DON contaminated diet numerically (P = 0.06) decreased the body weight gain (BWG) by 84 g compared to the control group. The bacterial compositions of the cecal contents were analyzed by sequencing the V3–V4 region of the 16S rRNA gene. Overall, microbial richness and diversity increased (P < 0.02) during the studied period (d 21–35). Cecal contents of birds in the FUM + DON group had greater (P < 0.05) microbial evenness and diversity (Shannon index) compared to the control group. FUM + DON exposure decreased (P = 0.001) the relative abundance of Proteobacteria in the cecal content, compared to the control group. The combined FUM + DON significantly increased the relative abundance of the Defluviitaleaceae and Lachnospiraceae families (P < 0.05) but decreased the abundances of the Moraxellaceae and Streptococcaceae (P < 0.05) compared to the control group birds. At the genus level, FUM + DON exposure decreased (P < 0.05) Acinetobacter and Pseudomonas abundance and had a tendency (P = 0.08) to decrease Thermincola abundance compared to the control group. In the ileum, no NE-specific microscopic abnormalities were found; however, the tip of the ileal villi were compromised. The present findings showed that dietary FUM and DON contamination, even at subclinical levels, altered cecal microbial composition, dysregulated intestinal functions, and impaired the gut immune response, potentially predisposing the birds to necrotic enteritis.

Deoxynivalenol (DON) is widely emerging in various grain crops, milk, and wine products, which can trigger different toxic effects on humans and animals by inhalation or ingestion. It also imposes a considerable financial loss on the agriculture and food industry each year. Previous studies have reported acute and chronic toxicity of DON in liver, and liver is not only the main detoxification organ for DON but also the circadian clock oscillator directly or indirectly regulates critical physiologically hepatic functions under different physiological and pathological conditions. However, researches on the association of circadian rhythm in DON-induced liver damage are limited. In the present study, mice were divided into four groups (CON, DON, Bmal1OE, and Bmal1OE + DON) and AAV8 was used to activate (Bmal1) expression in liver. Then mice were gavaged with 5 mg/kg bw/day DON or saline at different time points (ZT24 = 0, 4, 8, 12, 16, and 20 h) in 1 day and were sacrificed 30 min after oral gavage. The inflammatory cytokines, signal transducers, and activators of transcription Janus kinase/signal transducers and activator of transcription 3 (JAKs/STAT3) pathway and bile acids levels were detected by enzyme-linked immunosorbent assay (ELISA), western blotting, and target metabolomics, respectively. The DON group showed significantly elevated interleukin-1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) levels (P < 0.05 for both) and impaired liver function with rhythm disturbances compared to the CON and Bmal1OE groups. At the molecular level, expressions of some circadian clock proteins were significantly downregulated (P < 0.05 for both) and JAKs/STAT3 pathway was activated during DON exposure, accompanied by indicated circadian rhythm disturbance and inflammatory damage. Importantly, Bmal1 overexpression attenuated DON-induced liver damage, while related hepatic bile acids such as cholic acid (CA) showed a decreasing trend in the DON group compared with the CON group. Our study demonstrates a novel finding that Bmal1 plays a critical role in attenuating liver damage by inhibiting inflammatory levels and maintaining bile acids levels under the DON condition. Therefore, Bmal1 may also be a potential molecular target for reducing the hepatotoxic effects of DON in future studies. [ABSTRACT FROM AUTHOR]

Barley (Hordeum vulgare L.) is the fourth largest cereal crop in the world. One of the most devastating diseases in barley worldwide is Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe. Several mycotoxins are produced by FHB infection, and deoxynivalenol (DON) is one of them responsible for the deterioration of grain quality. The current limited number of reliable molecular markers makes the development of FHB-resistant cultivars rather difficult and laborious. Moreover, there is a limited number of designed specific biomarkers that could distinguish the FHB resistance and mycotoxin accumulation in barley cultivars. This study investigated the phenolic compounds of ten different Canadian barley cultivars, grown in artificially FHB-infected and non-infected field trials. The enzyme-linked immunosorbent assay (ELISA) was used to assess the presence of DON in the harvested infected grains of each tested variety. High-performance liquid chromatography (HPLC) analysis was performed using both infected and non-infected samples. We identified differences among cultivars tested in non-infected samples through quantitative analysis of free and bound phenolic compounds. The resistant cultivars showed higher amounts of major bound phenolic compounds compared to the susceptible check CDC Bold. Additionally, the FHB-infected cultivars produced significantly higher amounts of sinapic acid (SIN) () and catechin (CAT) in the soluble free form of phenolics in barley compared to the non-infected subjects. This study suggests that phenolic compounds in barley could allow barley breeders to precisely identify and develop FHB-resistant barley germplasm and cultivars.

