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Determination of six groups of mycotoxins in Chinese dark tea and the associated risk assessment
2020
Chinese dark tea is widely enjoyed for its multiple health-promoting effects and pleasant taste. However, its production involves fermentation by microbiota in raw tea, some of which are filamentous fungi and thus potential mycotoxin producers. Accordingly, whether mycotoxins pose health risk on dark tea consumption has become a public concern. In this study, a cleaning method of multi-functional column (MFC) and immunoaffinity column (IAC) in tandem combined to HPLC detection was developed and validated for determining ten mycotoxins of six groups (i.e., aflatoxins of B₁, B₂, G₁ and G₂, ochratoxin A, zearalenone, deoxynivalenol, fumonisins of B₁, B₂, and T-2) in dark teas. The interferences from secondary metabolites were effectively reduced, and the sensitivities and recoveries of the method were qualified for tea matrices. Six groups mycotoxins were determined in 108 samples representing the major Chinese dark teas by using the new method. Subsequently, the dietary exposure and health risks were evaluated for different age and gender groups in Kunming and Pu’er in China and Ulan Bator in Mongolia. The occurrence of zearalenone was 4.63% and that of ochratoxin A was 1.85%, with the other four groups mycotoxins were below the limits of quantification. The hazard index values for the five groups’ non-carcinogenic mycotoxins were far below 1.0. The deterministic risk assessment indicated no non-carcinogenic risks for dark tea consumption in the three areas. Probabilistic estimation showed that the maximum value of 95th percentile carcinogenic risk value for the aflatoxins was 2.12 × 10⁻⁸, which is far below the acceptable carcinogenic risk level (10⁻⁶). Hereby, six groups mycotoxins in Chinese dark tea showed no observed risk concern to consumers.
Показать больше [+] Меньше [-]Mycotoxins induce developmental toxicity and behavioural aberrations in zebrafish larvae
2018
Khezri, Abdolrahman | Herranz-Jusdado, Juan G. | Ropstad, Erik | Fraser, Thomas WK.
Mycotoxins are secondary metabolites produced by varieties of fungi that contaminate food and feed resources and are capable of inducing a wide range of toxicity. In the current study, we investigated developmental and behavioural toxicity in zebrafish larvae after exposure to six different mycotoxins; ochratoxin A (OTA), type A trichothecenes mycotoxin (T-2 toxin), type B trichothecenes mycotoxin (deoxynivalenol - DON), and zearalenone (ZEN) and its metabolites alpha-zearalenol (α-ZOL) and beta-zearalenol (β-ZOL). Developmental defects, hatching time, and survival were monitored until 96 h post fertilisation (hpf). The EC₅₀, LC₅₀, and IC₅₀ values were calculated. Subsequently, to assess behavioural toxicity, new sets of embryos were exposed to a series of non-lethal doses within the range of environmental and/or developmental concern. Results indicated that all the tested mycotoxins were toxic, they all induced developmental defects, and with the exception of OTA, all affected hatching time. Behavioural effects were only observed following exposure to OTA and ZEN and its metabolites, α ZOL and β ZOL. These results demonstrate that mycotoxins are teratogenic and can influence behaviour in a vertebrate model.
Показать больше [+] Меньше [-]3-Acetyldeoxynivalenol induces lysosomal membrane permeabilization-mediated apoptosis and inhibits autophagic flux in macrophages
2020
Liu, Ning | Yang, Ying | Chen, Jingqing | Jia, Hai | Zhang, Yunchang | Jiang, Da | Wu, Guoyao | Wu, Zhenlong
3-Acetyldeoxynivalenol (3-Ac-DON), the acetylated derivative of deoxynivalenol (DON), has been reported to be coexisted with DON in various cereal grains. Ingestion of grain-based food products contaminated by 3-Ac-DON might exert deleterious effects on the health of both humans and animals. However, the biological toxicity of 3-Ac-DON on macrophages and the underlying mechanisms remain largely unknown. In the present study, we showed that RAW 264.7 macrophages treated with 0.75 or 1.50 μg/mL of 3-Ac-DON resulted in DNA damage and the related cell cycle arrest at G1 phase and cell death, activation of the ribotoxic stress and the endoplasmic reticulum (ER) stress responses. The 3-Ac-DON-induced cell death was accompanied by a protective autophagy, because gene silencing of Atg5 using the small interfering RNA enhanced cell death. Results of further experiments revealed a role for lysosomal membrane permeabilization in the 3-Ac-DON triggered inhibition of autophagic flux. Additional work also showed that increased lysosomal biogenesis and leakage of cathepsin B (CTSB) from lysosomes to cytosol was critical for the 3-Ac-DON-induced cell death. Importantly, 3-Ac-DON-induced DNA damage and cell death were rescued by CA-074-me, a CTSB inhibitor. Collectively, these results indicated a critical role of lysosomal membrane permeabilization in the 3-Ac-DON-induced apoptosis of RAW 264.7 macrophages.
