Zearalenol Research Articles

Raman spectroscopic comparison of zearalenone and its derivatives for non-destructive rapid detection

Raman spectroscopy is used to investigate the absorption and dispersion of zearalenone (ZEN) and its derivatives. The C=C stretching vibration modes of ZEN and zearalenol (ZENOL) were appeared at 975–990 cm−1. The C=O vibration mode was present at 1680–1690 cm−1 for ZEN and zearalanone (ZAN), but it was absent for ZENOL and zearalanol (ZANOL) which have –OH group instead of –C=O group in ZEN and ZAN molecular structures. On the basis of this characterization, Raman spectra of specific chemical groups and linkages corresponding to the structural difference of ZEN and its derivatives were identified. These results indicated that Raman spectroscopy can apply for the identification of ZEN and its derivatives and has a potential for the non-destructive rapid detection of these compounds in food.

Recent progress on the detection of animal-derived food stimulants using mass spectrometry-based techniques.

The misuse of animal-derived stimulants in food is becoming increasingly common, and mass spectrometry (MS) is used extensively for their detection and analysis. There is a growing demand for abused-substances detection, highlighting the need for systematic studies on the advantages of MS-based methods in detecting animal-derived stimulants. We reviewed the application of chromatography-mass spectrometry to the screening and detection of food stimulants of animal origin. Specifically, we analyzed four common animal sources of synthetic steroids, β-receptor agonists, zearalenol (ZAL), and glucocorticoids. We also explored the potential of using chromatography-mass spectrometry to detect and analyze animal-derived foods. We searched and screened the Web of Science and Google Scholar databases until April 2023. Our inclusion criteria included a publication year within the last 5 years, publication language of English, and the research fields of food analysis, environmental chemistry, and polymer science. Our keywords were "mass spectrometry," "anabolic androgenic steroids," "β-2agonists," "glucocorticoids," "zearalenone," and "doping." Although traditional techniques such as thin-layer chromatography and enzyme-linked immunoassays are simple, fast, and suitable for the initial screening of bulk products, they are limited by their relatively high detection limits. Among the methods based on MS, gas chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry are the most widely used for detecting food doping agents of animal origin. However, a sensitive method with high repeatability and a short analysis time for a large number of samples is still required. Advances in MS have enabled the detection of extremely low concentrations of these substances. Combining different techniques, such as high-resolution mass spectrometry, ultra-high performance liquid chromatography-tandem mass spectrometry, gas chromatography-combustion-isotope ratio mass spectrometry, ultra-high performance liquid chromatography-high resolution mass spectrometry, and two-dimensional chromatography, offers significant advantages for detecting trace illicit drugs in animal-derived foods. Due to advances in assay technology and sample preparation methods, sample collection and storage methods such as dried blood spots, dried urine spots, and volumetric absorptive microsampling are increasingly accepted because of their increased stability and cost-effectiveness. MS significantly improves the efficiency of detecting doping agents of animal origin. With the continuous development of MS technology, its application in the fields of doping detection and the analysis of doping agents of animal origin is expected to become more extensive.

Effects of Intestinal Microorganisms on Metabolism and Toxicity Mitigation of Zearalenone in Broilers

Simple SummaryZearalenone (ZEN) widely contaminates all the feed crops, and ZEN may cause harmful damage to animals and humans. Different animals have different sensitivity to ZEN. Among these animals, chickens show a strong resistance. Intestinal microorganisms are essential in digestion and degradation. Therefore, we hypothesise whether intestinal microorganisms in chickens play an important role in digesting and degrading ZEN. In this study, we found that intestinal microorganisms could degrade ZEN to a certain degree by both vivo and vitro experiments. We concluded that the intestinal microbiota of broilers had metabolic effects on ZEN and alleviated antioxidant and liver damage caused by ZEN to broilers. Moreover, we found some key bacteria that are important in degrading ZEN.Zearalenone (ZEN) is an estrogenic mycotoxin, and chickens are relatively insensitive to it. In this study, the effects of intestinal microorganisms on ZEN metabolism and toxicity mitigation in broilers were studied by two experiments. Firstly, in vitro, ZEN was incubated anaerobically with chyme from each part of the chicken intestine to study its intestinal microbial metabolism. Then, in vivo, we explored the effects of intestinal microbiota on ZEN by inhibiting intestinal microorganisms. Broilers were fed a control diet, 2.5 mg/kg ZEN diet, microbial inhibition diet or ‘microbial inhibition +2.5 mg/kg ZEN’ diet. In vitro, the results showed that the rates of ZEN degradation by microorganisms in the duodenum, ileum, caecum, and colon were 56%, 12%, 15%, and 17%, respectively, and the microorganisms could convert ZEN into Zearalenol (ZOL). After microbial inhibition in vivo, the content of ZEN and its metabolites in excreta of broilers increased significantly, and antioxidant damage and liver damage were aggravated. 16S rRNA sequencing results showed that antioxidant indices and the content of ZEN and its metabolites in excreta were significantly correlated with the relative abundance of Streptococcus, Lactococcus and Enterococcus, etc. In conclusion, the intestinal microorganisms of broilers play an important role in ZEN metabolism and ZEN-induced antioxidant and liver injury mitigation, among which the key bacteria include Streptococcus, Lactococcus and Enterococcus, etc.

