Enniatin B1 Research Articles

Occurrence of mycotoxins of the beauvericin and enniatin groups in infant foods and risk assessment study

The aim of this study was the determination of mycotoxins belonging to beauvericin and enniatin groups in infant foods to be considered for the assessment of dietary exposure. The extraction phase with acetonitrile was followed by a purification step before liquid chromatography-mass spectrometry analysis. The method was suitable to detect these mycotoxins in different categories of infant foods (cereal-based creams, biscuits, and pasta) at concentrations between 0.30 μg/kg for beauvericin and enniatin A and 0.60 μg/kg for the remaining enniatins (A1, B and B1) up to a maximum of 20 μg/kg. Beauvericin was the most detected mycotoxin in pasta samples (15%) at a maximum concentration of 1.1 μg/kg, while enniatin B showed the highest presence (90%) in biscuits, at levels ranging from 0.63 to 11 μg/kg. Enniatin B1 was found in all matrices at comparable ranks of positivity, with the highest occurrence rate (30%) in biscuits. The highest estimated daily intake was observed in infant consumers aged 1–3 years and was associated with the consumption of biscuits, followed by samples of pasta and cereal-based creams. It was higher in infants than in children aged 4–9 years due to consumption data expressed per kg of body weight. The lowest estimated daily intake was obtained for beauvericin in all matrices as its concentrations were lower than those of the enniatin group. The results of this study highlighted the need to implement monitoring also for these mycotoxin groups, paying particular attention to infants as a high-risk category.

Potential for the Bio-Detoxification of the Mycotoxins Enniatin B and Deoxynivalenol by Lactic Acid Bacteria and Bacillus spp.

Mycotoxins, toxic compounds produced by fungi, pose significant risks to food safety and human health. This study investigates the bio-detoxification potential of 238 strains of lactic acid bacteria (LAB) and Bacillus spp., previously isolated from cereals (including mycotoxin-contaminated grains), against the emerging mycotoxin, enniatin B (ENB), and the prevalent mycotoxin, deoxynivalenol (DON). Out of the tested strains, 26 demonstrated notable mycotoxin reduction capabilities, including 2 Bacillus pumilus and 24 Bacillus licheniformis strains. B. licheniformis strains MA572, MA695, MA696, TR174a, TR284, TR363, and TR466a degraded ENB to levels below the detection limit, and six strains reduced DON by 30-35%; B. licheniformis TR251b and TR374 showed the highest DON reduction with 35.7%. The most promising strains for bio-detoxification were B. licheniformis TR284, which achieved a 100% reduction in ENB and a 28.6% reduction in DON and B. licheniformis TR388 with a 97.5% reduction in ENB and a 31.9% reduction in DON. None of the tested LAB strains significantly reduced either mycotoxin. These findings highlight the promising potential of B. licheniformis strains in bio-detoxifying mycotoxin-contaminated cereal products. Further research into the underlying detoxification mechanisms and safety aspects is essential to develop effective bio-detoxification strategies for enhancing food safety.

Cation Dependence of Enniatin B/Membrane-Interactions Assessed Using Surface-Enhanced Infrared Absorption (SEIRA) Spectroscopy.

Enniatins are mycotoxins with well-known antibacterial, antifungal, antihelmintic and antiviral activity, which have recently come to attention as potential mitochondriotoxic anticancer agents. The cytotoxicity of enniatins is traced back to ionophoric properties, in which the cyclodepsipeptidic structure results in enniatin:cation-complexes of various stoichiometries proposed as membrane-active species. In this work, we employed a combination of surface-enhanced infrared absorption (SEIRA) spectroscopy, tethered bilayer lipid membranes (tBLMs) and density functional theory (DFT)-based computational spectroscopy to monitor the cation-dependence (Mz+=Na+, K+, Cs+, Li+, Mg2+, Ca2+) on the mechanism of enniatin B (EB) incorporation into membranes and identify the functionally relevant EBn : Mz+ complexes formed. We find that Na+ promotes a cooperative incorporation, modelled via an autocatalytic mechanism and mediated by a distorted 2 : 1-EB2 : Na+ complex. K+ (and Cs+) leads to a direct but less efficient insertion into membranes due to the adoption of "ideal" EB2 : K+ sandwich complexes. In contrast, the presence of Li+, Mg2+, and Ca2+ causes a (partial) extraction of EB from the membrane via the formation of "belted" 1 : 1-EB : Mz+ complexes, which screen the cationic charge less efficiently. Our results point to a relevance of the cation dependence for the transport into the malignant cells where the mitochondriotoxic anticancer activity is exerted.