Mycotoxins are natural metabolites produced by fungi that contaminate food and feed worldwide. They can pose a threat to human and animal health, mainly causing chronic effects, e.g., immunotoxic and carcinogenic. Due to climate change, an increase in European population exposure to mycotoxins is expected to occur, raising public health concerns. This urges us to assess the current human exposure to mycotoxins in Europe to allow monitoring exposure and prevent future health impacts. The mycotoxins deoxynivalenol (DON) and fumonisin B1 (FB1) were considered as priority substances to be studied within the European Human Biomonitoring Initiative (HBM4EU) to generate knowledge on internal exposure and their potential health impacts. Several policy questions were addressed concerning hazard characterization, exposure and risk assessment. The present article presents the current advances attained under the HBM4EU, research needs and gaps. Overall, the knowledge on the European population risk from exposure to DON was improved by using new harmonised data and a newly derived reference value. In addition, mechanistic information on FB1 was, for the first time, organized into an adverse outcome pathway for a congenital anomaly. It is expected that this knowledge will support policy making and contribute to driving new Human Biomonitoring (HBM) studies on mycotoxin exposure in Europe. [ABSTRACT FROM AUTHOR]

【Objective】To explore the accumulation of deoxynivalenol (DON) in the kernels of different wheat varieties under natural disease conditions and the effect on Fuasrium diseased kernel rate (FDK) and test weight, so as to provide reference for breeding wheat varieties resistant to Fuasrium.【Methods】The content of DON, 3A-DON (3-acetyldeoxynivalenol) and 15A-DON (15-acetyldeoxynivalenol) in the kernels of 65 wheat varieties from different sources were tested by liquid chromatography-tandem mass spectrometry (LC-MS/MS) to analyze the difference in Fuasrium diseased kernel rate (FDK) and test weight (TW) under different DON content.【Results】The FDK of the 65 wheat varieties ranged from 0.20% to 7.80%. The content of DON was 0.11 to 4.26 mg/kg. The test weight was 703.00 to 791.70 g/L. With correlation coefficients of 0.514, 0.549, 0.529 and 0.521, respectively, FDK was significantly positively correlated with the contents of DON, 3A-DON, 15A-DON and DON-Total (P<0.01), and was significantly negatively correlated with test weight (P<0.05), with correlation coefficients of -0.254. With correlation coefficients of 0.999,0.911 and 0.899, respectively, the kernel contents of DON, 3A-DON, 15A-DON had significant positive correlation (P<0.01) with DON-Total, and significant negative correlation with test weight (P<0.01), with correlation coefficients of -0.502, -0.504 and -0.481, respectively. In terms of the source of the varieties, the kernel accumulation of DON was lowest in Jiangsu (1.51 mg/kg), followed by Anhui (2.06 mg/kg), Henan (2.75 mg/kg) and Shandong (2.98 mg/kg). The content of 3A-DON was lowest in Jiangsu (0.047 mg/kg), followed by Anhui (0.075 mg/kg), Shandong (0.111 mg/kg) and Henan (0.113 mg/kg). The content of 15A-DON was lowest in Jiangsu (0.059 mg/kg), followed by Anhui (0.090 mg/kg), Shandong (0.132 mg/kg) and Henan (0.141 mg/kg). The content of DON-Total was lowest in Jiangsu (1.61 mg/kg), followed by Anhui (2.23 mg/kg), Henan (3.00 mg/kg）and Shandong (3.23 mg/kg). The number and proportion of varieties with DON no more than 2 mg/kg in wheat kernels were as follows：Anhui (18,64.29%）>Jiangsu (13,61.90%）>Shandong (2,22.22%）>Henan (1,14.29%). The average test weight was highest in Jiangsu (758.57 g/L), followed by Anhui (751.90 g/L), Henan (739.71 g/L) and Shandong (735.27 g/L）. In terms of the characteristics of different varieties, eight wheat varieties with DON no more than 2 mg/kg and test weight no less than 770.00 g/L were selected. Among these, there were seven semi winter varieties including Huaimai 40, Huaimai 44, Nongmai 152,Quanmai 725, Womai No. 9, Xunong 029 and Huaimai 45, one spring variety, Guohong No.6.【Conclusion】The kernel FDK,DON contents and test weight of the 65 wheat varieties were quite different. With the increase of FDK, the kernel accumulation of 3A-DON, 15A-DON and DON-Total increased gradually, and the average test weight decreased. Huaimai 40, Huaimai 44, Nongmai 152, Quanmai 725, Womai No.9, Xunong 029, Huaimai 45, and Guohong No. 6 had better resistance to Fuasrium disease. [ABSTRACT FROM AUTHOR]

Fusarium crown rot (FCR) of wheat has become more and more serious in China in recent years, which not only threatens food security, but also has the potential threat of mycotoxin contamination, which endangers human and animal health. This paper summarizes the damage status of wheat crown rot and the dominant Fusarium spp. that cause the disease in different regions. It is clear that Fusarium pseudograminearum has gradually become the dominant pathogen of FCR in many major wheat producing areas in China. On this basis, the infection cycle and genetic diversity of F. pseudograminearum were further analyzed, revealing the serious occurrence of FCR and the accumulation of F. pseudograminearum in soil, agricultural measures and various environmental and climatic factors, especially drought environment is closely related. This paper also summarized the reported key proteins that regulate the pathogenicity of F. pseudograminearum, revealed the toxin production type of F. pseudograminearum, clarified the biochemical pathway of deoxynivalenol (DON) synthesis, effects of different fungicides on DON synthesis, and the mechanism by which fungicides stimulate or inhibit DON synthesis. With the purpose of "disease control and deoxynivalenol reduction", a variety of comprehensive prevention and control measures for coordinated disease prevention are proposed, which have important reference value for the realization of green prevention and control of FCR. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) (also called vomitoxin) is a mycotoxin caused by pathogens that periodically contaminate crops such as maize, wheat, barley, oats, and rye, making them unusable. We explored pyrolysis as a process for the decontamination of vomitoxin-corn grains and their transformation into value-added products. Pyrolysis was carried out in a bench-scale batch reactor at maximum temperatures between 450 and 650 °C. This resulted in the total destruction of DON, from 5–7 ppm in raw corn grains to non-detectable levels in the treated bio-char. The effect of pyrolysis conditions, including temperature and heating rate, on the conversion of toxic corn grains was investigated. The maximum bio-oil yield was achieved at 650 °C (47 wt.%). The co-products were bio-char (29 wt.%) and non-condensable gases (24 wt.%). Acetic acid and levoglucosan were the two major valuable components in the bio-oil, corresponding to 26 g/kg and 13 g/kg of bio-oil, respectively. The bio-chars were analyzed and upgraded by physical activation using CO2 at 900 °C. Activation increased the bio-char surface area to 419 m2g−1 and promoted pore development, which was verified by SEM. Proximate analysis illustrated that stable carbon increased to 88.8% after activation compared to 10.9% in the raw corn. FT-IR results showed that the thermally unstable functional groups had been completely eliminated after activation. All characterization results confirmed that the activated corn bio-char has excellent potential for adsorption processes. The HHV of the non-condensable gas stream was 16.46 MJ/Nm3, showing the potential application of this product as an energy source. [ABSTRACT FROM AUTHOR]

