Zearalenone Research Articles

Phosphorylation of Zearalenone Retains Its Toxicity.

Microbial biotransformation of Zearalenone (ZEN) is a promising deactivation approach. The residual toxicity and stability of Zearalenone-14-phosphate (ZEN-14-P) and Zearalenone-16-phosphate (ZEN-16-P), two novel microbial phosphorylation products of ZEN, remain unknown. We investigated the cytotoxicity, oxidative stress, proinflammatory, and estrogenic activity of phosphorylated ZENs using porcine intestinal cells, uterine explants, and human endometrial cells and traced their metabolic fate by liquid chromatography-tandem mass spectrometry (LC-MS)/MS analysis. The phosphorylated ZENs significantly decreased the viability of the IPEC-J2 and Ishikawa cells. Similar to ZEN, phosphorylation products induced significant oxidative stress, activated the expression of proinflammatory cytokines, and demonstrated estrogenic activity through upregulation of estrogen-responsive genes, activation of alkaline phosphatase, and proliferation of endometrial glands. LC-MS/MS analysis pointed out that although phosphorylated ZENs are partially hydrolyzed to ZEN, their respective metabolic pathways differ. We conclude that phosphorylation might not be sufficient to detoxify ZEN, leaving its cytotoxic, proinflammatory, and estrogenic properties intact.

Co-Occurrence of Mycotoxins in the Diet and in the Milk of Dairy Cows from the Southeast Region of Brazil

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.

Mycotoxin Challenge in Dairy Cows: Assessment of the Efficacy of an Anti-Mycotoxin Agent by Adopting an In Vitro Rumen Simulation Method

To protect ruminants from the harmful effects of mycotoxins, anti-mycotoxin agents can be added to the dietary ration, thus guaranteeing animal health and production. Therefore, the objective of this study was to evaluate the in vitro ruminal initial sequestration (weak binding) and subsequent desorption (strong binding) of an anti-mycotoxin agent based on a mixture of adsorbing material, turmeric and milk thistle extracts and yeast-based components to adsorb or bio-convert aflatoxins (AF), fumonisins B1 and B2 (FB), trichothecene deoxynivalenol (DON), T-2 and HT-2 toxins, and zearalenone (ZEN). Two doses were tested: Dose 1 simulated 30 mg/cow/d, while Dose 2 simulated 90 mg/cow/d of the anti-mycotoxin agent. Each treatment involved three analytical replicates at each of three incubation times (1, 4, and 24 h post-incubation), with two independent experimental runs providing experimental replicates. Analytical methods, including UHPLC-HRMS and multivariate analyses, were used to both quantify mycotoxin concentrations and reveal dose-dependent reductions, with statistical validations indicating significant changes in mycotoxin levels across both dose and time. The results indicated that the anti-mycotoxin agent was able to highly bind AFB1, T2, and HT-2 toxins since its concentration was always under the limit of detection (<1 ppb). Regarding ZEN (weak binding mean: 94.6%; strong binding mean: 62.4%) and FBs (weak binding mean: 58.7%; strong binding mean: 32.3%), orthogonal contrasts indicated that the anti-mycotoxin agent was able to effectively bind these toxins using Dose 1 (p < 0.05). This finding suggests that Dose 1 may be sufficient to achieve the targeted effect and that a further increase does not significantly improve the outcome. Regarding DON, a strong linear relationship was observed between dose and adsorption. However, the complex interactions between the mycotoxin, the ruminal environment, and the anti-mycotoxin agent made it difficult to establish a clear dose–effect relationship (p > 0.10). UHPLC-HRMS analysis identified over 1500 mass features in rumen samples, which were further analyzed to assess the effects of the anti-mycotoxin agent. Hierarchical clustering analysis (HCA) revealed significant changes in the untargeted metabolomic profiles of samples treated with mycotoxins compared to control samples, particularly after 24 h with the anti-mycotoxin treatments. Clear differences were noted between strong binding and weak binding samples. Further analysis using orthogonal partial least squares discriminant analysis (OPLS-DA) highlighted distinct metabolomic profiles, with stronger predictive ability in the strong binding group (Q2 cumulative value of 0.57) compared to the weak binding group (0.30). The analysis identified 44 discriminant compounds in the strong binding model and 16 in the weak binding model. Seven compounds were common to both groups, while silibinin, known for its antioxidant and anti-inflammatory properties, was found among the unique compounds in the weak binding group. Overall, the findings suggest that both doses of the anti-mycotoxin agent significantly influenced the chemical profiles in the rumen, particularly enhancing the binding of mycotoxins, thereby supporting the role of phytogenic extracts in mitigating mycotoxin effects.

