Sample records for aflatoxin b1 afb1-induced

  1. Aflatoxin B1-induced epigenetic alterations: An overview.

    PubMed

    Dai, Yaqi; Huang, Kunlun; Zhang, Boyang; Zhu, Liye; Xu, Wentao

    2017-11-01

    Aflatoxin B1 (AFB1) is widely distributed in nature, especially in a variety of food commodities. It is confirmed to be the most toxic of all the aflatoxins. The toxicity of AFB1 has been well investigated, and it may result in severe health problems including carcinogenesis, mutagenesis, growth retardation, and immune suppression. Epigenetic modifications including DNA methylation, histone modifications and regulation of non-coding RNA play an important role in AFB1-induced disease and carcinogenesis. To better understand the evidence for AFB1-induced epigenetic alterations and the potential mechanisms of the toxicity of AFB1, we conducted a review of published studies of AFB1-induced epigenetic alterations. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Protective Effect of Selenium on Aflatoxin B1-Induced Testicular Toxicity in Mice.

    PubMed

    Cao, Zheng; Shao, Bing; Xu, Feibo; Liu, Yunfeng; Li, Yanfei; Zhu, Yanzhu

    2017-12-01

    Aflatoxins have been considered as one of the major risk factors of male infertility, and aflatoxin B1 (AFB1) is the most highly toxic and prevalent member of the aflatoxins family. Selenium (Se), an essential nutritional trace mineral for normal testicular development and male fertility, has received extensive intensive on protective effects of male reproductive system due to its potential antioxidant and activating testosterone synthesis. To investigate the protective effect of Se on AFB1-induced testicular toxicity, the mice were orally administered with AFB1 (0.75 mg/kg) and Se (0.2 mg/kg or 0.4 mg/kg) for 45 days. We found that that Se elevated testes index, sperm functional parameters (concentration, malformation, and motility), and the level of serum testosterone in AFB1-exposed mice. Moreover, our results showed that Se attenuated the AFB1-induced oxidative stress and the reduction of testicular testosterone synthesis enzyme protein expression such as steroidogenic acute regulatory protein (StAR), P450 side-chain cleavage (P450scc), and 17β-hydroxysteroid dehydrogenase (17β-HSD) in AFB1-exposed mice. These results demonstrated that Se conferred protection against AFB1-induced testicular toxicity and can be attributed to its antioxidant and increased testosterone level by stimulating protein expression of StAR and testosterone synthetic enzymes.

  3. Aflatoxin B1 Induced Systemic Toxicity in Poultry and Rescue Effects of Selenium and Zinc.

    PubMed

    Mughal, Muhammad Jameel; Peng, Xi; Kamboh, Asghar Ali; Zhou, Yi; Fang, Jing

    2017-08-01

    Among many challenges, exposure to aflatoxins, particularly aflatoxin B 1 (AFB 1 ), is one of the major concerns in poultry industry. AFB 1 intoxication results in decreased meat/egg production, hepatotoxicity, nephrotoxicity, disturbance in gastrointestinal tract (GIT) and reproduction, immune suppression, and increased disease susceptibility. Selenium (Se) and zinc (Zn), in dietary supplementation, offer easy, cost-effective, and efficient ways to neutralize the toxic effect of AFB 1 . In the current review, we discussed the impact of AFB 1 on poultry industry, its biotransformation, and organ-specific noxious effects, along with the action mechanism of AFB 1 -induced toxicity. Moreover, we explained the biological and detoxifying roles of Se and Zn in avian species as well as the protection mechanism of these two trace elements. Ultimately, we discussed the use of Se and Zn supplementation against AFB 1 -induced toxicity in poultry birds.

  4. Transcriptome, antioxidant enzyme activity and histopathology analysis of hepatopancreas from the white shrimp Litopenaeus vannamei fed with aflatoxin B1(AFB1).

    PubMed

    Zhao, Wei; Wang, Lei; Liu, Mei; Jiang, Keyong; Wang, Mengqiang; Yang, Guang; Qi, Cancan; Wang, Baojie

    2017-09-01

    Aflatoxin produced by Aspergillus flavus or Aspergillus parasiticus fungi during grain and feed processing and storage. Aflatoxins cause severe health problems reducing the yield and profitability of shrimp cultures. We sought to understand the interaction between shrimp immunity and aflatoxin B1 (AFB1), analyzing transcriptome expression, antioxidant enzyme activity, and histological features of the hepatopancreas of shrimp fed with AFB1. From over 4 million high-quality reads, de novo unigene assembly produced 103,644 fully annotated genes. A total of 1024 genes were differentially expressed in shrimp fed with AFB1, being involved in functions, such as peroxidase metabolism, signal transduction, transcriptional control, apoptosis, proteolysis, endocytosis, and cell adhesion and cell junction. Upon AFB1 challenge, there were severe histological alterations in shrimp hepatopancreas. AFB1 challenge increased the activity of several antioxidant enzymes. Our data contribute to improve the current understanding of host-AFB1 interaction, providing an abundant source for identification of novel genes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Aflatoxin B1 can be complexed with oxidised tea polyphenols and the absorption of the complexed aflatoxin B1 is inhibited in rats.

    PubMed

    Lu, Hao; Liu, Feifei; Zhu, Qiangqiang; Zhang, Mengmeng; Li, Tong; Chen, Jiming; Huang, Yewei; Wang, Xuanjun; Sheng, Jun

    2017-04-01

    Aflatoxin B1 (AFB1) is the most prevalent and carcinogenic form of the aflatoxins. In this report, we explored the interaction between AFB1 and oxidised tea polyphenols (OTP). Then, the influence of OTP on the absorption and toxicity of AFB1 in rats was investigated. We found that AFB1 can be complexed with OTP, and a transmembrane bidirectional transport experiment verified the absorption of complexed AFB1 (C-AFB1) was inhibited by OTP dramatically (P < 0.001). Animal experiments results showed that the AFB1 plus OTP group had significantly (P < 0.05) decreased AFB1-albumin (AFB1-alb) compared to the AFB1 group at 4 h after ingestion. OTP could significantly (P < 0.01) promote the elimination of AFB1 in faeces. Moreover, the liver injury induced by AFB1 was significantly inhibited by OTP. Our results demonstrated AFB1 can be complexed with OTP and the absorption of the C-AFB1 is inhibited in rats. Consequently, the liver injury induced by AFB1 can be inhibited by OTP. These results provide insight that consuming OTP-containing products, like fermented Pu-er tea, can protect damage from AFB1, and OTP may be used as a kind of food additive. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  6. Aflatoxin B1 and M1: Biological Properties and Their Involvement in Cancer Development.

    PubMed

    Marchese, Silvia; Polo, Andrea; Ariano, Andrea; Velotto, Salvatore; Costantini, Susan; Severino, Lorella

    2018-05-24

    Aflatoxins are fungal metabolites found in feeds and foods. When the ruminants eat feedstuffs containing Aflatoxin B1 (AFB1), this toxin is metabolized and Aflatoxin M1 (AFM1) is excreted in milk. International Agency for Research on Cancer (IARC) classified AFB1 and AFM1 as human carcinogens belonging to Group 1 and Group 2B, respectively, with the formation of DNA adducts. In the last years, some epidemiological studies were conducted on cancer patients aimed to evaluate the effects of AFB1 and AFM1 exposure on cancer cells in order to verify the correlation between toxin exposure and cancer cell proliferation and invasion. In this review, we summarize the activation pathways of AFB1 and AFM1 and the data already reported in literature about their correlation with cancer development and progression. Moreover, considering that few data are still reported about what genes/proteins/miRNAs can be used as damage markers due to AFB1 and AFM1 exposure, we performed a bioinformatic analysis based on interaction network and miRNA predictions to identify a panel of genes/proteins/miRNAs that can be used as targets in further studies for evaluating the effects of the damages induced by AFB1 and AFM1 and their capacity to induce cancer initiation.

  7. The antioxidant effects of vitamin A, C, and E on aflatoxin B1-induced oxidative stress in human lymphocytes.

    PubMed

    Alpsoy, L; Yildirim, A; Agar, G

    2009-03-01

    The aim of this study was to investigate the possible protective role of vitamin A, C, and E on aflatoxin B(1)-induced in human lymphocytes using biochemical approaches. The control group received dimethyl sulfoxide, the second group of cultures were administered aflatoxin B(1) (AFB(1)) at a dose of 5 muM. The other group of cultures were treated with AFB(1)+vitamin A (0.5 and 1.0 and 1.5 microM) and AFB(1)+vitamin C (25, 50, and 100 microM) and AFB(1)+vitamin E (40, 100, and 200 microM). The results of this experiment show that AFB(1) significantly decreased the level of GSH and the activities of superoxide dismutase and GPx and increased level of malondialdehyde. Simultaneous supplementation with vitamin A, C, and E restored these parameters to that of normal range. In conclusion, vitamin A, C, and E exhibited protective effects in human lymphocytes by inhibiting AFB(1)-induced ROS generation.

  8. Solvent-dependent transformation of aflatoxin B1 in soil.

    PubMed

    Starr, James M; Rushing, Blake R; Selim, Mustafa I

    2017-08-01

    To date, all studies of aflatoxin B 1 (AFB 1 ) transformation in soil or in purified mineral systems have identified aflatoxins B 2 (AFB 2 ) and G 2 (AFG 2 ) as the primary transformation products. However, identification in these studies was made using thin layer chromatography which has relatively low resolution, and these studies did not identify a viable mechanism by which such transformations would occur. Further, the use of methanol as the solvent delivery vehicle in these studies may have contributed to formation of artifactual transformation products. In this study, we investigated the role of the solvent vehicle in the transformation of AFB 1 in soil. To do this, we spiked soils with AFB 1 dissolved in water (93:7, water/methanol) or methanol and used HPLC-UV and HPLC-MS to identify the transformation products. Contrasting previous published reports, we did not detect AFB 2 or AFG 2 . In an aqueous-soil environment, we identified aflatoxin B 2a (AFB 2a ) as the single major transformation product. We propose that AFB 2a is formed from hydrolysis of AFB 1 with the soil acting as an acid catalyst. Alternatively, when methanol was used, we identified methoxy aflatoxin species likely formed via acid-catalyzed addition of methanol to AFB 1 . These results suggest that where soil moisture is adequate, AFB 1 is hydrolyzed to AFB 2a and that reactive organic solvents should be avoided when replicating natural conditions to study the fate of AFB 1 in soil.

  9. Detection of aflatoxin B1 (AFB1) in individual maize kernels using short wave infrared (SWIR) hyperspectral imaging

    USDA-ARS?s Scientific Manuscript database

    Short wave infrared hyperspectral imaging (SWIR) (1000-2500 nm) was used to detect aflatoxin B1 (AFB1) in individual maize kernels. A total of 120 kernels of four varieties (or 30 kernels per variety) that had been artificially inoculated with a toxigenic strain of Aspergillus flavus and harvested f...

  10. The hepatoprotective effect of sea buckthorn (Hippophae rhamnoides) berries on induced aflatoxin B1 poisoning in chickens 1.

    PubMed

    Solcan, Carmen; Gogu, Mihaela; Floristean, Viorel; Oprisan, Bogdan; Solcan, Gheorghe

    2013-04-01

    The leaves and berries of sea buckthorn (SB; Hippophae rhamnoides; family Elaeagnaceae) are medically claimed as having phytoantioxidant, antiinflammatory, and anticancerous properties in humans. This study evaluated the hepatoprotective activity of oil from SB berries against toxicity induced by aflatoxin B1 (AFB1) in broiler chickens. The toxicity of AFB1 led to lower total serum proteins and specifically reduced albumin (P < 0.001). Serum aspartate aminotransferase increased from 191.14 ± 11.56 to 218.80 ± 13.68 (P < 0.001). When chickens were simultaneously dosed with AFB1 and an extract of SB berries, subsequent histology of the liver showed a significant reduction of necrosis and fatty formation compared with chickens treated with AFB1 alone. Immunohistochemical results indicated that COX2, Bcl-2, and p53 were highly expressed in the liver of AFB1-treated chickens and their expression was significantly reduced by SB oil supplementation. The levels of AFB1 residues in chickens livers were significantly reduced by SB oil from 460.92 ± 6.2 ng/mL in the AFB1 group to 15.59 ± 6.1 ng/mL in the AFB1 and SB oil group. These findings suggest that SB oil has a potent hepatoprotective activity, reducing the concentration of aflatoxins in liver and diminishing their adverse effects.

  11. The production and repair of aflatoxin B sub 1 -induced DNA damage

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Leadon, S.A.

    To investigate the influence of function or activity of a DNA sequence on its repair, we have studied excision repair of aflatoxin B{sub 1} (AFB{sub 1})-induced damage in the nontranscribed, heterochromatic alpha DNA of monkey cells and in the metallothionein genes of human cells. In confluent cells, AFB{sub 1} adducts are produced in similar frequencies in alpha and in the rest of the DNA, but removal from alpha DNA is severely deficient, however, removal of AFB{sub 1} adducts from alpha DNA is enhanced by small doses of UV. The repair deficiencies are not observed in actively growing cells. We havemore » also shown that there is preferential repair of AFB{sub 1} damage in active genes. AFB{sub 1} damage is efficiently repaired in the active human metallothionein (hMT) genes, but deficiently repaired in inactive hMT genes. 51 refs., 3 tabs.« less

  12. Effect of High Protein Diet and Probiotic Lactobacillus casei Shirota Supplementation in Aflatoxin B1-Induced Rats

    PubMed Central

    Nurul Adilah, Z.; Liew, Winnie-Pui-Pui; Amin, I.

    2018-01-01

    Probiotic Lactobacillus casei Shirota (LcS) is a potential decontaminating agent of aflatoxin B1 (AFB1). However, few studies have investigated the influence of diet, especially a high protein (HP) diet, on the binding of AFB1 by probiotics. This research was conducted to determine the effect of HP diet on the ability of LcS to bind AFB1 and reduce aflatoxin M1 (AFM1) in AFB1-induced rats. Sprague Dawley rats were randomly divided into three groups: A (HP only), B (HP + 108 CFU LcS + 25 μg AFB1/kg BW), and C (HP + 25 μg AFB1/kg BW). Levels of AST and ALP were higher in all groups but other liver function's biomarkers were in the normal range, and the liver's histology showed no structural changes. The urea level of rats in group B (10.02 ± 0.73 mmol/l) was significantly lower (p < 0.05) than that of rats in group A (10.82 ± 0.26 mmol/l). The presence of carcinoma in the small intestine and colon was more obvious in group C than in group B. Moreover, rats in group B had significantly (p < 0.05) lower AFM1 concentration (0.39 ± 0.01 ng/ml) than rats in group C (5.22 ± 0.28 ng/ml). Through these findings, LcS supplementation with HP diet alleviated the adverse effects of AFB1 by preventing AFB1 absorption in the small intestine and reducing urinary AFM1.

  13. Purification of aflatoxin B1 antibody for the development of aflatoxin biosensor

    NASA Astrophysics Data System (ADS)

    Prihantoro, E. A. B.; Saepudin, E.; Ivandini, T. A.

    2017-07-01

    Aflatoxin B1 (AFB1) is produced from agricultural products especially peanuts overgrown with aspergillus flavus during the post-harvest process. Aflatoxin is classified as a highly toxic and carcinogenic substance to humans by the International Agency for Research on Cancer (IARC), WHO. This research was conducted to develop the AFB1 biosensor using antibody that specifically binds to aflatoxin B1. This antibody was produced by injecting an AFB1 hapten-protein (immunogen) to a rabbit. Antibody was obtained from rabbit's blood serum and purified using Protein A affinity chromatography and precipitation at the isoelectric point. The result showed that purification using protein A contains antibody of 4.0 mg/mL, whereas purification using precipitation at isoelectric pH contains antibody of 0.3 mg/mL. The pure antibody was tested for its specificity against AFB1, tetrahydrofuran (THF), dimethyl formamide (DMF), bovine serum albumin (BSA), and ethanol. The result revealed that THF, BSA, and ethanol were bound to antibody, while DMF showed no interaction. It was concluded that the polyclonal antibody which have been successfully purified from rabbit's blood serum using protein A affinity chromatography and precipitation methods showed an unspecific identification.

  14. Hepatitis B virus x gene and cyanobacterial toxins promote aflatoxin B1-induced hepatotumorigenesis in mice

    PubMed Central

    Lian, Min; Liu, Ying; Yu, Shun-Zhang; Qian, Geng-Sun; Wan, Shu-Guang; Dixon, Kenneth R

    2006-01-01

    AIM: To assess the combinative role of aflatoxin B1 (AFB1), cyanobacterial toxins (cyanotoxins), and hepatitis B virus (HBV) x gene in hepatotumorigenicity. METHODS: One-week-old animals carrying HBV x gene and their wild-type littermates were intraperitoneally (ip) injected with either single-dose AFB1 [6 mg/kg body weight (bw)], repeated-dose cyanotoxins (microcystin-LR or nodularin, 10 μg/kg bw once a week for 15 wk), DMSO (vehicle control) alone, or AFB1 followed by cyanotoxins a week later, and were sacrificed at 24 and 52 wk post-treatment. RESULTS: AFB1 induced liver tumors in 13 of 29 (44.8%) transgenic mice at 52 wk post-treatment, significantly more frequent than in wild-type mice (13.3%). This significant difference was not shown in the 24-wk study. Compared with AFB1 exposure alone, MC-LR and nodularin yielded approximately 3-fold and 6-fold increases in the incidence of AFB1-induced liver tumors in wild-type animals at 24 wk, respectively. HBV x gene did not further elevate the risk associated with co-exposure to AFB1 and cyanotoxins. With the exception of an MC-LR-dosed wild-type mouse, no liver tumor was observed in mice treated with cyanotoxins alone at 24 wk. Neither DMSO-treated transgenic mice nor their wild-type littermates had pathologic alterations relevant to hepatotumorigenesis in even up to 52 wk. CONCLUSION: HBV x gene and nodularin promote the development of AFB1-induced liver tumors. Co-exposure to AFB1 and MC-LR tends to elevate the risk of liver tumors at 24 wk relative to exposure to one of them. The combinative effect of AFB1, cyanotoxins and HBVx on hepatotumorigenesis is weak at 24 wk. PMID:16718789

  15. Response of Intestinal Bacterial Flora to the Long-term Feeding of Aflatoxin B1 (AFB1) in Mice.

    PubMed

    Yang, Xiai; Liu, Liangliang; Chen, Jing; Xiao, Aiping

    2017-10-12

    In order to investigate the influence of aflatoxin B1 (AFB1) on intestinal bacterial flora, 24 Kunming mice (KM mice) were randomly placed into four groups, which were labeled as control, low-dose, medium-dose, and high-dose groups. They were fed intragastrically with 0.4 mL of 0 mg/L, 2.5 mg/L, 4 mg/L, or 10 mg/L of AFB1 solutions, twice a day for 2 months. The hypervariable region V3 + V4 on 16S rDNA of intestinal bacterial flora was sequenced by the use of a high-flux sequencing system on a Miseq Illumina platform; then, the obtained sequences were analyzed. The results showed that, when compared with the control group, both genera and phyla of intestinal bacteria in the three treatment groups decreased. About one third of the total genera and one half of the total phyla remained in the high-dose group. The dominant flora were Lactobacillus and Bacteroides in all groups. There were significant differences in the relative abundance of intestinal bacterial flora among groups. Most bacteria decreased as a whole from the control to the high-dose groups, but several beneficial and pathogenic bacterial species increased significantly with increasing dose of AFB1. Thus, the conclusion was that intragastric feeding with 2.5~10 mg/mL AFB1 for 2 months could decrease the majority of intestinal bacterial flora and induce the proliferation of some intestinal bacteria flora.

  16. Protective Effect of Korean Red Ginseng against Aflatoxin B1-Induced Hepatotoxicity in Rat

    PubMed Central

    Kim, Yong-Seong; Kim, Yong-Hoon; Noh, Jung-Ran; Cho, Eun-Sang; Park, Jong-Ho; Son, Hwa-Young

    2011-01-01

    Korean red ginseng (KRG), the steamed root of Panax ginseng Meyer, has a variety of biological properties, including anti-inflammatory, antioxidant and anticancer effects. Aflatoxin B1 (AFB1) produced by the Aspergillus spp. causes acute hepatotoxicity by lipid peroxidation and oxidative DNA damage, and induces liver carcinoma in humans and laboratory animals. This study was performed to examine the protective effects of KRG against hepatotoxicity induced by AFB1 using liver-specific serum marker analysis, histopathology, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling. In addition, to elucidate the possible mechanism of hepatoprotective effects, superoxide dismutase, catalase, glutathione peroxidase, and malondialdehyde were analyzed. Rats were treated with 250 mg/kg of KRG (KRG group) or saline (AFB1 group) for 4 weeks and then received 150 μg/kg of AFB1 intraperitoneally for 3 days. Rats were sacrificed at 12 h, 24 h, 48 h, 72 h, or 1 wk after AFB1 treatment. In the KRG pre-treatment group, serum alanine aminotransferase, aspartate aminotransferase, and malondialdehyde levels were low, but superoxide dismutase, catalase, and glutathione peroxidase activities were high as compared to the AFB1 alone group. Histopathologically, AFB1 treatment induced necrosis and apoptosis in hepatocytes, and led to inflammatory cells infiltration in the liver. KRG pre-treatment ameliorated these changes. These results indicate that KRG may have protective effects against hepatotoxicity induced by AFB1 that involve the antioxidant properties of KRG. PMID:23717067

  17. Vitamin E supplementation ameliorates aflatoxin B1-induced nephrotoxicity in rats.

    PubMed

    Abdel-Hamid, Ahmed A M; Firgany, Alaa El-Din L

    2015-10-01

    Fungal toxins in nutrition can cause organ dysfunction or even failure. Aflatoxin B1 (AFB1)-induced renal impairment is not sufficiently studied regarding its extent and prevention. The aim of this experiment was to study the effect of AFB1 on renal cortical tissue and whether its possible harmful effect could be prevented by the conventional economical antioxidant, vitamin E. Forty rats were divided into four groups; I-IV. Group I represented the control while the others received vitamin E (Vit E), AFB1 and AFB1+Vit E, respectively. Renal cortex specimens were taken from each group after 25 days. Then, specimens were prepared for histological study by hematoxlyin and eosin (H&E), Masson's trichrome, caspase-3 as well as for ultrastructural examination and oxidative stress parameters evaluation. Data were morphometrically and statistically analyzed. In AFB1-treated group, focal tubulo-interstitial affection in the form of tubular cytoplasmic vacuolation, mitochondrial disruption, numerous lysosomes, marked increase in collagen deposition and in caspase-3 expression were observed. Glomerular impairment in the form of fusion of podocytes enlarged foot processes and thickening of the glomerular basement membrane (GBM) with loss of its trilaminar appearance were detected. In the group treated by AFB1+Vit E, there were minimal affection of the histological structure of the renal cortex as well as significant increase in the anti-oxidative parameters which were significantly decreased in the AFB1-treated group. Therefore, Vit E could be considered in wide experimental studies to be a first choice antioxidant of high cost-effectiveness in prevention of fungal toxins pro-oxidant-induced renal impairment. Copyright © 2015 Elsevier GmbH. All rights reserved.

  18. Investigation of Non-Covalent Interactions of Aflatoxins (B1, B2, G1, G2, and M1) with Serum Albumin

    PubMed Central

    Poór, Miklós; Bálint, Mónika; Hetényi, Csaba; Gődér, Beatrix; Kunsági-Máté, Sándor; Lemli, Beáta

    2017-01-01

    Aflatoxins are widely spread mycotoxins produced mainly by Aspergillus species. Consumption of aflatoxin-contaminated foods and drinks causes serious health risks for people worldwide. It is well-known that the reactive epoxide metabolite of aflatoxin B1 (AFB1) forms covalent adducts with serum albumin. However, non-covalent interactions of aflatoxins with human serum albumin (HSA) are poorly characterized. Thus, in this study the complex formation of aflatoxins was examined with HSA applying spectroscopic and molecular modelling studies. Our results demonstrate that aflatoxins form stable complexes with HSA as reflected by binding constants between 2.1 × 104 and 4.5 × 104 dm3/mol. A binding free energy value of −26.90 kJ mol−1 suggests a spontaneous binding process between AFB1 and HSA at room-temperature, while the positive entropy change of 55.1 JK−1 mol−1 indicates a partial decomposition of the solvation shells of the interacting molecules. Modeling studies and investigations with site markers suggest that Sudlow’s Site I of subdomain IIA is the high affinity binding site of aflatoxins on HSA. Interaction of AFB1 with bovine, porcine, and rat serum albumins was also investigated. Similar stabilities of the examined AFB1-albumin complexes were observed suggesting the low species differences of the albumin-binding of aflatoxins. PMID:29068381

  19. Aflatoxin B1 impairs sperm quality and fertilization competence.

    PubMed

    Komsky-Elbaz, A; Saktsier, M; Roth, Z

    2018-01-15

    Aflatoxins are poisonous byproducts of the soilborne fungus Aspergillus, involved in the decomposition of plant materials. Aflatoxins can be found in various food products, such as maize, sorghum, millet, rice and wheat. AFB1 is the most toxic of these, classified as a carcinogen and mutagen for both humans and animals. AFB1 has been detected in human cord blood and placenta; however, its toxic effect on sperm is less known. The current study examines sperm responses associated with AFB1 exposure. These included acrosome integrity and function, mitochondrial polarity, DNA fragmentation, fertilization competence and early embryonic development. Spermatozoa were obtained from bull ejaculate and epididymis and capacitated in vitro for 4h with 0, 0.1, 1, 10 and 100μM AFB1. Following capacitation, acrosome reaction (AR) was induced by Ca 2+ ionophore. The integrity and functionality of sperm were examined simultaneously by florescent staining. A Halosperm DNA fragmentation kit was used to evaluate DNA integrity. An in-vitro culture system was used to evaluate fertilization competence and blastocyst formation rate, using bovine oocytes. Findings indicate dose-responsive variation among compartments to AFB1 exposure. Sperm viability, expressed by integrity of the plasma membrane, was lower in sperm isolated from ejaculate or epididymis after culturing with AFB1. Exposure to AFB1 reduced the proportion of sperm from the epididymis tail undergoing acrosome reaction induced by Ca 2+ ionophore. AFB1 impaired mitochondrial membrane potential (ΔYm) in sperm isolated from ejaculate and the epididymis tail. Exposing ejaculated sperm to AFB1 increased the proportion of sperm with fragmented DNA and reduced the proportion of embryos that cleaved to the 2- to 4-cell stage, 42h postfertilization, however, the proportion of embryos that developed to blastocysts, 7days postfertilization, did not differ among groups. The findings explore the harmful effects of AFB1 on sperm viability

  20. Probing the Characterization of the Interaction of Aflatoxins B1 and G1 with Calf Thymus DNA In Vitro

    PubMed Central

    Ma, Liang; Wang, Jiaman; Zhang, Yuhao

    2017-01-01

    The binding characterization of aflatoxins with calf thymus DNA (ctDNA) under physiological conditions was investigated. Multispectroscopic techniques, ctDNA melting, viscosity measurements, and molecular docking techniques were employed to elucidate the binding mechanism of the aflatoxins with DNA. The fluorescence results indicated that both aflatoxin B1 (AFB1) and aflatoxin G1 (AFG1) bound to the ctDNA, forming complexes through hydrogen bonding. The binding constants of AFB1 and AFG1 with ctDNA reached up to 103 L·mol−1 and 104 L·mol−1, respectively, and AFG1 exhibited a higher binding propensity than that of AFB1. Furthermore, both AFB1 and AFG1 bound to the ctDNA through groove binding, as evidenced by the results of the spectroscopic, iodide quenching effect, viscosity, and ctDNA melting measurements. Changes in the circular dichroism signal manifested that both AFB1 and AFG1 induced an increase in the right-handed helicity, but only minimally influenced the base stacking of the DNA. A molecular docking study of the aflatoxin’s binding with the DNA revealed a groove binding mode, which was driven mainly by hydrogen bonding. This study of aflatoxin–ctDNA interaction may provide novel insights into the toxicological effect of the mycotoxins. PMID:28671585