Incidence of fungal contamination in fresh ginseng samples and mycotoxigenic potential of representative fungal isolates.

Fresh ginseng is typically accompanied by soil after harvest, leading to contamination with harmful fungi during storage and distribution. In this study, we investigated the incidence of fungal contamination in fresh ginseng (5-6 years old) purchased from 22 different stores in Geumsan, Korea. The incidence of fungal contamination in the samples was 67.4-111.5%. Fusarium solani was the most abundant species in the head (38.5%) and fine root (19.3%) parts of the ginseng samples, whereas F. oxysporum was the most abundant in the main root (22.0%) part. We isolated Aspergillus, Fusarium and Penicillium spp. (total number of isolates: 395) from the ginseng samples, and 138 isolates were identified using phylogenetic analysis. Polymerase chain reaction-based screening of 65 mycotoxin-producing species revealed that two P. expansum isolates were positive for citrinin and/or patulin, and five F. oxysporum isolates were positive for fumonisin biosynthesis gene. One P. expansum isolate produced 738.0 mg kg-1 patulin, and the other produced 10.4 mg kg-1 citrinin and 12.0 mg kg-1 patulin on potato dextrose agar (PDA) medium. Among the 47 representative F. oxysporum isolates, 43 (91.5%) produced beauvericin (0.1-15.4 mg kg-1) and four of them (8.5%) produced enniatin B and enniatin B1 (0.1-1.8 mg kg-1) as well. However, none of these toxins was detected in fresh ginseng samples. Fusarium solani and F. oxysporum were the most abundant species in fresh ginseng samples. Most F. oxysporum (43) and P. expansum (2) strains isolated from fresh ginseng produced beauvericin and enniatins (B and B1), and patulin or citrinin, respectively, on PDA medium. This is the first report of the mycotoxigenic potential of P. expansum and F. oxysporum strains isolated from fresh ginseng. © 2024 Society of Chemical Industry.

Emerging mycotoxin occurrence in chicken feed and eggs from Algeria

Poultry farming has developed into one of Algeria’s most productive industrial farming because of the growing demand for sources of protein among Algerian society. Laying hen feed consists mainly of cereals, which can be contaminated with molds and subsequently with their secondary metabolites known as mycotoxins. These later can pose a serious danger to the production and quality of eggs in the commercial layer industry. This work focuses on the detection of emerging mycotoxins, mainly enniatins (ENNs) and beauvericin (BEA), in poultry feed and eggs from different locations in Algeria. Two different QuEChERS-based extractions were established to extract ENNs and BEA from chicken feed and eggs. The determination of mycotoxin occurrence was achieved by a UHPLC-MS/MS method using 0.1% (v/v) formic acid in water and MeOH as mobile phase, an ESI interface operating in positive mode, and a triple quadrupole mass spectrometer operating in MRM for the detection. Matrix-matched calibration curves were carried out for both matrices, obtaining good linearity (R2 > 0.99). The method performance was assessed in terms of extraction recovery (from 87 to 107%), matrix effect (from − 47 to − 86%), precision (RSD < 15%), and limits of quantitation (≤ 1.1 µg/kg for feed and ≤ 0.8 µg/kg for eggs). The analysis of 10 chicken feed samples and 35 egg samples composed of a 10-egg pool each showed that ENN B1 was the most common mycotoxin (i.e., found in 9 feed samples) with contamination levels ranging from 3.6 to 41.5 µg/kg, while BEA was detected only in one feed sample (12 µg/kg). However, eggs were not found to be contaminated with any mycotoxin at the detection limit levels. Our findings indicate that the searched mycotoxins are present in traces in feed and absent in eggs. This can be explained by the application of a mycotoxin binder. However, this does not put a stop on the conduction of additional research and ultimately setting regulations to prevent the occurrence of emerging mycotoxins.