IntroductionWheat is a staple food that is important to global food security, but in epidemic years, fungal pathogens can threaten production, quality, and safety of wheat grain. Globally, one of the most important fungal diseases of wheat is Fusarium head blight (FHB). This disease can be caused by several different Fusarium species with known differences in aggressiveness and mycotoxin-production potential, with the trichothecene toxin deoxynivalenol (DON) and its derivatives being of particular concern. In North America, the most predominant species causing FHB is F. graminearum, which has two distinct sub-populations that are commonly classified into two main chemotypes/genotypes based on their propensity to form trichothecene derivatives, namely 15-acetyldeoxynivalenol (15-ADON) and 3-acetyldeoxynivalenol (3-ADON).Materials and methodsWe used a panel of 13 DNA markers to perform species and ADON genotype identification for 55, 444 wheat kernels from 7, 783 samples originating from across Canada from 2014 to 2020.Results and discussionBased on single-seed analyses, we demonstrate the relationships between Fusarium species and trichothecene chemotype with sample year, sample location, wheat species (hexaploid and durum wheat), severity of Fusarium damaged kernels (FDK), and accumulation of DON. Results indicate that various Fusarium species are present across wheat growing regions in Canada; however, F. graminearum is the most common species and 3-ADON the most common genotype. We observed an increase in the occurrence of the 3-ADON genotype, particularly in the western Prairie regions. Our data provides important information on special-temporal trends in Fusarium species and chemotypes that can aid with the implementation of integrated disease management strategies to control the detrimental effects of this devastating disease.

This study aimed to investigate the antifungal and antimycotoxicogenic effect of binary and tertiary mixtures of Thymus vulgaris, Origanum sativum, and Coriandrum sativum essential oils (EOs), as well as emulsions based on EO mixtures, on fungi developed on wheat grains destined for the bakery industry. The chemical composition of the EO mixtures, the physical characteristics of the emulsions, and the influence of treatments on the proximate composition of wheat seeds were also studied. The methods used included the microbiological analysis of fungi developed on wheat seeds, the ELISA technique for determining the deoxynivalenol content (DON), gas chromatography coupled with mass spectrometry (GC-MS) to detect the chemical composition of the EOs, Zetasizer to analyse the particle sizes and their electric charge at the surface, and NIR analysis of the proximate composition of wheat. The chemical composition analysis revealed that thymol and o-cymene were the major components in the binary mixture of the EOs with thyme, linalool in the binary mixtures of the EOs with coriander and carvacrol, and o-cymene in the binary mixtures of the EOs with oregano. The results showed that, based on the zeta potential, the tertiary mixture ensured maximum emulsion stability, while the emulsion based on thyme and oregano was the less stable system. Regarding the antifungal and antimycotoxicogenic effect, the results showed that the highest inhibition potential on fungi was observed with the binary mixtures of the EOs based on thyme and oregano, and on deoxynivalenol (DON) when the binary emulsion based on the same EOs was applied to wheat seeds. The proximate composition of wheat seeds contaminated with DON showed an increase in protein content and mineral substances, and there were changes in the colour of the wheat seeds after treatment with the EOs. In conclusion, the results obtained in this study showed the possibility of using binary/tertiary mixtures of EOs and emulsions as healthy and environmentally friendly alternatives in the bakery industry. [ABSTRACT FROM AUTHOR]

Deoxynivalenol (DON) is a secondary fungal metabolite that is associated with many adverse toxicological effects in agriculture as well as human/animal nutrition. Bioremediation efforts in recent years have led to the discovery of numerous bacterial isolates that can transform DON to less toxic derivatives. Both 3-keto-DON and 3-epi-DON were recently shown to exhibit reduced toxicity, compared to DON, when tested using different cell lines and mammalian models. In the current study, the toxicological assessment of 3-keto-DON and 3-epi-DON using in planta models surprisingly revealed that 3-keto-DON, but not 3-epi-DON, retained its toxicity to a large extent in both duckweeds (Lemna minor L.) and common wheat (Triticum aestivum L.) model systems. RNA-Seq analysis revealed that the exposure of L. minor to 3-keto-DON and DON resulted in substantial transcriptomic changes and similar gene expression profiles, whereas 3-epi-DON did not. These novel findings are pivotal for understanding the environmental burden of the above metabolites as well as informing the development of future transgenic plant applications. Collectively, they emphasize the fundamental need to assess both plant and animal models when evaluating metabolites/host interactions. [ABSTRACT FROM AUTHOR]

