Simultaneous Detoxification of Aflatoxin B1, Zearalenone and Deoxynivalenol by Modified Montmorillonites.
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ABSTRACT:
SUBMITTER: Mao J
PROVIDER: S-EPMC8746407 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:To develop a new simple and simultaneous purification method for mycotoxins in feeds and grains, magnetic nanoparticles (MNPs) conjugated with monoclonal antibodies (mAbs) against mycotoxins were used to separate aflatoxin B1 (AFB1), zearalenone (ZEA) and deoxynivalenol (DON). For a single spike of each mycotoxin into the buffer solution (16% MeOH in PBS), mean recoveries were 93.1-95.0% for AFB1 (5-20 ng/mL spiked), 87.2-96.0% for ZEA (125-500 ng/mL spiked) and 75.2-96.9% for DON (250-1,000 ng/mL spiked) by HPLC and ELISA. Recoveries of AFB1 (20 ng/mL) and ZEA (500 ng/mL) simultaneously spiked into the buffer solution were 87.0 and 99.8%, respectively. Recovery rates of AFB1/DON and DON/ZEA spiked simultaneously were 86.2%/76.6% and 92.0%/86.7%, respectively, at concentrations of 20 ng/mL AFB1, 500 ng/mL ZEA, and 1,000 ng/mL DON. Recoveries using the novel mAb-MNP conjugated system in a buffer solution simultaneously spiked with AFB1, ZEA and DON were 82.5, 94.6 and 73.4%, respectively. Recoveries of DON in animal feed were 107.7-132.5% at concentrations of 250-1,000 ng/g spiked in feed. The immunoaffinity chromatography (IAC) clean-up method was compared with the purification method using novel mAb-MNP. After fortification of animal feed with AFB1 (5, 10 and 20 ng/g feed) and ZEA (125, 250 and 500 ng/g feed), AFB1 and ZEA were purified using both the methods. In the case of the novel mAb-MNP conjugated system, mean recoveries for AFB1 were 89.4, 73.1 and 88.3% at concentrations of 5, 10 and 20 ng/g feed, respectively. For ZEA, mean recoveries were 86.7, 85.9 and 79.1% at concentrations of 125, 250 and 500 ng/g, respectively. For IAC purification, recoveries were 42.9-45.1% for AFB1 and 96.8-103.2% for ZEA. In conclusion, the present purification method using monoclonal antibodies conjugated to MNPs can be used for simple and simultaneous purification of mycotoxins from feed and maize.Supplementary informationThe online version contains supplementary material available at 10.1007/s43188-020-00083-w.
| S-EPMC8476696 | biostudies-literature
Project description:Mycotoxins are secondary metabolites produced by fungi. These contaminate dried seafoods during processing and storage and represent a potential health hazard for consumers. A sensitive, selective and accurate liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was established for simultaneous quantification of four common mycotoxins (aflatoxin B1 (AFB1), T-2 toxin (T-2), ochratoxin A (OTA) and deoxynivalenol (DON)) in dried shrimp, dried fish and dried mussel products. Mycotoxins were extracted from dried seafood samples by acetonitrile/water (85/15, v/v), subjected to ultrasound for 60 min at 20 °C and cleaned up by defatting with n-hexane. The sample matrix affected the linearity of detection (R2 ? 0.9974). The limit of detection (LOD) and limit of quantification (LOQ) in dried seafood products varied from 0.1 to 2.0 µg·kg-1 and 0.3 to 5.0 µg·kg-1, respectively. The method was validated by spiking samples with specific mycotoxin levels, and the recoveries, intra-relative standard deviation (RSDs) and inter-RSDs ranged between 72.2-98.4%, 2.8-10.6%, and 5.5-15.4%, respectively. This method was used to analyze 40 dried seafood products purchased from the Zhanjiang seafood market. Results of this product sampling showed that while no DON was detected, AFB1, T-2 and OTA were detected in 30.8%, 17.5% and 33.3% of the samples, respectively. AFB1, T-2 and OTA concentrations varied at 0.58-0.89, 0.55-1.34 and 0.36-1.51 µg·kg-1, respectively. Relatively high frequency of contamination and the presence of AFB1, OTA and T-2 residues indicate the need to monitor mycotoxins in dried seafood products.