In Vitro Digestion and Intestinal Absorption of Mycotoxins Due to Exposure from Breakfast Cereals: Implications for Children's Health.

Breakfast cereals play a crucial role in children's diets, providing essential nutrients that are vital for their growth and development. Children are known to be more susceptible than adults to the harmful effects of food contaminants, with mycotoxins being a common concern in cereals. This study specifically investigated aflatoxin B1 (AFB1), enniatin B (ENNB), and sterigmatocystin (STG), three well-characterized mycotoxins found in cereals. The research aimed to address existing knowledge gaps by comprehensively evaluating the bioaccessibility and intestinal absorption of these three mycotoxins, both individually and in combination, when consumed with breakfast cereals and milk. The in vitro gastrointestinal method revealed patterns in the bioaccessibility of AFB1, ENNB, and STG. Overall, bioaccessibility increased as the food progressed from the stomach to the intestinal compartment, with the exception of ENNB, whose behavior differed depending on the type of milk. The ranking of overall bioaccessibility in different matrices was as follows: digested cereal > cereal with semi-skimmed milk > cereal with lactose-free milk > cereal with soy beverage. Bioaccessibility percentages varied considerably, ranging from 3.1% to 86.2% for AFB1, 1.5% to 59.3% for STG, and 0.6% to 98.2% for ENNB. Overall, the inclusion of milk in the ingested mixture had a greater impact on bioaccessibility compared to consuming the mycotoxins as a single compound or in combination. During intestinal transport, ENNB and STG exhibited the highest absorption rates when ingested together. This study highlights the importance of investigating the combined ingestion and transport of these mycotoxins to comprehensively assess their absorption and potential toxicity in humans, considering their frequent co-occurrence and the possibility of simultaneous exposure.

Investigating the individual and mixture cytotoxicity of co-occurring aflatoxin B1, enniatin B, and sterigmatocystin on gastric, intestinal, hepatic, and renal cellular models

This study investigates the individual and combined effects of the mycotoxins, Aflatoxin B1 (AFB1), Enniatin B (ENNB) and Sterigmatocystin (STG), on the cellular viability of gastric (NCI–N87), intestinal (Caco-2), hepatic (Hep-G2) and renal (Hek-293) cells, shedding light on synergistic or antagonistic effects using a constant ratio combination design proposed by Chou-Talalay. These toxins are prevalent in cereal-based foods, frequently consumed by children which raises concerns about their exposure to these mycotoxins. This population is particularly vulnerable to the effects of these toxins due to their underdeveloped organs and incompletely structured physiological processes. Results showed that ENB was the most toxic of the three mycotoxins across all cell lines, while STG and AFB1 showed lower toxicity. The combination of ENNB + STG was found to be the most potent in terms of binary mixtures. In regard to ternary combinations, Caco-2 cells are more sensitive to the tested mycotoxins, whereas NCI–N87 cells show lower levels of cell damage. Worrying dose reduction values (>10-fold) were found for ENNB in binary and ternary combinations at low exposure levels. These findings are significant for establishing initial reference values, which play a pivotal role in estimating reference doses that are subsequently incorporated into the broader risk assessment process.