Carvacrol modulates antioxidant enzymes, DNA integrity, and apoptotic markers in zearalenone-exposed fetal rat liver

Maternal exposure to zearalenone (ZEA), a mycotoxin, can impact fetal liver development. This study investigated the protective effects of carvacrol (CRV) against ZEA-induced fetal liver damage. Thirty-two pregnant rats were allocated to four groups (eight rats/group); control, CRV (75 mg/kg), ZEA (5 mg/kg), and co-treated group (ZEA + CRV). The animals were given their doses during the gestation period. Maternal exposure to ZEA revealed a significant increase in the malondialdehyde (MDA) level in the fetal liver. In contrast, glutathione S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) activities, besides glutathione (GSH) levels, were decreased in ZEA-intoxicated rats. Additionally, ZEA increased the expression of pro-apoptotic genes (P53, Bax, and caspase-9), elevated the immunoreactivity of caspase-3, decreased anti-apoptotic Bcl-2, and induced severe fatty degeneration, congestion, and necrosis in the fetal liver. The comet assays revealed significant DNA damage, as evidenced by reduced head DNA content and increased tail DNA content and tail moment in the ZEA-exposed rats. Surprisingly, co-treatment with CRV significantly mitigated fetal hepatic lipid peroxidation, antioxidant disturbance, apoptosis, and DNA damage after maternal exposure to ZEA. These findings highlight the potential of CRV as a promising approach to mitigate ZEA-associated developmental hepatotoxicity.

Reversing Zearalenone Toxicity: The Role of Hydroxytyrosol in Zebrafish.

Zearalenone (ZEA) is a widely distributed mycotoxin that presents a substantial worldwide health risk to animals. Several natural compounds have shown promise in mitigating the detrimental impacts of ZEA. This study examined the detoxification potential of previously identified compounds by utilizing zebrafish embryos as a model organism. Hydroxytyrosol stands out among these natural compounds. Our findings indicate that hydroxytyrosol effectively mitigated mortality, hatching delay, and phenotypic abnormalities induced by ZEA in the assessed embryos. Furthermore, hydroxytyrosol restored the frequency and intensity of tail coiling (TC) while decreasing the expression of heat shock proteins (HSPs) in the zebrafish embryos. Extended incubation with hydroxytyrosol demonstrated protective effects on zebrafish growth and morphology, muscle birefringence, and touch-evoked escape behavior. Subsequent investigations indicated that hydroxytyrosol reversed the expression of proapoptotic targets (e.g., bax and caspase8) and cell cycle regulators (e.g., p21, gadd45a, and rbl2), thereby mitigating apoptosis and G2 cell cycle arrest induced by ZEA in zebrafish embryos. Additionally, hydroxytyrosol decreased staining for senescence associated-β-galactosidase (SA-β-Gal). Notably, p53/FoxO pathway plays an important role in detoxification mechanisms. Overall, these novel findings highlight the potential of hydroxytyrosol to reverse ZEA-induced toxicity in multiple aspects. The mitigating effect of hydroxytyrosol on ZEA toxicity may have been underestimated.

Preparation and characterization of aptamer-based sorbent for the selective extraction of zearalenone and its derivatives from human urine.

