Search Results (62)

Zearalenone (ZEN), a non-steroidal estrogenic mycotoxin produced by Fusarium graminearum species, poses a significant threat to both human food safety and animal feed quality. In this study, we isolated a strain, designated as Bacillus licheniformis YJ25, from a contaminated moldy corn sample, demonstrating substantial effectiveness in removing ZEN. Our findings revealed that YJ25’s ZEN removal occurs primarily through cell wall adsorption, with enzymatic degradation representing a potential mechanism. In practical applications, enzymatic degradation may yield metabolites with heightened toxicity. However, liquid chromatography–mass spectrometry (LC–MS) analysis revealed that ZEN was not converted into α-/β-zearalenol (α-/β-ZEL) or α-/β-zearalanol (α-/β-ZAL) by YJ25, substantiating the safety profile of YJ25 in the removal of ZEN. Our mechanistic investigations revealed that the cell wall components peptidoglycan and teichoic acid serve as the primary binding sites for ZEN adsorption. Fourier-transform infrared spectroscopy (FTIR) analysis identified O-H, C-H, C=O, and C-O as the principal functional groups participating in the cell wall adsorption process. These investigations establish a scientific foundation for the prospective application of this strain as an efficient biological detoxification agent in food and feed safety management systems. Full article

Mycotoxins are frequent food contaminants posing health risk to humans and animals. Since these interactions have been barely studied yet, we examined the potential complex formation of mycotoxins with human α₁-acid glycoprotein (AGP) and with bovine milk proteins (including casein (CSN), β-lactoglobulin (LG), and α-lactalbumin (LA)) based on fluorescence spectroscopic and ultracentrifugation techniques. Only weak interactions (logK = 2.7 to 3.5) of certain mycotoxins were observed with CSN, LG, and/or LA. Ultracentrifugation experiments demonstrated that aflatoxin M1, zearalenone, and α-zearalenol form more stable complexes with CSN than with LG or LA. These mycotoxins bound to bovine serum albumin with more than a tenfold higher affinity compared to CSN; nevertheless, it has likely limited importance due to the relatively low levels of BSA in bovine milk. Zearalenone, zearalenols, and sterigmatocystin showed strong interactions with AGP (logK = 5.5 to 6.4), suggesting that AGP may play an important role in the plasma protein binding of these mycotoxins. Full article

Metarhizium fungi, essential for ecosystem function and commonly utilised in pest control, often occupy ecological niches contaminated by toxic compounds of both anthropogenic and microbiological origin. The present study reveals the potential of Metarhizium anisopliae for biodegradation of the Fusarium mycotoxin zearalenone (ZEN), a common contaminant of crops that poses a significant threat to human and animal health due to its oestrogenic potential and toxicity. A key aspect of the pathway described is the degradation of ZEN by cleaving the lactone bond, which results in a significant reduction in mycotoxin toxicity, highlighting the fungus’s bioremediation potential. Furthermore, this study provides the first evidence of subsequent degradation of ZEN metabolites through progressive shortening of the aliphatic chain, primarily via alternating oxidation and demethylation, ultimately yielding trihydroxybenzene. Significantly, lactone bond cleavage occurred not only in ZEN itself but also in its reduced forms, the zearalanols, formed through the initial reduction of ZEN to zearalenols. Elevated mRNA levels of cytochrome P450 (CYP450) monooxygenases in M. anisopliae exposed to ZEN indicate their significant involvement in degradation mechanisms. Intriguingly, the inhibition of CYP450 activity resulted in a substantial shift in the quantitative ratio of α- and β-epimers of zearalenols and zearalanols. The observed alteration towards β-form production likely stems from the inhibition of other CYP450-dependent reactions, indirectly influencing ZEN reduction pathways—a particularly noteworthy finding. These insights are crucial for developing strategies to utilise M. anisopliae in the bioremediation of ZEN-contaminated areas. Full article