Evaluating the human neurotoxicity and toxicological interactions impact of co-occurring regulated and emerging mycotoxins

Mycotoxins can inflict harmful effects on diverse organs, and mounting evidence indicates their potential involvement in human neurodegenerative diseases. Given the common occurrence of these toxins in food, there is an increasing demand for a comprehensive assessment of their combined toxicity to enhance our understanding of their potential hazards. This research investigates mycotoxin exposure from widely consumed cereal-based products, including enniatin B (ENNB), sterigmatocystin (STG), aflatoxin B1 (AFB1), cyclopiazonic acid (CPZ), citrinin (CIT), and ochratoxin A (OTA). Employing the median-effect equation based on Chou and Talalay's mass-action law, we assessed their cytotoxicity in human SH-SY5Y neuronal cells. Notably, ENNB displayed the highest neurotoxicity (IC50 = 3.72 µM), followed by OTA (9.10 µM) and STG (9.99 µM). The combination of OTA + STG exhibited the highest toxicity (IC50 = 3.77 µM), while CPZ + CIT showed the least detrimental effect. Approximately 70 % of tested binary combinations displayed synergistic or additive effects, except for ENNB + STG, ENNB + AFB1, and CPZ + CIT, which showed antagonistic interactions. Intriguingly, the senary combination displayed moderate antagonism at the lowest exposure and moderate synergism at higher doses. OTA exhibited predominantly synergistic interactions, comprising approximately 90 %, a noteworthy finding considering its prevalence in food. Conversely, ENNB interactions tended to be antagonistic. The most remarkable synergy occurred in the STG and CIT combination, enabling a 50-fold reduction in CIT dosage for an equivalent toxic effect. These findings highlight the biological relevance of robust synergistic interactions, emphasizing the need to assess human exposure hazards accurately, particularly considering frequent mycotoxin co-occurrence in environmental and food settings.

Effect of the mycotoxins enniatin B and deoxynivalenol on the wheat aphid Sitobion avenae and on the predatory lacewing Chrysoperla carnea.

Fusarium species are responsible for Fusarium head blight (FHB) in wheat, resulting in yield losses and mycotoxin contamination. Deoxynivalenol (DON) and enniatins (ENNs) are common mycotoxins produced by Fusarium, affecting plant, animal and human health. Although DON's effects have been widely studied, limited research has explored the impact of ENNs on insects. This study examines the influence of DON and enniatin B (ENB), both singularly and in combination, on the wheat aphid Sitobion avenae and one of its predators, the lacewing Chrysoperla carnea. When exposed to DON (100 mg L-1) or DON + ENB (100 mg L-1), S. avenae exhibited significantly increased mortality compared to the negative control. ENB (100 mg L-1) had no significant effect on aphid mortality. DON-treated aphids showed increasing mortality from 48 to 96 h. A dose-response relationship with DON revealed significant cumulative mortality starting at 25 mg L-1. By contrast, C. carnea larvae exposed to mycotoxins via cuticular application did not show significant differences in mortality when mycotoxins were dissolved in water but exhibited increased mortality with acetone-solubilized DON + ENB (100 mg L-1). Feeding C. carnea with aphids exposed to mycotoxins (indirect exposure) did not impact their survival or predatory activity. Additionally, the impact of mycotoxins on C. carnea was observed only with acetone-solubilized DON + ENB. These findings shed light on the complex interactions involving mycotoxins, aphids and their predators, offering valuable insights for integrated pest management strategies. Further research should explore broader ecological consequences of mycotoxin contamination in agroecosystems. © 2024 Society of Chemical Industry.

A tiered approach of hazard-prioritization and risk-ranking for chemical hazards in food commodities: Application for selected mycotoxins

Chemical hazards in foods, especially naturally occurring food contaminants like mycotoxins, are of serious public health concern. It is important to develop a practical framework to assess and rank health risks of chemical contaminants which can be further utilized by regulatory agencies to prioritize resources for risk assessment and management. In this study, a tiered hazard-prioritization and risk-ranking approach, which included two steps: exposure-based screening and margin of exposure (MOE)-based probabilistic risk ranking; was proposed to efficiently identify and rank chemicals of health concerns. Given the exposure-based hazard prioritization, chemicals with negligible or low health risks were first excluded. The remaining chemicals, imposing a higher health risk, were then ranked to facilitate risk-based decision making. The proposed approach was applied to identify and rank the mycotoxins with substantial health concerns in food commodities randomly sampled in China. A total of 19 mycotoxins were analyzed in 783 food commodities, including infant cookie, noodle, rice flour samples, wheat flour, millet, and rice. Results showed that the mycotoxins in infant foods with the highest health risk were Tenuazonic acid, Deoxynivalenol, and Enniatin B1, but as indicated by the probabilistic MOE estimation, the risks were still in the acceptable range and generally lower than the risks imposed by trace elements (e.g., Arsenic and Cadmium). The health risks of the other 16 mycotoxins were negligible mainly due to their low exposure levels. This study demonstrated that the proposed tiered approach was an efficient and effective tool to quantify and prioritize health risks in support of human health risk management.