The aim of this work is the development of a biomimetic strategy involving a molecular recognition mechanism using aptamers immobilized on a solid support for the analysis of the mycotoxin zearalenone (ZEA) and two of its derivatives in human urine: alpha-zearelenol (α-ZEL) and beta-zearelenol (β-ZEL). Three oligonucleotide sequences reported in theliterature as beingspecific to ZEA were thus covalently grafted onto activated sepharose, and a thorough study of the percolation and washing conditions was performed to promote the selective retention of the three targeted compounds. With the optimized extraction procedure, a strong and selective retention was obtained for ZEA and to a lesser extent α-ZEL and β-ZEL, with extraction recoveries of 88±9%, 77±15%, and 45±12% respectively, in standard solutions. Application of this procedure to spiked human urine strongly highlighted the efficiency of the clean-up effect resulting from the use of this selective sorbent. Limits of quantification of the whole analytical procedure including extraction on oligosorbent andLC-MS analysis were 0.18 and 0.24ngmL-1, for ZEA and α-ZEL, respectively, thus demonstrating clearly the potential of the developed method for monitoring human dietary exposure to these compounds.

The High-Efficiency Degradation of Multiple Mycotoxins by Lac-W Laccase in the Presence of Mediators

Mycotoxin cocontamination is a severe threat to health and economic security worldwide. The mycotoxins aflatoxin B1 (AFB1), zearalenone (ZEN), deoxynivalenol, T-2 toxin, fumonisin B1, and ochratoxin A are of particular concern due to their substantial toxicity. Lac-W is a laccase with the unique property of degrading these six mycotoxins in the absence of redox mediators. Nevertheless, their degradation rates are low. This work aims to improve the ability of Lac-W to degrade these six mycotoxins and to elucidate its detoxification mechanism. Including redox mediators increased the Lac-W degradation efficiency drastically, and completely degraded AFB1 and ZEN within one hour. Additionally, Lac-W-AS has good temperature, pH, and ions adaptability in ZEN degradation. Lac-W-AS reduced the ZEN toxicity because ZEN degradation products significantly restored the bioluminescence intensity of Vibrio fischeri. A Lac-W-AS-mediated oxidation product of ZEN was structurally characterized as 15-OH-ZEN by UHPLC-MS/MS. Linear sweep voltammetry showed that AS affected the potential of Lac-W and accelerated the oxidation of ZEN. Finally, the combination of mediators (acetosyringone and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate)) improved the degradation rate of mycotoxins. This work highlights that the combination of Lac-W with mediators serves as a good candidate for degrading multi-mycotoxin contaminants in food and feedstuff.

Visible light-activated signal amplification PEC aptasensor for ultrasensitive zearalenone detection based on double Z-type BiOI/g-C3N4@Au/Bi2S3 heterojunctions

Zearalenone (ZEN), also known as F-2 toxin, can enter the body through the food chain and accumulate, leading to DNA break and an abnormal number of chromosomes. Therefore, it is crucial to develop a facile and sensitive method for detecting ZEN. Herein, a visible light-activated signal amplification photoelectrochemical (PEC) aptasensor for ultrasensitive detection of ZEN has been successfully constructed, exploiting the synergistic effect of the Z-type BiOI/g-C3N4/Au heterojunctions, Bi2S3 as a signal enhancer, and the remarkable affinity and specificity of aptamers. The PEC properties and electron transfer mechanism of the double Z-type heterojunctions were thoroughly investigated. When the aptamers captured the ZEN molecules, Bi2S3 and cDNA departed from the electrode, resulting in a reduction in the photocurrent signal. Therefore, the concentration of ZEN can be quantitatively analyzed by measuring the change in the photocurrent response. The constructed PEC aptasensor exhibited exceptional sensitivity and selectivity for ZEN in the concentration range of 0.1 fg⋅mL−1 ––1.0 ng⋅mL−1, with a detection limit of 0.087 fg⋅mL−1 (S/N = 3). Finally, the PEC aptasensor was successfully applied to detect ZEN in actual coix seed samples, providing some guidance for the development of PEC aptasensors for practical medicine monitoring.