Abstract Background Intestinal epithelial cells are challenged by mycotoxins and many bacterial pathogens. It was previously shown that the mycotoxin deoxynivalenol (DON) as well as Campylobacter (C.) jejuni have a negative impact on gut integrity. Recently, it was demonstrated that DON increased the load of C. jejuni in the gut and inner organs. Based on this finding, it was hypothesized the DON metabolite (deepoxy-deoxynivalenol, DOM-1) should be able to reduce the negative effects of DON on colonization and translocation of C. jejuni in broilers, since it lacks the epoxide ring, which is responsible for the toxicity of DON. Methods A total of 180 broiler chickens were housed in floor pens on wood shavings with feed and water provided ad libitum. Birds were divided into six groups (n = 30 with 5 replicates/group): 1. Control, 2. DOM-1, 3. DON, 4. DOM-1 + C. jejuni, 5. DON + C. jejuni, 6. C. jejuni. At day 14, birds of groups 4, 5 and 6 were orally inoculated via feeding tube (gavage) with 1-ml of a PBS suspension containing 1 × 108 CFU of C. jejuni NCTC 12744. The performance parameters: body weight (BW), body weight gain (BWG), and feed intake of the birds were determined. At 7, 14, and 21 days post infection, samples from liver, spleen, duodenum, jejunum and cecum were aseptically collected and processed for bacteriological investigations. Finally, at each killing time point, segments of duodenum, jejunum and cecum were harvested and prepared for Ussing chamber studies to measure the paracellular mannitol fluxes. Results A significant decrease in body weight was observed for chickens receiving the DON diet with or without C. jejuni compared to the other groups. Furthermore, it was found that the co-exposure of birds to DON and C. jejuni resulted in a higher C. jejuni load not only in the gut but also in liver and spleen due to increased paracellular permeability of the duodenum, jejunum and cecum. On the contrary, DOM-1 supplementation in the feed improved the birds’ performance and led to a better feed conversion ratio throughout the trial. Furthermore, DOM-1 did not negatively affect gut permeability and decreased the C. jejuni counts in the intestine and internal organs. Conclusion Altogether, the presence of DOM-1 in the feed as a result of the enzymatic biotransformation of DON leads to a lower C. jejuni count in the intestine and better feed conversion ratio. Moreover, this study demonstrates that the detoxification product of DON, DOM-1, does not have negative effects on the gastrointestinal tract and reduces the Campylobacter burden in chickens and also the risk for human infection.

Deoxynivalenol (DON) is a common mycotoxin in nature, which is not only widely distributed, but also can cause human and animal poisoning, posing a serious threat to food safety and social economy. Therefore, it is very important to develop a sensitive and stable method to detect DON. In this research, the optimal aptamer D40 was obtained by molecular docking prediction and truncation based on molecular docking results. Then, we labeled the Cy3 Raman reporter molecule with D40, as a capture probe for the aptasensor. Finally, the porous π-rich structure Cu@C derived from the thermal decomposition of CuBTC is combined with AuNPs to generate abundant hot spots, and the Cy3-aptamers are adsorbed to the composite by the π-π to produce sharp and significant Raman signals. In the presence of DON, Cy3-aptamers bind to DON specifically and dissociate from the composite material, and the Raman signal decreases. The prepared SERS aptasensor was capable of detecting DON in a wide range from 0.01 to 10,000 ng/mL with a detection limit of 4.56 pg/mL, and the recovery rate of 94.42–107.04 % was obtained in two real samples, which indicated that the aptasensor proposed in this study exhibited outstanding practical performance and had potential application value in the field of food safety. • A widely used Deoxynivalenol aptamer containing 80 bases was successfully truncated, and elucidates the binding site. • The truncated aptamer D40 possesses improved binding affinity and specificity and lower cost. • A highly sensitive SERS aptasensor using a novel truncated aptamer was established for the detection of Deoxynivalenol. • The SERS aptasensor achieved sensitive detection of DON at 4.56 pg/mL, and exhibited satisfying uniformity, specificity and stability. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the impact of carboxymethylation on the structural and functional properties of β-glucan derived from the pathogenic oomycete Pythium insidiosum. β-Glucan was extracted and subjected to carboxymethylation (CM-glucan), with structural changes analyzed using 13C and 1H NMR spectroscopy. The modified β-glucan's ability to adsorb mycotoxins, specifically deoxynivalenol (DON) and T2 toxin, was assessed through in vitro adsorption assays. Results demonstrated that the adsorption of DON by CM-glucan increased from 0 % to 59.11 %, corresponding to the adsorption of approximately 1.18 μg of DON from the initial concentration (2 μg/mL). Similarly, the adsorption of T2 toxin increased slightly from 0 % to 4.54 %, corresponding to 0.09 μg of T2 toxin adsorbed from the initial concentration (2 μg/mL). These findings underscore the potential of chemical modifications to enhance the functional properties of natural polysaccharides, suggesting future applications in mycotoxin adsorption and other biological properties across various areas. [Display omitted] • Novel carboxymethylation of β-glucan from Pythium insidiosum enhances adsorption properties. • CM-Glucan shows a significant increase in DON adsorption efficiency, from 0 % to 59.11 %. • Structural changes confirmed by detailed 13C and 1H NMR spectroscopy. • Findings pave the way for future studies on optimizing β-glucan modifications. [ABSTRACT FROM AUTHOR]