Evaluation of distribution of emerging mycotoxins in human tissues: applications of dispersive liquid–liquid microextraction and liquid chromatography-mass spectrometry

In this work, a complete study of the distribution of emerging mycotoxins in the human body has been carried out. Specifically, the presence of enniatins (A, A1, B, B1) and beauvericin has been monitored in brain, lung, kidney, fat, liver, and heart samples. A unique methodology based on solid–liquid extraction (SLE) followed by dispersive liquid–liquid microextraction (DLLME) was proposed for the six different matrices. Mycotoxin isolation was performed by adding ultrapure water, acetonitrile, and sodium chloride to the tissue sample for SLE, while the DLLME step was performed using chloroform as extraction solvent. Subsequently, the analysis was carried out by high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC–MS/MS). The proposed method allowed limits of quantification (LOQs) to be obtained in a range of 0.001–0.150 ng g−1, depending on the tissue and mycotoxin. The precision was investigated intraday and interday, not exceeding of 9.8% of relative standard deviation. In addition, trueness studies achieved 75 to 115% at a mycotoxin concentration of 25 ng g−1 and from 82 to 118% at 5 ng g−1. The application of this methodology to 26 forensic autopsies demonstrated the bioaccumulation of emerging mycotoxins in the human body since all mycotoxins were detected in tissues. Enniatin B (ENNB) showed a high occurrence, being detected in 100% of liver (7 ± 13 ng g−1) and fat samples (0.2 ± 0.8 ng g−1). The lung had a high incidence of all emerging mycotoxins at low concentrations, while ENNB, ENNB1, and ENNA1 were not quantifiable in heart samples. Co-occurrence of mycotoxins was also investigated, and statistical tests were applied to evaluate the distribution of these mycotoxins in the human body.Graphical

Associated factors with mycotoxin exposure in Spanish population

Human exposure to mycotoxins is a global concern since filamentous fungi can contaminate food and feed from crops to ready-to-eat meals. Human urine biomonitoring is a widely used technique to evaluate mycotoxins exposure, as an alternative to food correlation studies. The aim of this study is to describe human exposure to mycotoxins and to investigate the associated sociodemographic, lifestyle and dietary variables. Participants were 540 women from the Valencia (Spain) cohort of the Spanish Childhood and Environment Project (INMA). A validated multi-mycotoxin method using HPLC-Q-TOF-MS was applied to determine the concentration of ten selected mycotoxins: Enniatin A, Enniatin B, Enniatin A1, Enniatin B1, Beauvericine, Aflatoxin B1, Aflatoxin B2, Aflatoxin G1, Aflatoxin G2 and Ochratoxin A. A simultaneous untargeted screening of mycotoxins and their metabolites has been performed. Mycotoxins associations were assessed by bivariate and multivariate regression models using participants' sociodemographic, lifestyle and dietary data collected through questionnaires. Mycotoxins were detected in 81% of urine samples. The method quantified mycotoxins concentrations in up to 151 samples. Most quantified mycotoxins were: Enniatin B [% of detection (concentration range)] = 26% (1.0–39.7 ng/mg) and Enniatin B1 = 7% (0.5–14.4 ng/mg). Besides the ten-targeted mycotoxins, other mycotoxins and metabolites were studied, and higher incidence was observed for Deepoxy-deoxynivalenol (45%), Ochratoxin B (18%) and Ochratoxin α (17%). Higher mycotoxins concentrations were associated with rural areas as well as with participants belonged to lower social class, beer, light sodas and fruit juice consumers. On the contrary, higher processed meat intake was related to lower mycotoxins’ levels. Studies are required to better evaluate the exposure to mycotoxins from food and their environmental relationships.