Individual and combined occurrences of the prevalent mycotoxins in commercial feline and canine food.

Mycotoxins, such as aflatoxin B1 (AFB1), deoxynivalenol (DON), fumonisins (FBs), ochratoxin A (OTA), T-2 toxin (T-2), and zearalenone (ZEN), can contaminate animal feeds and pose risks to animal health and production performance. These mycotoxins are commonly found in cereals and grains, with the increased use of cereals in pet food, there is a rising concern about mycotoxin contamination among pet owners. To address this, we analyzed imported brands of feline and canine food from the Chinese market produced in 2021-2022. Ninety-three samples were analyzed, comprising 45 feline food and 48 canine food samples. Among them, 14 were canned food and 79 were dry food. The results indicate that AFB1, DON, FBs, OTA, T-2, and ZEN occurred in 32.26%, 98.92%, 22.58%, 73.12%, 55.91%, and 7.53% of the samples, respectively. The most prevalent mycotoxin was DON, followed by OTA, T-2, AFB1, and FBs, whereas ZEN was less frequently detected. The mean concentrations of the six mycotoxins in pet feed samples were 3.17μg/kg for AFB1, 0.65mg/kg for DON, 2.15mg/kg for FBs, 6.27μg/kg for OTA, 20.00μg/kg for T-2, and 30.00μg/kg for ZEN. The levels of mycotoxins were generally below the limits of the Pet Feed Hygiene Regulations of China and the EU. Notably, a substantial majority of the pet food samples (88 out of 93) were contaminated by two or more mycotoxins. AFB1, FBs, OTA, and ZEN occurred slightly more often in feline food than in canine food. Except for OTA, the contamination rates for the other five mycotoxins in canned food were lower than those in dry food. Moreover, except for AFB1, the levels of the other five mycotoxins in canned foods were lower than those in dry foods. This study highlights the widespread contamination of pet foods with mycotoxins, which poses a significant risk to pets from continuous exposure to multiple mycotoxins.

Dual-signal ratiometric electrochemical aptasensor for Zearalenone detection based on magnetic-assisted enrichment and hybridization chain reaction

In this work, a dual-signal ratiometric electrochemical aptasensor based on the hybrid chain reaction (HCR) and streptavidin-modified magnetic beads (SA-MBs) was developed to rapidly detect zearalenone (ZEN). The HCR, as a powerful signal amplification technique to imporve the signal of sonser. When the target is present, they specifically bind with ZEN-Apt and release ZEN-cDNA to trigger HCR. Simultaneously, more double-stranded DNA causes the signal of Thi to be blocked. As a result, the two signals tend to change in the opposite direction as the ZEN concentration changes. Additionally, the peak current ratio of IThi/IFc showed a positive correlation with the ZEN concentration. Under optimal conditions, the constructed biosensor showed an excellent linear detection range (1.0 × 10−10 mol/L to 1.0 × 10−6 mol/L), a low detection limit (4.4 × 10−11 mol/L) and high specificity for ZEN. In addition, the detection method retains the characteristics of low cost and rapid detection of electrochemical detection, while improving the detection limit and detection accuracy via SA-MBs and internal reference signal. This provides a new idea for the practical detection of ZEN.

Mycotoxigenic Fusarium species and zearalenone concentration in commercial maize kernels in northern Ghana.