| S-EPMC7472416 | biostudies-literature
Project description:Multiple-mycotoxin contamination has been frequently found in the agro-food monitoring due to the coexistence of fungi. However, many determination methods focused on a single mycotoxin, highlighting the demand for on-site determination of multiple mycotoxins in a single run. We develop a multicolor-based immunochromatographic strip (ICS) for simultaneous determination of aflatoxin B1 (AFB1), zearalenone (ZEN) and T-2 toxin in maize- and cereal-based animal feeds. The nanoparticles with different colors are conjugated with three monoclonal antibodies, which serve as the immunoassay probes. The decrease in color intensity is observed by the naked eyes, providing simultaneous quantification of three mycotoxins. The visible limits of detection for AFB1, ZEN and T-2 are estimated to be 0.5, 2, and 30 ng/mL, respectively. The cut-off values are 1, 10, and 50 ng/mL, respectively. Considerable specificity and stability are found using real samples. The results are in excellent agreement with those from high-performance liquid chromatography/tandem mass spectrometry. The multi-color ICS boasts sensitive and rapid visual differentiation and simultaneous semi-quantification of aflatoxin B1, zearalenone and T-2 toxin in maize- and cereal-based feed samples within 20 min.
| S-EPMC5848188 | biostudies-literature
Project description:Aflatoxin B1 (AFB1) is one of the most hazardous mycotoxins contamination in food and feed products, which leads to hepatocellular carcinoma in humans and animals. In the present study, we isolated and characterized an AFB1 degrading bacteria CG1061 from chicken cecum, exhibited an 93.7% AFB1 degradation rate by HPLC. 16S rRNA gene sequence analysis and a multiplex PCR experiment demonstrated that CG1061 was a non-pathogenic Escherichia coli. The culture supernatant of E. coli CG1061 showed an 61.8% degradation rate, whereas the degradation rates produced by the intracellular extracts was only 17.6%, indicating that the active component was constitutively secreted into the extracellular space. The degradation rate decreased from 61.8 to 37.5% when the culture supernatant was treated with 1 mg/mL proteinase K, and remained 51.3% when that treated with 100°C for 20 min. We postulated that AFB1 degradation was mediated by heat-resistant proteins. The content of AFB1 decreased rapidly when it was incubated with the culture supernatant during the first 24 h. The optimal incubation pH and temperature were pH 8.5 and 55°C respectively. According to the UPLC Q-TOF MS analysis, AFB1 was bio-transformed to the product C16H14O5 and other metabolites. Based on the results of in vitro experiments on chicken hepatocellular carcinoma (LMH) cells and in vivo experiments on mice, we confirmed that CG1061-degraded AFB1 are less toxic than the standard AFB1. E. coli CG1061 isolated from healthy chicken cerum is more likely to colonize the animal gut, which might be an excellent candidate for the detoxification of AFB1 in food and feed industry.
| S-EPMC6344451 | biostudies-literature
Project description:Microbial degradation is considered as an attractive method to eliminate exposure to aflatoxin B1 (AFB1), the most toxic mycotoxin that causes great economic losses and brings a serious threat to human and animal health, in food and feed. In this study, Bacillus amyloliquefaciens WF2020, isolated from naturally fermented pickles, could effectively degrade AFB1 ranging from 1 to 8 μg/ml, and the optimum temperature and pH value were 37-45°C and 8.0, respectively. Moreover, B. amyloliquefaciens WF2020 was considered to be a potential probiotic due to the synthesis of active compounds, absence of virulence genes, susceptibility to various antibiotics, and enhanced lifespan of Caenorhabditis elegans. Extracellular enzymes or proteins played a major role in AFB1 degradation mediated by B. amyloliquefaciens WF2020 into metabolites with low or no mutagenicity and toxicity to C. elegans. AFB1 degradation by the cell-free supernatant was stable up to 70°C, with an optimal pH of 8.0, and the cell-free supernatant could still degrade AFB1 by 37.16% after boiling for 20 min. Furthermore, B. amyloliquefaciens WF2020 caused a slight defect in fungal growth and completely inhibited AFB1 production when co-incubated with Aspergillus flavus. Additionally, B. amyloliquefaciens WF2020 suppressed the expression of 10 aflatoxin pathway genes and 2 transcription factors (alfR and alfS), suggesting that B. amyloliquefaciens WF2020 might inhibit AFB1 synthesis in A. flavus. These results indicate that B. amyloliquefaciens WF2020 and/or its extracellular enzymes or proteins have a promising potential to be applied in protecting food and feed from AFB1 contamination.