Evaluation of Immunomodulatory Effects of Fusarium Mycotoxins Using Bacterial Endotoxin-Stimulated Bovine Epithelial Cells and Macrophages in Co-Culture.

Mycotoxins are secondary metabolites produced by a variety of fungi that contaminate animal food and feeds and are capable of inducing a wide range of toxicities. Predictive in vitro models represent valuable substitutes for animal experiments to assess the toxicity of mycotoxins. The complexities of the interactions between epithelial and innate immune cells, vital for upholding barrier integrity and averting infections, remain inadequately understood. In the current study, a co-culture model of bovine epithelial cells (MAC-T) and macrophages (BoMac) was used to investigate the impact of exposure to Fusarium mycotoxins, namely deoxynivalenol (DON), zearalenone (ZEN), enniatin B (ENB), and beauvericin (BEA), on the inflammatory response elicited by the bacterial lipopolysaccharide (LPS) endotoxin. The MAC-T cells and BoMac were seeded on the apical side of a Transwell membrane and in the lower chamber, respectively, and mycotoxin exposure on the apical side of the membrane was carried out with the different mycotoxins (LC20; concentrations that elicited 20% cytotoxicity) for 48 h followed by an LPS immunity challenge for 24 h. The culture supernatants were collected from the basolateral compartment and these samples were submitted for cytokine/chemokine multiplex analysis. RNA-Seq analysis was performed using total RNA extracted from the MAC-T cells to acquire a more detailed insight into their cellular functions. The multiplex analysis indicated that IFN-γ, IL-1α, IL-8, and MCP-1 were significantly induced post-DON treatment when compared to control cells, and levels of IL-1α and IL-8 were enhanced significantly in all mycotoxin-treated groups post-LPS challenge. Analysis of the sequencing results showed that there were 341, 357, and 318 differentially expressed MAC-T cell genes that were up-regulated in the DON, ENB, and BEA groups, respectively. Gene ontology and pathway analysis revealed that these DEGs were significantly enriched in various biological processes and pathways related to inflammation, apoptosis signaling, and Wnt signaling. These results provide a comprehensive analysis of the co-culture cytokine/chemokine production and MAC-T cells' gene expression profiles elicited by Fusarium mycotoxins, which further contributes to the understanding of early endotoxemia post-mycotoxin exposure.

Determination of beauvercin and enniatins in rice flour and wheat flour by solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry

To develop a method for the determination of beauvercin(BEA), enniatin A(ENNA), enniatin A1(ENNA1), enniatin B(ENNB) and enniatin B1(ENNB1) in rice flour and wheat flour by ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). Samples were extracted by acetonitrile-water, purified by Oasis Prime HLB solid-phase extraction column. The sample solution was separated by waters BEH C_(18) column(2.1 mm×100 mm, 1.8 μm). The detection was performed in the electrospray positive ionization(ESI+) under multiple reaction monitoring(MRM) mode. The internal standard method and the matrix-matched calibrations were used for quantification. The linear relationships of BEA and 4 kinds of enniatins(ENNs) were good in the range of 0.1-50.0 ng/mL(r&gt;0.999). The average recoveries of BEA and ENNs in rice flour and wheat flour were 96.4%-105.4% and 99.1%-109.2%, with the relative standard deviations(RSD) of 1.01%-7.42% and 1.09%-9.69%(n=6). The detection limits(LOD) of BEA and ENNs were 0.03 μg/kg. The quantitative limits(LOQ) of BEA and ENNs were 0.1μg/kg. The matrix induced suppression or enhancement effect were 72.7%-99.3% and 60.8%-100.4%, respectively. The levels of emerging BEA and ENNs in wheat flour were higher than rice flour. The detection rate of enniatin B was highest in wheat flour and rice flour, the contents were 0.03-9.57 μg/kg and 0.03-0.56 μg/kg, the positive percentage were 98.5% and 36.4%. The method is quick, easy, accurate and sensitive, which is suitable for the determination of BEA and 4 kinds of ENNs in rice flour and wheat flour.