The degradation of zearalenone (ZEN) in the rumen of dairy cows is influenced by rumen pH, which is a key factor affecting this process. The aim of this study was to investigate the variation of ZEN in interaction with other mycotoxins at different ruminal pH environments (physiological (pH 6.5) and acidic (pH 5.5)) using an in vitro rumen model. Rumen fluid was collected from the caudoventral part of the rumen of cows using a pharyngeal–esophageal probe. To determine the changes in different mycotoxins (ZEN; AFLB₁; DON; T-2) in the rumen of cows, a model rumen system was used, and mycotoxins concentrations were detected by HPLC. The study found that at pH 6.5, ZEN alone and in combination with other mycotoxins (DON; T-2; AFLB₁) significantly (p < 0.05) reduced ZEN levels compared to the rumen environment at pH 5.5. It was observed that α-zearalenol (α-ZEL) and β-zearalenol (β-ZEL) concentrations were generally higher at a rumen pH of 6.5 compared to pH 5.5, averaging 47.09 µg/L and 35.23 µg/L, respectively. Additionally, the frequency of detection for both α-ZEL and β-ZEL was greater at pH 6.5 than at pH 5.5. A comparison of α-ZEL concentrations in rumen samples at pH 5.5 showed a 20% increase from the 6th to the 9th hour of the test, while β-ZEL levels remained unchanged over the same period. Full article

Mycotoxins are toxic fungi secondary metabolites that develop on feedstuffs and can be transferred into milk, thus representing a public health risk. The objective of this study was to assess the co-occurrence of mycotoxins in the diet and in the milk of dairy cows from the southeast region of Brazil. Samples of total mixed ration (TMR, n = 70) and milk (n = 70) were collected in dairy farms and subjected to multi-mycotoxin analysis using liquid chromatography coupled to tandem mass spectrometry. The aflatoxins (AFs), ochratoxin A (OTA), and T-2 and HT-2 toxins were not detected in TMR samples. In contrast, fumonisins (FBs), zearalenone (ZEN), and deoxynivalenol (DON) were detected in 100, 93, and 24% of TMR samples at mean levels of 336.7 ± 36.98, 80.32 ± 16.06 µg/kg and 292.1 ± 85.68 µg/kg, respectively. Ninety-two percent of TMR samples exhibited co-occurring mycotoxins. In milk, 54% of samples (n = 38) had detectable levels of mycotoxin, while 43% (n = 30) contained two or more types of mycotoxins. DON, FB, and ZEN metabolites (α-zearalenol and β-zearalenol) were the most frequent mycotoxins detected in the milk samples analyzed, at mean concentrations of 0.562 ± 0.112, 2.135 ± 0.296 µg/kg, 2.472 ± 0.436 µg/kg, and 0.343 ± 0.062 µg/kg, respectively. However, none of the analyzed milk samples had levels higher than the maximum permitted limit for AFM₁ in Brazil (0.5 µg/L). The results of this trial highlight the concern about the co-occurrence of multiple mycotoxins in TMR and in milk, due to the possible additive or synergistic effects of these compounds. The presence of co-occurring mycotoxins in milk underscores the need for stringent preventive practices to avoid mycotoxin contamination in the diet of dairy cows in Brazil. Full article

Zearalenone (ZEN) is a toxic secondary metabolite produced by the Fusarium fungi, which widely contaminates grains, food, and feed, causing health hazards for humans and animals. Therefore, it is essential to find effective ZEN detoxification methods. Enzymatic degradation of ZEN is believed to be an eco-friendly detoxification strategy, specifically thermostable ZEN degradation enzymes are needed in the food and feed industry. In this study, a novel ZEN lactone hydrolase ZHRnZ from Rosellinia necatrix was discovered using bioinformatic and molecular docking technology. The recombinant ZHRnZ showed the best activity at pH 9.0 and 45 °C with more than 90% degradation for ZEN, α-zearalenol (α-ZOL), β-zearalenol (β-ZOL) and α-zearalanol (α-ZAL) after incubation for 15 min. We obtained 10 mutants with improved thermostability by single point mutation technology. Among them, mutants E122Q and E122R showed the best performance, which retained more than 30% of their initial activity at 50 °C for 2 min, and approximately 10% of their initial activity at 60 °C for 1 min. The enzymatic kinetic study showed that the catalytic efficiency of E122R was 1.3 times higher than that of the wild-type (WT). Comprehensive consideration suggests that mutant E122R is a promising hydrolase to detoxify ZEN in food and feed. Full article