  1. Sulforaphane, a cancer chemopreventive agent, induces pathways associated with membrane biosynthesis in response to tissue damage by aflatoxin B{sub 1}

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Techapiesancharoenkij, Nirachara; Fiala, Jeannette L.A.; Navasumrit, Panida

    Aflatoxin B{sub 1} (AFB{sub 1}) is one of the major risk factors for liver cancer globally. A recent study showed that sulforaphane (SF), a potent inducer of phase II enzymes that occurs naturally in widely consumed vegetables, effectively induces hepatic glutathione S-transferases (GSTs) and reduces levels of hepatic AFB{sub 1}-DNA adducts in AFB{sub 1}-exposed Sprague Dawley rats. The present study characterized the effects of SF pre-treatment on global gene expression in the livers of similarly treated male rats. Combined treatment with AFB{sub 1} and SF caused reprogramming of a network of genes involved in signal transduction and transcription. Changes inmore » gene regulation were observable 4 h after AFB{sub 1} administration in SF-pretreated animals and may reflect regeneration of cells in the wake of AFB{sub 1}-induced hepatotoxicity. At 24 h after AFB{sub 1} administration, significant induction of genes that play roles in cellular lipid metabolism and acetyl-CoA biosynthesis was detected in SF-pretreated AFB{sub 1}-dosed rats. Induction of this group of genes may indicate a metabolic shift toward glycolysis and fatty acid synthesis to generate and maintain pools of intermediate molecules required for tissue repair, cell growth and compensatory hepatic cell proliferation. Collectively, gene expression data from this study provide insights into molecular mechanisms underlying the protective effects of SF against AFB{sub 1} hepatotoxicity and hepatocarcinogenicity, in addition to the chemopreventive activity of this compound as a GST inducer. - Highlights: • This study revealed sulforaphane (SF)-deregulated gene sets in aflatoxin B{sub 1} (AFB{sub 1})-treated rat livers. • SF redirects biochemical networks toward lipid biosynthesis in AFB{sub 1}-dosed rats. • SF enhanced gene sets that would be expected to favor cell repair and regeneration.« less

  2. Dietary modulation of the biotransformation and genotoxicity of aflatoxin B(1).

    PubMed

    Gross-Steinmeyer, Kerstin; Eaton, David L

    2012-09-28

    Diet and its various components are consistently identified as among the most important 'risk factors' for cancer worldwide, yet great uncertainty remains regarding the relative contribution of nutritive (e.g., vitamins, calories) vs. non-nutritive (e.g., phytochemicals, fiber, contaminants) factors in both cancer induction and cancer prevention. Among the most potent known human dietary carcinogens is the mycotoxin, aflatoxin B(1) (AFB). AFB and related aflatoxins are produced as secondary metabolites by the molds Aspergillus flavus and Aspergillus parasiticus that commonly infect poorly stored foods including peanuts, pistachios, corn, and rice. AFB is a potent hepatocarcinogenic agent in numerous animal species, and has been implicated in the etiology of human hepatocellular carcinoma. Recent research has shown that many diet-derived factors have great potential to influence AFB biotransformation, and some efficiently protect from AFB-induced genotoxicity. One key mode of action for reducing AFB-induced carcinogenesis in experimental animals was shown to be the induction of detoxification enzymes such as certain glutathione-S-transferases that are regulated through the Keap1-Nrf2-ARE signaling pathway. Although initial studies utilized the dithiolthione drug, oltipraz, as a prototypical inducer of antioxidant response, dietary components such as suforaphane (SFN) are also effective inducers of this pathway in rodent models. However, human GSTs in general do not appear to be extensively induced by SFN, and GSTM1 - the only human GST with measurable catalytic activity toward aflatoxin B(1)-8,9-epoxide (AFBO; the genotoxic metabolite of AFB), does not appear to be induced by SFN, at least in human hepatocytes, even though its expression in human liver cells does appear to offer considerable protection against AFB-DNA damage. Although induction of detoxification pathways has served as the primary mechanistic focus of chemoprevention studies, protective effects of

  3. Effects of different sources of Saccharomyces cerevisiae biomass on milk production, composition, and aflatoxin M1 excretion in milk from dairy cows fed aflatoxin B1.

    PubMed

    Gonçalves, B L; Gonçalves, J L; Rosim, R E; Cappato, L P; Cruz, A G; Oliveira, C A F; Corassin, C H

    2017-07-01

    The aim of the present study was to evaluate the effect of different sources of Saccharomyces cerevisiae (SC) biomass (20.0 g/d) obtained from sugarcane (cell wall, CW; dried yeast, DY; autolyzed yeast, AY) and the beer industry (partially dehydrated brewery yeast, BY) on milk production, fat and protein percentages, and aflatoxin M 1 (AFM 1 ) excretion in milk from dairy cows receiving 480 µg aflatoxin B 1 (AFB 1 ) per day. A completely randomized design was used with 2 lactating cows assigned to each of 10 dietary treatments, as follows: negative controls (no AFB 1 or SC-based biomass), positive controls (AFB 1 alone), DY alone, DY + AFB 1 , BY alone, BY + AFB 1 , CW alone, CW + AFB 1 , AY alone, and AY + AFB 1 . The cows in the aflatoxin treatment group received AFB 1 from d 1 to 6, while the SC biomass was administered with the AFB 1 bolus from d 4 to 6. Aflatoxin B 1 or SC-based products did not affect milk production or milk composition during the experimental period. Aflatoxin M 1 was detected in the milk from all aflatoxin treatment group cows, reaching maximum levels at d 3 and varying from 0.52 ± 0.03 to 1.00 ± 0.04 µg/L. At end of the treatment period, CW, AY, DY, and BY removed 78%, 89%, 45%, and 50% of AFM 1 from the milk, respectively, based on the highest level found on d 3. Results indicate a potential application of industrial fermentation by-products, especially CW and AY, as a feed additive in the diets of dairy cows to reduce the excretion of AFM 1 in milk. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  4. Exposure to aflatoxins in Japan: risk assessment for aflatoxin B1.

    PubMed

    Sugita-Konishi, Y; Sato, T; Saito, S; Nakajima, M; Tabata, S; Tanaka, T; Norizuki, H; Itoh, Y; Kai, S; Sugiyama, K; Kamata, Y; Yoshiike, N; Kumagai, S

    2010-03-01

    The intake of total aflatoxins (AFT) and aflatoxin B(1) (AFB(1)) from food in Japan was estimated from AFT and AFB(1) concentration and frequency data in 24 foods (884 samples) from a 3-year retail market survey from the summer of 2004 to the winter of 2006, and by food consumption data from the National Health and Nutrition Survey performed in 2005. The AFT and AFB(1) survey revealed that peanut, peanut products, cocoa, chocolate, pistachio, white pepper, red pepper, almond, job's tears, buckwheat and corn grits are considered to be contributors of AFT (or AFB(1)) intake in Japan (maximum AFB(1) (AFT) levels ranged from 0.21 to 28.0 microg kg(-1) (from 0.21 to 9.0 microg kg(-1))) in AFT-contaminated food. A probabilistic approach using the Monte Carlo method was carried out to simulate an estimate of the AFT (or AFB(1)) intake distributions in each age group in Japan. In this study, AFB(1) intake ranged from 0.003 to 0.004 ng kg(-1) body weight day(-1) (from lower to upper limits), and the potential risk for cancer using a formula devised by the Joint Food and Agricultural Organization/World Health Organization (FAO/WHO) Expert Committee on Food Additives (JECFA) was estimated at 0.00004-0.00005 person/year/100,000 persons, even though this was in the higher levels (95.0th percentile) of the consumer population. The results suggest that the current dietary intake of AFB(1) in Japan has no appreciable effect on health.

  5. Protective effect of esculin against prooxidant aflatoxin B1-induced nephrotoxicity in mice.

    PubMed

    Naaz, Farah; Abdin, M Z; Javed, Saleem

    2014-02-01

    The study was designed to investigate the protective effect of esculin against pro-oxidant aflatoxin B1 (AFB1)-induced nephrotoxicity in mice. In this study toxicity was developed by oral administration of AFB1 at a dose of 66.60 μg/kg bw/day for 90 days in male Swiss albino mice. Esculin (150 mg/kg bw/0.2 ml/day) and standard compound ascorbic acid (300 mg/kg bw/0.2 ml/day) was given after 30 min of AFB1 administration for 90 days. Protective efficacy was assessed by measuring the levels of lipid peroxidation (LPO) and non-enzymatic antioxidants such as reduced glutathione (GSH) and also by measuring activities of enzymatic antioxidants such as glutathione peroxidase (GPX), glutathione-S-transferase (GST), glutathione reductase (GR), superoxide dismutase (SOD) and catalase (CAT) in kidney. Results were analysed at the 30(th), 60(th) and 90(th) day of the daily treatments, which showed a decrease in the level of LPO and an increase in the levels of enzymatic and non-enzymatic antioxidants. The protective effect of esculin was further proved by histopathological findings as it exhibited regenerative activities in mice renal tubules against AFB1-induced nephrotoxicity. The results obtained clearly demonstrate that the protective efficacy of esculin against pro-oxidant AFB1-induced nephrotoxicity in mice might be due to its antioxidants and free radical scavenging properties.

  6. Degradation kinetics of aflatoxin B1 and B2 in solid medium by using pulsed light irradiation.

    PubMed

    Wang, Bei; Mahoney, Noreen E; Khir, Ragab; Wu, Bengang; Zhou, Cunshan; Pan, Zhongli; Ma, Haile

    2018-04-10

    Pulsed light (PL) is a new potential technology to degrade aflatoxin. The objective of this study was to investigate the degradation characters of aflatoxin B 1 (AFB 1 ) and B 2 (AFB 2 ) treated under PL irradiation. A kinetic degradation study of AFB 1 and AFB 2 in solid medium was performed under PL irradiation at different initial concentrations of AFB 1 (229.9, 30.7 and 17.8 μg kg -1 ) and AFB 2 (248.2, 32.2 and 19.5 μg kg -1 ) and irradiation intensities (2.86, 1.60 and 0.93 W cm -2 ) of PL. A second-order reaction model was applied to describe degradation of AFB 1 and AFB 2 . The results showed that the degradation of AFB 1 and AFB 2 followed the second-order reaction kinetic model well (R 2  > 0.97). The degradation rate was proportional to the intensities of PL irradiation and the initial concentrations of aflatoxins. It is concluded that the degradation of AFB 1 and AFB 2 with the use of PL could be accurately described using the second-order reaction kinetic model. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  7. Residues of aflatoxin B1 in broiler meat: effect of age and dietary aflatoxin B1 levels.

    PubMed

    Hussain, Zahid; Khan, Muhammad Zargham; Khan, Ahrar; Javed, Ijaz; Saleemi, Muhammad Kashif; Mahmood, Sultan; Asi, Muhammad Rafique

    2010-12-01

    This study describes the effect of dietary levels of aflatoxin B1 (AFB1) and age of the birds upon the residue level in liver and muscles of broiler chicks. In three different experiments broiler chicks of 7, 14 and 28 days of age were kept for 7 days on contaminated rations having 1600, 3200 and 6400 μg/kg AFB1. AFB1 residues were detected earlier in younger birds and those fed high AFB1 dietary levels. The highest residue levels in liver and muscles of young chicks fed 6400 μg/kg AFB1 was 6.97±0.08 and 3.27±0.05 ng/g, respectively. Maximum residue concentration was high in birds of young age and those kept on high AFB1 ration. After withdrawal of AF contaminated rations, residues clearance was slow and AFB1 was detectable in liver and muscles of birds for longer duration in younger birds and those fed high AFB1 dietary levels. AFB1 residues in poultry tissues may buildup to high levels in areas with no regulatory limits on AFB1 levels of poultry feed and may pose a risk to consumers health. Copyright © 2010 Elsevier Ltd. All rights reserved.

  8. Protective effects of phenolics rich extract of ginger against Aflatoxin B1-induced oxidative stress and hepatotoxicity.

    PubMed

    A V, Vipin; K, Raksha Rao; Kurrey, Nawneet Kumar; K A, Anu Appaiah; G, Venkateswaran

    2017-07-01

    Aflatoxin B 1 (AFB 1 ) is one of the predominant mycotoxin contaminant in food and feed, causing oxidative stress and hepatotoxicity. Ginger phenolics have been reported for its antioxidant potential and hepatoprotective activity. The present study investigated the protective effects of phenolics rich ginger extract (GE) against AFB 1 induced oxidative stress and hepatotoxicity, in vitro and in vivo. The phenolic acid profiles of GE showed 6-gingerol and 6-shogaol as predominant components. Pretreatment of HepG2 cells with GE significantly inhibited the production of intracellular reactive oxygen species (ROS), DNA strand break, and cytotoxicity induced by AFB 1 . A comparable effect was observed in in vivo. Male Wistar rats were orally treated with GE (100 and 250mg/kg) daily, with the administration of AFB 1 (200μg/kg) every alternative day for 28days. Treatment with GE significantly reduced AFB 1 induced toxicity on the serum markers of liver damage. In addition, GE also showed significant hepatoprotective effect by reducing the lipid peroxidation and by enhancing the antioxidant enzymes activities. These results combined with liver histopathological observations indicated that GE has potential protective effect against AFB 1 induced hepatotoxicity. Additionally, administration of GE up-regulated Nrf2/HO-1 pathway, which further proved the efficiency of GE to inhibit AFB 1 induced hepatotoxicity. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

  9. Vaccination of heifers with anaflatoxin improves the reduction of aflatoxin b1 carry over in milk of lactating dairy cows.

    PubMed

    Giovati, Laura; Gallo, Antonio; Masoero, Francesco; Cerioli, Carla; Ciociola, Tecla; Conti, Stefania; Magliani, Walter; Polonelli, Luciano

    2014-01-01

    It was previously reported that injection of anaflatoxin B1 (AnAFB1) conjugated to keyhole limpet hemocyanin (KLH), together with Freund's adjuvant, was effective in inducing in cows a long lasting titer of anti-aflatoxin B1 (AFB1) antibodies (Abs), cross-reacting with other aflatoxins, which were able to hinder, proportionally to their titer, the secretion of aflatoxin M1 (AFM1) into the milk of cows continuously fed with AFB1. According to anti-AFB1 Ab titer, 50% of the vaccinated cows were recognized as high responder animals. In an attempt to prepare a more effective formulation for vaccination of cows, it was compared the immunogenicity, in Holstein Friesian heifers, of AnAFB1 covalently conjugated to KLH or to recombinant diphtheria toxin (CRM197) molecules, and injected together with various adjuvants. This study demonstrated that injection of AnAFB1 conjugated to KLH and mixed with complete (priming) and incomplete Freund's adjuvant (boosters), as in the previous schedule of immunization, was the most effective regimen for inducing Ab responses against AFB1, although pre-calving administration could increase the effectiveness of vaccination, resulting in 100% high responder animals. After one booster dose at the beginning of the milk production cycle, anti-AFB1 Ab titers were comparable to those recorded at the end of the immunization schedule, and proved to be effective in reducing significantly AFB1 carry over, as AFM1, from feed to milk. Pre-calving vaccination of dairy heifers with conjugated AnAFB1, adjuvated with complete and incomplete Freund's adjuvant, may represent the most effective tool for preventing the public health hazard constituted by milk and cheese contaminated with aflatoxins.

  10. Failure of catalase to protect against aflatoxin B{sub 1}-induced mouse lung tumorigenicity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Guindon, Katherine A.; Foley, Julie F.; Maronpot, Robert R.

    The carcinogenic mycotoxin aflatoxin B{sub 1} (AFB{sub 1}) induces 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation in mouse lung, an effect that can be prevented by treatment with polyethylene glycol-conjugated catalase (PEG-CAT). G {yields} T transversion mutation in K-ras, an early event in AFB{sub 1}-induced mouse lung carcinogenesis, is thought to result from AFB{sub 1}-8,9-exo-epoxide binding to DNA to form AFB{sub 1}-N{sup 7}-guanine, but may also result from formation of 8-OHdG. Therefore, oxidative DNA damage may be important in AFB{sub 1} carcinogenicity. The objective of this study was to determine whether PEG-CAT would prevent AFB{sub 1} tumorigenicity. Mouse lung tumorigenesis was assessed following treatmentmore » of female A/J mice with 300 kU/kg PEG-CAT ip and/or 50 mg/kg AFB{sub 1}. Mice were killed 7 months post-treatment and tumors greater than 1 mm in diameter were excised. Unexpectedly, the mean number of tumors per mouse in the PEG-CAT + AFB{sub 1} group (8.81 {+-} 3.64, n = 47) was greater than that of the group treated with AFB{sub 1} alone (7.05 {+-} 3.45, n = 42) (P < 0.05). The tumors obtained from mice treated with PEG-CAT + AFB{sub 1} were larger than those from mice treated with AFB{sub 1} alone (P < 0.05). There was no difference in K-ras exon 1 mutation spectrum or in the histological diagnosis of tumors between AFB{sub 1} and PEG-CAT + AFB{sub 1} groups (P > 0.05). In vitro incubation with mouse liver catalase (CAT) resulted in conversion of [{sup 3}H]AFB{sub 1} into a DNA-binding species, a possible explanation for the results observed in vivo. These results demonstrate that PEG-CAT is not protective against AFB{sub 1} carcinogenicity in mouse lung despite preventing DNA oxidation.« less

  11. Development of High Capacity Enterosorbents for Aflatoxin B1 and Other Hazardous Chemicals.

    PubMed

    Wang, Meichen; Maki, Cody R; Deng, Youjun; Tian, Yanan; Phillips, Timothy D

    2017-09-18

    Previously, a calcium montmorillonite clay (NovaSil) included in the diet of animals has been shown to bind aflatoxin B1 (AfB1) and reduce the symptoms of aflatoxicosis. To investigate and improve the capacity and efficacy of clay-based materials as aflatoxin sorbents, we developed and tested calcium and sodium montmorillonite clays amended with nutrients including l-carnitine and choline. Also, we determined the sorption of AfB1 by isothermal analysis and tested the ability of these amended sorbents to protect adult hydra from AfB1 toxicity. The results showed that exchanging montmorillonite clays with l-carnitine and choline inhibited swelling of the clays and increased the sorption capacity and efficacy of clay surfaces for AfB1. Results from dehydroxylated and heat-collapsed clays suggested that AfB1 was primarily adsorbed in the clay interlayer, as predicted from thermodynamic calculations and computational modeling. The hydra bioassay further indicated that the modified clays can significantly protect adult hydra from AfB1 with as low as 0.005% clay inclusion. This enterosorbent therapy may also be applied to screen hazardous chemicals such as pesticides and PAHs based on similar sorption mechanisms. Taken together, enterosorbent therapy could be delivered in nutritional supplements, foods that are vulnerable to aflatoxin contamination, flavored liquids and animal feeds during emergencies and outbreaks of acute aflatoxicosis, and as a screening model for hazardous environmental chemicals.

  12. Fluorescent sensor systems based on nanostructured polymeric membranes for selective recognition of Aflatoxin B1.

    PubMed

    Sergeyeva, Tetyana; Yarynka, Daria; Piletska, Elena; Lynnik, Rostyslav; Zaporozhets, Olga; Brovko, Oleksandr; Piletsky, Sergey; El'skaya, Anna

    2017-12-01

    Nanostructured polymeric membranes for selective recognition of aflatoxin B1 were synthesized in situ and used as highly sensitive recognition elements in the developed fluorescent sensor. Artificial binding sites capable of selective recognition of aflatoxin B1 were formed in the structure of the polymeric membranes using the method of molecular imprinting. A composition of molecularly imprinted polymer (MIP) membranes was optimized using the method of computational modeling. The MIP membranes were synthesized using the non-toxic close structural analogue of aflatoxin B1, ethyl-2-oxocyclopentanecarboxylate as a dummy template. The MIP membranes with the optimized composition demonstrated extremely high selectivity towards aflatoxin B1 (AFB1). Negligible binding of close structural analogues of AFB1 - aflatoxins B2 (AFB2), aflatoxin G2 (AFG2), and ochratoxin A (OTA) was demonstrated. Binding of AFB1 by the MIP membranes was investigated as a function of both type and concentration of the functional monomer in the initial monomer composition used for the membranes' synthesis, as well as sample composition. The conditions of the solid-phase extraction of the mycotoxin using the MIP membrane as a stationary phase (pH, ionic strength, buffer concentration, volume of the solution, ratio between water and organic solvent, filtration rate) were optimized. The fluorescent sensor system based on the optimized MIP membranes provided a possibility of AFB1 detection within the range 14-500ngmL -1 demonstrating detection limit (3Ϭ) of 14ngmL -1 . The developed technique was successfully applied for the analysis of model solutions and waste waters from bread-making plants. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. Metabolism of aflatoxin B-1 in cotton bolls

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mellon, J.E.; Lee, L.S.

    Aspergillus flavus is a fungus capable of producing the potent carcinogen aflatoxin (AFB-1) when it infects developing cotton seed. Although high levels of toxin can readily be isolated from internal tissues of infected seeds, very low toxin levels are observed in the fiber-linter matrix. In order to test the hypothesis that constituents associated with the lint of the host plant are metabolizing aflatoxin, {sup 14}C-AFB-1 was introduced into cotton bolls (30 days postanthesis). Other sets of bolls received inoculations of toxigenic or nontoxigenic strains of A. flavus plus exogenous {sup 14}C-AFB-1. In addition to the exogenously applied {sup 14}C-AFB-1, atmore » least two new labelled metabolites were recovered from the test bolls. One of these metabolites was very polar and remained on the origin of the thin layer analysis system. Test bolls which received both A. flavus and AFB-1 produced significantly lower levels of this polar metabolite. Results indicated that some constituent(s) associated with cotton fiber may metabolize fungal-produced aflatoxin, rather than inhibit its formation.« less

  14. Systems responses of rats to aflatoxin B1 exposure revealed with metabonomic changes in multiple biological matrices.

    PubMed

    Zhang, Limin; Ye, Yangfang; An, Yanpeng; Tian, Yuan; Wang, Yulan; Tang, Huiru

    2011-02-04

    Exposure to aflatoxins causes liver fibrosis and hepatocellular carcinoma posing a significant health risk for human populations and livestock. To understand the mammalian systems responses to aflatoxin-B1 (AFB1) exposure, we analyzed the AFB1-induced metabonomic changes in multiple biological matrices (plasma, urine, and liver) of rats using (1)H NMR spectroscopy together with clinical biochemistry and histopathologic assessments. We found that AFB1 exposure caused significant elevation of glucose, amino acids, and choline metabolites (choline, phosphocholine, and glycerophosphocholine) in plasma but reduction of plasma lipids. AFB1 also induced elevation of liver lipids, amino acids (tyrosine, histidine, phenylalanine, leucine, isoleucine, and valine), choline, and nucleic acid metabolites (inosine, adenosine, and uridine) together with reduction of hepatic glycogen and glucose. AFB1 further caused decreases in urinary TCA cycle intermediates (2-oxoglutarate and citrate) and elevation of gut microbiota cometabolites (phenylacetylglycine and hippurate). These indicated that AFB1 exposure caused hepatic steatosis accompanied with widespread metabolic changes including lipid and cell membrane metabolisms, protein biosynthesis, glycolysis, TCA cycle, and gut microbiota functions. This implied that AFB1 exposure probably caused oxidative-stress-mediated impairments of mitochondria functions. These findings provide an overview of biochemical consequences of AFB1 exposure and comprehensive insights into the metabolic aspects of AFB1-induced hepatotoxicity in rats.

  15. Aflatoxin B1 levels in groundnut products from local markets in Zambia.

    PubMed

    Njoroge, Samuel M C; Matumba, Limbikani; Kanenga, Kennedy; Siambi, Moses; Waliyar, Farid; Maruwo, Joseph; Machinjiri, Norah; Monyo, Emmanuel S

    2017-05-01

    In Zambia, groundnut products (milled groundnut powder, groundnut kernels) are mostly sold in under-regulated markets. Coupled with the lack of quality enforcement in such markets, consumers may be at risk to aflatoxin exposure. However, the level of aflatoxin contamination in these products is not known. Compared to groundnut kernels, milled groundnut powder obscures visual indicators of aflatoxin contamination in groundnuts such as moldiness, discoloration, insect damage or kernel damage. A survey was therefore conducted from 2012 to 2014, to estimate and compare aflatoxin levels in these products (n = 202), purchased from markets in important groundnut growing districts and in urban areas. Samples of whole groundnut kernels (n = 163) and milled groundnut powder (n = 39) were analysed for aflatoxin B 1 (AFB 1 ) by competitive enzyme-linked immunosorbent assay (cELISA). Results showed substantial AFB 1 contamination levels in both types of groundnut products with maximum AFB 1 levels of 11,100 μg/kg (groundnut kernels) and 3000 μg/kg (milled groundnut powder). However, paired t test analysis showed that AFB 1 contamination levels in milled groundnut powder were not always significantly higher (P > 0.05) than those in groundnut kernels. Even for products from the same vendor, AFB 1 levels were not consistently higher in milled groundnut powder than in whole groundnut kernels. This suggests that vendors do not systematically sort out whole groundnut kernels of visually poor quality for milling. However, the overall contamination levels of groundnut products with AFB 1 were found to be alarmingly high in all years and locations. Therefore, solutions are needed to reduce aflatoxin levels in such under-regulated markets.

  16. Cytochrome P450 2A13 enhances the sensitivity of human bronchial epithelial cells to aflatoxin B1-induced DNA damage

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yang, Xuejiao; Jiaojiang District Center for Disease Control and Prevention, 518 Jingdong Rd., Taizhou 318000; Zhang, Zhan

    Cytochrome P450 2A13 (CYP2A13) mainly expresses in human respiratory system and mediates the metabolic activation of aflatoxin B1 (AFB1). Our previous study suggested that CYP2A13 could increase the cytotoxic and apoptotic effects of AFB1 in immortalized human bronchial epithelial cells (BEAS-2B). However, the role of CYP2A13 in AFB1-induced DNA damage is unclear. Using BEAS-2B cells that stably express CYP2A13 (B-2A13), CYP1A2 (B-1A2), and CYP2A6 (B-2A6), we compared their effects in AFB1-induced DNA adducts, DNA damage, and cell cycle changes. BEAS-2B cells that were transfected with vector (B-vector) were used as a control. The results showed that AFB1 (5–80 nM) dose-more » and time-dependently induced DNA damage in B-2A13 cells. AFB1 at 10 and 80 nM significantly augmented this effect in B-2A13 and B-1A2 cells, respectively. B-2A6 cells showed no obvious DNA damage, similar to B-vector cells and the vehicle control. Similarly, compared with B-vector, B-1A2 or B-2A6 cells, B-2A13 cells showed more sensitivity in AFB1-induced γH2AX expression, DNA adduct 8-hydroxy-deoxyguanosine formation, and S-phase cell-cycle arrest. Furthermore, AFB1 activated the proteins related to DNA damage responses, such as ATM, ATR, Chk2, p53, BRCA1, and H2AX, rather than the proteins related to DNA repair. These effects could be almost completely inhibited by 100 μM nicotine (a substrate of CYP2A13) or 1 μM 8-methoxypsoralen (8-MOP; an inhibitor of CYP enzyme). Collectively, these findings suggest that CYP2A13 plays an important role in low-concentration AFB1-induced DNA damage, possibly linking environmental airborne AFB1 to genetic injury in human respiratory system. - Highlights: • CYP2A13 plays a critical role in low concentration of AFB1-induced DNA damage. • B-2A13 cells were more sensitive to AFB1 than B-1A2 cells and B-2A6 cells. • AFB1 dose- and time-dependently induced DNA damage in B-2A13 cells • AFB1-induced DNA adducts and damage can be inhibited by

  17. Study of aflatoxicosis reduction: effect of Alchornea cordifolia on biomarkers in an aflatoxin B1 exposed rats.

    PubMed

    Adépo, Jean-Baptiste Aholia; Manda, Pierre; Ngbé, Jean Verdier; Diakité, Aïssata; Tigori Sangaré, Béatrice; Dano, Sébastien Djédjé

    2018-01-17

    The toxicity of aflatoxins results in cancer and liver disease. Several natural substances such as plants exhibited their ability to inhibit the initiation of aflatoxin carcinogenesis. The aim of this study was to evaluate the effect of Alchornea cordifolia on biomarkers in an aflatoxin B1 (AFB1) exposed rats. The contents of polyphenols, flavonoids and the antioxidant activity of A. cordifolia ethanolic leaf extract (EELac) were assessed. Groups of rats were treated orally with a daily dose of a mixture of AFB1 at a dose of 150 μg/kg body weight and EELac (50, 100 and 300 mg/kg body weight) for 21 days. Biomarkers of AFB1, such as the AFB1-lysine adduct and aflatoxin M1 were assayed in blood and urine, respectively, using an HPLC system with a fluorescence detector. The contents of polyphenols and flavonoids were 6783.23 ± 272.76 μg EAG/g and 10.54 ± 3.15% of dry matter, respectively. EELac showed a good antioxidant activity (IC50 = 12.65 ± 0.13 μg/mL). The administration of the mixture (AFB1 + EELac) at different doses significantly reduced the level of AFB1-lysine adduct from 14.04 ± 2.1 to 4.13 ± 0.9 ng/mg albumin and that of Aflatoxin M1 (AFM1) from 456 ± 16 to 220 ± 24 ng/mL (p <0.05). The rate of reduction was 70.58% for AFB1-lysine adduct and 51.75% for AFM1. A. cordifolia could be used in the prevention of toxicity induced by AFB1 on account of its high content in phenolic compounds.