Pharmacokinetics and 28-day repeated-dose toxicity of enniatin B after oral administration in mice.

Enniatins are emerging mycotoxins that contaminate foods. The present study investigated the oral pharmacokinetics and 28-day repeated-dose oral toxicity of enniatin B (ENNB) in CD1 (ICR) mice. In the pharmacokinetic study, male mice received a single oral or intravenous dose of ENNB [30mg/kg body weight (BW) and 1mg/kg BW, respectively]. After oral dosing, ENNB exhibited 139.9% bioavailability, a 5.1-h elimination half-life, 5.26% fecal excretion from 4 to 24h post-dose, and upregulation of Cyp7a1, Cyp2a12, Cyp2b10, and Cyp26a1 in the liver 2h post-dosing. In the 28-day toxicity study, ENNB was administered to male and female mice by oral gavage at 0, 7.5, 15, and 30mg/kg BW/day. Females (7.5 and 30mg/kg) showed dose-unrelated decreased food consumption without accompanying changes in clinical parameters. Males (30mg/kg) showed low red blood cell counts and high blood urea nitrogen levels and absolute kidney weights; however, other related parameters including the histopathology of systemic organs/tissues were unchanged. These results suggest that ENNB may not induce toxicity after 28 days of oral administration in mice, despite high absorption. The no-observed-adverse-effect level of ENNB after 28 days of repeated oral doses was 30mg/kg BW/day for both sexes of mice.

Emerging mycotoxins induce hepatotoxicity in pigs’ precision-cut liver slices and HepG2 cells

Emerging mycotoxins are currently gaining more attention due to their high frequency of contamination in foods and grains. However, most data available in the literature are in vitro, with few in vivo results that prevent establishing their regulation. Beauvericin (BEA), enniatins (ENNs), emodin (EMO), apicidin (API) and aurofusarin (AFN) are emerging mycotoxins frequently found contaminating food and there is growing interest in studying their impact on the liver, a key organ in the metabolization of these components. We used an ex vivo model of precision-cut liver slices (PCLS) to verify morphological and transcriptional changes after acute exposure (4 h) to these mycotoxins. The human liver cell line HepG2 was used for comparison purposes. Most of the emerging mycotoxins were cytotoxic to the cells, except for AFN. In cells, BEA and ENNs were able to increase the expression of genes related to transcription factors, inflammation, and hepatic metabolism. In the explants, only ENN B1 led to significant changes in the morphology and expression of a few genes. Overall, our results demonstrate that BEA, ENNs, and API have the potential to be hepatotoxic.

Occurrence of pre- and postharvest multi-mycotoxins in durum wheat grains collected in 2020 and 2021 in two climatic regions of Tunisia

ABSTRACT Twenty two mycotoxins in 136 durum wheat collected from Tunisia in 2020 and 2021 were investigated. Mycotoxins were analyzed by UHPLCMS/MS. In 2020, 60.9% of the samples were contaminated with Aflatoxin B1 (AFB1) and/or enniatin. Whereas, in 2021, 34.4% were contaminated by enniatins. AFB1 was detected only in 2020, in the continental region (6/46) and all samples exceeded limits. AFB1 was detected in stored wheat (24-37.8 µg/kg) but also in pre-stored wheat (17-28.4 µg/kg) and in one sample collected in the field (21 µg/kg). Enniatin A1, enniatin B and enniatin B1 were detected in wheat collected in the field (30-7684 µg/kg), pre-storage (42-1266 µg/kg) and storage (65.8-498.2 µg/kg) from the continental region also, in sample collected in pre-storage (31.3-1410 µg/kg) and at harvest (48- 1060 µg/kg). Samples had a water activity less than 0.7 and moisture content ranged between 09-14%. AFB1 level represent a health risk to the Tunisian consumers.