Mycotoxins are toxic secondary metabolites produced by various fungi that can contaminate food crops, which, in turn, may lead to human exposure. Chronic exposure to mycotoxins can cause adverse health effects including reproductive and developmental toxicity. Pregnant women and their foetuses present a vulnerable group for exposure to mycotoxins that can cross the placenta. Human biomonitoring of mycotoxins provides a real-life approach to estimate internal exposure. In this pilot study, 24-h urine samples from 36 pregnant Dutch women were analysed for aflatoxin M1 (AFM1), total deoxynivalenol (DON), de-epoxy-deoxynivalenol (DOM-1), total zearalenone (ZEN), total α-zearalenol (α-ZEL), total β-zearalenol (β-ZEL) and total zearalanone (ZAN), where ‘total’ refers to mycotoxins and their conjugated forms. Serum samples from these women were analysed for fumonisin B1 (FB1) and ochratoxin A (OTA). All samples were measured using liquid chromatography–tandem mass spectrometry (LC-MS/MS). The most prevalent mycotoxins were total DON, total ZEN and OTA, with a detection frequency of 100%. DOM-1, total α-ZEL and total β-ZEL were detected but to a lesser extent, while AFM1, total ZAN and FB1 were undetected. Median concentrations were 4.75 μg total DON/L, 0.0350 μg DOM-1/L, 0.0413 μg total ZEN/L, 0.0379 μg total α-ZEL/L, 0.0189 μg total β-ZEL/L, and 0.121 μg OTA/L. The calculated median concentration for total ZEN and its metabolites was 0.105 μg/L. Based on two separate risk assessment approaches, total DON exposure in this group was considered to be of low concern. Similarly, exposure to total ZEN and its metabolites in this group was of low concern. For OTA, the risk of non-neoplastic effects was of low concern based on exposure in this group, and the risk of neoplastic effects was of low concern in the majority of participants in this group. The findings of this pilot study confirm the presence of mycotoxins in the urine and serum of pregnant Dutch women, with total DON, total ZEN, and OTA most frequently detected. Exposure to all measured mycotoxins was considered to be of low concern in this group, except for exposure to OTA, which was of low concern for the majority of participants. The study’s findings offer valuable insights but should be confirmed using a larger and more diverse sample of the Dutch general population. Full article

Zearalenone (ZEN) is a mycotoxin produced by various Fusarium strains, that is present in food and feed raw materials worldwide, causing toxicity effects in animals and humans. This research aimed to explore the toxicokinetics of ZEN on female Dezhou donkeys following a single oral exposure dosage of 2 mg/kg BW (body weight). The sample collection of donkeys plasma was carried out at 0, 5, 10, 15, 20, 30, 45, 60, 90 min, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 6 h, 9 h, 12 h, 24 h, 48 h, 72 h, 96 h and 120 h via intravenous catheter, and fecal and urinary samples were severally collected at 0 h and every 6 h until 120 h. The concentrations of ZEN, α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL), zearalanone (ZAN) in plasma, urine, and feces were detected by UPLC-MS/MS. Only ZEN was detected in plasma, and the maximum was 15.34 ± 5.12 µg/L occurred at 0.48 h after gavage. The total plasma clearance (Cl) of ZEN was 95.20 ± 8.01 L·kg·BW⁻¹·h⁻¹. In addition, the volume of distribution (Vd) was up to 216.17 ± 58.71 L/kg. The percentage of total ZEN (ZEN plus the main metabolites) excretion in feces and urine was 2.49% and 2.10%, respectively. In summary, ZEN was fast absorbed and relatively slowly excreted in female donkeys during 120 h after a single gavage, indicating a trend of wider tissue distribution and longer tissue persistence. Full article

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by the fungi of the Fusarium genera, and is a contaminant of cereals and plant products. ZEN and its metabolites are considered endocrine disruptors, and could have various toxic effects on animals and humans. In recent years, there has been a significant demographic increase in wild boar (Sus scrofa) in many mountainous and hilly areas of Italy, including the Campania region, mainly due to global climate change. The wild boar can be defined as a generalist and omnivorous species capable of varying its diet; therefore, it can play a role as an environmental bioindicator towards contaminants such as mycotoxins. This study was conducted to evaluate, for the first time, the concentrations of ZEN and its metabolites in the liver, kidney, and muscle of 82 wild boars shot in their habitat by hunters with hunting permits in different localities of Avellino province (Campania region, Southern Italy) from 2021 to 2022. The samples were collected and analyzed with an SPE clean-up and high-pressure liquid chromatography method with fluorescence detection. The results indicated that ZEN and α-Zearalenol were present in most of the samples, suggesting that a plan to monitor these mycoestrogens is essential to achieve the goals of “One Health”. Full article