  18. Effect of γ-radiation on the production of aflatoxin B1 by Aspergillus parasiticus in raisins (Vitis vinifera L.)

    NASA Astrophysics Data System (ADS)

    Kanapitsas, Alexandros; Batrinou, Anthimia; Aravantinos, Athanasios; Markaki, Panagiota

    2015-01-01

    Aflatoxin B1 (AFB1) mostly produced by Aspergillus flavus and Aspergillus parasiticus, is an extremely toxic and carcinogenic metabolite. The effect of gamma irradiation at dose of 10 kGy on the production of aflatoxin B1 (AFB1) inoculated by Aspergillus parasiticus in raisins (Vitis vinifera L.) and on AFB1 in contaminated samples, was investigated. Values of the amount of aflatoxin B1 produced on the 12th day of incubation, after irradiation, showed that gamma radiation exposure at 10 kGy decreased AFB1 production at 65% compared with the non-irradiated sample, on the same day. The application of 10 kGy gamma radiation directly on 100 ng of AFB1 which were spiked in raisins resulted in ~29% reduction of AFB1. According to the risk assessment analysis the Provisional Maximum Tolerable Daily Intake (PMTDI) of 1.0 ng AFB1 kg-1bw, indicates that consumers are less exposed to AFB1 from the irradiated raisins.

  19. Dual effects of phloretin on aflatoxin B1 metabolism: activation and detoxification of aflatoxin B1.

    PubMed

    Gao, Shang Shang; Chen, Xiao Yan; Zhu, Ri Zhe; Choi, Byung-Min; Kim, Sun Jun; Kim, Bok-Ryang

    2012-01-01

    Typically, chemopreventive agents involve either induction of phase II detoxifying enzymes and/or inhibition of cytochrome P450 enzymes (CYPs) that are required for the activation of procarcinogens. In this study, we investigated the protective effects of phloretin against aflatoxin B1 (AFB1) activation to the ultimate carcinogenic intermediate, AFB(1)-8, 9-epoxide (AFBO), and its subsequent detoxification. Phloretin markedly inhibited formation of the epoxide with human liver microsomes in a dose-dependent manner. Phloretin also inhibited the activities of nifedipine oxidation and ethoxyresorufin O-deethylase (EROD) in human liver microsomes. These data show that phloretin strongly inhibits CYP1A2 and CYP3A4 activities, which are involved in the activation of AFB1. Phloretin increased glutathione S-transferase (GST) activity of alpha mouse liver 12 (AML 12) cells in a dose-dependent manner. GST activity toward AFBO in cell lysates treated with 20 μM phloretin was 23-fold that of untreated control cell lysates. The expression of GSTA3, GSTA4, GSTM1, GSTP1 and GSTT1 was induced by phloretin in a dose-dependent manner in AML 12 cells. GSTP1, GSTM1, and GSTT1 were able to significantly increase the conjugation of AFBO with glutathione. Concurrently, induction of the GST isozyme genes was partially associated with the Nrf2/ARE pathway. Taken together, the results demonstrate that phloretin has a strong chemopreventive effect against AFB1 through its inhibitory effect on CYP1A2, CYP3A4, and its inductive effect on GST activity. Copyright © 2012 International Union of Biochemistry and Molecular Biology, Inc.

  20. Fisetin Modulates Antioxidant Enzymes and Inflammatory Factors to Inhibit Aflatoxin-B1 Induced Hepatocellular Carcinoma in Rats

    PubMed Central

    Maurya, Brajesh Kumar; Trigun, Surendra Kumar

    2016-01-01

    Fisetin, a known antioxidant, has been found to be cytotoxic against certain cell lines. However, the mechanism by which it inhibits tumor growth in vivo remains unexplored. Recently, we have demonstrated that Aflatoxin-B1 (AFB1) induced hepatocarcinogenesis is associated with activation of oxidative stress-inflammatory pathway in rat liver. The present paper describes the effect of in vivo treatment with 20 mg/kg b.w. Fisetin on antioxidant enzymes vis-a-vis oxidative stress level and on the profile of certain proinflammatory cytokines in the hepatocellular carcinoma (HCC) induced by two doses of 1 mg/kg b.w. AFB1 i.p. in rats. The reduced levels of most of the antioxidant enzymes, coinciding with the enhanced level of reactive oxygen species in the HCC liver, were observed to regain their normal profiles due to Fisetin treatment. Also, Fisetin treatment could normalize the enhanced expression of TNFα and IL1α, the two proinflammatory cytokines, reported to be involved in HCC pathogenesis. These observations were consistent with the regression of neoplastic lesion and declined GST-pi (placental type glutathione-S-transferase) level, a HCC marker, in the liver of the Fisetin treated HCC rats. The findings suggest that Fisetin attenuates oxidative stress-inflammatory pathway of AFB1 induced hepatocarcinogenesis. PMID:26682000

  1. Dietary vitamin E in White Leghorn layer breeder hens: a strategy to combat aflatoxin B1-induced damage.

    PubMed

    Khan, Wajid Arshad; Khan, Muhammad Zargham; Khan, Ahrar; Hassan, Zahoor Ul; Rafique, Shahid; Saleemi, Muhammad Kashif; Ahad, Abdul

    2014-01-01

    Mycotoxins are unavoidable contaminants of animal and human feed and food respectively. This study was designed to investigate the protective activity of vitamin E (Vit E) in White Leghorn breeder hens and their progeny against aflatoxin B1 (AFB1)-induced damage. The results indicated a significant decrease in egg production and quality in the groups exposed to dietary AFB1. A detectable amount of AFB1 residue appeared in the eggs during the first week of mycotoxin exposure at levels ≥ 2.5 mg kg(-1), which reached its peak (0.403 ± 0.04 ng/g [mean ± standard deviation]) during the second week of the experiment (in the group fed 10 mg kg(-1)). Feeding Vit E + AFB1 resulted in higher AFB1 residues (0.467 ± 0.03) when compared with the hens fed AFB1 alone. The resistance of red blood cells to oxidative damage was decreased, while embryonic mortalities and deformities were increased in the AFB1-fed groups. The protective effect of Vit E on these parameters was noted in the groups fed lower doses of AFB1. After the withdrawal of mycotoxin-contaminated feed, most of the parameters returned towards normal within 2 weeks, except AFB1 residues that were still detectable. From the findings of this study one can conclude that the addition of Vit E in the diet of hens provided only partial protection against AFB1-induced damage.

  2. In vitro interaction of actinomycetes isolates with Aspergillus flavus: impact on aflatoxins B1 and B2 production.

    PubMed

    Verheecke, C; Liboz, T; Darriet, M; Sabaou, N; Mathieu, F

    2014-06-01

    This work aimed to study the interaction between Actinomycetal isolates and Aspergillus flavus to promote mutual antagonism in contact. Thirty-seven soilborn Streptomyces spp. isolates were chosen as potential candidates. After a 10-day in vitro co-incubation period, 27 isolates respond to the criteria, that is, mutual antagonism in contact. Further aflatoxins B1 and B2 analysis revealed that those 27 isolates reduced aflatoxin B1 residual concentration from 38·6 to 4·4%, depending on the isolate. We selected 12 isolates and tested their capacity to reduce AFB1 in pure culture to start identifying the mechanisms involved in its reduction. AFB1 was reduced by eight isolates. The remaining AFB1 concentration varied between 82·2 and 15·6%. These findings led us to suggest that these eight isolates could be used as biocontrol agents against AFB1 and B2 with low risk of impacting the natural microbial equilibrium. Interaction between Aspergillus flavus and Actinomycetes isolates was conducted in vitro. Actinomycetes isolates having a mutual antagonism in contact with A. flavus were chosen for further aflatoxins production study. This is a new approach based to develop biocontrol against aflatoxins accumulation in maize while respecting natural microbial equilibrium. © 2014 The Society for Applied Microbiology.

  3. Cytoprotective effect of palm kernel cake phenolics against aflatoxin B1-induced cell damage and its underlying mechanism of action.

    PubMed

    Oskoueian, Ehsan; Abdullah, Norhani; Zulkifli, Idrus; Ebrahimi, Mahdi; Karimi, Ehsan; Goh, Yong Meng; Oskoueian, Armin; Shakeri, Majid

    2015-10-30

    Palm kernel cake (PKC), a by-product of the palm oil industry is abundantly available in many tropical and subtropical countries. The product is known to contain high levels of phenolic compounds that may impede the deleterious effects of fungal mycotoxins. This study focused on the evaluation of PKC phenolics as a potential cytoprotective agent towards aflatoxin B1 (AFB1)-induced cell damage. The phenolic compounds of PKC were obtained by solvent extraction and the product rich in phenolic compounds was labeled as phenolic-enriched fraction (PEF). This fraction was evaluated for its phenolic compounds composition. The antioxidant activity of PEF was determined by using 1,1-diphenyl-2-picryl-hydrazil scavenging activity, ferric reducing antioxidant power, inhibition of ß-carotene bleaching, and thiobarbituric acid reactive substances assays. The cytotoxicity assay and molecular biomarkers analyses were performed to evaluate the cytoprotective effects of PEF towards aflatoxin B1 (AFB1)-induced cell damage. The results showed that PEF contained gallic acid, pyrogallol, vanillic acid, caffeic acid, syringic acid, epicatechin, catechin and ferulic acid. The PEF exhibited free radical scavenging activity, ferric reducing antioxidant power, ß-carotene bleaching inhibition and thiobarbituric acid reactive substances inhibition. The PEF demonstrated cytoprotective effects in AFB1-treated chicken hepatocytes by reducing the cellular lipid peroxidation and enhancing antioxidant enzymes production. The viability of AFB1-treated hepatocytes was improved by PEF through up-regulation of oxidative stress tolerance genes and down-regulation of pro-inflammatory and apoptosis associated genes. The present findings supported the proposition that the phenolic compounds present in PKC could be a potential cytoprotective agent towards AFB1 cytotoxicity.

  4. Reduction of aflatoxin level in aflatoxin-induced rats by the activity of probiotic Lactobacillus casei strain Shirota.

    PubMed

    Nikbakht Nasrabadi, E; Jamaluddin, R; Abdul Mutalib, M S; Khaza'ai, H; Khalesi, S; Mohd Redzwan, S

    2013-05-01

    Aflatoxin B1 (AFB1 ) is considered as the most toxic food contaminant, and microorganisms, especially bacteria, have been studied for their potential to reduce the bioavailability of mycotoxins including aflatoxins. Therefore, this research investigated the efficacy of oral administration of Lactobacillus casei Shirota (LcS) in aflatoxin-induced rats. Sprague Dawley rats were divided into three groups of untreated control, the group induced with AFB1 only, and the group given probiotic in addition to AFB1. In the group induced with AFB1 only, food intake and body weight were reduced significantly. The liver and kidney enzymes were significantly enhanced in both groups induced with AFB1 , but they were lower in the group given LcS. AFB1 was detected from all serum samples except for untreated control group's samples. Blood serum level of AFB1 in the group induced with AFB1 only was significantly higher than the group which received probiotic as a treatment (P < 0·05), and there was no significant difference between the control group and the group treated with probiotic. LcS supplementation could improve the adverse effect of AFB1 induction on rats' body weight, plasma biochemical parameters and also could reduce the level of AFB1 in blood serum. This study's outcomes contribute to better understanding of the potential of probiotic to reduce the bioavailability ofAFB1 . Moreover, it can open an opportunity for future investigations to study the efficacy of oral supplementation of probiotic LcS in reducing aflatoxin level in human. © 2013 The Society for Applied Microbiology.

  5. Effects of feeding corn naturally contaminated with aflatoxin B1 and B2 on hepatic functions of broilers.

    PubMed

    Yang, J; Bai, F; Zhang, K; Bai, S; Peng, X; Ding, X; Li, Y; Zhang, J; Zhao, L

    2012-11-01

    The purpose of this study was to evaluate the effects of feeding corn naturally contaminated with aflatoxin B(1) (AFB(1)) and aflatoxin B(2) (AFB(2)) on serum biochemical parameters, hepatic antioxidant enzyme activities, and pathological lesions of broilers. In total, 1,200 Cobb male broilers were randomly allocated into 5 treatments, with 8 replicates per treatment and 30 birds per replicate, in a 42-d experiment. The dietary treatments were as follows: control, 25, 50, 75, and 100% contaminated corn groups. Results showed that serum aspartate aminotransferase activity in the 75 and 100% contaminated groups were higher than that in the control group on d 21 (P < 0.05). Decreased content of hepatic total protein and increased activities of hepatic glutathione reductase and glutathione-S-transferase were observed as the percentage of contaminated corn increased (P < 0.05). The activity of superoxide dismutase and the content of hepatic malondialdehyde increased when the broilers were fed with more than 50% contaminated corn (P < 0.05). A reduction in glutathione peroxidase level was observed in the AFB(1)- and AFB(2)-contaminated groups on d 21 (P < 0.05). The average pathological lesion scores and apoptosis rate of liver cells increased as the concentration of dietary AFB(1) and AFB(2) increased. Ultrastructural changes were found in the livers of broilers fed 100% contaminated corn. In conclusion, diets containing AFB(1) and AFB(2) could induce pathological lesions in the livers, slightly change the serum biochemical parameters, and damage the hepatic antioxidant functions when the inclusion of AFB(1)- and AFB(2)-contaminated corn reached or exceeded 50%.

  6. Determination of the aflatoxin AFB1 from corn by direct analysis in real time-mass spectrometry (DART-MS).

    PubMed

    Busman, Mark; Liu, Jihong; Zhong, Hongjian; Bobell, John R; Maragos, Chris M

    2014-01-01

    Direct analysis in real time (DART) ionisation coupled to a high-resolution mass spectrometer (MS) was used for screening of aflatoxins from a variety of surfaces and the rapid quantitative analysis of a common form of aflatoxin, AFB1, extracted from corn. Sample preparation procedure and instrument parameter settings were optimised to obtain sensitive and accurate determination of aflatoxin AFB1. 84:16 acetonitrile water extracts of corn were analysed by DART-MS. The lowest calibration level (LCL) for aflatoxin AFB1 was 4 μg kg⁻¹. Quantitative analysis was performed with the use of matrix-matched standards employing the ¹³C-labelled internal standard for AFB1. DART-MS of spiked corn extracts gave linear response in the range 4-1000 μg kg⁻¹. Good recoveries (94-110%) and repeatabilities (RSD = 0.7-6.9%) were obtained at spiking levels of 20 and 100 μg kg⁻¹ with the use of an isotope dilution technique. Trueness of data obtained for AFB1 in maize by DART-MS was demonstrated by analysis of corn certified reference materials.

  7. Aflatoxin B1 and sterigmatocystin survey in beer sold in China.

    PubMed

    Zhao, Yarong; Huang, Jianxiang; Ma, Liyan; Liu, Shuai; Wang, Fuhua

    2017-03-01

    A total of 101 samples of beer from the Chinese market were analysed for the presence of aflatoxin B 1 (AFB 1 ) and sterigmatocystin (STC), using methods based on liquid chromatography-tandem mass spectrometry. The limit of quantification and the limit of detection in beer were 0.1 and 0.03 µg/kg, respectively. Recoveries of AFB 1 and STC from spiked beer samples were 97.8-103.6% and 92.7-102.1%, respectively. None of the beer purchased samples were contaminated with AFB 1 or STC.

  8. Effects of Aflatoxin B1 and Fumonisin B1 on Blood Biochemical Parameters in Broilers

    PubMed Central

    Tessari, Eliana N. C.; Kobashigawa, Estela; Cardoso, Ana Lúcia S. P.; Ledoux, David R.; Rottinghaus, George E.; Oliveira, Carlos A. F.

    2010-01-01

    The individual and combined effects of dietary aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on liver pathology, serum levels of aspartate amino-transferase (AST) and plasma total protein (TP) of broilers were evaluated from 8 to 41 days of age. Dietary treatments included a 3 × 3 factorial arrangement with three levels of AFB1 (0, 50 and 200 μg AFB1/kg), and three levels of FB1 (0, 50 and 200 mg FB1/kg). At 33 days post feeding, with the exception of birds fed 50 mg FB1 only, concentrations of AST were higher (p < 0.05) in all other treatment groups when compared with controls. Plasma TP was lower (p < 0.05) at six days post feeding in groups fed 200 μg AFB1/kg alone or in combination with FB1. At day 33 days post feeding, with the exception of birds fed the highest combination of AFB1 and FB1 which had higher plasma TP than control birds, plasma TP of birds fed other dietary treatments were similar to controls. Broilers receiving the highest levels of AFB1 and FB1 had bile duct proliferation and trabecular disorder in liver samples. AFB1 singly or in combination with FB at the levels studied, caused liver damage and an increase in serum levels of AST. PMID:22069595

  9. Broad-spectrum immunoaffinity cleanup for the determination of aflatoxins B1, B2, G1, G2, M1, M2 in Ophiocordyceps sinensis and its pharmaceutical preparations by ultra performance liquid chromatography tandem mass spectrometry.

    PubMed

    Sun, Shujuan; Xie, Jie; Peng, Tao; Shao, Bing; Zhu, Kui; Sun, Yuanze; Yao, Kai; Gu, Qiang; Zhang, Jing; Fan, Chunlin; Chen, Ying; Jiang, Haiyang

    2017-11-15

    An ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed and validated for the simultaneous determination of aflatoxins B 1 , B 2 , G 1 , G 2 , M 1 and M 2 (AFB 1 , AFB 2 , AFG 1 , AFG 2 , AFM 1 and AFM 2 ) in Ophiocordyceps sinensis and its pharmaceutical preparations. A rapid and reliable immunoaffinity column containing a broad-spectrum monoclonal antibody for six aflatoxins was used for sample cleanup. Under the optimized conditions, the home-made immunoaffinity column capacity were about 315, 319, 292, 102, 444 and 369ng/mL gel for AFB 1 , AFB 2 , AFG 1 , AFG 2 , AFM 1 and AFM 2 , respectively. Recoveries for all tested aflatoxins ranged from 79.28% to 103.42% with relative standard deviation less than 8%. The limits of quantitation were in the range of 0.008-0.045μg/kg. Among 31 real samples analyzed, one sample was contaminated with AFB 1 , AFB 2 and AFM 1 at levels of 0.483, 0.068 and 0.104μg/kg, respectively. The established method is simple, accurate, and can be effectively used to determine the aflatoxins in Ophiocordyceps sinensis and its pharmaceutical preparations. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Absence of the aflatoxin biosynthesis gene, norA, allows accumulation of deoxyaflatoxin B1 in Aspergillus flavus cultures.

    PubMed

    Ehrlich, Kenneth C; Chang, Perng-Kuang; Scharfenstein, Leslie L; Cary, Jeffrey W; Crawford, Jason M; Townsend, Craig A

    2010-04-01

    Biosynthesis of the highly toxic and carcinogenic aflatoxins in select Aspergillus species from the common intermediate O-methylsterigmatocystin has been postulated to require only the cytochrome P450 monooxygenase, OrdA (AflQ). We now provide evidence that the aryl alcohol dehydrogenase NorA (AflE) encoded by the aflatoxin biosynthetic gene cluster in Aspergillus flavus affects the accumulation of aflatoxins in the final steps of aflatoxin biosynthesis. Mutants with inactive norA produced reduced quantities of aflatoxin B(1) (AFB(1)), but elevated quantities of a new metabolite, deoxyAFB(1). To explain this result, we suggest that, in the absence of NorA, the AFB(1) reduction product, aflatoxicol, is produced and is readily dehydrated to deoxyAFB(1) in the acidic medium, enabling us to observe this otherwise minor toxin produced in wild-type A. flavus.

  11. The metabolism of aflatoxin B1 by hepatocytes isolated from rats following the in vivo administration of some xenobiotics

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Metcalfe, S.A.; Neal, G.E.

    Isolated rat hepatocytes, an intact cellular system capable of performing phase I and phase II metabolism, have been used to investigate metabolism of aflatoxin B1. These cells were found to metabolise (/sup 14/C)aflatoxin B1 to aflatoxins M1 and Q1, and to radiolabelled polar material, presumably conjugates, as analysed by h.p.l.c., t.l.c. and radioactive determination. In vivo administration of the mixed function oxidase inducers, phenobarbitone and 3-methylcholanthrene, resulted in enhanced hepatocyte phase I (microsomal) metabolism of aflatoxin B1. In contrast to metabolism of AFB1 by in vitro subcellular systems increased production of polar material (conjugated metabolites) derived from (/sup 14/C)aflatoxin B1more » was also detected in hepatocytes isolated from these pretreated animals. Formation of aflatoxin Q1 by isolated hepatocytes appeared to be mediated by cytochrome P450-linked enzymes whereas cytochrome P448-linked enzymes were apparently involved in aflatoxin M1 production. Chronic feeding of aflatoxin B1 to rats enhanced hepatocyte production of conjugated material only and did not elevate cellular cytochrome P450 levels, thus suggesting that aflatoxin B1 is not an inducer of its own primary metabolism.« less

  12. Carry-Over of Aflatoxin B1 to Aflatoxin M1 in High Yielding Israeli Cows in Mid- and Late-Lactation

    PubMed Central

    Britzi, Malka; Friedman, Shmulik; Miron, Joshua; Solomon, Ran; Cuneah, Olga; Shimshoni, Jakob A.; Soback, Stefan; Ashkenazi, Rina; Armer, Sima; Shlosberg, Alan

    2013-01-01

    The potent hepatotoxin and carcinogen aflatoxin B1 (AFB1) is a common mycotoxin contaminant of grains used in animal feeds. Aflatoxin M1 (AFM1) is the major metabolite of AFB1 in mammals, being partially excreted into milk, and is a possible human carcinogen. The maximum permitted concentration of AFM1 in cows’ milk is 0.05 μg/kg in Israel and the European Union. Since milk yield and the carry-over of AFB1 in the feed to AFM1 in the milk are highly correlated, it was considered important to determine the AFM1 carry-over in Israeli-Holstein dairy cows, distinguished by world record high milk production. Twelve such cows were used to determine AFM1 carry-over following daily oral administration of feed containing ~86 μg AFB1 for 7 days. The mean carry-over rate at steady-state (Days 3–7) was 5.8% and 2.5% in mid-lactation and late-lactation groups, respectively. The carry-over appears to increase exponentially with milk yield and could be described by the equation: carry-over% = 0.5154 e0.0521 × milk yield, with r2 = 0.6224. If these data truly reflect the carry-over in the national Israeli dairy herd, the maximum level of AFB1 in feed should not exceed 1.4 μg/kg, a value 3.6 times lower than the maximum residue level currently applied in Israel. PMID:23325299

  13. The Protective Role of Selenium in AFB1-Induced Tissue Damage and Cell Cycle Arrest in Chicken's Bursa of Fabricius.

    PubMed

    Hu, Ping; Zuo, Zhicai; Wang, Fengyuan; Peng, Xi; Guan, Ke; Li, Hang; Fang, Jing; Cui, Hengmin; Su, Gang; Ouyang, Ping; Zhou, Yi

    2018-03-06

    Aflatoxin B 1 (AFB 1 ) is a naturally occurring secondary metabolites of Aspergillus flavus and Aspergillus parasiticus, and is the most toxic form of aflatoxins. Selenium (Se) with antioxidant and detoxification functions is one of the essential trace elements for human beings and animals. This study aims to evaluate the protective effects of Se on AFB 1 -induced tissue damage and cell cycle arrest in bursa of Fabricius (BF) of chickens. The results showed that a dietary supplement of 0.4 mg·kg -1 Se alleviated the histological lesions induced by AFB 1 , as demonstrated by decreasing vacuoles and nuclear debris, and relieving oxidative stress. Furthermore, flow cytometry studies showed that a Se supplement protected AFB 1 -induced G 2 M phase arrest at 7 days and G 0 G 1 phase arrest at 14 and 21 days. Moreover, the mRNA expression results of ATM, Chk2, p53, p21, cdc25, PCNA, cyclin D 1 , cyclin E 1 , cyclin B 3 , CDK6, CDK2, and cdc2 indicated that Se supplement could restore these parameters to be close to those in the control group. It is concluded that a dietary supplement of 0.4 mg kg -1 Se could diminish AFB 1 -induced immune toxicity in chicken's BF by alleviating oxidative damage and cell cycle arrest through an ATM-Chk2-cdc25 route and the ATM-Chk2-p21 pathway.

  14. Microbial degradation of aflatoxin B1: Current status and future advances.

    PubMed

    Verheecke, C; Liboz, T; Mathieu, F

    2016-11-21

    Aflatoxin B1 (AFB1) is a natural toxin produced by many food-contaminant fungi and is a threat to human and animal health. This review summarizes current knowledge of the different ways to limit AFB1 in the food chain. We start by introducing current data and reviews available on the prevention of AFB1 occurrence, on AFB1 non-biological decontamination and biological adsorption. We then focus on microbial AFB1-degradation. The latter has already been well studied using living organisms, supernatants or purified enzymes. This review compiles information on the variety of protocols and the efficacy of the different sub-kingdoms or classes of microorganisms or their enzymes. We pay particular attention to publications closest to in vivo applications of microbial AFB1-degradation. In addition, this review also provides a summary of the currently known microbial degradation metabolites of AFB1 and their levels of toxicity, and provides recommendations on the most promising techniques to pursue the aim of minimizing ABF1 in the food supply. Copyright © 2016. Published by Elsevier B.V.