The fungal genus Fusarium contains many toxigenic pathogens of maize with associated yield losses, reduction of grain quality, and accumulation of mycotoxins in harvested grains. To determine zearalenone (ZEN) concentration and identify the various Fusarium species in commercial maize grains, a survey of 75 maize samples, collected from 11 market centers in the five regions in northern Ghana was identified based on morphological characteristics, sequence analysis of the internal transcribed spacer region, and polymerase chain reaction using species-specific primers. ZEN levels were determined using HPLC. ZEN contamination was recorded in 33.3% of the maize samples, with concentrations ranging from 0.61 to 3.05µg/kg. Based on VERT1/2 and TEF 1-α sequencing, F. verticillioides was the most prevalent species in the studied samples: 40.35% from the Upper East Region, 28.07% from the North East Region, 19.30% from the Upper West Region, 10.53% from the Savannah Region, and 1.75% for the Northern Region. Other fungal species found were F. equiseti and F. solani. A higher number of the Fusarium isolates were found in white maize (609 isolates from 27 samples) compared to yellow maize (225 isolates from 23 samples).

Determination of some mycotoxins from poultry feed in Baghdad city

The aims of this study were to determine some mycotoxins and to isolate the fungi from chicken feed samples from broiler and layer farms in Baghdad city. The isolation rate (percentage) of fungi from poultry feed samples was Aspergillus niger 17.7%, Aspergillus flavus 14.7%, Aspergillus fumigatus 12.4%, Aspergillus ochraceus 10.7%, Penicillium spp. 9.3%, Fusarium spp. 9.3%, Rhizopus spp. 8.4%, Mucar spp. 8.4%, Absidia spp. 7.6% and Chrysonilia sitophila 1.5%, while the most common yeast genera were Candida spp. 44.8%, Candida albicans 20.7%, Cryptococcus neoformans 17.3%, Geotricum candidum 12% and Trichosorom spp. 5.2%. The mycotoxins ochratoxin A (OTA), deoxynivalenol (DON), fumonisin B1 (FB1) and zearalenone (ZON) were detected, and the percentages of these mycotoxins were 43.1%, 27.2%, 21.1% and 8.6%, respectively. This study indicates that the presence of mold that produce mycotoxins in poultry feed increases the risk of mycotoxins in feed, meat, eggs, etc. of animals and therefore humans, and we need to raise awareness of the Ministry of Agriculture and Health to prevent these measures to reduce the mycotoxin levels in poultry products.

Determination of mycotoxins in breakfast cereals by LC-MS/MS

Breakfast cereals, attributed to low microbiological risk due to their low water activity, are not very innocent due to their components such as cereal, dried fruit and cocoa. Secondary fungal metabolites called mycotoxins are one of the most important risks that these foods can carry. The multimycotoxin level of 140 breakfast cereal samples sold in local markets in Istanbul, Turkiye, was analysed.Samples were tested using the LC-MS/MS technique. Contamination levels of aflatoxin B1, B2, G1, G2, ochratoxin (OTA), zearalenone (ZEN), deoxynivalenol (DON), fumonisin B1, B2, citrinin, HT-2 and T-2 toxins were determined in breakfast cereal.As a result, 71% (n = 100) of the samples were contaminated with at least one mycotoxin. Aflatoxin B1 levels in 17% of the samples and total aflatoxin levels in 31% were determined to exceed the limits defined in the “Turkish Food Codex Contaminants Regulation”. The sample rate containing all toxins (except CIT) above the detection limit was 24%. ZEN + DON + OTA was found in 24% of the samples, total aflatoxins (TAF) + OTA + ZEN in 25%, fumonisins (FUM) + OTA + DON in 29%, and TAF + FUM + OTA + DON + ZEN + HT-2+T-2 in 24% of the samples.Our findings indicate that breakfast cereal samples are susceptible to mycotoxin contamination and necessitate consistent monitoring to ensure safety. The presence of other mycotoxins not included in the legal regulation is noteworthy. Action is needed to prepare a legal regulation on the co-occurrence of mycotoxins.

Simultaneous Degradation of AFB1 and ZEN by CotA Laccase from Bacillus subtilis ZJ-2019-1 in the Mediator-Assisted or Immobilization System.