Fridericia chica (Bignoniaceae) is a Colombian Caribbean plant with numerous health benefits, including properties such as wound healing, immune system stimulation, and antioxidant capacity, among others. Mycotoxins alpha-zearalenol (α-ZEL) and beta-zearalenol (β-ZEL) are phase I metabolites of zearalenone, a natural product involved in endocrine disruption and cell proliferation processes. This study aimed to investigate the cytotoxic potential of the hydroethanolic extract of F. chica leaves (HEFc) and determine their protective effects against proliferation induced by α-ZEL and β-ZEL on human hepatoma HepG2, lung cancer Calu-1, and primary normal human epidermal keratinocytes, neonatal (HEKn). The cytotoxicity of HEFc was measured in a range from 4 to 1000 µg/mL and from 0.4 to 100 μM for both α-ZEL and β-ZEL. Cell production of intracellular ROS was monitored using the H₂-DCFDA probe. The cells exposed to HEFc presented IC₅₀ of 128, 249, and 602 µg/mL for the HepG2, Calu-1, and HEKn cells, respectively. A greater selectivity was seen in HepG2 cells [selectivity index (SI) = 3.5] than in Calu-1 cells (SI = 2.4). Cells treated with mycotoxins remained viable during the first day, and cell proliferation increased at low tested concentrations (0.4-6.3 µM) in all three cell lines. However, after 48 h treatment, cells exposed to 50 and 100 µM of α-ZEL and β-ZEL displayed decreased viability. HEFc at 16 µg/mL was able to give some protection against cytotoxicity induced by high concentrations of β-ZEL in HepG2, reducing also cell proliferation elicited at low levels of α-ZEL and β-ZEL. ROS production was not observed in cells treated with this HEFc concentration; however, it prevented ROS formation induced by treatment with 50 µM α-ZEL or β-ZEL. In summary, HEFc isolated from plants grown in northern Colombia displayed promising results against cell proliferation and oxidative stress caused by mycotoxins. Full article

This study evaluated the ability of selected strains of Trichoderma viride, T. viridescens, and T. atroviride to inhibit mycelium growth and the biosynthesis of mycotoxins deoxynivalenol (DON), nivalenol (NIV), zearalenone (ZEN), α-(α-ZOL) and β-zearalenol (β-ZOL) by selected strains of Fusarium culmorum and F. cerealis. For this purpose, an in vitro experiment was carried out on solid substrates (PDA and rice). After 5 days of co-culture, it was found that all Trichoderma strains used in the experiment significantly inhibited the growth of Fusarium mycelium. Qualitative assessment of pathogen–antagonist interactions showed that Trichoderma colonized 75% to 100% of the medium surface (depending on the species and strain of the antagonist and the pathogen) and was also able to grow over the mycelium of the pathogen and sporulate. The rate of inhibition of Fusarium mycelium growth by Trichoderma ranged from approximately 24% to 66%. When Fusarium and Trichoderma were co-cultured on rice, Trichoderma strains were found to inhibit DON biosynthesis by about 73% to 98%, NIV by about 87% to 100%, and ZEN by about 12% to 100%, depending on the pathogen and antagonist strain. A glycosylated form of DON was detected in the co-culture of F. culmorum and Trichoderma, whereas it was absent in cultures of the pathogen alone, thus suggesting that Trichoderma is able to glycosylate DON. The results also suggest that a strain of T. viride is able to convert ZEN into its hydroxylated derivative, β-ZOL. Full article

This study was undertaken to analyze whether prolonged exposure to low-dose zearalenone (ZEN) mycotoxicosis affects the concentrations of ZEN, α-zearalenol (α-ZEL), and β-zearalenol (β-ZEL) in selected reproductive system tissues (ovaries, uterine horn—ovarian and uterine sections, and the middle part of the cervix), the hypothalamus, and pituitary gland, or the concentrations of selected steroid hormones in pre-pubertal gilts. For 42 days, gilts were administered per os different ZEN doses (MABEL dose [5 µg/kg BW], the highest NOAEL dose [10 µg/kg BW], and the lowest LOAEL dose [15 µg/kg BW]). Tissue samples were collected on days seven, twenty-one, and forty-two of exposure to ZEN (exposure days D1, D2, and D3, respectively). Blood for the analyses of estradiol and progesterone concentrations was collected in vivo on six dates at seven-day intervals (on analytical dates D1–D6). The analyses revealed that both ZEN and its metabolites were accumulated in the examined tissues. On successive analytical dates, the rate of mycotoxin accumulation in the studied tissues decreased gradually by 50% and proportionally to the administered ZEN dose. A hierarchical visualization revealed that values of the carry-over factor (CF) were highest on exposure day D2. In most groups and on most exposure days, the highest CF values were found in the middle part of the cervix, followed by the ovaries, both sections of the uterine horn, and the hypothalamus. These results suggest that ZEN, α-ZEL, and β-ZEL were deposited in all analyzed tissues despite exposure to very low ZEN doses. The presence of these undesirable compounds in the examined tissues can inhibit the somatic development of the reproductive system and compromise neuroendocrine coordination of reproductive competence in pre-pubertal gilts. Full article