| S-EPMC9127598 | biostudies-literature
Project description:Water kefir grains (WKGs), the starter used to develop a traditional beverage named water kefir, consist of a symbiotic mixture of probiotics with diverse bioactivities, but little is known about their abilities to remove mycotoxins that have serious adverse effects on humans and animals. This study investigated the ability of WKGs to remove aflatoxin B1 (AFB1), one of the most toxic mycotoxins, under different settings, and determined the mechanism of absorption mediated by WKGs and the effect of WKGs on the toxicity induced by AFB1 and the reduction in AFB1 in cow milk and tea soups. The results showed the WKGs used herein were dominated by Lactobacillus, Acetobacter, Phenylobacterium, Sediminibacterium, Saccharomyces, Issatchenkia, and Kodamaea. HPLC analysis demonstrated that the WKGs effectively removed AFB1 at concentrations ranging from 1 to 5 µg/mL, pH values ranging from 3 to 9, and temperatures ranging from 4 to 45 °C. Additionally, the removal of AFB1 mainly depended on absorption, which was consistent with the Freundlich and pseudo-second-order kinetic models. Moreover, only 49.63% of AFB1 was released from the AFB1-WKG complex after four washes when the release of AFB1 was non-detectable. Furthermore, WKG treatment caused a dramatic reduction in the mutagenicity induced by AFB1 according to an Ames test and reduced more than 54% of AFB1 in cow milk and three tea soups. These results suggested that WKGs can act as a potential bio-absorbent with a high binding ability to detoxify AFB1 in food and feed via a chemical action step and multi-binding sites of AFB1 absorption in a wide range of scenarios.
| S-EPMC10893553 | biostudies-literature
Project description:Here, we have found for the first time that the catalytic activity of "naked" DNAzyme, a single-stranded G-quadruplex DNAzyme (S.DNAzyme), can be modulated by aflatoxin B1 (AFB1) and zearalenone (ZEN). In fact, S.DNAzyme can mimic the activity of horseradish peroxidase to perform oxidation of 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) to colored ABTS˙+ (dark green). This catalytic activity can be inhibited upon addition of AFB1, leading to color fade of the solution. But with ZEN, the solution color will change to a yellowish-brown or yellow color. Thus, we have exploited this finding to achieve label-free, naked-eye detection of AFB1 or ZEN, and have explored the possible detection mechanisms. This approach is able to detect AFB1 and ZEN concentrations as low as 0.18 μM and 0.29 μM, respectively. Even in real samples, the limit of detection values of these two analytes are lower than 3 μM. Notably, S.DNAzyme cross-reacted with three other tested aflatoxins, including aflatoxin B2, aflatoxin G1 and aflatoxin M1, revealing the potential for a broadly applicable method to identify the family of aflatoxins. The AFB1- or ZEN-triggered new catalytic reaction between "naked" DNAzyme and ABTS may offer new opportunities for on-site visual detection of mycotoxins.