Enniatin B1: Emerging Mycotoxin and Emerging Issues

Although over the last 10 years several studies have focused on the emerging mycotoxins known as enniatins (ENNs), there is still a lack of knowledge regarding their toxicological effects and the development of a correct risk assessment. This is especially true for enniatin B1 (ENN B1), considered the younger sister of the widely studied enniatin B (ENN B). ENN B1 has been found in several food commodities and, as with other mycotoxins, presents antibacterial and antifungal properties. On the other hand, ENN B1 has shown cytotoxic activity, impairment of the cell cycle, the induction of oxidative stress, and changes in mitochondrial membrane permeabilization, as well as negative genotoxic and estrogenic effects. Overall, considering the paucity of information available regarding ENN B1, further studies are necessary to perform a risk assessment. This review summarizes information on the biological characteristics and toxicological effects of ENN B1 as well as the future challenges that this mycotoxin could present.

Occurrence of Fusarium mycotoxins in freshly harvested highland barley (qingke) grains from Tibet, China.

Highland barley, also called "qingke" in Tibetan, is mainly cultivated in the Tibetan Plateau of China and has been used as a major staple food for Tibetans. Recently, Fusarium head blight (FHB) of qingke was frequently observed around the Brahmaputra River in Tibet. Considering the importance of qingke for Tibetans, the assessment of Fusarium mycotoxin contamination is essential for food safety. In this study, a total of 150 freshly harvested qingke grain samples were obtained from three regions around the Brahmaputra River in Tibet (China) in 2020. The samples were investigated for the occurrence of 20 Fusarium mycotoxins using high-performance liquid chromatography-tandem mass spectrometry (HPLC‒MS/MS). The most frequently occurring mycotoxin was enniatin B (ENB) (46%), followed by enniatin B1 (ENB1) (14.7%), zearalenone (ZEN) (6.0%), enniatin A1 (ENA1) (3.3%), enniatin A (ENA) (1.3%), beauvericin (BEA) (0.7%), and nivalenol (NIV) (0.7%). Due to the increase in altitude, the cumulative precipitation level and average temperature decreased from the downstream to the upstream of the Brahmaputra River; this directly correlated to the contamination level of ENB in qingke, which gradually decreased from downstream to upstream. In addition, the level of ENB in qingke obtained from qingke-rape rotation was significantly lower than that from qingke-wheat and qingke-qingke rotations (p < 0.05). These results disseminated the occurrence of Fusarium mycotoxins and provided further understanding of the effect of environmental factors and crop rotation on Fusarium mycotoxins.

Impact of Combined Processes Involving Ultrasound and Pulsed Electric Fields on ENNs, and OTA Mitigation of an Orange Juice-Milk Based Beverage.

In recent years, several innovative food processing technologies such as ultrasound (USN) and pulsed electric fields (PEF) have emerged in the market, showing a great potential both alone and in combination for the preservation of fresh and processed products. Recently, these technologies have also shown promising applications to reduce mycotoxin levels in food products. Therefore, the objective of this study is to investigate the potential of the combined treatments USN + PEF and PEF + USN on the reduction of Ochratoxin A (OTA) and Enniatins (ENNs) of an orange juice mixed with milk beverage. For this purpose, the beverages were elaborated in the laboratory and individually spiked with mycotoxins at a concentration of 100 µg/L. They were then treated by PEF (30 kV, 500 kJ/Kg) and USN (20 kHz, 100 W, at a maximum power for 30 min). Finally, mycotoxins were extracted using dispersive liquid-liquid microextraction (DLLME), and liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS-IT) was employed to determine them. The results showed promising applications, with reductions up to 50% for OTA and up to 47% for Enniatin B (ENNB) after the PEF + USN treatment combination. Lower reduction rates, up to 37%, were obtained with the USN + PEF combination. In conclusion, the combination of USN and PEF technologies could be a useful tool to reduce mycotoxins in fruit juices mixed with milk.