Zearalenone (ZON), zearalanone (ZAN) and their phase I metabolites: α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), α-zearalalanol (α-ZAL) and β-zearalalanol (β-ZAL) are compounds with estrogenic activity that are metabolized and distributed by the circulatory system in animals and can access the food chain through meat products from livestock. Furthermore, biomonitoring of zearalenones in biological matrices can provide useful information to directly assess mycotoxin exposure; therefore, their metabolites may be suitable biomarkers. The aim of this study was to determine the presence of ZON, ZAN and their metabolites in alternative biological matrices, such as liver, from three different animals: chicken, pig and lamb, in order to evaluate their exposure. A solid–liquid extraction procedure coupled to a GC-MS/MS analysis was performed. The results showed that 69% of the samples were contaminated with at least one mycotoxin or metabolite at varying levels. The highest value (max. 152.62 ng/g of β-ZOL) observed, and the most contaminated livers (42%), were the chicken liver samples. However, pig liver samples presented a high incidence of ZAN (33%) and lamb liver samples presented a high incidence of α-ZOL (40%). The values indicate that there is exposure to these mycotoxins and, although the values are low (ranged to 0.11–152.6 ng/g for α-ZOL and β-ZOL, respectively), analysis and continuous monitoring are necessary to avoid exceeding the regulatory limits and to control the presence of these mycotoxins in order to protect animal and human health. Full article

As one of the most prevalent estrogenic mycotoxins in cereals and animal feed, zearalenone (ZEN) can cause serious reproductive disorders. ZEN control in food and feed commodities has been an imperative area of research. In this study, 87 lactic acid bacteria (LAB) were isolated from pickles and their ZEN (5 mg/L) removal abilities ranged from 0% to 68.4%. Then, five strains with potent ZEN removal ability (>50%) were identified: Lactobacillus plantarum 22, L. plantarum 37, L. plantarum 47, L. paracasei 85, and L. buchneri 93. Under optimization conditions (48 h, pH 4.0, 37 °C, and 5 mg/L), the highest ZEN removal abilities of L. paracasei 85 and L. buchneri 93 reached 77.7% and 72.8%, respectively. Moreover, the two lactic acid bacteria decreased the toxicity of ZEN, because the levels of β-zearalenol (β-ZOL) transformed from ZEN were more than two-fold higher than α-zearalenol (α-ZOL). Additionally, cell free supernatant and pellet biotransformation of ZEN to α-ZOL and β-ZOL in LAB were detected for the first time. Furthermore, chemical and enzymatical treatments combined with Fourier-transform infrared spectroscopy analysis indicated that exopolysaccharides, proteins, and lipids on the cell wall could bond to ZEN through hydrophobic interactions. Scanning electron microscopy indicated that cell structure damage occurred during the ZEN clearance to L. buchneri 93, but it did not with L. paracasei 85. In addition, various organic acids, alcohols, and esters of the two LAB participated in ZEN removal. Hence, L. paracasei 85 and L. buchneri 93 can be considered as potential detoxification agents for ZEN removal for food and feedstuff. Full article

In this study, a gas chromatography–mass spectrometry (GC-MS) method was established for the determination of zearalenone and its five derivatives in feed, including zearalanone, α-zearalanol, β-zearalanol, α-zearalenol, and β-zearalenol. An effective immunoaffinity column was prepared for sample purification, which was followed by the silane derivatization of the eluate after an immunoaffinity chromatography analysis for target compounds by GC-MS. Matrix effects were corrected by an isotope internal standard of zearalenone in this method. The six analytes had a good linear relationship in the range of 2–500 ng/mL, and the correlation coefficients were all greater than 0.99. The limits of detection (LODs) and limits of quantification (LOQs) were less than 1.5 μg/kg and 5.0 μg/kg, respectively. The average spike recoveries for the six feed matrices ranged from 89.6% to 112.3% with relative standard deviations (RSDs) less than 12.6%. Twenty actual feed samples were analyzed using the established method, and at least one target was detected. The established GC-MS method was proven to be reliable and suitable for the determination of zearalenone and its derivatives in feed. Full article