  15. NEIL1 protects against aflatoxin-induced hepatocellular carcinoma in mice.

    PubMed

    Vartanian, Vladimir; Minko, Irina G; Chawanthayatham, Supawadee; Egner, Patricia A; Lin, Ying-Chih; Earley, Lauriel F; Makar, Rosemary; Eng, Jennifer R; Camp, Matthew T; Li, Liang; Stone, Michael P; Lasarev, Michael R; Groopman, John D; Croy, Robert G; Essigmann, John M; McCullough, Amanda K; Lloyd, R Stephen

    2017-04-18

    Global distribution of hepatocellular carcinomas (HCCs) is dominated by its incidence in developing countries, accounting for >700,000 estimated deaths per year, with dietary exposures to aflatoxin (AFB 1 ) and subsequent DNA adduct formation being a significant driver. Genetic variants that increase individual susceptibility to AFB 1 -induced HCCs are poorly understood. Herein, it is shown that the DNA base excision repair (BER) enzyme, DNA glycosylase NEIL1, efficiently recognizes and excises the highly mutagenic imidazole ring-opened AFB 1 -deoxyguanosine adduct (AFB 1 -Fapy-dG). Consistent with this in vitro result, newborn mice injected with AFB 1 show significant increases in the levels of AFB 1 -Fapy-dG in Neil1 -/- vs. wild-type liver DNA. Further, Neil1 -/- mice are highly susceptible to AFB 1 -induced HCCs relative to WT controls, with both the frequency and average size of hepatocellular carcinomas being elevated in Neil1 -/- The magnitude of this effect in Neil1 -/- mice is greater than that previously measured in Xeroderma pigmentosum complementation group A (XPA) mice that are deficient in nucleotide excision repair (NER). Given that several human polymorphic variants of NEIL1 are catalytically inactive for their DNA glycosylase activity, these deficiencies may increase susceptibility to AFB 1 -associated HCCs.

  16. MicroRNA Responses to the Genotoxic Carcinogens Aflatoxin B1 and Benzo[a]pyrene in Human HepaRG Cells.

    PubMed

    Marrone, April K; Tryndyak, Volodymyr; Beland, Frederick A; Pogribny, Igor P

    2016-02-01

    Recent advances in toxicogenomics present an opportunity to develop new in vitro testing methodologies to identify human carcinogens. We have investigated microRNA expression responses to the treatment of human liver HepaRG cells with the human genotoxic carcinogens aflatoxin B1 (AFB1) and benzo[a]pyrene (B[a]P), and the structurally similar compounds aflatoxin B2 (AFB2) and benzo[e]pyrene (B[e]P) that exhibit minimal carcinogenic potential. We demonstrate that treatment of HepaRG cells with AFB1 or B[a]P resulted in specific changes in the expression of miRNAs as compared with their non-carcinogenic analogues, particularly in a marked over-expression of miR-410. An additional novel finding is the dose- and time-dependent inhibition of miR-122 in AFB1-treated HepaRG cells. Mechanistically, the AFB1-induced down-regulation of miR-122 was attributed to inhibition of the HNF4A/miR-122 regulatory pathway. These results demonstrate that HepaRG cells can be used to investigate miRNA responses to xenobiotic exposure, and illustrate the existence of early non-genotoxic events, in addition to a well-established genotoxic mode of action changes, in the mechanism of AFB1 and B[a]P carcinogenicity. Published by Oxford University Press on behalf of the Society of Toxicology 2015. This work is written by US Government employees and is in the public domain in the US.

  17. Association of reduction of AFB1-induced liver tumours by antioxidants with increased activity of microsomal enzymes.

    PubMed

    Nyandieka, H S; Wakhis, J; Kilonzo, M M

    1990-10-01

    The influence of nutritional factors on aflatoxin B1 (AFB1)-induced liver tumours was investigated in rats. When a dose of 500 micrograms AFB1/kg body weight was given to rats in the absence of any anticarcinogen, 80 per cent of the rats developed liver tumours as compared to 0 to 40 per cent in those which received anticarcinogens. While beta-carotene totally inhibited the development of liver tumours ascorbic acid, selenium, and uric acid reduced the percentages of tumour-bearing rats to 13 per cent each. GSH and vitamin E also reduced these percentages to 20 and 40 per cent respectively. The reduction of tumour incidence by each anticarcinogen was associated with induction of increased microsomal enzyme activity. Inhibition of AFB1-induced liver cancer development thus seems to occur through microsomal enzyme induction and AFB1 activation.

  18. Aflatoxin B1 Tolerance and Accumulation in Black Soldier Fly Larvae (Hermetia illucens) and Yellow Mealworms (Tenebrio molitor).

    PubMed

    Bosch, Guido; Fels-Klerx, H J van der; Rijk, Theo C de; Oonincx, Dennis G A B

    2017-06-02

    Crops contaminated with fungal mycotoxins such as aflatoxin B1 (AFB1) are often downgraded or removed from the food chain. This study aimed to evaluate the tolerance and accumulation of AFB1 in two insect species to determine whether they could be used to retain condemned mycotoxin contaminated crops in the food chain. First, instar black soldier fly larvae ( Hermetia illucens , BSF) and yellow mealworm ( Tenebrio molitor , YMW) were fed poultry feed spiked with AFB1 and formulated to contain levels of 0.01, 0.025, 0.05, 0.10, 0.25, and up to 0.5 mg/kg dry feed. Poultry feed without any additions and feed with only the solvent added served as controls. The AFB1 in the feed did not affect survival and body weight in the BSF and YMW larvae ( p > 0.10), indicating a high tolerance to aflatoxin B1 in both species. Furthermore, AFB1 and aflatoxin M1 (AFM1) were below the detection limit (0.10 µg/kg) in BSF larvae, whereas the YMW had AFB1 levels that were approximately 10% of the European Union's legal limit for feed materials and excreted AFM1. It is concluded that both BSF larvae and YMW have a high AFB1 tolerance and do not accumulate AFB1.

  19. Aflatoxin B1 Tolerance and Accumulation in Black Soldier Fly Larvae (Hermetia illucens) and Yellow Mealworms (Tenebrio molitor)

    PubMed Central

    Bosch, Guido; van der Fels-Klerx, H. J.; de Rijk, Theo C.; Oonincx, Dennis G. A. B.

    2017-01-01

    Crops contaminated with fungal mycotoxins such as aflatoxin B1 (AFB1) are often downgraded or removed from the food chain. This study aimed to evaluate the tolerance and accumulation of AFB1 in two insect species to determine whether they could be used to retain condemned mycotoxin contaminated crops in the food chain. First, instar black soldier fly larvae (Hermetia illucens, BSF) and yellow mealworm (Tenebrio molitor, YMW) were fed poultry feed spiked with AFB1 and formulated to contain levels of 0.01, 0.025, 0.05, 0.10, 0.25, and up to 0.5 mg/kg dry feed. Poultry feed without any additions and feed with only the solvent added served as controls. The AFB1 in the feed did not affect survival and body weight in the BSF and YMW larvae (p > 0.10), indicating a high tolerance to aflatoxin B1 in both species. Furthermore, AFB1 and aflatoxin M1 (AFM1) were below the detection limit (0.10 µg/kg) in BSF larvae, whereas the YMW had AFB1 levels that were approximately 10% of the European Union’s legal limit for feed materials and excreted AFM1. It is concluded that both BSF larvae and YMW have a high AFB1 tolerance and do not accumulate AFB1. PMID:28574433

  20. Changes in serum cytokine levels, hepatic and intestinal morphology in aflatoxin B1-induced injury: modulatory roles of melatonin and flavonoid-rich fractions from Chromolena odorata.

    PubMed

    Akinrinmade, Fadeyemi Joseph; Akinrinde, Akinleye Stephen; Amid, Adetayo

    2016-05-01

    Aflatoxins are known to produce chronic carcinogenic, mutagenic, and teratogenic effects, as well as acute inflammatory effects, especially in the gastrointestinal tract. The potentials of the flavonoid-rich extract from Chromolena odorata (FCO) and melatonin (a standard anti-oxidant and anti-inflammatory agent) against aflatoxin B1 (AFB1)-induced alterations in pro-inflammatory cytokine levels and morphology of liver and small intestines were evaluated in this study. We utilized Wistar albino rats (200-230 g) randomly divided into five groups made up of group A, control rats; group B, rats given AFB1 (2.5 mg/kg, intraperitoneal) twice on days 5 and 7; rats in groups C, D, and E were treated with melatonin (10 mg/kg, intraperitoneal) or oral doses of FCO1 (50 mg/kg) and FCO2 (100 mg/kg) for 7 days, respectively, along with AFB1 injection on days 5 and 7. Serum levels of interleukin 1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) were determined using commercial ELISA kits and histopathological evaluation of the liver, duodenum, and ileum were also carried out. We observed significant elevation (p < 0.05) in serum IL-1β correlating with hemorrhages and leucocytic and lymphocytic infiltration in the liver and intestines as evidences of an acute inflammatory response to AFB1 administration. All treatments yielded significant reduction (p < 0.05) in IL-1β levels, although TNF-α levels were not significantly altered in all rats that received AFB1, irrespective of the treatments. Melatonin and FCO2 produced considerable protection of hepatic tissues, although melatonin was not quite effective in protecting the intestinal lesions. Our findings suggest a modulation of cytokine expression that may, in part, be responsible for the abilities of C. odorata or melatonin in amelioration of hepatic and intestinal lesions associated with aflatoxin B1 injury.

  1. Determination of aflatoxin B1 levels in organic spices and herbs.

    PubMed

    Tosun, Halil; Arslan, Recep

    2013-01-01

    Organically produced spices and herbs were analyzed for determination of aflatoxin B1 (AFB1) by ELISA using immunoaffinity column. For this purpose 93 organic spices and 37 organic herbs were randomly selected from organic markets and organic shops in Turkey. AFB1 was detected in 58 organic spice and 32 organic herb samples. Among organic spice samples, the maximum value was detected in cinnamon sample (53 μg/kg). AFB1 was not detected in thyme samples. AFB1 levels of 41 organic spice samples were above the EU regulatory limit (5 μg/kg). Among organic herb samples the highest concentration of AFB1 (52.5 μg/kg) was detected in a rosehip sample. AFB1 levels of 21 organic herb samples were above the regulatory limits of the European Union. These results showed that more stringent measures must be taken for the prevention of mold contamination in the production of organic spices and herbs.

  2. Determination of Aflatoxin B1 Levels in Organic Spices and Herbs

    PubMed Central

    Tosun, Halil; Arslan, Recep

    2013-01-01

    Organically produced spices and herbs were analyzed for determination of aflatoxin B1 (AFB1) by ELISA using immunoaffinity column. For this purpose 93 organic spices and 37 organic herbs were randomly selected from organic markets and organic shops in Turkey. AFB1 was detected in 58 organic spice and 32 organic herb samples. Among organic spice samples, the maximum value was detected in cinnamon sample (53 μg/kg). AFB1 was not detected in thyme samples. AFB1 levels of 41 organic spice samples were above the EU regulatory limit (5 μg/kg). Among organic herb samples the highest concentration of AFB1 (52.5 μg/kg) was detected in a rosehip sample. AFB1 levels of 21 organic herb samples were above the regulatory limits of the European Union. These results showed that more stringent measures must be taken for the prevention of mold contamination in the production of organic spices and herbs. PMID:23766719

  3. Aflatoxin B1 affects apoptosis and expression of death receptor and endoplasmic reticulum molecules in chicken spleen.

    PubMed

    Zhu, Panpan; Zuo, Zhicai; Zheng, Zhixiang; Wang, Fengyuan; Peng, Xi; Fang, Jing; Cui, Hengmin; Gao, Caixia; Song, Hetao; Zhou, Yi; Liu, Xici

    2017-11-21

    Aflatoxin B 1 (AFB 1 ) is a natural product of the Aspergillus genus of molds, which grow on several foodstuffs stored in hot moist conditions, and is among the most potent hepatocarcinogens and immunosuppression presently known. The latter was related to the up-regulated apoptosis of immune organs. However, the effect of expression of death receptor and endoplasmic reticulum molecules in AFB 1 -induced apoptosis of chicken splenocytes was largely unknown. The objective of this study was to investigate this unknown field. One hundred and forty four one-day-old chickens were randomly divided into control group (0 mg/kg AFB 1 ) and AFB 1 group (0.6 mg/kg AFB 1 ), respectively and fed with AFB 1 for 21 days. Histological observation demonstrated that AFB 1 caused slight congestion and lymphocytic depletion in the spleen. TUNEL and flow cytometry assays showed the excessive apoptosis of splenocytes provoked by AFB 1 . Moreover, quantitative real-time PCR analysis revealed that AFB 1 induced the elevated mRNA expression of Fas, FasL, TNF-α, TNF-R 1 , Caspase-3, Caspase-8, Caspase-10, Grp78 and Grp94 in the spleen. These findings suggested that AFB 1 could lead the excessive apoptosis and alter the expression of death receptor and endoplasmic reticulum molecules in chicken spleen.

  4. The aflatoxin B1 -fumonisin B1 toxicity in BRL-3A hepatocytes is associated to induction of cytochrome P450 activity and arachidonic acid metabolism.

    PubMed

    Mary, Verónica S; Arias, Silvina L; Otaiza, Santiago N; Velez, Pilar A; Rubinstein, Héctor R; Theumer, Martín G

    2017-06-01

    Human oral exposure to aflatoxin B 1 (AFB 1 ) and fumonisin B 1 (FB 1 ) is associated with increased hepatocellular carcinoma. Although evidence suggested interactive AFB 1 -FB 1 hepatotoxicity, the underlying mechanisms remain mostly unidentified. This work was aimed at evaluating the possible AFB 1 -FB 1 interplay to induce genetic and cell cycle toxicities in BRL-3A rat hepatocytes, reactive oxygen species (ROS) involvement, and the AFB 1 metabolizing pathways cytochrome P450 (CYP) and arachidonic acid (ArAc) metabolism as ROS contributors. Flow cytometry of stained BRL-3A hepatocytes was used to study the cell cycle (propidium iodide), ROS intracellular production (DCFH-DA, HE, DAF-2 DA), and phospholipase A activity (staining with bis-BODIPY FL C11-PC). The CYP1A activity was assessed by the 7-ethoxyresorufin-O-deethylase (EROD) assay. Despite a 48-h exposure to FB 1 (30 μM) not being genotoxic, the AFB 1 (20 μM)-induced micronucleus frequency was overcome by the AFB 1 -FB 1 mixture (MIX), presumably showing toxin interaction. The mycotoxins blocked G1/S-phase, but only MIX caused cell death. Overall, the oxidative stress led these alterations as the pretreatment with N-acetyl-l-cysteine reduced such toxic effects. While AFB 1 had a major input to the MIX pro-oxidant activity, with CYP and ArAc metabolism being ROS contributors, these pathways were not involved in the FB 1 -elicited weak oxidative stress. The MIX-induced micronucleus frequency in N-acetyl-l-cysteine pretreated cells was greater than that caused by AFB 1 without antioxidants, suggesting enhanced AFB 1 direct genotoxicity probably owing to the higher CYP activity and ArAc metabolism found in MIX. The metabolic pathways modulation by AFB 1 -FB 1 mixtures could raise its hepatocarcinogenic properties. © 2017 Wiley Periodicals, Inc.

  5. Reduction of aflatoxin B1 to aflatoxicol: a comprehensive DFT study provides clues to its toxicity.

    PubMed

    Karabulut, Sedat; Paytakov, Guvanchmyrat; Leszczynski, Jerzy

    2014-12-01

    Aflatoxicol (AFL) is one of most the important metabolites of aflatoxin B1 (AFB1). AFL can be formed through enzymatic or synthetic reduction of AFB1. Various experimental and theoretical studies have been focused on the AFB1 due to its high toxicity and carcinogenicity. The selective reduction of AFB1 carbonyls, molecular structure of AFL and its effect on toxicity has been studied here by the density functional theory (DFT) method. Although the toxicity of AFL is 18 times lower than that of AFB1, it has been concluded that both molecular structures have similar potency to form an exo-epoxide (AFEP) analogue which can bind to DNA. Calculations revealed that only one of the three possible tautomers of AFL is stable, both in the gas phase and water. The electronic properties of aflatoxicol are calculated as similar to aflatoxin B1 and this may be an explanation of similar carcinogenicity and toxicity of these compounds, which has been proved by experimental results. © 2014 Society of Chemical Industry.

  6. Effect of dietary resveratrol in ameliorating aflatoxin B1-induced changes in broiler birds.

    PubMed

    Sridhar, M; Suganthi, R U; Thammiaha, V

    2015-12-01

    Consumption of aflatoxin B1 (AFB1) contaminated feed by poultry affects the health of broiler birds causing severe economic losses. The use of phytochemicals is a safe, effective, alternative and practical approach to combat the toxic effect of AF in broilers. Resveratrol, a polyphenol derived from red grapes, berries and peanuts, exerts anti-inflammatory, antioxidant and immunomodulatory effects. Our study was aimed at evaluating the possible protective effects of resveratrol against the adverse effects of AFB1 in broiler birds. A feeding trial of 42 days of duration was undertaken in a completely randomized design with five dietary treatments: G1-AFB1(1.0 ppm); G2-CTR (basal diet alone); G3-AFB1(1.0 ppm)+Resv 0.5%; G4-AFB1(1.0 ppm)+Resv 1%; and G5-Resv 1%. Gain in body weight (BWG) and feed intake (FI) was observed to be highest (p < 0.05) in the AFB1 birds followed by the control group. Feed conversion ratio was lowest in G2-CTR birds and failed to record any significant variation (p > 0.05) between groups as well as within groups. Birds fed resveratrol at both 0.5% and 1.0% levels in combination with AFB1 as well as alone along with basal diet had lower BWG and FI between the fourth and fifth week and also at the fifth week (p < 0.05). No variation (p > 0.05) was obtained in the FCR of AFB1 and resveratrol group of broiler birds. AFB1 feeding significantly increased the activities of aspartate-(AST) and alanine-(ALT) amino transferase, superoxide dismutase (SOD) and catalase (CAT) activities (p < 0.05) but lowered glucose, cholesterol and triglyceride levels in serum. Supplementation of resveratrol helped in increasing the activities of the oxidative enzymes and in improving the plasma total antioxidant capacity (TAOC) and total protein (TP) significantly (p < 0.05) and protein values. The livers of AFB1 group showed degeneration of hepatocytes, bile duct hyperplasia and microgranuloma formation. In resveratrol supplemented birds, the severity and degree of the

  7. Degradation of Aflatoxin B1 during the Fermentation of Alcoholic Beverages

    PubMed Central

    Inoue, Tomonori; Nagatomi, Yasushi; Uyama, Atsuo; Mochizuki, Naoki

    2013-01-01

    Aflatoxin B1 (AFB1) is a contaminant of grain and fruit and has one of the highest levels of carcinogenicity of any natural toxin. AFB1 and the fungi that produce it can also contaminate the raw materials used for beer and wine manufacture, such as corn and grapes. Therefore, brewers must ensure strict monitoring to reduce the risk of contamination. In this study, the fate of AFB1 during the fermentation process was investigated using laboratory-scale bottom and top beer fermentation and wine fermentation. During fermentation, cool wort beer samples and wine must samples were artificially spiked with AFB1 and the levels of AFB1 remaining after fermentation were analyzed. AFB1 levels were unchanged during both types of fermentation used for beer but were reduced to 30% of their initial concentration in wine. Differential analysis of the spiked and unspiked wine samples showed that the degradation compound was AFB2a, a hydrated derivative of AFB1. Thus, the results showed that the risk of AFB1 carryover was still present for both types of beer fermentation but was reduced in the case of wine fermentation because of hydration. PMID:23812408

  8. A non-toxic enzyme-linked immunosorbent assay for aflatoxin B1 using anti-idiotypic antibodies as substitutes.

    PubMed

    Hu, Li; Liu, Aiping; Chen, Weifeng; Yang, Hongxiu; Wang, Xiaohong; Chen, Fusheng

    2017-03-01

    Immunoassays are widely employed techniques to detect aflatoxins since they are rapid, selective and sensitive. One common disadvantage of them is using aflatoxins as standard substances, which may trigger exposure risks to operators and environmental contamination without proper handling. Anti-idiotypic antibodies (anti-Ids or Ab2s), also named as internal-image anti-Ids, are able to mimic and function as antigens, so a non-toxic enzyme-linked immunosorbent assay (ELISA) for aflatoxin B 1 (AFB 1 ) is developed and validated using anti-Ids as substitutes. Mouse monoclonal anti-idiotypic antibody (McAb2) to AFB 1 was generated by the hybridoma technique using Fab fragments of rabbit anti-AFB 1 idiotype antibody (Ab1) as immunogen. As indicated by indirect competitive ELISA, McAb2, represented an internal-image of antigen AFB 1 , was able to bind Fab with competition to AFB 1 . Then, analysis of AFB 1 in spiked samples by non-toxic ELISA using anti-Ids as substitutes was developed, and it showed no significant differences with comparison to AFB 1 as competitive antigens. Our work demonstrated that anti-Ids could be used as internal-image mimicry of AFB 1 , and it had potential applications in immunoassays for antigen substitution to reduce operational risk for operators and environmental contamination. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  9. Associations of serum aflatoxin B1-lysine adduct level with socio-demographic factors and aflatoxins intake from nuts and related nut products in Malaysia.

    PubMed

    Leong, Yin-Hui; Rosma, Ahmad; Latiff, Aishah A; Izzah, A Nurul

    2012-04-01

    Aflatoxins are one of the major risk factors in the multi-factorial etiology of human hepatocellular carcinoma. Therefore, the information on aflatoxins exposure is very important in the intervention planning in order to reduce the dietary intake of aflatoxins, especially among the children. This study investigated the relationship between aflatoxin B(1) (AFB(1)) lysine adduct levers in serum and socio-demographic factors and dietary intake of aflatoxins from nuts and nut products in Penang, Malaysia. A cross-sectional field study was conducted in five districts of Penang. A survey on socio-demographic characteristics was administered to 364 healthy adults from the three main ethnic groups (Malay, Chinese and Indian). A total of 170 blood samples were successfully collected and tested for the level of AFB(1)-lysine adduct. 97% of the samples contained AFB(1)-lysine adduct above the detection limit of 0.4 pg/mg albumin and ranged from 0.20 to 23.16 pg/mg albumin (mean±standard deviation=7.67±4.54 pg/mg albumin; median=7.12 pg/mg albumin). There was no significant association between AFB(1)-lysine adduct levels with gender, district, education level, household number and occupation when these socio-demographic characteristics were examined according to high or low levels of AFB(1)-lysine. However, participants in the age group of 31-50 years were 3.08 times more likely to have high AFB(1) levels compared to those aged between 18 and 30 years (P=0.026). Significant difference (P=0.000) was found among different ethnic groups. Chinese and Indian participants were 3.05 and 2.35 times more likely to have high AFB(1) levels than Malay. The result of AFB(1)-lysine adduct suggested that Penang adult population is likely to be exposed to AFB(1) but at a level of less than that needed to cause direct acute illness or death. Copyright © 2011 Elsevier GmbH. All rights reserved.

  10. Metabolism of aflatoxin B{sub 1} in Turkey liver microsomes: The relative roles of cytochromes P450 1A5 and 3A37

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rawal, Sumit; Coulombe, Roger A., E-mail: roger@usu.edu

    The extreme sensitivity of turkeys to aflatoxin B{sub 1} (AFB{sub 1}) is associated with efficient epoxidation by hepatic cytochromes P450 (P450) 1A5 and 3A37 to exo-aflatoxin B{sub 1}-8,9-epoxide (exo-AFBO). The combined presence of 1A5 and 3A37, which obey different kinetic models, both of which metabolize AFB{sub 1} to the exo-AFBO and to detoxification products aflatoxin M{sub 1} (AFM{sub 1}) and aflatoxin Q{sub 1} (AFQ{sub 1}), respectively, complicates the kinetic analysis of AFB{sub 1} in turkey liver microsomes (TLMs). Antisera directed against 1A5 and 3A37, thereby individually removing the catalytic contribution of these enzymes, were used to identify the P450 responsiblemore » for epoxidating AFB{sub 1} in TLMs. In control TLMs, AFB{sub 1} was converted to exo-AFBO in addition to AFM{sub 1} and AFQ{sub 1} confirming the presence of functional 1A5 and 3A37. Pretreatment with anti-1A5 inhibited exo-AFBO formation, especially at low, submicromolar ({approx} 0.1 {mu}M), while anti-3A37, resulted in inhibition of exo-AFBO formation, but at higher (> 50 {mu}M) AFB{sub 1} concentrations. Metabolism in immunoinhibited TLMs resembled that of individual enzymes: 1A5 produced exo-AFBO and AFM{sub 1}, conforming to Michaelis-Menten, while 3A37 produced exo-AFBO and AFQ{sub 1} following the kinetic Hill equation. At 0.1 {mu}M AFB{sub 1}, close to concentrations in livers of exposed animals, 1A5 contributed to 98% of the total exo-AFBO formation. At this concentration, 1A5 accounted for a higher activation:detoxification (50:1, exo-AFBO: AFM{sub 1}) compared to 3A37 (0.15: 1, exo-AFBO: AFQ{sub 1}), suggesting that 1A5 is high, while 3A4 is the low affinity enzyme in turkey liver. The data support the conclusion that P450 1A5 is the dominant enzyme responsible for AFB{sub 1} bioactivation and metabolism at environmentally-relevant AFB{sub 1} concentrations in turkey liver. - Graphical abstract: Display Omitted Highlights: > Efficient bioactivation by P450s 1A5 and 3A

  11. Antigenotoxic Effect of Piperine in Broiler Chickens Intoxicated with Aflatoxin B1

    PubMed Central

    da Silva Cardoso, Verônica; Vermelho, Alane Beatriz; Ribeiro de Lima, Cristina Amorim; Mendes de Oliveira, Jéssica; Freire de Lima, Marco Edilson; Pinto da Silva, Lúcia Helena; Direito, Glória Maria; Miranda Danelli, Maria das Graças

    2016-01-01

    Piperine is an abundant amide extracted from black pepper seeds which has been shown to have protective effects against cytotoxic and genotoxic carcinogenesis induced by certain chemical carcinogens and aflatoxin B1 (AFB1) in vitro. The aim of this work was to study, in vivo, the antigenotoxic potential of feed-added piperine on broiler chickens experimentally intoxicated with AFB1, using micronucleus and comet assays. The antigenotoxicity assessment of 9-day-old chicks was performed on a total of 60 chickens divided into four groups of 15 broilers each: (C) control, (P) 60 mg·piperine kg−1 feed, (A) 0.5 mg·AFB1·kg−1 body weight, (daily by oral route), and (P + A) co-treatment with piperine and AFB1. The experiment was conducted for 26 days. Chicks intoxicated with AFB1 showed significant genotoxic effects in the first 24 h post intoxication, and the effects remained in the other periods analyzed (48, 72, and 96 h and 26 days of treatment). The DNA damage in peripheral blood cells, the number of erythrocytes with micronuclei, and polychromatic-to-normochromatic erythrocyte ratio were significantly reduced or absent in the piperine/AFB1 group. No significant differences were observed between the group piperine/AFB1 and the control and piperine-alone groups. The addition 60 mg·kg−1 of piperine to the diet of the broiler chicks was safe, promoting beneficial effects in poultry health with respect to the toxic effects 0.5 mg·AFB1·kg−1 body weight. PMID:27809242

  12. [Determination of aflatoxin B1, B2, G1, G2 in armeniacae semen amarum by high-performance liquid chromatography-tandem mass spectrometry].

    PubMed

    Zheng, Run-Sheng; Xu, Hui; Wang, Wen-Li; Zhan, Ruo-Ting; Chen, Wei-Wen

    2013-10-01

    A simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/ MS) method was established for simultaneous determination of aflatoxins (AFB1, AFB2, AFG1, AFG2) in Armeniacae Semen Amarum and the application was performance in 11 samples collected from different markets, medical stores and hospitals. The sample was extracted with 84% acetonitrile/water and 250 microL extraction was directly injected into a LC-MS/MS system without further purification procedure after being redissolved with methanol. The LC separation was performed on a C18 column with a linear gradient elution program of 4 mmol x L(-1) NH4 Ac-0.1% formic acid solution and menthol as the mobile phase. Selected reaction monitoring (SRM) was used for selective determination of the four aflatoxins on a triple quadruple mass spectrometer, which was operated in positive ionization modes. All the four aflatoxins showed a good linear relationship with r > 0.999 0, the average recoveries were between 87.88% and 102.9% and the matrix effect was ranged from 90.71% to 99.30% in low, intermediate and high levels. Furthermore, the higher recovery was obtained by the method reported in this study, comparing to the cleanup procedure with the Mycosep 226 purification column. Eleven samples collected were detected and the contamination levels of the AFB1 were between 1.590-2.340 microg x kg(-1) and the AF (B1 + B2 + G1 + G2) was ranged from 2.340 to 3.340 microg x kg(-1). In summary, the developed method was suitable to detect and screen AFB1, AFB2, AFG1, AFG2 in Armeniacae Semen Amarum.