Natural incidence of aspergilli, penicilli, fusaria and their multiple toxins in the seeds of dried red chillies (Capsicum annuum Linn.) from Jammu and Kashmir (UT)

An investigation was carried out to isolate and enumerate the species of three mycotoxin producing fungal genera (Aspergillus, Penicillium and Fusarium) and determine their mycotoxin profile from the sampled chilli seeds. Both household and market samples of chilli pods were randomly collected from the two divisions of Jammu and Kashmir (UT) and their seeds were removed aseptically for analysis. Mycoflora assessment indicates that chilli seeds are capable of harbouring several fungal species belonging to these three genera and their perfect states. Overall Aspergillus and Penicillium were represented by 10 and 13 species respectively followed by decreasing order by the perfect states, Eurotium (4) and Emericella (2). In addition to this, Fusarium was represented by 2 species. Results obtained from HPLC analysis showed that the chilli seed samples were contaminated with aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), sterigmatocystin (STER), cyclopiazonic acid (CPA), citrinin (CIT), patulin (PAT), zearalenone (ZEN) and zearalenol (ZOL). The two fusarial toxins, ZEN and ZOL were detected to be quite high up to 40.2 μg/g of seed sample, which shows that these mycotoxins might have been produced both in the field and storage. Presence of multiple toxins in the chilli seeds is of concern as it forms a substantial part of the chilli powder that is consumed in every Indian household.

Precise Modeling of the Protective Effects of Quercetin against Mycotoxin via System Identification with Neural Networks.

Cell cytotoxicity assays, such as cell viability and lactate dehydrogenase (LDH) activity assays, play an important role in toxicological studies of pharmaceutical compounds. However, precise modeling for cytotoxicity studies is essential for successful drug discovery. The aim of our study was to develop a computational modeling that is capable of performing precise prediction, processing, and data representation of cell cytotoxicity. For this, we investigated protective effect of quercetin against various mycotoxins (MTXs), including citrinin (CTN), patulin (PAT), and zearalenol (ZEAR) in four different human cancer cell lines (HeLa, PC-3, Hep G2, and SK-N-MC) in vitro. In addition, the protective effect of quercetin (QCT) against various MTXs was verified via modeling of their nonlinear protective functions using artificial neural networks. The protective model of QCT is built precisely via learning of sparsely measured experimental data by the artificial neural networks (ANNs). The neuromodel revealed that QCT pretreatment at doses of 7.5 to 20 μg/mL significantly attenuated MTX-induced alteration of the cell viability and the LDH activity on HeLa, PC-3, Hep G2, and SK-N-MC cell lines. It has shown that the neuromodel can be used to predict the protective effect of QCT against MTX-induced cytotoxicity for the measurement of percentage (%) of inhibition, cell viability, and LDH activity of MTXs.