Показать больше [+] Меньше [-]Determination of multiple mycotoxins in paired plasma and urine samples to assess human exposure in Nanjing, China
2019
Fan, Kai | Xu, Juanjuan | Jiang, Keqiu | Liu, Xing | Meng, Jiajia | Di Mavungu, José Diana | Guo, Wenbo | Zhang, Zhiqi | Jing, Jun | Li, Hongru | Yao, Bing | Li, Hui | Zhao, Zhihui | Han, Zheng
This study was conducted to investigate mycotoxin exposure in 260 rural residents (age 18–66 years) in Nanjing, China. Paired plasma and first morning urine samples were analyzed for 26 mycotoxin biomarkers, including 12 parent mycotoxins and 14 mycotoxin metabolites, by an ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method. Mycotoxins and their metabolites were detected in 95/260 (36.5%) plasma samples and 144/260 (55.4%) urine samples. The most prevalent mycotoxin in plasma was ochratoxin A (OTA), with the incidence of 27.7% (range 0.312–9.18 μg/L), while aflatoxin B₁-lysine (AFB₁-lysine) (incidence 19.6%, range 10.5–74.5 pg/mg albumin), fumonisin B₁ (FB₁) (incidence 2.7%, range 0.305–0.993 μg/L), deoxynivalenol (DON) (incidence 2.3%, range 1.39–5.53 μg/L), zearalenone (ZEN) (incidence 6.5%, range 0.063–0.418 μg/L) and zearalanone (ZAN) (incidence 1.2%, range 0.164–0.346 μg/L) were also detected in plasma samples. Deoxynivalenol-15-glucuronide (DON-15-GlcA) was the most frequently detected urinary mycotoxin, with the incidence of 43.8% (range 0.828–37.7 μg/L). DON (incidence 10.0%, range 1.39–14.7 μg/L), DON-3-glucuronide (DON-3-GlcA) (incidence 15.8%, range 0.583–5.84 μg/L), aflatoxin M₁ (AFM₁) (incidence 10.4%, range 0.125–0.464 μg/L), ZAN (incidence 7.7%, range 0.106–1.82 μg/L), ZEN (incidence 6.9%, range 0.056–0.311 μg/L), FB₁ (incidence 3.1%, range 0.230–1.33 μg/L), T-2 toxin (incidence 2.3%, range 0.248–3.61 μg/L) and OTA (incidence 1.2%, range 0.153–0.557 μg/L) were also found in urine samples. Based on the plasma or urinary levels, the daily intakes of AFB₁, FB₁, ZEN, DON and OTA were estimated. The results showed that the investigated rural dwellers were exposed to multiple mycotoxins, especially to carcinogenic mycotoxin AFB₁ with a mean daily intake of 0.41 μg/kg·bw/day, thereby underlining a potential public health concern. To the best of our knowledge, this is the first study to evaluate human exposure to mycotoxins with direct measurements of multiple mycotoxins in paired plasma and urine samples for over 200 subjects of a single population.
Показать больше [+] Меньше [-]3-Acetyldeoxynivalenol induces cell death through endoplasmic reticulum stress in mouse liver
2021
Jia, Hai | Liu, Ning | Zhang, Yunchang | Wang, Chao | Yang, Ying | Wu, Zhenlong
Ingestion of food or cereal products contaminated by deoxynivalenol (DON) and related derivatives poses a threat to the health of humans and animals. However, the toxicity and underlying mechanisms of 3-acetyldeoxynivalenol (3-Ac-DON), an acetylated form of deoxynivalenol, have not been fully elucidated. In the present study, we showed that 3-Ac-DON caused significant oxidative damage, as shown by elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactic dehydrogenase (LDH) in serum, increased lipid peroxidation products, such as hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), decreased activities of antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD). In addition, 3-Ac-DON exposure led to elevated infiltrations of immune cell, increased apoptosis and autophagy in the liver. Interestingly, 3-Ac-DON-resulted apoptosis and liver injury were partially reduced by autophagy inhibitors. Further study showed that 3-Ac-DON-treated mice had altered ultrastructural changes of endoplasmic reticulum (ER), as well as enhanced protein levels of p-IRE1α, p-PERK, and downstream targets, indicating activation of unfolded protein response (UPR) in the liver. Importantly, 3-Ac-DON induced ER stress, oxidative damage, cell death, infiltration of immune cells, and increased mRNA levels of inflammatory cytokines were significantly abolished by 4-phenylbutyric acid (4-PBA), an ER stress inhibitor, indicating a critical role of UPR signaling for the cellular damage of the liver in response to 3-Ac-DON exposure. In conclusion, using mice as an animal model, we showed that 3-Ac-DON exposure impaired the function of liver, as shown by oxidative damage, cell death, and infiltration of immune cell, in which ER stress played an important role. Restoration of the ER function might be a preventive strategy to reduce the deleterious effect of 3-Ac-DON on the liver of animals.