The global prevalence of aflatoxin B1 (AFB1) and zearalenone (ZEN) contamination in food and feed poses a serious health risk to humans and animals. Recently, enzymatic detoxification has received increasing attention, yet most enzymes are limited to degrading only one type of mycotoxin, and free enzymes often exhibit reduced stability and activity, limiting their practicality in real-world applications. In this study, the laccase CotA gene from ZEN/AFB1-degrading Bacillus subtilis ZJ-2019-1 was cloned and successfully expressed in Escherichia coli BL21, achieving a protein yield of 7.0 mg/g. The recombinant CotA (rCotA) completely degraded AFB1 and ZEN, with optimal activity at 70 °C and pH 7.0. After rCotA treatment, neither AFB1 nor ZEN showed significantly cytotoxicity to mouse macrophage cell lines. Additionally, the AFB1/ZEN degradation efficiency of rCotA was significantly enhanced by five natural redox mediators: acetosyringone, syringaldehyde, vanillin, matrine, and sophoridin. Among them, the acetosyringone-rCotA was the most effective mediator system, which could completely degrade 10 μg of AFB1 and ZEN within 1 h. Furthermore, the chitosan-immobilized rCotA system exhibited higher degradation activity than free rCotA. The immobilized rCotA degraded 27.95% of ZEN and 41.37% of AFB1 in contaminated maize meal within 12 h, and it still maintained more than 40% activity after 12 reuse cycles. These results suggest that media-assisted or immobilized enzyme systems not only boost degradation efficiency but also demonstrate remarkable reusability, offering promising strategies to enhance the degradation efficiency of rCotA for mycotoxin detoxification.

Protective and Detoxifying Effects of Resveratrol on Zearalenone-Mediated Toxicity: A Review

Zearalenone (ZEA) is a mycotoxin produced by Fusarium spp. fungi and is widely found in moldy corn, wheat, barley, and other grains. ZEA is distributed to the whole body via blood circulation after metabolic transformation in animals. Through oxidative stress, immunosuppression, apoptosis, autophagy, and mitochondrial dysfunction, ZEA leads to hepatitis, neurodegenerative diseases, cancer, abortion, and stillbirth in female animals, and decreased sperm motility in male animals. In recent years, due to the influence of climate, storage facilities, and other factors, the problem of ZEA pollution in global food crops has become particularly prominent, resulting in serious problems for the animal husbandry and feed industries, and threatening human health. Resveratrol (RSV) is a natural product with therapeutic activities such as anti-inflammatory, antioxidant, and anticancer properties. RSV can alleviate ZEA-induced toxic effects by targeting signaling pathways such as NF-κB, Nrf2/Keap1, and PI3K/AKT/mTOR via attenuating oxidative damage, inflammatory response, and apoptosis, and regulating cellular autophagy. Therefore, this paper provides a review of the protective effect of RSV against ZEA-induced toxicity and its molecular mechanism, and discusses the safety and potential clinical applications of RSV in the search for natural mycotoxin detoxification agents.

Effect of zearalenone in sugar beet products on zootechnical and reproductive performance and lesions of sows and piglets.

Following the use of sugar beet pulp that was retrospectively found to be predominantly contaminated with zearalenone (ZEN) in diets of reproducing sows largely exceeding the EU-guidance value for critical ZEN concentration of 0.25mg/kg, farmers did not report any changes in the reproductive performance of sows. Thus, the aim of the study was to verify this guidance value in a dose-response setup by using sugar beet pulp as a ZEN source hitherto not considered a risky feedstuff additionally characterized by comparatively low levels of deoxynivalenol. A total of 90 sows was equally allocated to one of the three feed groups during experimental lactation 1 and up to 40days after insemination: CON with a minimal ZEN concentration, ZEN1 with a target concentration of 250µg ZEN/kg feed, and ZEN2 with a target concentration of 500µg ZEN/kg feed. Thereafter, all sows received the same feed without ZEN for the rest of gestation, and the following lactation for testing of putative carry-over effects resulting from previous ZEN exposure. Exposure of sows to ZEN with blood serum as an indicator was linearly related to dietary ZEN concentrations. Reproductive and zootechnical performances of sows were only affected by ZEN exposure at weaning weight. Clinical-chemical parameters indicated no clear effect of ZEN exposure. An influence of ZEN on the occurrence of tail and ear injuries (not necrosis) in piglets and lesions on the mammary complexes in sows is possible. The influence of a ZEN concentration above the EU guidance value on the study farm can therefore not be neglected.