  13. Feasibility of detecting Aflatoxin B1 in single maize kernels using hyperspectral imaging

    USDA-ARS?s Scientific Manuscript database

    The feasibility of detecting Aflatoxin B1 (AFB1) in single maize kernel inoculated with Aspergillus flavus conidia in the field, as well as its spatial distribution in the kernels, was assessed using near-infrared hyperspectral imaging (HSI) technique. Firstly, an image mask was applied to a pixel-b...

  14. Occurrence of Aspergillus spp. and aflatoxin B1 in Malaysian foods used for human consumption.

    PubMed

    Reddy, Kasa R N; Farhana, Nazira I; Salleh, Baharuddin

    2011-05-01

    Malaysian population widely consumes the cereal-based foods, oilseeds, nuts, and spices in their daily diet. Mycotoxigenic fungi are well known to invade food products under storage conditions and produce mycotoxins that have threat to human and animal health. Therefore, determining toxigenic fungi and aflatoxin B(1) (AFB1) in foods used for human consumption is of prime importance to develop suitable management strategies and to minimize risk. Ninety-five food products marketed in Penang, Malaysia were randomly collected from different supermarkets and were analyzed for presence of Aspergillus spp. by agar plate assay and AFB1 by enzyme-linked immunosorbent assay (ELISA). A. flavus was the dominant fungi in all foods followed by A. niger. Fifty-five A. flavus strains were tested for their ability to produce aflatoxins on rice grain substrate. Thirty-six (65.4%) strains out of 55 produced AFB1 ranging from 1700 to 4400 μg/kg and 17 strains (31%) produced AFB2 ranging from 620 to 1670 μg/kg. Natural occurrence of AFB1 could be detected in 72.6% food products ranging from 0.54 to 15.33 μg/kg with a mean of 1.95 μg/kg. Maximum AFB1 levels were detected in peanut products ranging from 1.47 to 15.33 μg/kg. AFB1 levels detected in all food products were below the Malaysian permissible limits (<35 μg/kg). Aspergillus spp. and AFB1 was not detected in any cookies tested. Although this survey was not comprehensive, it provides valuable information on aflatoxin levels in foods marketed in Malaysia. © 2011 Institute of Food Technologists®

  15. Fisetin Attenuates AKT Associated Growth Promoting Events in AflatoxinB1 Induced Hepatocellular Carcinoma.

    PubMed

    Maurya, Brajesh Kumar; Trigun, Surendra Kumar

    2017-12-29

    Recently we have reported that Fisetin, a natural flavonol, is able to regress Aflatoxin-B1 (AFB1) induced hepatocellular carcinoma (HCC) by suppressing reactive oxygen species (ROS) led pro-inflammatory factors in rats. In the current study, we aimed to delineate whether Fisetin does so by modulating the cell growth promoting signaling cascade in HCC. The reciprocal interplay of 3-phosphoinositol kinase (PI3K) vs phosphatase and tensin homologue deleted on chromosome 10 (PTEN) displays Akt, a protein kinase B, to get phosphorylated at Thr308 by a 3-phosphoinositol dependent kinase 1 (PDK1). This commits cells of neoplastic niche to undergo rapid proliferation by p-Akt thr308 dependent phosphorylation of glycogen synthase kinase 3β (GSK3β) at Ser 9 position. In this study, the effect of in vivo treatment of 20 mg/kg b.w. Fisetin on relative profile of all these factors were studied in the liver from the HCC rats induced by two doses of 1mg/kg b.w. AFB1 i.p. As compared to the untreated HCC liver, liver from Fisetin treated HCC group rats showed a significant decline in the activity and level of p-Aktthr308 which was consistent with a similar decline in PDK1 level. Concordantly, the level of p-GSK3βSer 9 was also found to be declined significantly in those Fisetin-treated HCC livers. A concomitant decline in immunohistochemically detected number of the proliferating cell nuclear antigen (PCNA), a cell proliferation marker, in the HCC liver, further confirmed anti-cell proliferative role of Fisetin during HCC growth in vivo. This findings suggest that Fisetin is able to suppress Akt dependent cell growth signaling mechanisms in HCC mainly by down regulating PDK1 dependent Akt phosphorylation. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  16. miR-34a screened by miRNA profiling negatively regulates Wnt/β-catenin signaling pathway in Aflatoxin B1 induced hepatotoxicity

    PubMed Central

    Zhu, Liye; Gao, Jing; Huang, Kunlun; Luo, Yunbo; Zhang, Boyang; Xu, Wentao

    2015-01-01

    Aflatoxin-B1 (AFB1), a hepatocarcinogenic mycotoxin, was demonstrated to induce the high rate of hepatocellular carcinoma (HCC). MicroRNAs (miRNAs) participate in the regulation of several biological processes in HCC. However, the function of miRNAs in AFB1-induced HCC has received a little attention. Here, we applied Illumina deep sequencing technology for high-throughout profiling of microRNAs in HepG2 cells lines after treatment with AFB1. Analysis of the differential expression profile of miRNAs in two libraries, we identified 9 known miRNAs and 1 novel miRNA which exhibited abnormal expression. KEGG analysis indicated that predicted target genes of differentially expressed miRNAs are involved in cancer-related pathways. Down-regulated of Drosha, DGCR8 and Dicer 1 indicated an impairment of miRNA biogenesis in response to AFB1. miR-34a was up-regulated significantly, down-regulating the expression of Wnt/β-catenin signaling pathway by target gene β-catenin. Anti-miR-34a can significantly relieved the down-regulated β-catenin and its downstream genes, c-myc and Cyclin D1, and the S-phase arrest in cell cycle induced by AFB1 can also be relieved. These results suggested that AFB1 might down-regulate Wnt/β-catenin signaling pathway in HepG2 cells by up-regulating miR-34a, which may involve in the mechanism of liver tumorigenesis. PMID:26567713

  17. Identification of early target genes of aflatoxin B1 in human hepatocytes, inter-individual variability and comparison with other genotoxic compounds

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Josse, Rozenn; Dumont, Julie; Fautrel, Alain

    Gene expression profiling has recently emerged as a promising approach to identify early target genes and discriminate genotoxic carcinogens from non-genotoxic carcinogens and non-carcinogens. However, early gene changes induced by genotoxic compounds in human liver remain largely unknown. Primary human hepatocytes and differentiated HepaRG cells were exposed to aflatoxin B1 (AFB1) that induces DNA damage following enzyme-mediated bioactivation. Gene expression profile changes induced by a 24 h exposure of these hepatocyte models to 0.05 and 0.25 μM AFB1 were analyzed by using oligonucleotide pangenomic microarrays. The main altered signaling pathway was the p53 pathway and related functions such as cellmore » cycle, apoptosis and DNA repair. Direct involvement of the p53 protein in response to AFB1 was verified by using siRNA directed against p53. Among the 83 well-annotated genes commonly modulated in two pools of three human hepatocyte populations and HepaRG cells, several genes were identified as altered by AFB1 for the first time. In addition, a subset of 10 AFB1-altered genes, selected upon basis of their function or tumor suppressor role, was tested in four human hepatocyte populations and in response to other chemicals. Although they exhibited large variable inter-donor fold-changes, several of these genes, particularly FHIT, BCAS3 and SMYD3, were found to be altered by various direct and other indirect genotoxic compounds and unaffected by non-genotoxic compounds. Overall, this comprehensive analysis of early gene expression changes induced by AFB1 in human hepatocytes identified a gene subset that included several genes representing potential biomarkers of genotoxic compounds. -- Highlights: ► Gene expression profile changes induced by aflatoxin B1 in human hepatocytes. ► AFB1 modulates various genes including tumor suppressor genes and proto-oncogenes. ► Important inter-individual variations in the response to AFB1. ► Some genes also altered

  18. Reduction of Aflatoxin B1 Toxicity by Lactobacillus plantarum C88: A Potential Probiotic Strain Isolated from Chinese Traditional Fermented Food "Tofu".

    PubMed

    Huang, Li; Duan, Cuicui; Zhao, Yujuan; Gao, Lei; Niu, Chunhua; Xu, Jingbo; Li, Shengyu

    2017-01-01

    In this study, we investigated the potential of Lactobacillus plantarum isolated from Chinese traditional fermented foods to reduce the toxicity of aflatoxin B1 (AFB1), and its subsequent detoxification mechanism. Among all the investigated L. plantarum strains, L. plantarum C88 showed the strongest AFB1 binding capacity in vitro, and was orally administered to mice with liver oxidative damage induced by AFB1. In the therapy groups, the mice that received L. plantarum C88, especially heat-killed L. plantarum C88, after a single dose of AFB1 exposure, showed an increase in unabsorbed AFB1 in the feces. Moreover, the effects of L. plantarum C88 on the enzymes and non-enzymes antioxidant abilities in serum and liver, histological alterations of liver were assayed. The results indicated that compared to the control group, L. plantarum C88 alone administration induced significant increase of antioxidant capacity, but did not induce any significant changes in the histological picture. Compared to the mice that received AFB1 only, L. plantarum C88 treatment could weaken oxidative stress by enhancing the activity of antioxidant enzymes and elevating the expression of Glutathione S-transferase (GST) A3 through Nuclear factor erythroid (derived factor 2) related factor 2 (Nrf2) pathway. Furthermore, cytochrome P450 (CYP 450) 1A2 and CYP 3A4 expression was inhibited by L. plantarum C88, and urinary aflatoxin B1-N7-guanine (AFB-N7-guanine), a AFB1 metabolite formed by CYP 1A2 and CYP 3A4, was significantly reduced by the presence of viable L. plantarum C88. Meanwhile, the significant improvements were showed in histological pictures of the liver tissues in mice orally administered with viable L. plantarum C88. Collectively, L. plantarum C88 may alleviate AFB1 toxicity by increasing fecal AFB1 excretion, reversing deficits in antioxidant defense systems and regulating the metabolism of AFB1.

  19. Competitive horseradish peroxidase-linked aptamer assay for sensitive detection of Aflatoxin B1.

    PubMed

    Sun, Linlin; Zhao, Qiang

    2018-03-01

    Aflatoxin B1 (AFB1) is one of highly toxic mycotoxins and a known human carcinogen. The frequent contamination of AFB1 in food products and large health risk of AFB1 have raised global concerns. Sensitive detection of AFB1 is of vital importance and highly demanded. Herein, we reported a competitive horseradish peroxidase (HRP)-linked aptamer assay for AFB1, combining the advantages of aptamer for affinity binding and enzyme label for signal amplification. In this assay, free AFB1 in solution competed with a covalent conjugate of bovine serum albumin-AFB1 (BSA-AFB1) coated on the wells of microplate in binding to the HRP-labeled aptamer probe. HRP attached on BSA-AFB1 in the wells catalyzed the conversion of substrates into products, allowing the final detection of AFB1 through measurement of the generated products. When TMB (3,3',5,5'-tetramethylbenzidine dihydrochloride) was used as substrate, absorbance analysis of the product of enzyme reaction enabled the detection of AFB1 at 0.2nM. We further lowered the detection limit of AFB1 to 0.01nM through chemiluminescence analysis by using chemiluminescence substrate of HRP. This assay enabled the detection of AFB1 in complex sample matrix, such as diluted white wine and maize flour. This assay provides a simple, sensitive and rapid method for AFB1 determination. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Evaluation of STAT5A Gene Expression in Aflatoxin B1 Treated Bovine Mammary Epithelial Cells

    PubMed Central

    Forouharmehr, Ali; Harkinezhad, Taher; Qasemi-Panahi, Babak

    2013-01-01

    Purpose: Aflatoxin B1 (AFB1) is a potent mycotoxin which has been produced by fungi such as Aspergillus flavus and Aspergillus parasiticus as secondary metabolites due to their growth on food stuffs and induces hepatocellular carcinoma in many animal species, including humans. In the present study, the effect of AFB1 on STAT5A gene expression was investigated in bovine mammary epithelial cells using real time RT-PCR. Methods: Bovine mammary epithelial cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, cells were treated with AFB1 and incubated for 8 h. For real time PCR reaction, total RNA from the cultured and treated cells was extracted and used for complementary DNA synthesis. Results: The expression of STAT5A gene was significantly down regulated by AFB1 in dose- dependent manner and led to the reduction of proliferation and differentiation of epithelial cells, which has direct effect in milk protein quantity and quality. Conclusion: According to the results, it seems that down regulation of STAT5A gene in AFB1-treated cells maybe due to DNA damage induced by AFB1 in bovine mammary epithelial cells. PMID:24312879

  1. Aflatoxin B1 inhibition in Aspergillus flavus by Aspergillus niger through down-regulating expression of major biosynthetic genes and AFB1 degradation by atoxigenic A. flavus.

    PubMed

    Xing, Fuguo; Wang, Limin; Liu, Xiao; Selvaraj, Jonathan Nimal; Wang, Yan; Zhao, Yueju; Liu, Yang

    2017-09-01

    Twenty Aspergillus niger strains were isolated from peanuts and 14 strains were able to completely inhibit AFB 1 production with co-cultivation. By using a Spin-X centrifuge system, it was confirmed that there are some soluble signal molecules or antibiotics involved in the inhibition by A. niger, although they are absent during the initial 24h of A. flavus growth when it is sensitive to inhibition. In A. flavus, 19 of 20 aflatoxin biosynthetic genes were down-regulated by A. niger. Importantly, the expression of aflS was significantly down-regulated, resulting in a reduction of AflS/AflR ratio. The results suggest that A. niger could directly inhibit AFB 1 biosynthesis through reducing the abundance of aflS to aflR mRNAs. Interestingly, atoxigenic A. flavus JZ2 and GZ15 effectively degrade AFB 1 . Two new metabolites were identified and the key toxic lactone and furofuran rings both were destroyed and hydrogenated, meaning that lactonase and reductase might be involved in the degradation process. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. A novel method for determination of aflatoxin B1 mediated by FCLA + BSA

    NASA Astrophysics Data System (ADS)

    Chen, WenLi; Xing, Da

    2005-02-01

    As a chemiluminescence (CL) probe, 3,7-dihydro-6-{4-{2-(N"-(5-fluoresceinyl) thioureido)ethoxy}phenyl}-2-met -hylimi-dazo{1,2-a}pyrazin-3-one dosium salt (FCLA) can sensitively and specifically react with singlet oxygen (1O2 ) and superoxide(O2""). BSA (Bovine Serum Albumin) can enlarge the CL intensity of FCLA to 860%. This report presents a novel method for determination of Aflatoxin B1 (AfB1) mediated by FCLA+BSA. The concentration of AFB1 showed an obvious positive correlation with the CL intensity mediated by FCLA+BSA. This method could measure accurately ng/ml of AfB1 concentration. At the same time, the fluorescence spectrum of FCLA+BSA and FCLA+BSA+AfB1 were measured respectively, which showed that the fluorescence intensity of FCLA+BSA+AfB1 was higher than FCLA+BSA. Comparing the peak value of FCLA, FCLA+BSA and FCLA+BSA+AfB1 had a 6nm Einstein shift (red shift). The study suggested that CL method mediated by FCLA+BSA might be applicable to the determination of AfB1 concentration.

  3. Non-linear relationships between aflatoxin B1 levels and the biological response of monkey kidney vero cells

    USDA-ARS?s Scientific Manuscript database

    Aflatoxin (AF)-producing fungi contaminate food and feed during preharvest, storage and processing periods. Once consumed, AF accumulates in tissues, causing illnesses in animals and humans. At least 20 different types of AFs have been identified, and of these, aflatoxin B1 (AFB1) is the most ubiqui...

  4. Effect of Various Compounds Blocking the Colony Pigmentation on the Aflatoxin B1 Production by Aspergillus flavus.

    PubMed

    Dzhavakhiya, Vitaly G; Voinova, Tatiana M; Popletaeva, Sofya B; Statsyuk, Natalia V; Limantseva, Lyudmila A; Shcherbakova, Larisa A

    2016-10-28

    Aflatoxins and melanins are the products of a polyketide biosynthesis. In this study, the search of potential inhibitors of the aflatoxin B1 (AFB1) biosynthesis was performed among compounds blocking the pigmentation in fungi. Four compounds-three natural (thymol, 3-hydroxybenzaldehyde, compactin) and one synthetic (fluconazole)-were examined for their ability to block the pigmentation and AFB1 production in Aspergillus flavus . All compounds inhibited the mycelium pigmentation of a fungus growing on solid medium. At the same time, thymol, fluconazole, and 3-hydroxybenzaldehyde stimulated AFB1 accumulation in culture broth of A. flavus under submerged fermentation, whereas the addition of 2.5 μg/mL of compactin resulted in a 50× reduction in AFB1 production. Moreover, compactin also suppressed the sporulation of A. flavus on solid medium. In vivo treatment of corn and wheat grain with compactin (50 μg/g of grain) reduced the level of AFB1 accumulation 14 and 15 times, respectively. Further prospects of the compactin study as potential AFB1 inhibitor are discussed.

  5. Biochemical and molecular aspects of mammalian susceptibility to aflatoxin B{sub 1} carcinogenicity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Massey, T.E.; Stewart, R.K.; Daniels, J.M.

    Aflatoxin B{sub 1} (AFB{sub 1}) is a fungal toxin that has been implicated as a causative agent in human hepatic and extrahepatic carcinogenesis. In this review, the mechanisms involved in AFB{sub 1} toxicity are delineated, in order to describe the features that make a specific cell, tissue, or species susceptible to the mycotoxin. Important considerations include: (i) different mechanisms for bioactivation of AFB{sub 1} to its ultimate carcinogenic epoxide metabolite; (ii) the balance between bioactivation to and detoxification of the epoxide; (iii) the interaction of AFB{sub 1} epoxide with DNA and the mutational events leading to neoplastic transformation; (iv) themore » role of cytotoxicity in AFB{sub 1} carcinogenesis; (v) the significance of nonepoxide metabolites in toxicity; and (vi) the contribution of mycotoxin-unrelated disease processes. Although considerable controversy remains about the importance of specific events, a great deal has been learned about biochemical and molecular actions of AFB{sub 1}. 157 refs., 4 figs., 1 tab.« less

  6. Detoxification of Aflatoxin B1 by Zygosaccharomyces rouxii with Solid State Fermentation in Peanut Meal

    PubMed Central

    Zhou, Guanghui; Chen, Yujie; Kong, Qing; Ma, Yunxiao; Liu, Yang

    2017-01-01

    Aflatoxins are highly carcinogenic, teratogenetic, and morbigenous secondary metabolites of Aspergillus flavus and A. parasiticus that can contaminate multiple staple foods, such as peanut, maize, and tree nuts. In this study, Zygosaccharomyces rouxii was screened out and identified from fermented soy paste—one kind of traditional Chinese food—to detoxify aflatoxin B1 (AFB1) by aerobic solid state fermentation in peanut meal. The optimal degradation condition was chosen from single factor experiment, and the most effective detoxification rate was about 97%. As for liquid fermentation, we tested the binding ability of Z. rouxii, and the highest binding rate reached was 74.3% (nonviable cells of Z. rouxii) in phosphate-buffered saline (PBS). Moreover, the biotransformation of AFB1 through fermentation of Z. rouxii in peanut meal was further verified by liquid chromatography/mass spectrometry (LC/MS). According to TIC scan, after fermentation by Z. rouxii, the AFB1 in peanut meal was prominently degraded to the lowering peaks of AFB1. Additionally, m/s statistics demonstrated that AFB1 may be degraded to some new products whose structural properties may be different from AFB1, or the degradation products may be dissolved in the aqueous phase rather than the organic phase. As far as we know, this is the first report indicating that the safe strain of Z. rouxii has the ability to detoxify AFB1. PMID:28117705

  7. Low cost quantitative digital imaging as an alternative to qualitative in vivo bioassays for analysis of active aflatoxin B1

    USDA-ARS?s Scientific Manuscript database

    Aflatoxin B1 (AFB1) producing fungi contaminate food and feed and are a major health concern. To minimize the sources and incidence of AFB1 illness there is a need to develop affordable, sensitive mobile devices for detection of active AFB1. In the present study we used a low cost fluorescence detec...

  8. Aflatoxin B1 and M1 Degradation by Lac2 from Pleurotus pulmonarius and Redox Mediators

    PubMed Central

    Loi, Martina; Fanelli, Francesca; Zucca, Paolo; Liuzzi, Vania C.; Quintieri, Laura; Cimmarusti, Maria T.; Monaci, Linda; Haidukowski, Miriam; Logrieco, Antonio F.; Sanjust, Enrico; Mulè, Giuseppina

    2016-01-01

    Laccases (LCs) are multicopper oxidases that find application as versatile biocatalysts for the green bioremediation of environmental pollutants and xenobiotics. In this study we elucidate the degrading activity of Lac2 pure enzyme form Pleurotus pulmonarius towards aflatoxin B1 (AFB1) and M1 (AFM1). LC enzyme was purified using three chromatographic steps and identified as Lac2 through zymogram and LC-MS/MS. The degradation assays were performed in vitro at 25 °C for 72 h in buffer solution. AFB1 degradation by Lac2 direct oxidation was 23%. Toxin degradation was also investigated in the presence of three redox mediators, (2,2′-azino-bis-[3-ethylbenzothiazoline-6-sulfonic acid]) (ABTS) and two naturally-occurring phenols, acetosyringone (AS) and syringaldehyde (SA). The direct effect of the enzyme and the mediated action of Lac2 with redox mediators univocally proved the correlation between Lac2 activity and aflatoxins degradation. The degradation of AFB1 was enhanced by the addition of all mediators at 10 mM, with AS being the most effective (90% of degradation). AFM1 was completely degraded by Lac2 with all mediators at 10 mM. The novelty of this study relies on the identification of a pure enzyme as capable of degrading AFB1 and, for the first time, AFM1, and on the evidence that the mechanism of an effective degradation occurs via the mediation of natural phenolic compounds. These results opened new perspective for Lac2 application in the food and feed supply chains as a biotransforming agent of AFB1 and AFM1. PMID:27563923

  9. DNA BINDING AND ADDUCT FORMATION OF AFLATOXIN B1 IN CULTURED HUMAN AND ANIMAL TRACHEOBRONCHIAL AND BLADDER TISSUES

    EPA Science Inventory

    DNA binding and adduct formation of aflatoxin B1 (AFB1) was studied in cultured bladder and tracheobronchial explants from human, monkey, dog, hamster and rat. Explants were exposed to (3H)AFB1 (1 micrometer final concentration) in PFHR-4 medium (pH 7.4) without serum for 24 h, a...

  10. Protective effect of Bacillus subtilis ANSB060 on egg quality, biochemical and histopathological changes in layers exposed to aflatoxin B1.

    PubMed

    Ma, Q G; Gao, X; Zhou, T; Zhao, L H; Fan, Y; Li, X Y; Lei, Y P; Ji, C; Zhang, J Y

    2012-11-01

    Bacillus subtilis ANSB060 from the fish gut had strong ability to detoxify aflatoxins. The aim of this research was to investigate the protective effect of B. subtilis ANSB060 (ANSB060) on egg quality and biochemical and histopathological changes of liver and kidney in laying hens when exposed to aflatoxin B(1). Treatments (C20, C40, and C60) were prepared by substituting corn contaminated by aflatoxin B(1) (AFB1) at different proportions (20, 40, and 60%) for normal corn in basic diets. The aflatoxin degradation enzyme (E) treatments (E20, E40, and E60) were mixed with the fermentation liquor of ANSB060 with C20, C40, and C60, respectively. The results showed that ANSB060 can improve the eggshell strength in E60 compared with C60 (P ≤ 0.05), and toxin reduced the content of total protein (in groups C20, C40, and C60) and albumin (in C20 and C40; P < 0.05) and heightened the activities of GPT (in C60) and GOT (in C40 and C60) in serum (P < 0.05). In the liver, AFB1 inhibited the activity of superoxide dismutase and glutathione peroxidase (C40 and C60; P < 0.05) and increased the content of malonaldehyde (in C40 and C60), which induced the damage in the liver and kidney as shown in the photomicrographs of hematoxylin and eosin-stained sections. The addition of ANSB060 can enhance the activity of antioxidant enzymes, and it recovered the protein synthesis in liver. Moreover, ANSB060 also ameliorated the damage of liver and kidney tissue and restored them to normal. Hence, ANSB060 had the ability to inhibit the damage induced by AFB1; it will have a great potential in industrial applications.

  11. Comparative Efficacy of Silymarin and Choline Chloride (Liver Tonics) in Preventing the Effects of Aflatoxin B1 in Bovine Calves.

    PubMed

    Naseer, O; Khan, J A; Khan, M S; Omer, M O; Chishti, G A; Sohail, M L; Saleem, M U

    2016-09-01

    Aflatoxins are secondary metabolites produced by Aspergillus spp. which are injurious to animals and humans The aim of this study was to determine the effects of aflatoxin B1 (AFB1) on Average Daily Feed Intake (ADFI), Average Daily Weight Gain (ADWG), haematological and serum biochemical responses of Bovine Calves and to determine the comparative efficacy of two different liver tonics against AFB1. Twenty seven calves were selected from herd and divided into 3 groups. All calves were fed with 1.0 mg/kg AFB1 for a period of 10 days. After that they were fed with liver tonics: Silymarin fed at a rate of 600 mg/kg and Choline chloride 500 mg/kg for 7 days. The results indicate that the ADFI and ADWG of AFB1 treated calves decreased significantly. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN) and creatinine significantly increased due to AFB1. In haematology the total erythrocyte count (TEC), total leukocyte count (TLC), haemoglobin concentration (HGB), haematocrit levels (HCT), mean corpuscular haemoglobin (MCH), mean corpuscular volume (MCV) and mean corpuscular haemoglobin concentration (MCHC), lymphocyte %, neutrophil % and monocyte % significantly decreased in AFB1 treated calves after 10 days of feeding. Both liver tonics significantly (p<0.05) improved all the parameters, including ADFI, ADWG, hematologial and serum biochemical test. However, Silymarin comparatively more efficiently ameliorate the effects induced by AFB1 than choline chloride.

  12. Affinity capture of aflatoxin B1 and B2 by aptamer-functionalized magnetic agarose microspheres prior to their determination by HPLC.

    PubMed

    Liu, Hongmei; Lu, Anxiang; Fu, Hailong; Li, Bingru; Yang, Meihua; Wang, Jihua; Luan, Yunxia

    2018-06-12

    A novel adsorbent is described for magnetic solid-phase extraction (MSPE) of the aflatoxins AFB 1 and AFB 2 (AFBs). Magnetic agarose microspheres (MAMs) were functionalized with an aptamer to bind the AFBs which then were quantified by HPLC and on-line post-column photochemical derivatization with fluorescence detection. Streptavidin-conjugated MAMs were synthesized first by a highly reproducible strategy. They possess strong magnetism and high surface area. The MAMs were characterized by transmission electron microscopy, scanning electron microscopy, optical microscopy, laser diffraction particle size analyzer, Fourier transform infrared spectrometry, vibrating sample magnetometry and laser scanning confocal microscopy. Then, the AFB-aptamers were immobilized on MAMs through biotin-streptavidin interaction. Finally, the MSPE is performed by suspending the aptamer-modified MAMs in the sample. They are then collected by an external magnetic field and the AFBs are eluted with methanol/buffer (20:80). Several parameters affecting the coupling, capturing and eluting efficiency were optimized. Under the optimized conditions, the method is fast, has good linearity, high selectivity, and sensitivity. The LODs are 25 pg·mL -1 for AFB 1 and 10 pg·mL -1 for AFB 2 . The binding capacity is 350 ± 8 ng·g -1 for AFB 1 and 384 ± 8 ng·g -1 for AFB 2 , and the precision of the assay is <8%. The method was successfully applied to the analysis of AFBs in spiked maize samples. Graphical abstract Schematic of novel aptamer functionalized magnetic agarose microspheres (Apt-MAM) as magnetic adsorbents for simultaneous and specific affinity capture of aflatoxins B 1 and B 2 (AFBs).