Показать больше [+] Меньше [-]Zearalenone and deoxynivalenol reduced Th1-mediated cellular immune response after Listeria monocytogenes infection by inhibiting CD4+ T cell activation and differentiation
2021
Cai, Guodong | Xia, Sugan | Zhong, Fang | Liu, Shuangshuang | Gu, Jianhong | Yuan, Yan | Zhu, Guoqiang | Zou, Hui | Liu, Zongping | Bian, Jianchun
Based on the fact that mycotoxins and the food-borne bacteria coexist in the natural environment and pose a significant health hazard to humans and animals, it is important to investigate the immunosuppressive mechanism of ZEA (zearalenone), DON (deoxynivalenol), and their combination in bacterial infections. In this study, we established a mouse model of mycotoxin low-dose exposure combined with Listeria monocytogenes infection and investigated the effects of ZEA, DON and their combination on Th1-mediated anti-intracellular bacterial infection based on CD4⁺ T cell activation and differentiation using both in vitro and in vivo analyses. The present study showed that both ZEA and DON aggravated Listeria monocytogenes infection in mice and affected the activation of CD4⁺ T cells and Th1 differentiation, including the effects on costimulatory molecules CD28 and CD152 and on cross-linking of IL-12 and IL-12R, by inhibiting T cell receptor (TCR) signaling. When compared with ZEA, DON was found to have a greater impact on many related indicators. Surprisingly, the combined effects of ZEA and DON did not appear to enhance toxicity compared to treatment with the individual mycotoxins. Our findings more clearly revealed that exposure to low-dose ZEA and DON caused immunosuppression in the body by mechanisms including inhibition of CD4⁺ T cells activation and reduction of Th1 cell differentiation, thus exacerbating infection of animals by Listeria monocytogenes.
Показать больше [+] Меньше [-]Mycotoxin occurrence in breast milk and exposure estimation of lactating mothers using urinary biomarkers in São Paulo, Brazil
2021
Coppa, Carolina F.S.C. | Cirelli, Amanda C. | Gonçalves, Bruna L. | Barnabé, Eliana M.B. | Petta, Tânia | Franco, Larissa T. | Javanmardi, Fardin | Khaneghah, Amin Mousavi | Lee, Sarah H.I. | Corassin, Carlos H. | Oliveira, Carlos A.F.
In this study, the occurrence of aflatoxins (AFs), fumonisins (FBs), ochratoxin A (OTA), deoxynivalenol (DON), zearalenone (ZEN) and some of their metabolites were assessed in breast milk and urine of lactating women (N = 74) from Pirassununga, São Paulo, Brazil. Exposure estimations through urinary mycotoxin biomarkers was also performed. Samples were collected in four sampling times (May and August 2018, February and July 2019) and analyzed by liquid chromatography coupled to tandem mass spectrometry. Aflatoxin M₁ (AFM₁) was not detected in breast milk. However, two samples (3%) presented FB₁ at 2200 and 3400 ng/L, while 4 samples (5%) had OTA at the median level of 360 ng/L. In urine, AFM₁ and aflatoxin P₁ (AFP₁) were found in 51 and 11% of samples, respectively (median levels: 0.16 and 0.07 ng/mg creatinine, respectively). Urinary DON (median level: 38.59 ng/mg creatinine), OTA (median level: 2.38 ng/mg creatinine) and ZEN (median level: 0.02 ng/mg of creatinine) were quantified in 18, 8 and 10% of the samples, respectively. Mean probable daily intake (PDI) values based on urinary biomarkers were 1.58, 1.09, 5.07, and 0.05 μg/kg body weight/day for AFM₁, DON, OTA, and ZEN, respectively. Although a low mycotoxin occurrence was detected in breast milk, the PDI for the genotoxic AFs was much higher than those reported previously in Brazil, while PDI values obtained for OTA and DON were higher than recommended tolerable daily intakes. These outcomes warrant concern on the exposure of lactating women to these mycotoxins in the studied area.