Blood levels of zearalenone, thyroid-stimulating hormone, and thyroid hormones in patients with colorectal cancer

Mycotoxins are secondary metabolites produced by various species of mold fungi commonly found in plant materials. Zearalenone (ZEN) adversely affects the endocrine system. This study aimed to determine whether thyroid-stimulating hormone (TSH), procalcitonin (PCT), free triiodothyronine (fT3), and free thyroxine (fT4) levels are altered during natural zearalenone mycotoxicosis in patients diagnosed with sigmoid colon cancer (SCC) or colorectal cancer (CRC). A study was conducted on women and men diagnosed with SCC or CRC accompanied by the presence or absence (Patients Without ZEN – PWZ group) of ZEN in the blood. The PWZ group consisted of 17 patients with symptoms of SCC and CRC in whom ZEN and its metabolites were not detected in peripheral blood. The experimental (empirical) groups included a total of 16 SCC and CRC patients who tested positive for ZEN, but not its metabolites. TSH values in both sexes were within the upper limit of the reference range (0.27 – 4.2 μIU/mL) adopted by the hospital laboratory and corresponded to the upper second tertile and the lower third tertile. PCT values demonstrated that SCC and CRC were accompanied by a systemic or local bacterial infection. All mean values of fT3 were in the middle of the reference range, and the mean values of fT4 were within the upper reference limit. The fT3/fT4 prognostic marker was somewhat above the cut-off point of 0.22. These results indicate that in postmenopausal women and andropausal men who were diagnosed with SCC and CRC and were exposed to food-borne ZEN, higher values of the prognostic marker (fT3/fT4) were associated with an unfavorable prognosis. The study also revealed that the more distal the neoplastic lesions in the colon, the higher the percentage of both thyroid hormones, regardless of the patient's sex. The presence of ZEN in the diet alters thyroid activity in patients diagnosed with SCC and CRC.

Cascade Amplifying Electrochemical Bioanalysis for Zearalenone Detection in Agricultural Products: Utilizing a Glucose-Fenton-HQ System on Bimetallic-ZIF@CNP Nanocomposites.

The integration of advanced materials and signaling cascade strategies is a promising and highly relevant topic for enhancing the performance of bioanalysis. In this study, a three-stage cascade amplification electrochemical bioanalysis (TCAE-bioanalysis) was developed and evaluated for the detection of zearalenone (ZEN). This method couples immunoreaction with a glucose-Fenton-hydroquinone (HQ) system on bimetallic-ZIF (B-ZIF)@CNP nanocomposites. The B-ZIF@CNP-modified gold electrode (AuE) was prepared, offering high conductivity and an excellent reaction interface. The immunoreaction introduced glucose oxidase (GOx) into the glucose-Fenton-HQ system, generating an abundant electron signal. The method achieved an ultrasensitive limit of detection (LOD) as low as 0.87 pg/mL, with an IC50 value of 30.8 pg/mL, representing a 229-fold enhancement in sensitivity compared to ELISA using the same monoclonal antibody (McAb). The specificity, reliability, and practicality of this approach were thoroughly demonstrated for agricultural product samples. Additionally, the TCAE-bioanalysis offers several advantages, including simplified preparation for advanced B-ZIF@CNP, a convenient detection system, and the use of common and environmentally friendly reagents. This study presents a comprehensive approach to improving electrochemical bioanalysis and may also expand the application of signaling cascades and environmentally friendly techniques in other biosensing or diagnostic contexts.