  13. Development of an ultrasensitive aptasensor for the detection of aflatoxin B1.

    PubMed

    Guo, Xiaodong; Wen, Fang; Zheng, Nan; Luo, Qiujiang; Wang, Haiwei; Wang, Hui; Li, Songli; Wang, Jiaqi

    2014-06-15

    Contamination of feed and food by aflatoxin B1 (AFB1), one of the most toxic of the mycotoxins, is a global concern. To prevent food safety scares, and avoid subsequent economic losses due to the recall of contaminated items, methods for the rapid, sensitive and specific detection of AFB1 at trace levels are much in demand. In this work, a simple, ultrasensitive, and reliable aptasensor is described for the detection of AFB1. An AFB1 aptamer was used as a molecular recognition probe, while its complementary DNA played a role as a signal generator for amplification by real-time quantitative polymerase chain reaction (PCR). Under optimal conditions, a wide linear detection range (5.0 × 10(-5) to 5.0 ng mL(-1)) was achieved, with a high sensitivity (limit of detection (LOD)=25 fg mL(-1)). In addition, the proposed aptasensor exhibited excellent specificity for AFB1 compared with eight other mycotoxins, with no obvious Ct value change. This aptasensor can also be used in quantifying AFB1 levels in Chinese wild rye hay samples and infant rice cereal samples, demonstrating satisfactory recoveries in the range of 88-127% and 94-119%, respectively. This detection technique has a significant potential for high-throughput, quantitative determination of mycotoxin levels in a large range of feeds and foods. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Effects of dietary AflaDetox on performance, serum biochemistry, histopathological changes, and aflatoxin residues in broilers exposed to aflatoxin B(1).

    PubMed

    Denli, M; Blandon, J C; Guynot, M E; Salado, S; Perez, J F

    2009-07-01

    The aim of this study was to evaluate the ability of AflaDetox (Adiveter, Agro-Reus, Reus, Tarragona, Spain) in counteracting the deleterious effects of aflatoxin B(1) (AFB(1)) in broiler chicks. A total of 120 Ross 308 one-day-old male broiler chicks were assigned to 8 treatments for 42 d. The experiment had a 2 x 4 factorial arrangement of treatments involving 0 and 1 mg of AFB(1)/kg feed and 0, 1, 2, and 5 g of AflaDetox/kg feed. Chicks were fed on the ground during the first 7 d and in cages (3 chicks/cage; 5 cages/treatment) from 7 to 42 d. Growth performance was measured from d 7 to 42 and whole-tract digestibility of gross energy and protein on d 40 to 41. Serum biochemical parameters, organ weights, histopathological examination of liver, and AFB(1) residues in liver and breast muscle tissues were determined on d 42. Aflatoxin B(1) significantly decreased the BW gain, feed intake, and impaired feed conversion rate (P < 0.05). The addition of AflaDetox in the contaminated diets significantly diminished the inhibitory effects of dietary AFB(1) (P < 0.05) on the growth performance with no differences compared to the control diet. Feeding AFB(1) alone decreased serum protein concentration, increased the serum activity of alkaline phosphatase, and caused significant increases in the relative weights of livers. Treatment with AflaDetox significantly alleviated the negative effects of AFB(1) on these parameters (P < 0.05) with no effect on uncontaminated diets. Liver tissue of broilers receiving AFB(1) alone had perilobular inflammation and vacuolar degeneration of hepatocytes as compared with the tissue from the control group (P < 0.05). Residues of AFB(1) were detected in the liver tissues of broilers fed on the AFB(1) diet (0.166 microg/kg). Supplementation of AflaDetox reduced the incidence and severity of the hepatic histopathology changes associated with aflatoxicosis and the amount of AFB(1) residue in liver. In conclusion, our results showed that addition

  15. Exposure to aflatoxin B1 in Thailand by consumption of brown and color rice.

    PubMed

    Panrapee, Iamtaweejaroen; Phakpoom, Kooprasertying; Thanapoom, Maneeboon; Nampeung, Anukul; Warapa, Mahakarnchanakul

    2016-02-01

    This study assessed the aflatoxin B1 (AFB1) intake of the Thai population through consumption of contaminated brown and color rice. A total of 240 rice samples from two harvesting periods were collected in June/July 2012 (period I) and in December 2012/January 2013 (period II) and analyzed for AFB1 by HPLC with fluorescence detection (limit of detection (LOD) = 0.093 ng/g). Exposure assessment was based on AFB1 levels in rice and food intake data for rice according to Thai National Consumption. Frequency and levels of AFB1 were higher in period I (59%, 1)) than in period II (10%, 1)). Only one sample exceeded the Thai standard limit for total aflatoxin of 20 μg kg(-1), but 12 out of 240 rice samples exceeded the European Union maximum level for AFB1 of 2 μg kg(-1). The data showed that the quality and safety of Thai rice largely comply with the requirement for both exports and domestic consumption. According to the Thai National Consumption data, the estimated AFB1 intake via rice consumption in period I and period II was 0.80 and 0.12 μg kg(-1) bw day(-1), respectively. The potential risk for cancer, based on the recommendation of the JECFA, was estimated to be 0.011 person/year/100,000 people at a mean consumption. Although the risk via consumption of Thai rice seems to be low, the maximum levels of AFB1 in this staple food suggest that careful monitoring and surveillance of AFB1 contamination in rice is essential to ensure the safety of rice.

  16. Determination of the aflatoxin AFB1 from corn by direct analysis in real time-mass spectrometry (DART-MS)

    USDA-ARS?s Scientific Manuscript database

    Direct analysis in real time (DART) ionization coupled to a high resolution mass spectrometer (MS) was used for screening of aflatoxins from a variety of surfaces and the rapid quantitative analysis of a common form of aflatoxin, AFB1, extracted from corn. Sample preparation procedure and instrument...

  17. Modulatory Effect of the Intracellular Content of Lactobacillus casei CRL 431 Against the Aflatoxin B1-Induced Oxidative Stress in Rats.

    PubMed

    Aguilar-Toalá, J E; Astiazarán-García, H; Estrada-Montoya, M C; Garcia, H S; Vallejo-Cordoba, B; González-Córdova, A F; Hernández-Mendoza, A

    2018-06-03

    It has been recognized that lactic acid bacteria exhibit antioxidant properties, which have been mainly endorsed to the intact viable bacteria. However, recent studies have shown that intracellular content (IC) may also be good sources of antioxidative metabolites, which may potentially contribute to oxidative homeostasis in vivo. Hence, the modulatory effect of the intracellular content of Lactobacillus casei CRL 431 (IC431) on aflatoxin B 1 (AFB 1 )-induced oxidative stress in rats was evaluated on the basis of its influence on hepatic lipid peroxidation (LPO), antioxidant status-antioxidant capacity (TAC), catalase (CAT), and glutathione peroxidase (GPx) activities; and on the oxidative stress index (OSi). Results demonstrated that CAT and GPx activities, and TAC, determined in plasma samples, were significantly (P < 0.05) higher in rats treated with AFB 1 plus IC431 (3.98 μM/min/mg protein, 1.88 μM/min/mg protein, and 238.7 μM Trolox equivalent, respectively) than AFB 1 -treated rats (3.47 μM/min/mg protein, 1.46 μM/min/mg protein, and 179.7 μM Trolox equivalent, respectively). Furthermore, plasma and liver tissue samples from rats treated with AFB 1 plus IC431 showed significantly (P < 0.05) lower LPO values (52 and 51%, respectively) and OSi (59 and 51%, respectively) than AFB 1 -treated rats. Hence, our results proved that the intracellular content of Lact. casei CRL 431 contains metabolites that are capable to modulate the antioxidant defense systems in living organism, which may help to ameliorate the damage associated to AFB 1 -induced oxidative stress.

  18. Mass spectrometric identification and toxicity assessment of degraded products of aflatoxin B1 and B2 by Corymbia citriodora aqueous extracts

    PubMed Central

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen

    2015-01-01

    This study explores the detoxification potential of Corymbia citriodora plant extracts against aflatoxin B1 and B2 (AFB1; 100 μg L−1 and AFB2; 50 μg L−1) in In vitro and In vivo assays. Detoxification was qualitatively and quantitatively analyzed by TLC and HPLC, respectively. The study was carried out by using different parameters of optimal temperature, pH and incubation time period. Results indicated that C. citriodora leaf extract(s) more effectively degrade AFB1 and AFB2 i.e. 95.21% and 92.95% respectively than C. citriodora branch extract, under optimized conditions. The structural elucidation of degraded toxin products was done by LCMS/MS analysis. Ten degraded products of AFB1 and AFB2 and their fragmentation pathways were proposed based on molecular formulas and MS/MS spectra. Toxicity of these degraded products was significantly reduced as compared to that of parent compounds because of the removal of double bond in the terminal furan ring. The biological toxicity of degraded toxin was further analyzed by brine shrimps bioassay, which showed that only 17.5% mortality in larvae was recorded as compared to untreated toxin where 92.5% mortality was observed after 96hr of incubation. Therefore, our finding suggests that C. citriodora leaf extract can be used as an effective tool for the detoxification of aflatoxins. PMID:26423838

  19. Structural Analysis and Biological Toxicity of Aflatoxins B1 and B2 Degradation Products Following Detoxification by Ocimum basilicum and Cassia fistula Aqueous Extracts

    PubMed Central

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen; Khan, Abdul Muqeet

    2016-01-01

    This study showed the comparison between Ocimum basilicum and Cassia fistula (leaves and branch) aqueous extracts for their ability to detoxify of aflatoxins B1 and B2 (AFB1; 100 μg L-1 and AFB2; 50 μg L-1) by In Vitro assays and decontamination studies. Results indicated that O. basilicum leaves extract was found to be highly significant (P < 0.05) in degrading AFB1 and AFB2, i.e., 90.4 and 88.6%, respectively. However, O. basilicum branch, C. fistula leaves and branch extracts proved to be less efficient in degrading these aflatoxins, under optimized conditions, i.e., pH 8, temperature 30°C and incubation period of 72 h. Moreover the antifungal activity of these plants extracts were also tested. The findings depicted that O. basilicum leaves extract showed maximum growth inhibition of aflatoxigenic isolates, i.e., 82–87% as compared to other tested plants extracts. The structural elucidation of degraded toxin products by LCMS/MS analysis showed that nine degraded products of AFB1 and AFB2 were formed. MS/MS spectra showed that most of the products were formed by the removal of double bond in the terminal furan ring and modification of lactone group indicating less toxicity as compared to parent compounds. Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that O. basilicum leaves extract can be used as an effective tool for the detoxification of aflatoxins. PMID:27471501

  20. Aflatoxin B1 Detection Using a Highly-Sensitive Molecularly-Imprinted Electrochemical Sensor Based on an Electropolymerized Metal Organic Framework

    PubMed Central

    Jiang, Mengjuan; Braiek, Mohamed; Florea, Anca; Chrouda, Amani; Farre, Carole; Bonhomme, Anne; Bessueille, Francois; Vocanson, Francis; Zhang, Aidong; Jaffrezic-Renault, Nicole

    2015-01-01

    A sensitive electrochemical molecularly-imprinted sensor was developed for the detection of aflatoxin B1 (AFB1), by electropolymerization of p-aminothiophenol-functionalized gold nanoparticles in the presence of AFB1 as a template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognize and bind AFB1 through π-π interactions between AFB1 molecules and aniline moities. The performance of the developed sensor for the detection of AFB1 was investigated by linear sweep voltammetry using a hexacyanoferrate/hexacyanoferrite solution as a redox probe, the electron transfer rate increasing when the concentration of AFB1 increases, due to a p-doping effect. The molecularly-imprinted sensor exhibits a broad linear range, between 3.2 fM and 3.2 µM, and a quantification limit of 3 fM. Compared to the non-imprinted sensor, the imprinting factor was found to be 10. Selectivity studies were also performed towards the binding of other aflatoxins and ochratoxin A, proving good selectivity. PMID:26371042

  1. Immunotoxicity of aflatoxin B1: Impairment of the cell-mediated response to vaccine antigen and modulation of cytokine expression

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Meissonnier, Guylaine M.; Alltech-France, European Regulatory Department, F-92593 Levallois-Perret; Pinton, Philippe

    Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus flavus or A. parasiticus, is a frequent contaminant of food and feed. This toxin is hepatotoxic and immunotoxic. The present study analyzed in pigs the influence of AFB1 on humoral and cellular responses, and investigated whether the immunomodulation observed is produced through interference with cytokine expression. For 28 days, pigs were fed a control diet or a diet contaminated with 385, 867 or 1807 {mu}g pure AFB1/kg feed. At days 4 and 15, pigs were vaccinated with ovalbumin. AFB1 exposure, confirmed by an observed dose-response in blood aflatoxin-albumin adduct, had no majormore » effect on humoral immunity as measured by plasma concentrations of total IgA, IgG and IgM and of anti-ovalbumin IgG. Toxin exposure did not impair the mitogenic response of lymphocytes but delayed and decreased their specific proliferation in response to the vaccine antigen, suggesting impaired lymphocyte activation in pigs exposed to AFB1. The expression level of pro-inflammatory (TNF-{alpha}, IL-1{beta}, IL-6, IFN-{gamma}) and regulatory (IL-10) cytokines was assessed by real-time PCR in spleen. A significant up-regulation of all 5 cytokines was observed in spleen from pigs exposed to the highest dose of AFB1. In pigs exposed to the medium dose, IL-6 expression was increased and a trend towards increased IFN-{gamma} and IL-10 was observed. In addition we demonstrate that IL-6 impaired in vitro the antigenic- but not the mitogenic-induced proliferation of lymphocytes from control pigs vaccinated with ovalbumin. These results indicate that AFB1 dietary exposure decreases cell-mediated immunity while inducing an inflammatory response. These impairments in the immune response could participate in failure of vaccination protocols and increased susceptibility to infections described in pigs exposed to AFB1.« less

  2. Aflatoxin B1-induced DNA adduct formation and p53 mutations in CYP450-expressing human liver cell lines.

    PubMed

    Macé, K; Aguilar, F; Wang, J S; Vautravers, P; Gómez-Lechón, M; Gonzalez, F J; Groopman, J; Harris, C C; Pfeifer, A M

    1997-07-01

    Epidemiological evidence has been supporting a relationship between dietary aflatoxin B1 (AFB1) exposure, development of human primary hepatocellular carcinoma (HCC) and mutations in the p53 tumor suppressor gene. However, the correlation between the observed p53 mutations, the AFB1 DNA adducts and their activation pathways has not been elucidated. Development of relevant cellular in vitro models, taking into account species and tissue specificity, could significantly contribute to the knowledge of cytotoxicity and genotoxicity mechanisms of chemical procarcinogens, such as AFB1, in humans. For this purpose a non-tumorigenic SV40-immortalized human liver epithelial cell line (THLE cells) which retained most of the phase II enzymes, but had markedly reduced phase I activities was used for stable expression of the human CYP1A2, CYP2A6, CYP2B6 and CYP3A4 cDNA. The four genetically engineered cell lines (T5-1A2, T5-2A6, T5-2B6 and T5-3A4) produced high levels of the specific CYP450 proteins and showed comparable or higher catalytic activities related to the CYP450 expression when compared to human hepatocytes. The T5-1A2, T5-2A6, T5-2B6 and T5-3A4 cell lines exhibited a very high sensitivity to the cytotoxic effects of AFB1 and were approximately 125-, 2-, 2- and 15-fold, respectively, more sensitive than the control T5-neo cells, transfected with an expressing vector which does not contain CYP450 cDNA. In the CYP450-expressing cells, nanomolar doses of AFB1-induced DNA adduct formation including AFB1-N7-guanine, -pyrimidyl and -diol adducts. In addition, the T5-1A2 cells showed AFM1-DNA adducts. At similar levels of total DNA adducts, both the T5-1A2 and T5-3A4 cells showed, at codon 249 of the p53 gene, AGG to AGT transversions at a relative frequency of 15x10(-6). In contrast, only the T5-3A4 cells showed CCC to ACC transversion at codon 250 at a high frequency, whereas the second most frequent mutations found in the T5-1A2 cells were C to T transitions at the first

  3. Rapid screening of aflatoxin B1 in beer by fluorescence polarization immunoassay.

    PubMed

    Beloglazova, N V; Eremin, S A

    2015-09-01

    This manuscript describes the development of a sensitive, fast and easily-performed fluorescence polarization immunoassay (FPIA) for the mycotoxin aflatoxin B1 (AFB1) in various beer samples, both lager and dark. The highest sensitivity was determined for six poly- and monoclonal antibodies selective towards aflatoxins. The sample pretreatment design was emphasized since beer samples are characterized by extremely diverse matrices. Herein, the choice of sorbent for effective removal of matrix interferences prior to analysis was crucial. The samples were diluted with a borate buffer solution containing 1% PEG 6000 and passed through the clean-up column packed with NH2-derivated silica. This sample pretreatment technique was perfectly suitable for the FPIA of lager beer samples, but for dark beer and ale it did not suffice. An artificial matrix was constructed to plot a calibration curve and quantify the results of the latter samples. The developed immunoassay was characterized by a limit of detection of 1 ng mL(-1). Apparent recovery values of 89-114% for lager and 80-125% for dark beer were established. The FPIA data for AFB1 was characterized by elevated linear regression coefficients, 0.9953 for spiked lager and 0.9895 for dark beer samples respectively. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Study of an Educational Hand Sorting Intervention for Reducing Aflatoxin B1 in Groundnuts in Rural Gambia.

    PubMed

    Xu, Y A; Doel, Andrew; Watson, Sinead; Routledge, Michael N; Elliott, Christopher T; Moore, Sophie E; Gong, Yun Yun

    2017-01-01

    Aflatoxin, a human liver carcinogen, frequently contaminates groundnuts, maize, rice, and other grains, especially in Africa. The aim of this study was to evaluate the effectiveness of an educational intervention that involved training rural Gambian women on how to identify and remove moldy groundnuts to reduce aflatoxin B 1 (AFB 1 ) contamination. In total, 25 women, recruited from the West Kiang region of The Gambia, were trained on how to recognize and remove moldy groundnuts. Market-purchased groundnuts were hand sorted by the women. Groundnuts were sampled at baseline (n =5), after hand sorting ("clean," n =25 and "moldy," n =25), and after roasting (n =5). All samples were analyzed for AFB 1 by enzyme-linked immunosorbent assay. A reduction of 42.9% was achieved based on the median AFB 1 levels at baseline and after hand sorting (clean groundnuts), whereas an alternative estimate, based on the total AFB 1 in moldy and clean groundnuts, indicated a reduction of 96.7%, with a loss of only 2% of the groundnuts. By roasting the already clean sorted groundnuts, the AFB 1 reduction achieved (based on median levels) was 39.3%. This educational intervention on how to identify and remove moldy groundnuts was simple and effective in reducing AFB 1 contamination.

  5. Aflatoxin B1 Contamination in Chicken Livers and Gizzards from Industrial and Small Abattoirs, Measured by ELISA Technique in Maputo, Mozambique

    PubMed Central

    Sineque, Alberto Romão; Macuamule, Custódia Lina; Dos Anjos, Filomena Rosa

    2017-01-01

    Aflatoxins are the most toxic and carcinogenic mycotoxins produced by Aspergillus species. Aflatoxin B1 (AFB1) contamination in industrial and local chicken livers and gizzards in Maputo was investigated. One hundred boiler livers and 80 boiler gizzards were collected from industrial and local cutting poultry production sectors. The samples were analyzed by the ELISA method (MaxSignal®, Bioo Scientific Corporation). AFB1 was found in 39% of liver samples and 13.8% of gizzards, with mean levels of 1.73 µg/kg and 1.07 µg/kg, respectively. The frequency of contamination and AFB1 levels in samples from local sector producers was not significantly higher than those from industrial sector producers (p > 0.05). No correlation was found (p = 0.493; r2 = 0.013) between AFB1 levels in livers and hepatic weights. The AFB1 levels were lower than the allowed limits, suggesting that these products do not pose high risk to consumers. Notwithstanding, there is a need to implement aflatoxin residue monitoring and controls in all chicken meat products; this economic and efficient technique appears to be valuable for improved food safety in Mozambique. PMID:28832541

  6. Aflatoxin B1 in eggs and chicken livers by dispersive liquid-liquid microextraction and HPLC.

    PubMed

    Amirkhizi, Behzad; Arefhosseini, Seyed Rafie; Ansarin, Masoud; Nemati, Mahboob

    2015-01-01

    A rapid, low-cost and simple technique has been developed for the determination of aflatoxin B1 (AFB1) in eggs and livers using high-performance liquid chromatography (HPLC) with UV detection. In this study, the presence of AFB1 was investigated in 150 eggs and 50 chicken livers from the local market of Tabriz, Iran. AFB1 was extracted with a mixture of acetonitrile:water (80:20) and cleaned up by dispersive liquid-liquid microextraction which is a very economical, fast and sensitive method. AFB1 was quantified by HPLC-UV without need for any complex derivatisation in samples to enhance the detection. The results showed that 72% of the liver and 58% of the egg samples were contaminated with AFB1 ranging from 0.30 to 16.36 µg kg (̶1). limit of detection and limit of quantification for AFB1 were 0.08 and 0.28 µg kg (̶ 1), respectively. The proposed method is suitable for fast analysing of AFB1 in egg and liver samples.

  7. Evidence for involvement of multiple forms of cytochrome P-450 in aflatoxin B sup 1 metabolism in human liver

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Forrester, L.M.; Wolf, C.R.; Neal, G.E.

    Liver cancer is a major cause of premature death in many areas of Africa and Asia and its incidence is strongly correlated with exposure to aflatoxin B{sub 1} (AFB{sub 1}). Because AFB{sub 1} requires metabolic activation to achieve a biological response, there is a need for detailed knowledge of the mechanism of activation to assess individual risk. The authors carried out an extensive study using a total of 19 human liver samples to determine the individual variability in the metabolism of the toxin to mutagenic or detoxification products and to identify the specific cytochrome P-450 forms involved in these processes.more » Metabolism to the toxic 8,9-epoxide or to products mutagenic in the Ames test was found to exhibit very large individual variation. These data demonstrate that, although P450IIIA probably plays an important role in AFB{sub 1} activation, several other cytochrome P-450 forms have the capacity to activate the toxin. Similar considerations apply to detoxifying metabolism to aflatoxin Q{sub 1} and aflatoxin M{sub 1}. The levels of expression of many of the forms of cytochrome P-450 involved in AFB{sub 1} metabolism are known to be highly sensitive to environmental factors. This indicates that such factors will be an important determinant in individual susceptibility to the tumorigenic action of AFB{sub 1}.« less

  8. Interactions between water activity and temperature on the Aspergillus flavus transcriptome and aflatoxin B1 production

    USDA-ARS?s Scientific Manuscript database

    The objectives of this study were to examine the effects of Aspergillus flavus colonization of maize kernels under different water activity (aw; 0.99 and 0.91) and temperature (30 and 37°C) conditions on (a) aflatoxin B1 (AFB1) production and (b) impacts on the transcriptome using RNAseq. This study...

  9. Regression of Aflatoxin B1-Induced Hepatocellular Carcinomas by Reduced Glutathione

    NASA Astrophysics Data System (ADS)

    Novi, Anna M.

    1981-05-01

    Reduced glutathione administered to rats bearing aflatoxin B1-induced liver tumors caused regression of tumor growth and resulted in survival of the animals. Since glutathione is a harmless natural product, it merits further investigation as a potential antitumor drug for humans.

  10. Aflatoxin B1 albumin adducts in plasma and aflatoxin M1 in urine are associated with plasma concentrations of vitamins A and E

    PubMed Central

    Obuseh, Francis A.; Jolly, Pauline E.; Jiang, Yi; Shuaib, Faisal M. B.; Waterbor, John; Ellis, William O.; Piyathilake, Chandrika J.; Desmond, Renee A.; Afriyie-Gyawu, Evans; Phillips, Timothy D.

    2011-01-01

    Background Although aflatoxin exposure has been shown to be associated with micronutrient deficiency in animals, there are few investigations on the effects of aflatoxin exposure on micronutrient metabolism in humans. Objective To examine the relationship between aflatoxin B1 (AFB1) albumin adducts (AF-ALB) in plasma and the aflatoxin M1 (AFM1) metabolite in urine and plasma concentrations of retinol (vitamin A) and α-tocopherol (vitamin E) in Ghanaians. Methods A cross-sectional study of 147 adult participants was conducted. Blood and urine samples were tested for aflatoxin and vitamins A and E levels. Results Multivariable analysis showed that participants with high AF-ALB (≥ 0.80 pmol/mg albumin) had increased odds of having vitamin A deficiency compared to those with lower AF-ALB [Odds Ratio (OR) = 2.61; CI = 1.03 – 6.58; p=0.04]. Participants with high AF-ALB also showed increased odds of having vitamin E deficiency but this was not statistically significant (OR = 2.4; CI = 0.96–6.05; p = 0.06). Conversely, those with higher AFM1 values had a statistically nonsignificant reduced odds of having vitamin A deficiency (OR = 0.31; CI = 1.15–0.09; p=0.05) and statistically significant reduced odds of having vitamin E deficiency (OR = 0.31; CI = 0.10 – 0.97; p = 0.04). Participants with high AF-ALB or high AFM1 (≥ 437.95 pg/dL creatinine) were almost 6 times more likely to be hepatitis B virus surface antigen (HBsAg)- positive (OR = 5.88; CI = 1.71–20.14; p = 0.005) and (OR = 5.84; CI = 1.15–29.54; p = 0.03) respectively. Conclusions These data indicate that aflatoxin may modify plasma micronutrient status. Thus, preventing aflatoxin exposure may greatly reduce vitamins A and E deficiencies. PMID:21792816

  11. Low levels of aflatoxin B1, ricin and milk enhance recombinant protein production in mammalian cells

    USDA-ARS?s Scientific Manuscript database

    Changing the optimal tissue culture medium by adding low levels of environmental stress such as 1 µM of the fungal toxin aflatoxin B1 (AFB1), 1 ng of the castor bean protein toxin ricin in transduced mammalian cells or 1% reconstituted milk enhances transcription and increases production of the foll...

  12. Grapefruit juice intake does not enhance but rather protects against aflatoxin B1-induced liver DNA damage through a reduction in hepatic CYP3A activity.

    PubMed

    Miyata, Masaaki; Takano, Hiroki; Guo, Lian Q; Nagata, Kiyoshi; Yamazoe, Yasushi

    2004-02-01

    Influence of grapefruit juice intake on aflatoxin B1 (AFB1)-induced liver DNA damage was examined using a Comet assay in F344 rats given 5 mg/kg AFB1 by gavage. Rats allowed free access to grapefruit juice for 5 days prior to AFB1 administration resulted in clearly reduced DNA damage in liver, to 65% of the level in rats that did not receive grapefruit juice. Furthermore, rats treated with grapefruit juice extract (100 mg/kg per os) for 5 days prior to AFB1 treatment also reduced the DNA damage to 74% of the level in rats that did not receive grapefruit juice. No significant differences in the portal blood and liver concentrations of AFB1 were observed between grapefruit juice intake rats and the controls. In an Ames assay with AFB1 using Salmonella typhimurium TA98, lower numbers of revertant colonies were detected with hepatic microsomes prepared from rats administered grapefruit juice, compared with those from control rats. Microsomal testosterone 6beta-hydroxylation was also lower with rats given grapefruit juice than with control rats. Immunoblot analyses showed a significant decrease in hepatic CYP3A content, but not CYP1A and CYP2C content, in microsomes of grapefruit juice-treated rats than in non-treated rats. No significant difference in hepatic glutathione S-transferase (GST) activity and glutathione content was observed in the two groups. GSTA5 protein was not detected in hepatic cytosol of the two groups. In microsomal systems, grapefruit juice extract inhibited AFB1-induced mutagenesis in the presence of a microsomal activation system from livers of humans as well as rats. These results suggest that grapefruit juice intake suppresses AFB1-induced liver DNA damage through inactivation of the metabolic activation potency for AFB1 in rat liver.