Enhanced α-Zearalenol Hydrolyzing Activity of a Mycoestrogen-Detoxifying Lactonase by Structure-Based Engineering

The enzyme ZHD101 from Clonostachys rosea hydrolyzes and deactivates the mycotoxin zearalenone (ZEN) and its zearalenol (ZOL) derivatives. ZHD101 prefers ZEN to ZOL as its substrate, but ZOL, especially the α-form, shows higher estrogenic toxicity than ZEN. To enhance α-ZOL selectivity, we solved the complex structures of ZHD101 with both ZOLs and modified several lactone-surrounding residues. Among the mutants, V153H maintained activity for ZEN but showed a 3.7-fold increase in specific activity against α-ZOL, with an ∼2.7-fold reduction in substrate affinity but a 5.2-fold higher turnover rate. We then determined two V153H/ZOL complex structures. Here, the α-ZOL lactone ring is hydrogen-bonded to the H153 side chain, yielding a larger space for H242 to reconstitute the catalytic triad. In conclusion, structure-based engineering was successfully employed to improve the ZHD101 activity toward the more toxic α-ZOL, with great potential in further industrial applications.

Occurrence of Zearalenone, Zearalenol and Deoxynivalenol in Some Market Samples of Dried Vegetables in J&K State

In Jammu and Kashmir state, there is a trend of sun-drying the surplus vegetables and marketing them for consumption in off-seasons. Unfortunately, warm and humid weather conditions prevailing during dehydration, followed by faulty storage practices may allow proliferation of such microbes, which could be degradative and toxigenic. In view of this, an investigation was conducted on market samples of J&K state to determine the association of toxigenic fusarial species with the surface of commonly consumed sun-dried vegetables viz., brinjal, tomato, cauliflower, bottle gourd, bitter gourd and turnip. All the samples of dried vegetables excepting that of bottle gourd were found to be contaminated with two species of Fusarium viz., F. solani and F. subglutinans. HPLC analysis conducted for estimating natural incidence of fusarial toxins revealed the presence of zearalenone (ZEN), zearalenol (ZOL) and deoxynivalenol (DON). Contamination of ZEN was detected in market samples of dried brinjal, tomato, cauliflower, turnip and bitter gourd with highest concentration (4.75 ppm) in one of the tomato samples. However, while investigating the marketed dried vegetables for ZOL contamination, only bitter gourd samples were found to be positive with contamination varying from 1.79 to 2.16 ppm. Similarly, contamination of DON was detected from market samples of dried brinjal, turnip and cauliflower. Its concentration was detected to be maximum in a cauliflower sample (2.12 ppm), whereas least contamination (0.09 ppm) was detected from a turnip sample. Some market samples of dried brinjal, bitter gourd, turnip and cauliflower even showed co-occurrence of fusarial toxins.

Combination of LC-MS2 and GC-MS as a Tool to Differentiate Oxidative Metabolites of Zearalenone with Different Chemical Structures

Recent studies on the mammalian and fungal metabolism of the mycotoxin zearalenone (ZEN) have disclosed the formation of six regioisomers of monohydroxy-ZEN and its reductive metabolite zearalenol (ZEL). Hydroxylation occurs at the aromatic ring or at one of four positions of the aliphatic macrocycle. In addition, an aliphatic ZEN epoxide, its hydrolysis product, and other products were identified in fungal cultures. In this paper, we report the product ion spectra of the [M-H]− ions of 22 oxidative metabolites of ZEN and ZEL, obtained by LC-MS2 analysis using a linear ion trap mass spectrometer with negative electrospray ionization. The MS2 spectra exhibit qualitative and quantitative differences which allow a clear distinction of most metabolites. Moreover, GC-MS analysis of the trimethylsilylated metabolites yields electron impact mass spectra with numerous fragment ions which can be used as fingerprint to confirm the chemical structure derived by LC-MS2 analysis.

Effect of an esterified glucomannan on laying hens exposed to combined mycotoxins (aflatoxin B1, zearalenone and fumonisin B1)

Mycotoxin toxicity depends on species, exposure time, age, sex, health and possible synergistic effects of other mycotoxins present in feed. In poultry, aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are associated with poor growth performance, lowered feed utilization efficiency, liver damage and immunosuppression; metabolites may persist in tissues and eggs. Exposure of mature hens to zearalenone (ZEN) apparently does not cause adverse effects, but ZEN residues and α and β zearalenol persist in liver and muscle (Kuiper-Goodman et al., 1987) and may be transmitted to egg yolk (Dailey et al., 1990). Various treatments and dietary strategies have been tried to reduce mycotoxin levels in contaminated commodities...