The disease severity and mycotoxin DON content in grains caused by fusarium head blight (FHB) have been two prioritized economical traits in wheat. Reliable phenotyping is a prerequisite for genetically improving wheat resistances to these two traits. In this study, three inoculation methods: upper bilateral floret injection (UBFI), basal bilateral floret injection (BBFI), and basal rachis internode injection (BRII), were applied in a panel of 22 near-isogenic lines (NILs) contrasting in Fhb1 alleles. The results showed that inoculation methods had significant influence on both disease severity and mycotoxin accumulation in grains, and the relationship between them. UBFI method caused chronic FHB symptom characterized as slow progress of the pathogen downward from the inoculation site, which minimized the difference in disease severity of the NILs, but, unexpectedly, maximized the difference in DON content between them. The BBFI method usually caused an acute FHB symptom in susceptible lines characterized as premature spike death (PSD), which maximized the difference in disease severity, but minimized the difference in DON content in grains between resistant and susceptible lines. The BRII method occasionally caused acute FHB symptoms for susceptible lines and had relatively balanced characteristics of disease severity and DON content in grains. Therefore, two or more inoculation methods are recommended for precise and reliable evaluation of the overall resistance to FHB, including resistances to both disease spread within a spike and DON accumulation in grains. [ABSTRACT FROM AUTHOR]

This article assesses the occurrence of Fusarium-damaged kernels (FDKs) in common wheat (Triticum aestivum) under the influence of environmental factors and extreme weather events in Romania (exceptionally high air temperatures and extreme pedological drought produced by a dipole block in summer 2015, and extreme precipitation and floods produced by an omega block in spring 2016). Wheat samples (N = 272) were analyzed for FDKs via visual estimation and manual weighing according to ISO 7970 and are statistically evaluated using SPSS. The dipole block in 2015 reduced the effects of environmental factors to non-significant correlations with FDKs, while the omega block in 2016 was non-significantly to very significantly correlated with FDKs in the northwestern and western regions. The occurrence of FDKs was favored for wheat cultivation in acidic soils and inhibited in alkaline soils. Wheat samples with FDKs ≥ 1% were sampled from crops grown in river meadows with high and very high risks of flooding. Knowing the contaminants' geographical and spatial distributions under the influence of regular and extreme weather events is important for establishing measures to mitigate the effects of climate change and to ensure human and animal health. [ABSTRACT FROM AUTHOR]

Fusarium mycotoxins in cereals constitute major problems for animal and human health worldwide. A range of plant pathogenic Fusarium species that can infect cereal plants in the field are considered the most important source of mycotoxins, such as deoxynivalenol (DON), zearalenone (ZEA), T-2 toxin, and HT-2 toxin, in small-grain cereal crops in temperate climates. In this article, we (i) critically review the available knowledge on the impact of contrasting production systems (organic versus conventional) and specific agronomic parameters on the occurrence and concentrations of DON, ZEA, and T-2/HT-2 in small-grain cereals (wheat, oats, barley, and rye), and (ii) discuss Fusarium mycotoxin risks in the context of the need to develop more sustainable cereal production systems. Overall, the available evidence from studies of acceptable scientific quality suggests that the incidence and concentrations of Fusarium mycotoxin are lower in organic compared with conventional cereals. Specifically, 24 comparisons showed lower mycotoxin levels in organic production, 16 detected no significant difference, and only 2 showed higher levels in organic production. When the mean concentrations from all studies were compared, conventionally produced cereals had 62%, 110%, and 180% higher concentrations of DON, ZEA, and T-2/HT-2, respectively, than organic cereals. Overall, published studies on the effects of specific agronomic practices on mycotoxin levels suggest that diverse crop rotations and high soil organic matter content/biological activity are associated with a lower risk of Fusarium mycotoxin contamination, whereas (i) high mineral nitrogen fertiliser inputs, (ii) some fungicides and herbicides, and (iii) minimum or no tillage may increase the risks of Fusarium mycotoxin contamination in cereals. The management of Fusarium head blight and mycotoxins, therefore, requires a preventative, integrated, holistic agronomic approach. [ABSTRACT FROM AUTHOR]

Fusarium head blight (FHB) is a disease of small grains caused by the fungus Fusarium graminearum. In this study, we explored the use of hyperspectral imaging (HSI) to evaluate the damage caused by FHB in wheat kernels. We evaluated the use of HSI for disease classification and correlated the damage with the mycotoxin deoxynivalenol (DON) content. Computational analyses were carried out to determine which machine learning methods had the best accuracy to classify different levels of damage in wheat kernel samples. The classes of samples were based on the DON content obtained from Gas Chromatography–Mass Spectrometry (GC-MS). We found that G-Boost, an ensemble method, showed the best performance with 97% accuracy in classifying wheat kernels into different severity levels. Mask R-CNN, an instance segmentation method, was used to segment the wheat kernels from HSI data. The regions of interest (ROIs) obtained from Mask R-CNN achieved a high mAP of 0.97. The results from Mask R-CNN, when combined with the classification method, were able to correlate HSI data with the DON concentration in small grains with an R2 of 0.75. Our results show the potential of HSI to quantify DON in wheat kernels in commercial settings such as elevators or mills.