| S-EPMC9644268 | biostudies-literature
Project description:There are limited data on exposure to mycotoxins in Pakistan. Here, we measured exposure to deoxynivalenol (DON), a common contaminant of wheat, and aflatoxin B1 (AFB1), a known contaminant of rice, using biomarkers of exposure. Wheat (n = 195) and rice (n = 62) samples were analyzed for AFB1 and DON levels, and the corresponding urinary biomarkers were analyzed in urine samples from a rural population (n = 264, aged 4-80 years, male 58%) using ultra-sensitive liquid chromatography-tandem mass spectrometry. AFB1 was detected in 66% of rice (5.04 ± 11.94 µg/kg) and 3% of wheat samples. AFM1 (hydroxylated form of AFB1)was detected in 69% of urine samples, mean 0.023 ± 0.048 ng/mL and DON was detected in 20% of urine samples, mean 0.170 ± 0.129 ng/mL. The maximum probable daily intake for DON derived from the urinary biomarker was 59.8 ng/kg b.w./day, which is below the Joint Food and Agriculture Organization/World Health Organization Expert Committee on Food Additives' tolerable daily intake (1000 ng/kg b.w./day). However, for aflatoxin, the derived margin of exposure (MoE) of (13.2) was well below the safe MoE (10,000) suggested by the European Food Safety Authority. The calculated aflatoxin-associated cancer risk of 0.514/105 individuals/year suggests that measures should be taken to reduce the AFB1 contamination in food, particularly rice, in Pakistan.
| S-EPMC7551319 | biostudies-literature
Project description:This study aimed to investigate the potential accumulation of mycotoxins in the lesser mealworm (Alphitobius diaperinus, LMW) and black soldier fly (Hermetia illucens, BSF) larvae. Feed was spiked with aflatoxin B₁, deoxynivalenol (DON), ochratoxin A or zearalenone, and as a mixture of mycotoxins, to concentrations of 1, 10, and 25 times the maximum limits set by the European Commission for complete feed. This maximum limit is 0.02 mg/kg for aflatoxin B₁, 5 mg/kg for DON, 0.5 mg/kg for zearalenone and 0.1 mg/kg for ochratoxin A. The mycotoxins and some of their metabolites were analysed in the larvae and residual material using a validated and accredited LC-MS/MS-based method. Metabolites considered were aflatoxicol, aflatoxin P₁, aflatoxin Q₁, and aflatoxin M₁, 3-acetyl-DON, 15-acetyl-DON and DON-3-glycoside, and α- and β-zearalenol. No differences were observed between larvae reared on mycotoxins individually or as a mixture with regards to both larvae development and mycotoxin accumulation/excretion. None of the mycotoxins accumulated in the larvae and were only detected in BSF larvae several orders of magnitude lower than the concentration in feed. Mass balance calculations showed that BSF and LMW larvae metabolized the four mycotoxins to different extents. Metabolites accounted for minimal amounts of the mass balance, except for zearalenone metabolites in the BSF treatments, which accounted for an average maximum of 86% of the overall mass balance. Both insect species showed to excrete or metabolize the four mycotoxins present in their feed. Hence, safe limits for these mycotoxins in substrates to be used for these two insect species possibly could be higher than for production animals. However, additional analytical and toxicological research to fully understand the safe limits of mycotoxins in insect feed, and thus the safety of the insects, is required.
| S-EPMC5848191 | biostudies-literature
Project description:Zearalenone (ZEA) has long been recognized as a xenoestrogen, while the endocrine disrupting effects of aflatoxin B1 (AFB1) have been identified recently. Due to co-occurrence and endocrine disrupting potentials of ZEA and AFB1, it was hypothesized that co-exposure to ZEA and AFB1 might affect breast cancer cell growth. Consequently, the aim of this study was to evaluate the combined effects of ZEA and AFB1 (1nM-100nM) on cell growth and cell cycle progression, using a human breast cancer cell line MCF-7. Our results showed that ZEA and AFB1 produced significant interactive effects on cell growth, DNA synthesis and cell cycle progression. While ZEA promoted growth, DNA synthesis and cell cycle progression, AFB1 was cytotoxic and counteracted the effects of ZEA. ZEA altered the expression of several breast cancer related genes, whereas AFB1 had minimal effects on gene expression. With the use of specific inhibitors, ERα, GPER and MAPK pathways were found to be responsible for ZEA's effects on cell growth; while MAPK pathways might be involved in cytotoxic effects by AFB1. This study is first to report the effects of co-exposure of ZEA and AFB1 on breast cancer cell growth, possibly through ER dependent pathway. This suggested that endocrine-disrupting mycotoxins that co-occur in human food can interact and influence human health. Future work on interactive effects of endocrine-disrupting mycotoxins or other xenoestrogens is warranted, which will contribute to improved risk assessments.
| S-EPMC6436804 | biostudies-literature