Показать больше [+] Меньше [-]Deoxynivalenol induced spermatogenesis disorder by blood-testis barrier disruption associated with testosterone deficiency and inflammation in mice
2020
Cao, Zheng | Huang, Wanyue | Sun, Yiran | Li, Yanfei
Deoxynivalenol (DON) is an unavoidable cereal crops contaminants and environmental pollutants, which seriously threated the health of human and animal. DON has been reported to exert significant toxicity effects on spermatogenesis, but the underlying mechanisms remain largely inconclusive. The blood-testis barrier (BTB) provides a specialized biochemical microenvironment for maintaining spermatogenesis. Thus, we hypothesized that DON could impair BTB and lead to spermatogenesis disorder. To confirm this hypothesis, sixty male mice were intragastrically administered with 0, 1.2, 2.4 and 4.8 mg/kg body weight DON for 28 days, and several important observations were obtained in present study. First, we found that DON induced spermatogenesis disorder, reflected by the declines of sperm concentration and quality, sperm ultrastructural damage as well as seminiferous tubular damage. Then, we proved that DON induced BTB disruption as well as decreased the expressions of BTB junction proteins, including Occludin, Connexin 43 and N-cadherin. Finally, the present study showed that DON induced inflammation and inhibited T biosynthesis in testis of mice. These results revealed that DON induced spermatogenesis disorder by BTB disruption associated with testosterone deficiency and inflammation in mice, which shed a new light on the potential mechanisms of reproductive toxicity induced by DON.
Показать больше [+] Меньше [-]Deoxynivalenol induced apoptosis and inflammation of IPEC-J2 cells by promoting ROS production
2019
Kang, Ruifen | Li, Ruonan | Dai, Pengyuan | Li, Zhaojian | Li, Yansen | Li, Chunmei
Deoxynivalenol (DON) frequently detected in a wide range of foods and feeds, inducing cytotoxicity to animals and humans. To investigate the underlying mechanism of DON-induced apoptosis and inflammation in porcine small intestinal epithelium, intestinal porcine epithelial cells (IPEC-J2 cells) were chosen as objects, and were treated by different concentrations (0 μg/mL, 0.2 μg/mL, 0.5 μg/mL, 1.0 μg/mL, 2.0 μg/mL, 4.0 μg/mL, 6.0 μg/mL) of DON. The results showed that DON induced cytotoxicity of IPEC-J2 cells in a dose-dependent manner, which is demonstrated by decreasing cell viability. Compared with the control group, DON treatment increased the expressions of genes associated with inflammation and apoptosis, such as interleukin-1 beta (IL-1β), cyclooxgenase-2 (COX-2), interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-α), caspase-3, caspase-8, caspase-9, and decreased the cell anti-oxidative status. Protein immunofluorescence showed increased expression of caspase-3, nuclear factor kB (NF-κB) and phosphorylated NF-κB in IPEC-J2 cells. DON increased the content of intracellular reactive oxygen species (ROS) of IPEC-J2 cells. N-Acetyl-L-cysteine (NAC), a commonly used antioxidant, blocked DON-induced ROS generation, alleviated the DON-induced apoptosis and inflammation. These results suggested that DON-induced impairment of IPEC-J2 cells is possibly due to increased ROS production, and expressions of genes and proteins associated with apoptosis and inflammation.
Показать больше [+] Меньше [-]Testing the extraction of 12 mycotoxins from aqueous solutions by insoluble beta-cyclodextrin bead polymer
2022
Mohos, Violetta | Faisal, Zelma | Fliszár-Nyúl, Eszter | Szente, Lajos | Poór, Miklós
Mycotoxins are toxic metabolites of filamentous fungi; they are common contaminants in numerous foods and beverages. Cyclodextrins are ring-shaped oligosaccharides, which can form host-guest type complexes with certain mycotoxins. Insoluble beta-cyclodextrin bead polymer (BBP) extracted successfully some mycotoxins (e.g., alternariol and zearalenone) from aqueous solutions, including beverages. Therefore, in this study, we aimed to examine the ability of BBP to remove other 12 mycotoxins (including aflatoxin B1, aflatoxin M1, citrinin, dihydrocitrinone, cyclopiazonic acid, deoxynivalenol, ochratoxin A, patulin, sterigmatocystin, zearalanone, α-zearalanol, and β-zearalanol) from different buffers (pH 3.0, 5.0, and 7.0). Our results showed that BBP can effectively extract citrinin, dihydrocitrinone, sterigmatocystin, zearalanone, α-zearalanol, and β-zearalanol at each pH tested. However, for the removal of ochratoxin A, BBP was far the most effective at pH 3.0. Based on these observations, BBP may be a suitable mycotoxin binder to extract certain mycotoxins from aqueous solutions for decontamination and/or for analytical purposes.
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