Sociodemographic and dietary predictors of maternal and placental mycoestrogen concentrations in a US pregnancy cohort.

Zearalenone (ZEN) is a mycotoxin contaminating grains and processed foods. ZEN alters nuclear estrogen receptor α/β signaling earning its designation as a mycoestrogen. Experimental evidence demonstrates that mycoestrogen exposure during pregnancy is associated with altered maternal sex steroid hormones, changes in placental size, and decreases in fetal weight and length. While mycoestrogens have been detected in human biospecimens worldwide, exposure assessment of ZEN in US populations, particularly during pregnancy, is lacking. To characterize urinary and placental concentrations of ZEN and its metabolites in healthy US pregnant people and examine demographic, perinatal, and dietary predictors of exposure. Urine samples were collected in each trimester from pregnant participants in the UPSIDE study and placenta samples were collected at delivery (Rochester, NY, n = 317). We used high performance liquid chromatography and high-resolution tandem mass spectrometry to measure total urinary (ng/ml) and placental mycoestrogens (ng/g). Using linear regression and linear mixed effect models, we examined associations between mycoestrogen concentrations and demographic, perinatal, and dietary factors (Healthy Eating Index [HEI], ultra-processed food [UPF] consumption). Mycoestrogens were detected in 97% of urines (median 0.323 ng/ml) and 84% of placentas (median 0.012 ng/g). Stability of urinary mycoestrogens across pregnancy was low (ICC: 0.16-0.22) and did not correlate with placental levels. In adjusted models, parity (multiparous) and pre-pregnancy BMI (higher) predicted higher urinary concentrations. Birth season (fall) corresponded with higher placental mycoestrogens. Dietary analyses indicated that higher HEI (healthier diets) predicted lower exposure (e.g., Σmycoestrogens %∆ -2.03; 95%CI -3.23, -0.81) and higher percent calories from UPF predicted higher exposure (e.g., Σmycoestrogens %∆ 1.26; 95%CI 0.29, 2.24). The mycotoxin, zearalenone (ZEN), has been linked to adverse health and reproductive impacts in animal models and livestock. Despite evidence of widespread human exposure, relatively little is known about predictors of exposure. In a pregnant population, we observed that maternal ZEN concentrations varied by maternal pre-pregnancy BMI and parity. Consumption of ultra-processed foods, added sugars, and refined grains were linked to higher ZEN concentrations while healthier diets were associated with lower levels. Our research suggests disparities in exposure that are likely due to diet. Further research is needed to understand the impacts of ZEN on maternal and offspring health.

An impedimetric determination of zearalenone on MIP-modified carboceramic electrode

Zearalenone (ZEN) is a mycotoxin that poses significant risks to human and animal health due to its mutagenic, immunosuppressive, and carcinogenic properties. This study presents a novel analytical method for detecting ZEN using electrochemical impedance spectroscopy (EIS) combined with a molecularly imprinted polymer (MIP). ZEN, used as the template molecule, was incorporated into polypyrrole on screen-printed electrodes (SPE), and a ZEN-sensitive MIP sensor was created through template removal. The modified sensor surfaces were characterized by EIS and scanning electron microscopy (SEM). An impedimetric MIP sensor for ZEN was developed, offering a detection range from 1 pM to 500 pM. The method's limit of detection (LOD) was established at 1 pM (0.3 pg/mL) with a signal-to-noise ratio of 3 (S/N = 3). The method demonstrated high precision and accuracy, with a maximum relative standard deviation (RSD) of less than 4.4% at a 95% confidence level, and relative error (RE) values ranging from −0.8% to −2.7%. The selectivity of the developed MIP sensor was evaluated using ochratoxin A, ochratoxin B, and aflatoxin B1, with no significant interference observed. ZEN recovery from spiked samples was between 95% and 105%, indicating that the method was successfully applied to grain samples, including corn, rice, and wheat.