  13. Effect of Aflatoxin B1 on Growth of Bovine Mammary Epithelial Cells in 3D and Monolayer Culture System.

    PubMed

    Forouharmehr, Ali; Harkinezhad, Taher; Qasemi-Panahi, Babak

    2013-01-01

    Many studies have been showed transfer of aflatoxins, toxins produced by Aspergillus flvaus and Aspergillus parasiticus fungi, into milk. These toxins are transferred into the milk through digestive system by eating contaminated food. Due to the toxicity of these materials, it seems that it has side effects on the growth of mammary cells. Therefore, the present work aimed to investigate possible toxic effects of aflatoxin B1 (AFB1) on bovine mammary epithelial cells in monolayer and three-dimensional cultures. Specimens of the mammary tissue of bovine were sized out in size 2×2 cm in slaughterhouse. After disinfection and washing in sterile PBS, primary cell culture was performed by enzymatic digestion of tissue with collagenase. When proper numbers of cells were achieved in monolayer culture, cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, the concentrations of 15, 25 and 35 µL of AFB1 were added to the culture in quadruplicate and incubated for 8 hours. Cellular cytotoxicity was examined using standard colorimetric assay and finally, any change in the morphology of the cells was studied by microscopic technique. Microscopic investigations showed necrosis of the AFB1-exposed cells compared to the control cells. Also, bovine mammary epithelial cells were significantly affected by AFB1 in dose and time dependent manner in cell viability assays. According to the results, it seems that AFB1 can induce cytotoxicity and necrosis in bovine mammary epithelial cells.

  14. Nanoparticle based bio-bar code technology for trace analysis of aflatoxin B1 in Chinese herbs.

    PubMed

    Yu, Yu-Yan; Chen, Yuan-Yuan; Gao, Xuan; Liu, Yuan-Yuan; Zhang, Hong-Yan; Wang, Tong-Ying

    2018-04-01

    A novel and sensitive assay for aflatoxin B1 (AFB1) detection has been developed by using bio-bar code assay (BCA). The method that relies on polyclonal antibodies encoded with DNA modified gold nanoparticle (NP) and monoclonal antibodies modified magnetic microparticle (MMP), and subsequent detection of amplified target in the form of bio-bar code using a fluorescent quantitative polymerase chain reaction (FQ-PCR) detection method. First, NP probes encoded with DNA that was unique to AFB1, MMP probes with monoclonal antibodies that bind AFB1 specifically were prepared. Then, the MMP-AFB1-NP sandwich compounds were acquired, dehybridization of the oligonucleotides on the nanoparticle surface allows the determination of the presence of AFB1 by identifying the oligonucleotide sequence released from the NP through FQ-PCR detection. The bio-bar code techniques system for detecting AFB1 was established, and the sensitivity limit was about 10 -8  ng/mL, comparable ELISA assays for detecting the same target, it showed that we can detect AFB1 at low attomolar levels with the bio-bar-code amplification approach. This is also the first demonstration of a bio-bar code type assay for the detection of AFB1 in Chinese herbs. Copyright © 2017. Published by Elsevier B.V.

  15. Ameliorative Effects of Grape Seed Proanthocyanidin Extract on Growth Performance, Immune Function, Antioxidant Capacity, Biochemical Constituents, Liver Histopathology and Aflatoxin Residues in Broilers Exposed to Aflatoxin B1

    PubMed Central

    Sun, Lvhui; Zhang, Niya; Ling, Zhao; Zhu, Luoyi; Khan, Farhan Anwar; Zhang, Jiacai; Qi, Desheng

    2017-01-01

    Aflatoxicosis is a grave threat to the poultry industry. Dietary supplementation with antioxidants showed a great potential in enhancing the immune system; hence, protecting animals against aflatoxin B1-induced toxicity. Grape seed proanthocyanidin extract (GSPE) one of the most well-known and powerful antioxidants. Therefore, the purpose of this research was to investigate the effectiveness of GSPE in the detoxification of AFB1 in broilers. A total of 300 one-day-old Cobb chicks were randomly allocated into five treatments of six replicates (10 birds per replicate), fed ad libitum for four weeks with the following dietary treatments: 1. Basal diet (control); 2. Basal diet + 1 mg/kg AFB1 contaminated corn (AFB1); 3. Basal diet + GSPE 250 mg/kg; (GSPE 250 mg/kg) 4. Basal diet + AFB1 (1 mg/kg) + GSPE 250 mg/kg; (AFB1 + GSPE 250 mg/kg) 5. Basal diet + AFB1 (1mg/kg) + GSPE 500 mg/kg, (AFB1 + GSPE 500 mg/kg). When compared with the control group, feeding broilers with AFB1 alone significantly reduced growth performance, serum immunoglobulin contents, negatively altered serum biochemical contents, and enzyme activities, and induced histopathological lesion in the liver. In addition, AFB1 significantly increased malondialdehyde content and decreased total superoxide dismutase, catalase, glutathione peroxide, glutathione-S transferase, glutathione reductase activities, and glutathione concentration within the liver and serum. The supplementation of GSPE (250 and 500 mg/kg) to AFB1 contaminated diet reduced AFB1 residue in the liver and significantly mitigated AFB1 negative effects. From these results, it can be concluded that dietary supplementation of GSPE has protective effects against aflatoxicosis caused by AFB1 in broiler chickens. PMID:29140290

  16. A Structure Identification and Toxicity Assessment of the Degradation Products of Aflatoxin B1 in Peanut Oil under UV Irradiation

    PubMed Central

    Mao, Jin; He, Bing; Zhang, Liangxiao; Li, Peiwu; Zhang, Qi; Ding, Xiaoxia; Zhang, Wen

    2016-01-01

    Aflatoxins, a group of extremely hazardous compounds because of their genotoxicity and carcinogenicity to human and animals, are commonly found in many tropical and subtropical regions. Ultraviolet (UV) irradiation is proven to be an effective method to reduce or detoxify aflatoxins. However, the degradation products of aflatoxins under UV irradiation and their safety or toxicity have not been clear in practical production such as edible oil industry. In this study, the degradation products of aflatoxin B1 (AFB1) in peanut oil were analyzed by Ultra Performance Liquid Chromatograph-Thermo Quadrupole Exactive Focus mass spectrometry/mass spectrometry (UPLC-TQEF-MS/MS). The high-resolution mass spectra reflected that two main products were formed after the modification of a double bond in the terminal furan ring and the fracture of the lactone ring, while the small molecules especially nitrogen-containing compound may have participated in the photochemical reaction. According to the above results, the possible photodegradation pathway of AFB1 in peanut oil is proposed. Moreover, the human embryo hepatocytes viability assay indicated that the cell toxicity of degradation products after UV irradiation was much lower than that of AFB1, which could be attributed to the breakage of toxicological sites. These findings can provide new information for metabolic pathways and the hazard assessment of AFB1 using UV detoxification. PMID:27845743

  17. Feasibility of detecting aflatoxin B1 on inoculated maize kernels surface using Vis/NIR hyperspectral imaging

    USDA-ARS?s Scientific Manuscript database

    The feasibility of using a visible/near-infrared hyperspectral imaging system with a wavelength range between 400 and 1000 nm to detect and differentiate different levels of aflatoxin B1 (AFB1) artificially titrated on maize kernel surface was examined. To reduce the color effects of maize kernels, ...

  18. Liposome-coated mesoporous silica nanoparticles loaded with L-cysteine for photoelectrochemical immunoassay of aflatoxin B1.

    PubMed

    Lin, Youxiu; Zhou, Qian; Zeng, Yongyi; Tang, Dianping

    2018-06-02

    The authors describe a photoelectrochemical (PEC) immunoassay for determination of aflatoxin B 1 (AFB 1 ) in foodstuff. The competitive immunoreaction is carried out on a microplate coated with a capture antibody against AFB 1 using AFB 1 -bovine serum albumin (BSA)-liposome-coated mesoporous silica nanoparticles (MSN) loaded with L-cysteine as a support. The photocurrent is produced by a photoactive material consisting of cerium-doped Bi 2 MoO 6 . Initially, L-cysteine acting as the electron donor is gated in the pores by interaction between mesoporous silica and liposome. Thereafter, AFB 1 -BSA conjugates are covalently bound to the liposomes. Upon introduction of the analyte (AFB 1 ), the labeled AFB 1 -BSA complex competes with the analyte for the antibody deposited on the microplate. Accompanying with the immunocomplex, the liposomes on the MSNs are lysed upon addition of Triton X-100. This results in the opening of the pores and in a release of L-cysteine. Free cysteine then induces the electron-hole scavenger of the photoactive nanosheets to increase the photocurrent. The photocurrent (relative to background signal) increases with increasing AFB 1 concentration. Under optimum conditions, the photoactive nanosheets display good photoelectrochemical responses, and allow the detection of AFB 1 at a concentration as low as 0.1 pg·mL -1 within a linear response in the 0.3 pg·mL -1 to 10 ng·mL -1 concentration range. Accuracy was evaluated by analyzing naturally contaminated and spiked peanut samples by using a commercial AFB 1 ELISA kit as the reference, and well-matching results were obtained. Graphical abstract Schematic presentation of a photoelectrochemical immunoassay for AFB 1 . It is based on the use of Ce-doped Bi 2 MoO 6 nanosheets and of liposome-coated mesoporous silica nanoparticles loaded with L-cysteine.

  19. Impact of casing damaging on aflatoxin B1 concentration during the ripening of dry-fermented meat sausages.

    PubMed

    Pleadin, Jelka; Kovačević, Dragan; Perković, Irena

    2015-01-01

    The aim of this article is to investigate the impact of casing damaging on the formation of aflatoxin B1 (AFB1) during the ripening of dry-fermented meat sausages. The level of AFB1 contamination was determined in 24 samples using the ELISA immunoassay throughout a six-month production period. While with intact casing samples no contamination was observed throughout the whole production process, in damaged casing samples AFB1 was detected in the ripening end-stages in the range of 1.62-4.49 μg/kg. The results showed that casing damaging occurring during long-term ripening of dry-fermented sausages can cause AFB1 contamination, possibly arising on the grounds of diffusion of this mycotoxin from the product surface to its interior.

  20. Thermal treatment of bentonite reduces aflatoxin b1 adsorption and affects stem cell death.

    PubMed

    Nones, Janaína; Nones, Jader; Riella, Humberto Gracher; Poli, Anicleto; Trentin, Andrea Gonçalves; Kuhnen, Nivaldo Cabral

    2015-10-01

    Bentonites are clays that highly adsorb aflatoxin B1 (AFB1) and, therefore, protect human and animal cells from damage. We have recently demonstrated that bentonite protects the neural crest (NC) stem cells from the toxicity of AFB1. Its protective effects are due to the physico-chemical properties and chemical composition altered by heat treatment. The aim of this study is to prepare and characterize the natural and thermal treatments (125 to 1000 °C) of bentonite from Criciúma, Santa Catarina, Brazil and to investigate their effects in the AFB1 adsorption and in NC cell viability after challenging with AFB1. The displacement of water and mineralogical phases transformations were observed after the thermal treatments. Kaolinite disappeared at 500 °C and muscovite and montmorillonite at 1000 °C. Slight changes in morphology, chemical composition, and density of bentonite were observed. The adsorptive capacity of the bentonite particles progressively reduced with the increase in temperature. The observed alterations in the structure of bentonite suggest that the heat treatments influence its interlayer distance and also its adsorptive capacity. Therefore, bentonite, even after the thermal treatment (125 to 1000 °C), is able to increase the viability of NC stem cells previously treated with AFB1. Our results demonstrate the effectiveness of bentonite in preventing the toxic effects of AFB1. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo).

    PubMed

    Monson, Melissa S; Settlage, Robert E; McMahon, Kevin W; Mendoza, Kristelle M; Rawal, Sumit; El-Nezami, Hani S; Coulombe, Roger A; Reed, Kent M

    2014-01-01

    Dietary exposure to aflatoxin B1 (AFB1) is detrimental to avian health and leads to major economic losses for the poultry industry. AFB1 is especially hepatotoxic in domestic turkeys (Meleagris gallopavo), since these birds are unable to detoxify AFB1 by glutathione-conjugation. The impacts of AFB1 on the turkey hepatic transcriptome and the potential protection from pretreatment with a Lactobacillus-based probiotic mixture were investigated through RNA-sequencing. Animals were divided into four treatment groups and RNA was subsequently recovered from liver samples. Four pooled RNA-seq libraries were sequenced to produce over 322 M reads totaling 13.8 Gb of sequence. Approximately 170,000 predicted transcripts were de novo assembled, of which 803 had significant differential expression in at least one pair-wise comparison between treatment groups. Functional analysis linked many of the transcripts significantly affected by AFB1 exposure to cancer, apoptosis, the cell cycle or lipid regulation. Most notable were transcripts from the genes encoding E3 ubiquitin-protein ligase Mdm2, osteopontin, S-adenosylmethionine synthase isoform type-2, and lipoprotein lipase. Expression was modulated by the probiotics, but treatment did not completely mitigate the effects of AFB1. Genes identified through transcriptome analysis provide candidates for further study of AFB1 toxicity and targets for efforts to improve the health of domestic turkeys exposed to AFB1.

  2. Potential for amelioration of aflatoxin B1-induced immunotoxic effects in progeny of White Leghorn breeder hens co-exposed to vitamin E.

    PubMed

    Khan, Wajid Arshad; Khan, Muhammad Zargham; Khan, Ahrar; Ul Hassan, Zahoor; Saleemi, Muhammad Kashif

    2014-01-01

    This study was designed to evaluate the protective activity of Vitamin E (Vit E) on the immunotoxic effects induced by aflatoxin B1 (AFB1) in the progeny of breeder hens. For this purpose, 192 White Leghorn (WL) layer breeder hens were divided into 12 groups (A-L) and then fed test diets for either 1, 2 or 3 weeks. Group A was kept on basal feed (2900 Kcal/kg metabolizable energy) and served as control, while group B was offered a feed supplemented with Vit E at 100 mg/Kg. Groups C-G were offered feed containing 0.1, 0.5, 2.5, 5.0, and 10.0 mg/Kg AFB1, respectively, whereas groups H-L were offered the same dietary levels of AFB1 along with 100 mg/Kg Vit E supplementation. Hatching eggs were shifted to an incubator on a weekly basis to get progeny chicks. Hatched chicks in each group were maintained on basal ration and then subjected to different immunological assays. Lymphoproliferative responses (against PHA-P), antibody titers (against SRBC), oxidative damage to RBC, as well as phagocytic and nitrite production potential of the peritoneal macrophages from the chicks, were all adversely impacted by hen exposure to the higher doses of AFB1 or by increased intake (time) by the hens at a given dose of the toxin. No consistent ameliorative effects from Vit E were noted in these studies, i.e. effects seen against lower AFB1 doses were no longer apparent with the highest doses of AFB1. As such, for now it can be concluded that, with this particular single dose level of Vit E, AFB1-associated immunotoxic effects in progeny chicks can potentially be mitigated by dietary intake of Vit E by their hen dams. However, this is clearly an outcome that is driven by the level of the mycotoxin present in the feed. Future studies need to examine what impact higher Vit E doses than those employed herein might have in these ameliorative outcomes.

  3. Effect of the Egyptian propolis on the hepatic antioxidant defense and pro-apoptotic p53 and anti-apoptotic bcl2 expressions in aflatoxin B1 treated male mice.

    PubMed

    Alm-Eldeen, Abeer A; Basyony, Mohammed A; Elfiky, Nabil K; Ghalwash, Mohamed M

    2017-03-01

    Aflatoxins are potent hepatotoxic due to their role in producing reactive oxygen species and consequently peroxidative damage. Propolis is a honey bee product known for its antioxidant capacity. The aim of this study was to verify the antioxidant effect of the Egyptian propolis extract (EPE) against aflatoxin B1 (AFB1)-induced hepatotoxicity in mice. Forty eight male mice were divided: first, second and third groups were used as control receiving saline, olive oil and EPE respectively, fourth was AFB1 group, fifth and sixth received EPE post or pre AFB1 treatment, respectively. EPE was given as (0.2mg/kg) 3 times a week. AFB1 was given as a single dose (0.25μg/kg). After 2 weeks, the mice were scarified and biochemical, histopathological and immunohistochemical investigations were assessed. EPE has a high content of total phenolics and alkaloids. The inhibitory concentration 50 (IC50) value for DPPH radical scavenging was 1353.8μg/mL. Pretreatment with EPE improved AFB1-induced hepatotoxicity represented in lowering alanine transaminase, aspartate aminotransferase, alkaline phosphatase, cholesterol, triglycerides, lipid peroxidation and pro-apoptotic p53 expression to 33.48±1.98 IU/ml, 53.00±2.37 IU/ml, 123.50±2.02 IU/ml, 76.50±2.66mg/dl, 54.00±3.03mg/dl, 2.22±0.14 nmol/g and 4.31±2.1 cells/field and raising the reduced glutathione, catalase, superoxide dismutase and anti-apoptotic bcl2 expression to 3.37±1.65 nmol/g, 4.92±0.25 nmol/g, 57±0.91UI/g and 39.7±5.9 cells/field which all had non-significant differences with the control, respectively. In conclusion, EPE can attenuate aflatoxin B1-induced hepatotoxicity in mice. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  4. Effect of Aflatoxin B1 on Growth of Bovine Mammary Epithelial Cells in 3D and Monolayer Culture System

    PubMed Central

    Forouharmehr, Ali; Harkinezhad, Taher; Qasemi-Panahi, Babak

    2013-01-01

    Purpose: Many studies have been showed transfer of aflatoxins, toxins produced by Aspergillus flvaus and Aspergillus parasiticus fungi, into milk. These toxins are transferred into the milk through digestive system by eating contaminated food. Due to the toxicity of these materials, it seems that it has side effects on the growth of mammary cells. Therefore, the present work aimed to investigate possible toxic effects of aflatoxin B1 (AFB1) on bovine mammary epithelial cells in monolayer and three-dimensional cultures. Methods: Specimens of the mammary tissue of bovine were sized out in size 2×2 cm in slaughterhouse. After disinfection and washing in sterile PBS, primary cell culture was performed by enzymatic digestion of tissue with collagenase. When proper numbers of cells were achieved in monolayer culture, cells were seeded in a 24-well culture plate for three-dimensional (3D) culture in Matrigel matrix. After 21 days of 3D culture and reaching the required number of cells, the concentrations of 15, 25 and 35 µL of AFB1 were added to the culture in quadruplicate and incubated for 8 hours. Cellular cytotoxicity was examined using standard colorimetric assay and finally, any change in the morphology of the cells was studied by microscopic technique. Results: Microscopic investigations showed necrosis of the AFB1-exposed cells compared to the control cells. Also, bovine mammary epithelial cells were significantly affected by AFB1 in dose and time dependent manner in cell viability assays. Conclusion: According to the results, it seems that AFB1 can induce cytotoxicity and necrosis in bovine mammary epithelial cells. PMID:24312827

  5. Effects of milk thistle seed against aflatoxin B1 in broiler model.

    PubMed

    Amiridumari, Halimeh; Sarir, Hadi; Afzali, Nazar; Fanimakki, Omid

    2013-09-01

    Consumption of aflatoxin B1 (AFB1) contaminated products can pose a risk of development of various diseases in human and animals due to radical production. The scope of this work is to evaluate the efficacy of milk thistle seed (MTS), as a radical scavenger, on serum biochemistry, lipid profile and liver enzymes against AFB1 in broiler chickens contaminated with AFB1. The effect of nine experimental treatments (3 × 3 factorial design) was assessed using 216 one-d-old Ross 308 male broiler chicks in a randomized complete design with four replicates of six birds for each dietary treatments: Control (T1), 250 ppb AFB1 (T2), 500 ppb AFB1 (T3), 0.5% MTS (T4), 0.5% MTS Plus 250 ppb AFB1 (T5), 0.5% MTS Plus 500 ppb AFB1 (T6), 1.0% MTS (T7), 1.0% MTS Plus 250 ppb AFB1 (T8), and 1.0% MTS Plus 500 ppb AFB1 (T9). The individual and combined effects of dietary AFB1 and MTS on serum biochemistry factors (Glucose, Calcium, Phosphorus, Iron, Creatinine, and Uric acid), lipid profile (Triglyceride, Cholesterol, Low density lipoprotein (LDL), and High density lipoprotein (HDL)) and liver enzymes aspartate amino-transferase and alanine amino-transaminase (ALT) in broilers were evaluated at 21 days of age. Also, statistical packages Macros-1.002 (2010) were used to perform the above analysis on computer. Consumption of 500 ppb AFB1 in to the diet significantly decreased HDL (58.13 ± 2.65), Calcium (7.11 ± 0.13), and Glucose (197.1 ± 7.42) compared to the control group (85.12 ± 1.95, 9.45 ± 0.17 and 223.1 ± 6.61, respectively), (P < 0.05). In contrast, it significantly increased creatinine (2.25 ± 0.011) and AST (244.51 ± 4.91). Using MTS together with AFB1 significantly reduced the effect of AFB1 on the above parameters. MTS can provide protection against the negative effects of AFB1 on broiler chicks.

  6. Biotransformation of aflatoxin B1 and aflatoxin G1 in peanut meal by anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus.

    PubMed

    Chen, Yujie; Kong, Qing; Chi, Chen; Shan, Shihua; Guan, Bin

    2015-10-15

    The purpose of this study was to explore the ability of anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to biotransform aflatoxins in peanut meal. The pH of the peanut meal was adjusted above 10, and then heated for 10 min at 100 °C, 115 °C and 121 °C. The S. thermophilus and L. delbrueckii subsp. bulgaricus were precultured together in MRS broth for 48 h at 37 °C. The heated peanut meal was mixed with precultured MRS broth containing 7.0×10(8) CFU/mL of S. thermophilus and 3.0×10(3) CFU/mL of L. delbrueckii subsp. bulgaricus with the ratio of 1 to 1 (weight to volume) and incubated in anaerobic jars at 37 °C for 3 days. The aflatoxin content in the peanut meal samples was determined by HPLC. The results showed that the peanut meal contained mainly aflatoxin B1 (AFB1) (10.5±0.64 μg/kg) and aflatoxin G1 (AFG1) (18.7±0.55 μg/kg). When heat treatment was combined with anaerobic solid fermentation, the biotransformation rate of aflatoxins in peanut meal could attain 100%. The cytotoxicity of fermented peanut meal to L929 mouse connective tissue fibroblast cells was determined by MTT assay and no significant toxicity was observed in the fermented peanut meal. Furthermore, heat treatment and anaerobic solid fermentation did not change the amino acid concentrations and profile in peanut meal. Copyright © 2015 Elsevier B.V. All rights reserved.

  7. Aflatoxin B1 contamination in feed from Puglia and Basilicata regions (Italy): 5 years monitoring data.

    PubMed

    Vita, V; Clausi, M T; Franchino, C; De Pace, R

    2016-11-01

    During a 5-year period from 2010 to 2014, n = 919 samples of feed and raw materials were analyzed for aflatoxin B 1 (AFB 1 ) contamination using accredited ELISA screening methods. Only 0.76 % of these samples were non-compliant with maximum levels set by the European Union Regulation 32/2002. Non-compliant samples were mainly from the province of Bari (n = 3 samples, mean AFB 1 value 7.03 μg/kg), although the highest AFB 1 levels were found in two samples from the provinces of Foggia and Brindisi, at 32.6 ± 3.6 μg/kg and 31.0 ± 4.0 μg/kg, respectively. Mean AFB 1 levels in samples contaminated but compliant with the limits ranged from 1.4 to 2.2 μg/kg. Considering the great importance of climate conditions in mycotoxins production, during crops production and during the critical phases of materials storage and/or transport, to better understand the variability in contamination levels, the analytical results were reviewed in term of temperature and relative environmental humidity in the sampling areas. Correlations between aflatoxin B 1 levels in feed and these climate factors might explain seasonal and annual variations in contamination levels. The data from the present study provide useful suggestions for the organization of targeted monitoring plans and the protection of consumers, as well as for improvement in the quality standards of zootechnological activities and feed industry.

  8. Mapping the binding site of aflatoxin B/sub 1/ in DNA: systematic analysis of the reactivity of aflatoxin B/sub 1/ with guanines in different DNA sequences

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Benasutti, M.; Ejadi, S.; Whitlow, M.D.

    The mutagenic and carcinogenic chemical aflatoxin B/sub 1/ (AFB/sub 1/) reacts almost exclusively at the N(7)-position of guanine following activation to its reactive form, the 8,9-epoxide (AFB/sub 1/ oxide). In general N(7)-guanine adducts yield DNA strand breaks when heated in base, a property that serves as the basis for the Maxam-Gilbert DNA sequencing reaction specific for guanine. Using DNA sequencing methods, other workers have shown that AFB/sub 1/ oxide gives strand breaks at positions of guanines; however, the guanine bands varied in intensity. This phenomenon has been used to infer that AFB/sub 1/ oxide prefers to react with guanines inmore » some sequence contexts more than in others and has been referred to as sequence specificity of binding. Herein, data on the reaction of AFB/sub 1/ oxide with several synthetic DNA polymers with different sequences are presented, and (following hydrolysis) adduct levels are determine by high-pressure liquid chromatography. These results reveal that for AFB/sub 1/ oxide (1) the N(7)-guanine adduct is the major adduct found in all of the DNA polymers, (2) adduct levels vary in different sequences, and, thus, sequence specificity is also observed by this more direct method, and (3) the intensity of bands in DNA sequencing gels is likely to reflect adduct levels formed at the N(7)-position of guanine. Knowing this, a reinvestigation of the reactivity of guanines in different DNA sequences using DNA sequencing methods was undertaken. Methods are developed to determine the X (5'-side) base and the Y (3'-side) base are most influential in determining guanine reactivity. These rules in conjunction with molecular modeling studies were used to assess the binding sites that might be utilized by AFB/sub 1/ oxide in its reaction with DNA.« less

  9. Use of yeast (Pichia kudriavzevii) as a novel feed additive to ameliorate the effects of aflatoxin B1 on broiler chicken performance.

    PubMed

    Magnoli, A P; Rodriguez, M C; González Pereyra, M L; Poloni, V L; Peralta, M F; Nilson, A J; Miazzo, R D; Bagnis, G; Chiacchiera, S M; Cavaglieri, L R

    2017-11-01

    The aim of this study was to evaluate the efficacy of autochthonous Pichia kudriavzevii as a novel bioadsorbent for aflatoxin B 1 (AFB 1 ). The selection of this yeast was based on the AFB 1 adsorption capacity previously demonstrated in vitro (Magnoli et al. 2016). One-day-old Cobb broilers (n = 160) were randomly assigned to four dietary treatments (T1: basal diet (B); T2: B + 0.1% yeast; T3: B + AFB 1 , 100 μg/kg; T4: B + 0.1% yeast + AFB 1 , 100 μg/kg). Performance parameters (average daily weight gain body, average daily consumption, feed conversion ratio, carcass weight, and dead weight), biochemical parameters (albumin, globulin, and albumin/globulin), liver pathological changes, and AFB 1 residual levels in the liver and excreta were evaluated. Significant differences (P < 0.05) in performance parameters were observed among treatments and controls: T3 group showed the lowest average daily body weight gain value while in T4 group, the value of this parameter increased significantly (P < 0.05). T3 and T4 groups showed the lowest and highest values for average daily feed consumption, respectively. The feed conversion ratio (FC) showed no significant differences among treatments. T3 group showed the lowest dead weight and carcass weight compared with T1 group. The biochemical parameters showed no significant differences among treatments. T3 group showed macroscopic and microscopic liver changes compared to the control. Aflatoxin B 1 levels (μg/g) were detected in broiler livers and showed significant differences among treatments (P < 0.05). In conclusion, native P. kudriavzevii incorporation (0.1%) in broiler diets containing AFB 1 was shown to be effective in ameliorating the adverse effects of AFB 1 on production.