Summary: Fusarium graminearum produces the mycotoxin deoxynivalenol (DON) which promotes its expansion during infection on its plant host wheat. Conditional expression of DON production during infection is poorly characterized.Wheat produces the defense compound putrescine, which induces hypertranscription of DON biosynthetic genes (FgTRIs) and subsequently leads to DON accumulation during infection. Further, the regulatory mechanisms of FgTRIs hypertranscription upon putrescine treatment were investigated.The transcription factor FgAreA regulates putrescine‐mediated transcription of FgTRIs by facilitating the enrichment of histone H2B monoubiquitination (H2B ub1) and histone 3 lysine 4 di‐ and trimethylations (H3K4 me2/3) on FgTRIs. Importantly, a DNA‐binding domain (bZIP) specifically within the Fusarium H2B ub1 E3 ligase Bre1 othologs is identified, and the binding of this bZIP domain to FgTRIs depends on FgAreA‐mediated chromatin rearrangement. Interestingly, H2B ub1 regulates H3K4 me2/3 via the methyltransferase complex COMPASS component FgBre2, which is different from Saccharomyces cerevisiae.Taken together, our findings reveal the molecular mechanisms by which host‐generated putrescine induces DON production during F. graminearum infection. Our results also provide a novel insight into the role of putrescine during phytopathogen–host interactions and broaden our knowledge of H2B ub1 biogenesis and crosstalk between H2B ub1 and H3K4 me2/3 in eukaryotes. [ABSTRACT FROM AUTHOR]

Mycotoxins are natural metabolites produced by fungi that contaminate food and feed worldwide. They can pose a threat to human and animal health, mainly causing chronic effects, e.g., immunotoxic and carcinogenic. Due to climate change, an increase in European population exposure to mycotoxins is expected to occur, raising public health concerns. This urges us to assess the current human exposure to mycotoxins in Europe to allow monitoring exposure and prevent future health impacts. The mycotoxins deoxynivalenol (DON) and fumonisin B1 (FB1) were considered as priority substances to be studied within the European Human Biomonitoring Initiative (HBM4EU) to generate knowledge on internal exposure and their potential health impacts. Several policy questions were addressed concerning hazard characterization, exposure and risk assessment. The present article presents the current advances attained under the HBM4EU, research needs and gaps. Overall, the knowledge on the European population risk from exposure to DON was improved by using new harmonised data and a newly derived reference value. In addition, mechanistic information on FB1 was, for the first time, organized into an adverse outcome pathway for a congenital anomaly. It is expected that this knowledge will support policy making and contribute to driving new Human Biomonitoring (HBM) studies on mycotoxin exposure in Europe.

Background: Intestinal epithelial cells are challenged by mycotoxins and many bacterial pathogens. It was previously shown that the mycotoxin deoxynivalenol (DON) as well as Campylobacter (C.) jejuni have a negative impact on gut integrity. Recently, it was demonstrated that DON increased the load of C. jejuni in the gut and inner organs. Based on this finding, it was hypothesized the DON metabolite (deepoxy-deoxynivalenol, DOM-1) should be able to reduce the negative effects of DON on colonization and translocation of C. jejuni in broilers, since it lacks the epoxide ring, which is responsible for the toxicity of DON. Methods: A total of 180 broiler chickens were housed in floor pens on wood shavings with feed and water provided ad libitum. Birds were divided into six groups (n = 30 with 5 replicates/group): 1. Control, 2. DOM-1, 3. DON, 4. DOM-1 + C. jejuni, 5. DON + C. jejuni, 6. C. jejuni. At day 14, birds of groups 4, 5 and 6 were orally inoculated via feeding tube (gavage) with 1-ml of a PBS suspension containing 1 × 108 CFU of C. jejuni NCTC 12744. The performance parameters: body weight (BW), body weight gain (BWG), and feed intake of the birds were determined. At 7, 14, and 21 days post infection, samples from liver, spleen, duodenum, jejunum and cecum were aseptically collected and processed for bacteriological investigations. Finally, at each killing time point, segments of duodenum, jejunum and cecum were harvested and prepared for Ussing chamber studies to measure the paracellular mannitol fluxes. Results: A significant decrease in body weight was observed for chickens receiving the DON diet with or without C. jejuni compared to the other groups. Furthermore, it was found that the co-exposure of birds to DON and C. jejuni resulted in a higher C. jejuni load not only in the gut but also in liver and spleen due to increased paracellular permeability of the duodenum, jejunum and cecum. On the contrary, DOM-1 supplementation in the feed improved the birds' performance and led to a better feed conversion ratio throughout the trial. Furthermore, DOM-1 did not negatively affect gut permeability and decreased the C. jejuni counts in the intestine and internal organs. Conclusion: Altogether, the presence of DOM-1 in the feed as a result of the enzymatic biotransformation of DON leads to a lower C. jejuni count in the intestine and better feed conversion ratio. Moreover, this study demonstrates that the detoxification product of DON, DOM-1, does not have negative effects on the gastrointestinal tract and reduces the Campylobacter burden in chickens and also the risk for human infection. [ABSTRACT FROM AUTHOR]

Background: Several immunoassay methods have been developed for the detection of DON in cereal. However, there has been no method based on Eu-chelate antibody-conjugated magnetic nanoparticles established. Methods: Indirect competition for DON detection was established by combining Eu-chelate and immunomagnetic separation. The immunocomplex was separated and washed by exposure to a magnetic field and treatment with enhancement solution; then, fluorescences were measured according to the number of europium ions dissociated. Results: The working concentrations ranged from 0.05 to 100 ng/mL, and the sensitivity was 0.5 μg/Kg in cereal. The intra–assay CVs of corn and wheat were 5.7% and 7.1%, respectively. The recovery rates obtained using cereal sample were 83.2%–120.0%. Conclusion: This study suggested that the Eu-chelate antibody-conjugated magnetic nanoparticles method was fast and sensitive in screening large quantities of samples, and could be used for detection of DON in cereal.