Evaluation of mycotoxins contamination in intensive beef cattle production system

Feed samples from 15 intensive beef cattle farms located in Northern Italy (provinces of Verona and Mantova) were checked for mycotoxin contamination [aflatoxins (AF), fumonisins (FB), ochratoxin A (OTA), deoxynivalenol (DON) and zearalenol (ZEN)]. Total mixed rations (TMR) resulted positive for AF and FB contamination. Among feedstuffs corn and corn gluten feed were the only responsible of TMR contamination. Level of contamination was positively related to corn moisture content.

Development and application of analytical methods for the determination of mycotoxins in organic and conventional wheat

The aim of this study was to develop a multicomponent analytical method for the determination of deoxynivalenol (DON), ochratoxin A (OTA) and zearalenone (ZEN), nivalenol (NIV), 3-acetyl-DON (3-acDON), 15-acetyl-DON (15-acDON), zearalenol (ZOL) and citrinin (CIT) in wheat. It also aimed to survey the presence and amounts of DON, OTA and ZEN in Belgian conventionally and organically produced wheat grain and in wholemeal wheat flours. After solvent extraction, an anion-exchange column (SAX) was used to fix the acidic mycotoxins (OTA, CIT), whilst the neutral mycotoxins flowing through the SAX column were further purified by filtration on a MycoSep cartridge. OTA and CIT were then analysed by high-performance liquid chromatography (HPLC) using an isocratic flow and fluorescence detection, while the neutral mycotoxins were separated by a linear gradient and detected by double-mode (ultraviolet light fluorescence) detection. The average DON, ZEN and OTA recovery rates from spiked blank wheat flour were 92, 83 and 73% (RSDR = 12, 10 and 9%), respectively. Moreover, this method offered the respective detection limits of 50, 1.5 and 0.05 µg kg−1 and good agreement with reference methods and inter-laboratory comparison exercises. Organic and conventional wheat samples harvested in 2002 and 2003 in Belgium were analysed for DON, OTA and ZEN, while wholemeal wheat flour samples were taken from Belgian retail shops and analysed for OTA and DON. Conventional wheat tended to be more frequently contaminated with DON and ZEN than organic samples, the difference being more significant for ZEN in samples harvested in 2002. The mean OTA, DON and ZEA concentrations were 0.067, 675 and 75 µg kg−1 in conventional samples against 0.063, 285 and 19 µg kg−1 in organically produced wheat in 2002, respectively. Wheat samples collected in 2003 were less affected by DON and ZEN than the 2002 harvest. Organic wholemeal wheat flours were more frequently contaminated by OTA than conventional samples (p < 0.10). The opposite pattern was shown for DON, organic samples being more frequently contaminated than conventional flours (p < 0.10).

Species differences in zearalenone-reductase activity

Thin-layer and high-pressure liquid chromatography (HPLC) were used to investigate in several animal species the hepatic metabolites of zearalenone (ZEN), a non-steroidal oestrogenic fungal toxin produced by species of Fusarium. ZEN was reduced to zearalenol (ZEL) by the S-9 fraction of rat-liver homogenates in the presence of NADH or NADPH. In this species ZEN reductase showed two peaks of activity at pH 4.5 and 7.4. Of the species tested, cows showed the highest activity of NADH-dependent ZEN reductase in the hepatic S-9 fraction, followed in decreasing order by mice, pigs, rats, rabbits and guinea-pigs. The S-9 fraction of hamster liver showed optimal activity at pH 5.5 and 8.0 with NADH and at pH 6.0 and 8.5 with NADPH; NADPH was more efficient than NADH only in this species. HPLC showed that α-ZEL was a major hepatic metabolite in the rat, mouse, pig, cow and rabbit in incubations at pH 4.5 with either NADH or NADPH and at pH 7.4 with NADH, although at pH 7.4 with NADPH, β-ZEL was the predominant metabolite. In guinea-pigs, both α- and β-ZELs were produced in roughly similar amounts irrespective of the pH and cofactor, while in hamsters β-ZEL was the major metabolite. These findings indicate that there are two types of ZEN reductase differing in optimum pH and that the stereospecific reduction of ZEN depends on the source of S-9 and cofactors. Since the oestrogenic activity of α-ZEL is about ten times greater than that of ZEN, the possible presence of ZEN metabolites in edible tissues of livestock fed on ZEN-contaminated feeds is an important mycotoxicological problem.