This study aimed to investigate the antifungal and antimycotoxicogenic effect of binary and tertiary mixtures of Thymus vulgaris, Origanum sativum, and Coriandrum sativum essential oils (EOs), as well as emulsions based on EO mixtures, on fungi developed on wheat grains destined for the bakery industry. The chemical composition of the EO mixtures, the physical characteristics of the emulsions, and the influence of treatments on the proximate composition of wheat seeds were also studied. The methods used included the microbiological analysis of fungi developed on wheat seeds, the ELISA technique for determining the deoxynivalenol content (DON), gas chromatography coupled with mass spectrometry (GC-MS) to detect the chemical composition of the EOs, Zetasizer to analyse the particle sizes and their electric charge at the surface, and NIR analysis of the proximate composition of wheat. The chemical composition analysis revealed that thymol and o-cymene were the major components in the binary mixture of the EOs with thyme, linalool in the binary mixtures of the EOs with coriander and carvacrol, and o-cymene in the binary mixtures of the EOs with oregano. The results showed that, based on the zeta potential, the tertiary mixture ensured maximum emulsion stability, while the emulsion based on thyme and oregano was the less stable system. Regarding the antifungal and antimycotoxicogenic effect, the results showed that the highest inhibition potential on fungi was observed with the binary mixtures of the EOs based on thyme and oregano, and on deoxynivalenol (DON) when the binary emulsion based on the same EOs was applied to wheat seeds. The proximate composition of wheat seeds contaminated with DON showed an increase in protein content and mineral substances, and there were changes in the colour of the wheat seeds after treatment with the EOs. In conclusion, the results obtained in this study showed the possibility of using binary/tertiary mixtures of EOs and emulsions as healthy and environmentally friendly alternatives in the bakery industry.

The disease severity and mycotoxin DON content in grains caused by fusarium head blight (FHB) have been two prioritized economical traits in wheat. Reliable phenotyping is a prerequisite for genetically improving wheat resistances to these two traits. In this study, three inoculation methods: upper bilateral floret injection (UBFI), basal bilateral floret injection (BBFI), and basal rachis internode injection (BRII), were applied in a panel of 22 near-isogenic lines (NILs) contrasting in Fhb1 alleles. The results showed that inoculation methods had significant influence on both disease severity and mycotoxin accumulation in grains, and the relationship between them. UBFI method caused chronic FHB symptom characterized as slow progress of the pathogen downward from the inoculation site, which minimized the difference in disease severity of the NILs, but, unexpectedly, maximized the difference in DON content between them. The BBFI method usually caused an acute FHB symptom in susceptible lines characterized as premature spike death (PSD), which maximized the difference in disease severity, but minimized the difference in DON content in grains between resistant and susceptible lines. The BRII method occasionally caused acute FHB symptoms for susceptible lines and had relatively balanced characteristics of disease severity and DON content in grains. Therefore, two or more inoculation methods are recommended for precise and reliable evaluation of the overall resistance to FHB, including resistances to both disease spread within a spike and DON accumulation in grains.

This article assesses the occurrence of Fusarium-damaged kernels (FDKs) in common wheat (Triticum aestivum) under the influence of environmental factors and extreme weather events in Romania (exceptionally high air temperatures and extreme pedological drought produced by a dipole block in summer 2015, and extreme precipitation and floods produced by an omega block in spring 2016). Wheat samples (N = 272) were analyzed for FDKs via visual estimation and manual weighing according to ISO 7970 and are statistically evaluated using SPSS. The dipole block in 2015 reduced the effects of environmental factors to non-significant correlations with FDKs, while the omega block in 2016 was non-significantly to very significantly correlated with FDKs in the northwestern and western regions. The occurrence of FDKs was favored for wheat cultivation in acidic soils and inhibited in alkaline soils. Wheat samples with FDKs ≥ 1% were sampled from crops grown in river meadows with high and very high risks of flooding. Knowing the contaminants’ geographical and spatial distributions under the influence of regular and extreme weather events is important for establishing measures to mitigate the effects of climate change and to ensure human and animal health.

Deoxynivalenol (DON) is a type B trichothecene mycotoxin with worldwide high incidence in feed which is produced by Fusarium species. Strategies are needed to eliminate its health risk for livestock and to minimise its economic impact on production. In order to assess the efficacy of potential physical, chemical and biological DON detoxifying agents, a good in vitro model is necessary to perform a fast and high-throughput screening of new compounds before in vivo trials are set up. In this paper, an in vitro model was developed to screen potential commercial products for DON degradation and detoxification. Contaminated feed with potential detoxifying agents are first applied to a simulated gastrointestinal tract (GIT) of a pig, after which detoxification is assessed through a robust, inexpensive and readily applicable Lemna minor L. aquatic plant bioassay which enables evaluation of the residual toxicity of possible metabolites formed by DON detoxifying agents. The GIT simulation enables taking matrix and incubation parameters into account as they can affect the binding, removal or degradation of DON. One product could reduce DON in feed in the GIT model for almost 100% after 6 h. DON metabolites were tentatively identified with LC-MS/MS. This GIT simulation coupled to a detoxification bioassay is a valuable model for in vitro screening and assessing compounds for DON detoxification, and could be expanded towards other mycotoxins. [ABSTRACT FROM AUTHOR]

Fusarium crown rot (FCR) is an important and devastating disease of wheat ( Triticum aestivum ) caused by the fungus Fusarium pseudograminearum and related pathogens. Using two distinct susceptible cultivars, we investigated the isolation frequencies of F. pseudograminearum and quantified its biomass accumulation and the levels of the associated toxins deoxynivalenol (DON) and DON-3-glucoside (D3G) in inoculated field-grown wheat plants. We detected F. pseudograminearum in stem, peduncle, rachis, and husk tissues, but not in grains, whereas DON and D3G accumulated in stem, rachis, husk, and grain tissues. Disease severity was positively correlated with the frequency of pathogen isolation, F. pseudograminearum biomass, and mycotoxin levels. The amount of F. pseudograminearum biomass and mycotoxin contents in asymptomatic tissue of diseased plants were associated with the distance of the tissue from the diseased internode and the disease severity of the plant. Thus, apparently healthy tissue may harbor F. pseudograminearum and contain associated mycotoxins. This research helps clarify the relationship between F. pseudograminearum occurrence, F. pseudograminearum biomass, and mycotoxin accumulation in tissues of susceptible wheat cultivars with or without disease symptoms, providing information that can lead to more effective control measures., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Xu, Shi, Liu, Li, Wang, Han, Liu, Wang, Liu, Fan, Wang, Feng, Song, Zhou and Xu.)

Fusarium head blight (FHB) has become a limiting factor in soft red winter wheat production in the southeast United States. Recent epidemics have occurred in Georgia, but genetic information on the Fusarium species responsible for FHB is unknown. This study aimed to assess pathogen population structure and genetic diversity, trichothecene profiles, and representative pathogenicity of 196 Fusarium isolates collected from 44 wheat ( n = 85) and 53 corn ( n = 111) fields in Georgia. Phylogenetic analysis using the translation elongation factor 1-alpha (635 bp) and RNA polymerase second largest subunit (930 bp) sequence data resolved isolates into 185 haplotypes, which represented 12 Fusarium species grouped under five species complexes. F. graminearum with 15-acetyl-deoxynivalenol (15ADON) chemotype (75.6%) and F. incarnatum (57.7%) predominated in wheat and corn, respectively, with a surprisingly higher frequency of nivalenol (NIV) F. graminearum (21.8%). Using nine variable numbers of tandem repeat markers, 82 multilocus genotypes out of 86 F. graminearum isolates were identified and grouped into two genetic clusters, pop1fg ( n = 29) and pop2fg ( n = 32), as part of the North American populations (NA1 and NA2) but with no chemotype differentiation. F. graminearum populations in Georgia are mostly clonal and might have evolved through at least two introductions from the northeast United States and Canada and local adaptation to maintain high genetic diversity. Pathogenicity of F. graminearum isolates from wheat and corn had high FHB severity (>60%) in wheat, which depicted the risk they can pose towards future FHB outbreaks. Overall, this baseline study provided important information on Fusarium species diversity including F. graminearum associated with FHB in Georgia that will be useful to formulate integrated disease management incorporating improved host resistance and fungicide spray programs., Competing Interests: The author(s) declare no conflict of interest.

Previous results from our lab have shown that using an optical sorter to identify Fusarium head blight (FHB) resistant breeding lines was effective at reducing the toxin deoxynivalenol (DON) and FHB-associated kernel damage. In this paper we quantified the proportion of desirable genotypes at FHB resistance QTL in lines from three selection cycles of optical sorting. Breeding lines were genotyped at loci on chromosomes 3BS, 2DL, and 5A using the following DNA markers: TaHRC, CFD233, and GWM304. TaHRC is a KASP marker for Fhb1, a major FHB resistance QTL on chromosome 3BS. CFD233 is an SSR marker for Qfhs.nau-2DL on chromosome 2DL. GWM304 is an SSR marker for Qfhs.ifa-5A on chromosome 5A. Sorter selection was effective at identifying lines that had the resistant genotype at TaHRC; in other words, the sorter was able to identify lines with resistance alleles at Fhb1. The sorter was less effective at selecting for the resistant genotype at CFD233 and GWM304. However, the proportion of lines with resistant genotypes at GWM304 did increase with additional sorter selection, just not to the degree that was observed for the Fhb1-associated marker. The proportion of lines with resistant alleles at CFD233 did not show a consistent trend. In addition to increasing the proportion of lines with Fhb1 and Qfhs.ifa-5A each selection cycle, optical sorter-based mass selection enhanced FHB resistance in different marker genotype combinations evaluated in this study. For example, there were net reductions in DON and kernel damage after two cycles of sorter selection in 15X110601S07002, a line with Fhb1, with Qfhs.nau-2DL, and with Qfhs.ifa-5A; final C3 DON levels were 63% of the resistant check (KY02C-3005-25). Kernel damage was also reduced in 15X110601A08221 a line without Fhb1, without Qfhs.nau-2DL, and without Qfhs.ifa-5A. Our findings suggest the increased resistance observed in different marker genotype combinations was conferred by QTL other than Fhb1, QFhs.nau-2DL, and Qfhs.ifa-5, and validate our previous results that the optical sorter is effective at selecting FHB-resistant breeding material.