  10. Effects of chlorophyll and chlorophyllin on low-dose aflatoxin B1 pharmacokinetics in human volunteers: A pilot study

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jubert, C; Mata, J; Bench, G

    Chlorophyll (Chla) and chlorophyllin (CHL) were shown previously to reduce carcinogen bioavailability, biomarker damage, and tumorigenicity in trout and rats. These findings were partially extended to humans (Proc Natl Acad Sci USA 98, 14601-14606 (2001)), where CHL reduced excretion of aflatoxin B{sub 1} (AFB{sub 1})-DNA repair products in Chinese unavoidably exposed to dietary AFB{sub 1}. However, neither AFB{sub 1} pharmacokinetics nor Chla effects were examined. We conducted a small unblinded crossover study to establish AFB{sub 1} pharmacokinetic parameters in human volunteers, and to explore possible effects of CHL or Chla co-treatment on those parameters. For protocol 1, fasted subjects receivedmore » an IRB-approved dose of 14C-AFB{sub 1} (30 ng, 5 nCi) by capsule with 100 ml water, followed by normal eating and drinking after hr 2. Blood and cumulative urine samples were collected over 72 hr, and {sup 14}C-AFB{sub 1} equivalents were determined by Accelerator Mass Spectrometry. Protocols 2 and 3 were similar except capsules also contained 150 mg of purified Chla, or CHL, respectively. All protocols were repeated 3 times for each of three volunteers. The study revealed rapid human AFB{sub 1} uptake (plasma ka 5.05 {+-} 1.10 hr-1, Tmax 1.0 hr) and urinary elimination (95% complete by 24 hr) kinetics. Chla and CHL treatment each significantly impeded AFB{sub 1} absorption and reduced Cmax and AUC's (plasma and urine) in one or more subjects. These initial results provide AFB{sub 1} pharmacokinetic parameters previously unavailable for humans, and suggest that Chla or CHL co-consumption may limit the bioavailability of ingested aflatoxin in humans, as they do in animal models.« less

  11. Development of the custom polymeric materials specific for aflatoxin B1 and ochratoxin A for application with the ToxiQuant T1 sensor tool.

    PubMed

    Piletska, Elena; Karim, Kal; Coker, Raymond; Piletsky, Sergey

    2010-04-16

    Two polymers were computationally designed with affinity to two of the most abundant mycotoxins aflatoxin B1 (AFB1) and ochratoxin A (OTA) for application in the ToxiQuant T1 System. The principle of quantification of AFB1 and OTA using the ToxiQuant T1 instrument comprised of a fluorimetric analysis of mycotoxins adsorbed on the polymer upon exposure to UV light. High affinity of the developed resins allowed the adsorption of both toxins as discrete bands on the top of the cartridge with detection limit as low as 1ng quantity of mycotoxins. Copyright 2009 Elsevier B.V. All rights reserved.

  12. Up-regulation of nucleotide excision repair in mouse lung and liver following chronic exposure to aflatoxin B{sub 1} and its dependence on p53 genotype

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mulder, Jeanne E.; Bondy, Genevieve S.; Mehta, Rekha

    Aflatoxin B{sub 1} (AFB{sub 1}) is biotransformed in vivo into an epoxide metabolite that forms DNA adducts that may induce cancer if not repaired. p53 is a tumor suppressor gene implicated in the regulation of global nucleotide excision repair (NER). Male heterozygous p53 knockout (B6.129-Trp53{sup tm1Brd}N5, Taconic) and wild-type mice were exposed to 0, 0.2 or 1.0 ppm AFB{sub 1} for 26 weeks. NER activity was assessed with an in vitro assay, using AFB{sub 1}-epoxide adducted plasmid DNA as a substrate. For wild-type mice, repair of AFB{sub 1}–N7-Gua adducts was 124% and 96% greater in lung extracts from mice exposedmore » to 0.2 ppm and 1.0 ppm AFB{sub 1} respectively, and 224% greater in liver extracts from mice exposed to 0.2 ppm AFB{sub 1} (p < 0.05). In heterozygous p53 knockout mice, repair of AFB{sub 1}–N7-Gua was only 45% greater in lung extracts from mice exposed to 0.2 ppm AFB{sub 1} (p < 0.05), and no effect was observed in lung extracts from mice treated with 1.0 ppm AFB{sub 1} or in liver extracts from mice treated with either AFB{sub 1} concentration. p53 genotype did not affect basal levels of repair. AFB{sub 1} exposure did not alter repair of AFB{sub 1}-derived formamidopyrimidine adducts in lung or liver extracts of either mouse genotype nor did it affect XPA or XPB protein levels. In summary, chronic exposure to AFB{sub 1} increased NER activity in wild-type mice, and this response was diminished in heterozygous p53 knockout mice, indicating that loss of one allele of p53 limits the ability of NER to be up-regulated in response to DNA damage. - Highlights: • Mice are chronically exposed to low doses of the mycotoxin aflatoxin B{sub 1} (AFB{sub 1}). • The effects of AFB{sub 1} and p53 status on nucleotide excision repair are investigated. • AFB{sub 1} increases nucleotide excision repair in wild type mouse lung and liver. • This increase is attenuated in p53 heterozygous mouse lung and liver. • Results portray the role of p53 in

  13. Protective effect of Ocimum sanctum on 3-methylcholanthrene, 7,12-dimethylbenz(a)anthracene and aflatoxin B1 induced skin tumorigenesis in mice

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rastogi, Shipra; Shukla, Yogeshwer; Paul, Bhola N.

    A study on the protective effect of alcoholic extract of the leaves of Ocimum sanctum on 3-mthylcholanthrene (MCA), 7,12-dimethylbenzanthracene (DMBA) and aflatoxin B{sub 1} (AFB{sub 1}) induced skin tumorigenesis in a mouse model has been investigated. The study involved pretreatment of mice with the leaf extract prior to either MCA application or tetradecanoyl phorbol acetate (TPA) treatment in a two-stage tumor protocol viz a viz, DMBA/TPA and AFB1/TPA. The results of the present study indicate that the pretreatment with alcoholic extract of the leaves of O. sanctum decreased the number of tumors in MCA, DMBA/TPA and AFB1/TPA treated mice. Themore » skin tumor induced animals pretreated with alcoholic extract led to a decrease in the expression of cutaneous {gamma}-glutamyl transpeptidase (GGT) and glutathione-S-transferase-P (GST-P) protein. The histopathological examination of skin tumors treated with leaf extract showed increased infiltration of polymorphonuclear, mononuclear and lymphocytic cells, decreased ornithine decarboxylase activity with concomitant enhancement of interleukin-1{beta} (IL-1{beta}) and tumor necrosis factor-{alpha} (TNF-{alpha}) in the serum, implying the in vivo antiproliferative and immunomodulatory activity of leaf extract. The decrease in cutaneous phase I enzymes and elevation of phase II enzymes in response to topical application of leaf extract prior to MCA, AFB1, DMBA/TPA and AFB1/TPA treatment indicate the possibility of impairment in reactive metabolite(s) formation and thereby reducing skin carcinogenicity. Furthermore, pretreatment of leaf extract in the carcinogen induced animals resulted in elevation of glutathione levels and decrease in lipid peroxidation along with heat shock protein expression, indicating a scavenging or antioxidant potential of the extract during chemical carcinogenesis. Thus it can be concluded that leaf extract of O. sanctum provides protection against chemical carcinogenesis in one or more of the

  14. Testing an aflatoxin B1 gene signature in rat archival tissues.

    PubMed

    Merrick, B Alex; Auerbach, Scott S; Stockton, Patricia S; Foley, Julie F; Malarkey, David E; Sills, Robert C; Irwin, Richard D; Tice, Raymond R

    2012-05-21

    Archival tissues from laboratory studies represent a unique opportunity to explore the relationship between genomic changes and agent-induced disease. In this study, we evaluated the applicability of qPCR for detecting genomic changes in formalin-fixed, paraffin-embedded (FFPE) tissues by determining if a subset of 14 genes from a 90-gene signature derived from microarray data and associated with eventual tumor development could be detected in archival liver, kidney, and lung of rats exposed to aflatoxin B1 (AFB1) for 90 days in feed at 1 ppm. These tissues originated from the same rats used in the microarray study. The 14 genes evaluated were Adam8, Cdh13, Ddit4l, Mybl2, Akr7a3, Akr7a2, Fhit, Wwox, Abcb1b, Abcc3, Cxcl1, Gsta5, Grin2c, and the C8orf46 homologue. The qPCR FFPE liver results were compared to the original liver microarray data and to qPCR results using RNA from fresh frozen liver. Archival liver paraffin blocks yielded 30 to 50 μg of degraded RNA that ranged in size from 0.1 to 4 kB. qPCR results from FFPE and fresh frozen liver samples were positively correlated (p ≤ 0.05) by regression analysis and showed good agreement in direction and proportion of change with microarray data for 11 of 14 genes. All 14 transcripts could be amplified from FFPE kidney RNA except the glutamate receptor gene Grin2c; however, only Abcb1b was significantly upregulated from control. Abundant constitutive transcripts, S18 and β-actin, could be amplified from lung FFPE samples, but the narrow RNA size range (25-500 bp length) prevented consistent detection of target transcripts. Overall, a discrete gene signature derived from prior transcript profiling and representing cell cycle progression, DNA damage response, and xenosensor and detoxication pathways was successfully applied to archival liver and kidney by qPCR and indicated that gene expression changes in response to subchronic AFB1 exposure occurred predominantly in the liver, the primary target for AFB1-induced

  15. Bioremediation of aflatoxin B1-contaminated maize by king oyster mushroom (Pleurotus eryngii).

    PubMed

    Branà, Maria Teresa; Cimmarusti, Maria Teresa; Haidukowski, Miriam; Logrieco, Antonio Francesco; Altomare, Claudio

    2017-01-01

    Aflatoxin B1 (AFB1) is the most harmful mycotoxin that occurs as natural contaminant of agricultural commodities, particularly maize. Practical solutions for detoxification of contaminated staples and reduction of agricultural wastes are scarce. We investigated the capability of the white-rot and edible fungus Plerotus eryngii (king oyster mushroom) to degrade AFB1 both in vitro and in a laboratory-scale mushroom cultivation, using a substrate similar to that routinely used in mushroom farms. In malt extract broth, degradation of AFB1 (500 ng/mL) by nine isolates of P. eryngii ranged from 81 to 99% after 10 days growth, and reached 100% for all isolates after 30 days. The growth of P. eryngii on solid medium (malt extract-agar, MEA) was significantly reduced at concentrations of AFB1 500 ng/mL or higher. However, the addition of 5% wheat straw to the culture medium increased the tolerance of P. eryngii to AFB1 and no inhibition was observed at a AFB1 content of 500 ng/mL; degradation of AFB1 in MEA supplemented with 5% wheat straw and 2.5% (w/v) maize flour was 71-94% after 30 days of growth. Further, AFB1 degradation by P. eryngii strain ITEM 13681 was tested in a laboratory-scale mushroom cultivation. The mushroom growth medium contained 25% (w/w) of maize spiked with AFB1 to the final content of 128 μg/kg. Pleurotus eryngii degraded up to 86% of the AFB1 in 28 days, with no significant reduction of either biological efficiency or mushroom yield. Neither the biomass produced on the mushroom substrate nor the mature basidiocarps contained detectable levels of AFB1 or its metabolite aflatoxicol, thus ruling out the translocation of these toxins through the fungal thallus. These findings make a contribution towards the development of a novel technology for remediation of AFB1- contaminated corn through the exploitation of the degradative capability of P. eryngii and its bioconversion into high nutritional value material intended for feed production.

  16. In vitro ability of beer fermentation residue and yeast-based products to bind aflatoxin B1.

    PubMed

    Bovo, Fernanda; Franco, Larissa Tuanny; Rosim, Roice Eliana; Barbalho, Ricardo; de Oliveira, Carlos Augusto Fernandes

    2015-06-01

    This study aimed to verify the in vitro ability of beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells and five commercial products that differed in the viability and integrity of S. cerevisiae cells to remove aflatoxin B1 (AFB1) from a citrate-phosphate buffer solution (CPBS). BFR was collected at a microbrewery and prepared by drying and milling. The commercial yeast-based products were as follows: inactive intact yeast cells from beer alcoholic fermentation, inactive intact yeast cells from sugarcane alcoholic fermentation, hydrolyzed yeast cells, yeast cell walls and active yeast cells. Adsorption assays were performed in CPBS spiked with 1.0 μg AFB1/mL at pH 3.0 and 6.0 for a contact time of 60 min at room temperature. Analysis of AFB1 in the samples was performed by high performance liquid chromatography. AFB1 adsorption by the products ranged from 45.5% to 69.4% at pH 3.0 and from 24.0% to 63.8% at pH 6.0. The higher percentages (p < 0.05) of AFB1 binding at both pH values were achieved with products containing hydrolyzed yeast cells or yeast cell walls rather than intact cells. The AFB1 binding percentages of BFR were 55.0 ± 5.0% at pH 3.0 and 49.2 ± 4.5% at pH 6.0, which was not significantly different (p > 0.05) from commercial products containing inactive intact yeast cells. The results of this trial indicate that the yeast-based products tested, especially the BFR, have potential applications in animal feeds as a suitable biological method for reducing the adverse effects of aflatoxins.

  17. In vitro ability of beer fermentation residue and yeast-based products to bind aflatoxin B1

    PubMed Central

    Bovo, Fernanda; Franco, Larissa Tuanny; Rosim, Roice Eliana; Barbalho, Ricardo; de Oliveira, Carlos Augusto Fernandes

    2015-01-01

    This study aimed to verify the in vitro ability of beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells and five commercial products that differed in the viability and integrity of S. cerevisiae cells to remove aflatoxin B1 (AFB1) from a citrate-phosphate buffer solution (CPBS). BFR was collected at a microbrewery and prepared by drying and milling. The commercial yeast-based products were as follows: inactive intact yeast cells from beer alcoholic fermentation, inactive intact yeast cells from sugarcane alcoholic fermentation, hydrolyzed yeast cells, yeast cell walls and active yeast cells. Adsorption assays were performed in CPBS spiked with 1.0 μg AFB1/mL at pH 3.0 and 6.0 for a contact time of 60 min at room temperature. Analysis of AFB1 in the samples was performed by high performance liquid chromatography. AFB1 adsorption by the products ranged from 45.5% to 69.4% at pH 3.0 and from 24.0% to 63.8% at pH 6.0. The higher percentages (p < 0.05) of AFB1 binding at both pH values were achieved with products containing hydrolyzed yeast cells or yeast cell walls rather than intact cells. The AFB1 binding percentages of BFR were 55.0 ± 5.0% at pH 3.0 and 49.2 ± 4.5% at pH 6.0, which was not significantly different (p > 0.05) from commercial products containing inactive intact yeast cells. The results of this trial indicate that the yeast-based products tested, especially the BFR, have potential applications in animal feeds as a suitable biological method for reducing the adverse effects of aflatoxins. PMID:26273277

  18. Aflatoxin B1 in common Egyptian foods.

    PubMed

    Selim, M I; Popendorf, W; Ibrahim, M S; el Sharkawy, S; el Kashory, E S

    1996-01-01

    Samples of common Egyptian foods (17 nuts and seeds, 10 spices, 31 herbs and medicinal plants, 12 dried vegetables, and 28 cereal grains) were collected from markets in Cairo and Giza. A portion of each sample was extracted with chloroform, and the concentrated extract was cleaned by passing through a silica gel column. Aflatoxin B1 was determined by reversed-phase liquid chromatography with UV detection. The highest prevalence of aflatoxin B1 was in nuts and seeds (82%), followed by spices (40%), herbs and medicinal plants (29%), dried vegetables (25%), and cereal grains (21%). The highest mean concentration of aflatoxin B1 was in herb and medicinal plants (49 ppb), followed by cereals (36 ppb), spices (25 ppb), nuts and seeds (24 ppb), and dried vegetables (20 ppb). Among nuts and seeds, the prevalence of aflatoxin B1 was highest (100%) in watermelon seeds, inshell peanuts, and unshelled peanuts. The lowest prevalence and concentrations were in hommos (garbanzo beans). The highest concentrations of aflatoxin B1 were detected in foods that had no potential for field contamination but required drying during processing and storage, such as pomegranate peel, watermelon seeds, and molokhia.

  19. Occurrence of aflatoxin B1 in natural products

    PubMed Central

    Prado, Guilherme; Altoé, Aline F.; Gomes, Tatiana C. B.; Leal, Alexandre S.; Morais, Vanessa A. D.; Oliveira, Marize S.; Ferreira, Marli B.; Gomes, Mateus B.; Paschoal, Fabiano N.; von S. Souza, Rafael; Silva, Daniela A.; Cruz Madeira, Jovita E. G.

    2012-01-01

    The media claims for the consumption of natural resource-based food have gradually increased in both developing and developed countries. The interest in the safety of these products is partially due to the possible presence of toxigenic fungi acting as mycotoxin producers, such as aflatoxins produced during the secondary metabolism of Aspergillus flavus, A. parasiticus and A. nomius. Aflatoxins, mainly aflatoxin B1, are directly associated with liver cancer in human beings. This paper is aimed at evaluating the presence of aflatoxin B1 in a few vegetable drugs, dried plant extracts and industrialized products traded in 2010 in the city of Belo Horizonte, State of Minas Gerais, Brazil. The method used for the quantification of aflatoxin B1 was based on extraction through acetone:water (85:15), immunoaffinity column purification followed by separation and detection in high efficiency liquid chromatography. Under the conditions of analysis, the Limits of Detection and Quantification were 0.6 µg kg-1 and 1.0 µg kg-1 respectively. The complete sets of analyses were carried out in duplicate. Aflatoxin B1 was noticed in a single sample (< 1.0 µg kg-1). The results revealed low aflatoxin B1 contamination in the products under analysis. However, it is required to establish a broad monitoring program in order to obtain additional data and check up on the actual extension of contamination. PMID:24031973

  20. In vitro metabolism of the pro-carcinogen aflatoxin B1 by liver preparations of the calf, nurse shark and clearnose skate.

    PubMed

    Bodine, A B; Luer, C A; Gangjee, S A; Walsh, C J

    1989-01-01

    1. Liver postmitochondrial supernatant preparations of calf, clearnose skate, and nurse shark were able to metabolize the fungal toxin aflatoxin B1 to various metabolites. 2. Calf liver produced aflatoxin M1 and Q1 as the major chloroform soluble metabolites, with small amounts of aflatoxicol formed during incubation. 3. Liver preparations of the elasmobranchs, however, produced aflatoxicol as the major chloroform soluble metabolite with no other metabolite being detected. 4. The water soluble metabolite profiles for the three species were also quite different with the tris diol adduct being produced to a much greater extent in calf liver preparations. 5. Aflatoxicol production by the elasmobranch liver homogenates was reversible with the skate reconverting a large amount (30%) of aflatoxicol to AFB1. The nurse shark, however, appeared to convert a portion of aflatoxicol to an unknown metabolite more polar than AFB1. 6. Calf liver DNA bound approximately 3 x more 3H-AFB1 than shark liver DNA.

  1. Electrochemical immunosensor based on poly (3,4-ethylenedioxythiophene) modified with gold nanoparticle to detect aflatoxin B1.

    PubMed

    Sharma, Arati; Kumar, A; Khan, Raju

    2017-07-01

    Gold nanoparticles (Au-NPs) functionalized Poly (3,4-ethylenedioxythiophene) (PEDOT) bioelectrodes were fabricated layer by layer deposition on ITO electrode for detection of aflatoxin B 1 . The monoclonal anti-aflatoxin antibodies (Anti-AFB 1 ) were immobilized over the surface of PEDOT/Au-NPs/ITO using EDC/NHS coupling. The surface morphology and characteristics of the modified electrodes were investigated by scanning electron microscope and contact angle measurements. The electrochemical analysis of the fabricated immunoelectrode and the immobilization of the antibodies have been evaluated and confirmed by performing Cyclic Voltammetry (CV), Electrochemical Impedance Spectroscopy (EIS) and Fourier Transform Infrared Spectroscopy (FTIR). Decrease in the value of electron transfer resistance (R et ) and increase in the peak current values after incorporation of Au-NPs signifies the enhanced properties of PEDOT embedded Au-NPs. The heterogeneous rate constant (k s ) and transfer coefficient (α) have been determined by using Laviron's method. The fabricated immunosensor exhibits high sensitive amperometric response of 3.72μAngmL -1 towards AFB 1 concentration in a linear range of 1-25ngmL -1 with detection limit (LOD) of 0.0045ngmL -1 and limit of quantification (LOQ) of 0.0156ngmL -1 . The fabricated immunoelectrode shows a reproducibility of 96.13% and 94.5% towards real maize sample spiked with AFB 1 of concentration 30ngmL -1 and 50ngmL -1 , respectively. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Assessment of workers' exposure to aflatoxin B1 in a Portuguese waste industry.

    PubMed

    Viegas, Susana; Veiga, Luisa; Figueiredo, Paula; Almeida, Ana; Carolino, Elisabete; Viegas, Carla

    2015-03-01

    Aflatoxin B1 (AFB1) is considered by different International Agencies as a genotoxic and potent hepatocarcinogen. However, despite the fact that the fungi producing this compound are detected in some work environments, AFB1 is rarely monitored in occupational settings. The aim of the present investigation was to assess exposure to AFB1 of workers from one Portuguese waste company located in the outskirt of Lisbon. Occupational exposure assessment to AFB1 was done with a biomarker of internal dose that measures AFB1 in the serum by enzyme-linked immunosorbent assay. Forty-one workers from the waste company were enrolled in this study (26 from sorting; 9 from composting; 6 from incineration). A control group (n = 30) was also considered in order to know the AFB1 background levels for the Portuguese population. All the workers showed detectable levels of AFB1 with values ranging from 2.5ng ml(-1) to 25.9ng ml(-1) with a median value of 9.9±5.4ng ml(-1). All of the controls showed values below the method's detection limit. Results obtained showed much higher (8-fold higher) values when compared with other Portuguese settings already studied, such as poultry and swine production. Besides this mycotoxin, other mycotoxins are probably present in this occupational setting and this aspect should be taken into consideration for the risk assessment process due to possible synergistic reactions. The data obtained suggests that exposure to AFB1 occurs in a waste management setting and claims attention for the need of appliance of preventive and protective safety measures. © The Author 2014. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.

  3. Structures of Degradation Products and Degradation Pathways of Aflatoxin B1 by High-Voltage Atmospheric Cold Plasma (HVACP) Treatment.

    PubMed

    Shi, Hu; Cooper, Bruce; Stroshine, Richard L; Ileleji, Klein E; Keener, Kevin M

    2017-08-02

    High-voltage atmospheric cold plasma (HVACP) is a novel nonthermal decontamination technology that has potential for use in the food industry. In this study, HVACP was applied to treat pure aflatoxin B 1 (AFB 1 ) powder on a glass slide. AFB 1 was degraded by 76% using a 5 min HVACP treatment in air having 40% relative humidity. The degradation products of AFB 1 were separated, and their molecular formulas were elucidated using liquid-chromatography time-of-flight mass spectrometry (HPLC-TOF-MS). Six main degradation products were observed. The structures of the degradation products were further clarified via orbitrap mass spectrometry by means of fragmentation of the parental ions. Two degradation pathways were proposed on the basis of the structure of the degradation products. Among the six degradation products, two were ozonolysis products of AFB 1 . The appearance of the other four degradation products indicates that AFB 1 was degraded by other reactive species besides ozone that were generated during HVACP treatment. Reactive oxygen gas species are suggested as the major agents for aflatoxin degradation during HVACP treatment. Two degradation pathways of AFB 1 by HVACP treatment were proposed. One pathway involves reactions in which H • , OH • , CHO • radicals are added. The other involves epoxidation by HO 2 • radicals and oxidation of AFB 1 by the combined effects of the oxidative species OH • , H 2 O 2 , and O 3 . According to the structure-bioactivity relationship of AFB 1 , the bioactivity of the AFB 1 samples subjected to HVACP treatment is significantly reduced because of the disappearance of the C8═C9 double bond in the furofuran ring in all of the major degradation products as well as the modification of the lactone ring, cyclopentanone, and the methoxyl group.

  4. Prenatal exposure to aflatoxin B1: developmental, behavioral, and reproductive alterations in male rats

    NASA Astrophysics Data System (ADS)

    Supriya, Ch.; Reddy, P. Sreenivasula

    2015-06-01

    Previous studies have shown that aflatoxin B1 (AfB1) inhibits androgen biosynthesis as a result of its ability to form a high-affinity complex with the steroidogenic acute regulatory protein. The results of the present study demonstrate the postnatal effects of in utero exposure to AfB1 in the rat. Pregnant Wistar rats were given 10, 20, or 50 μg AfB1/kg body weight daily from gestation day (GD) 12 to GD 19. At parturition, newborns were observed for clinical signs and survival. All animals were born alive and initially appeared to be active. Male pups from control and AfB1-exposed animals were weaned and maintained up to postnatal day (PD) 100. Litter size, birth weight, sex ratio, survival rate, and crown-rump length of the pups were significantly decreased in AfB1-exposed rats when compared to controls. Elapsed time (days) for testes to descend into the scrotal sac was significantly delayed in experimental pups when compared to control pups. Behavioral observations such as cliff avoidance, negative geotaxis, surface rightening activity, ascending wire mesh, open field behavior, and exploratory and locomotory activities were significantly impaired in experimental pups. Body weights and the indices of testis, cauda epididymis, prostate, seminal vesicles, and liver were significantly reduced on PD 100 in male rats exposed to AfB1 during embryonic development when compared with controls. Significant reduction in the testicular daily sperm production, epididymal sperm count, and number of viable, motile, and hypo-osmotic tail coiled sperm was observed in experimental rats. The levels of serum testosterone and activity levels of testicular hydroxysteroid dehydrogenases were significantly decreased in a dose-dependent manner with a significant increase in the serum follicle-stimulating hormone and luteinizing hormone in experimental rats. Deterioration in the testicular and cauda epididymal architecture was observed in experimental rats. The results of fertility

  5. Mycotoxin Contamination in Sugarcane Grass and Juice: First Report on Detection of Multiple Mycotoxins and Exposure Assessment for Aflatoxins B1 and G1 in Humans

    PubMed Central

    Abdallah, Mohamed F.; Krska, Rudolf; Sulyok, Michael

    2016-01-01

    This study was conducted to investigate the natural co-occurrence of multiple toxic fungal and bacterial metabolites in sugarcane grass and juice intended for human consumption in Upper Egypt. Quantification of the target analytes has been done using the “dilute and shoot” approach followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total number of 29 and 33 different metabolites were detected in 21 sugarcane grass and 40 juice samples, respectively, with a trend of concentrations being higher in grass than in juice. Among the regulated mycotoxins, only aflatoxin B1 (AFB1) and aflatoxin G1 (AFG1) were detected. The prevalence of AFB1 was in 48% of grass samples and in 58% of juice with a maximum concentration of 30.6 μg/kg and 2.10 μg/kg, respectively. AFG1 was detected in 10% of grass samples (7.76 μg/kg) and 18% of juice samples (34 μg/kg). Dietary exposure was assessed using a juice frequency questionnaire of adult inhabitants in Assiut City. The assessment revealed different levels of exposure to AFB1 between males and females in winter and summer seasons. The estimated seasonal exposure ranged from 0.20 to 0.40 ng/kg b.w./day in winter and from 0.38 to 0.90 ng/kg b.w./day in summer. PMID:27869706

  6. The efficacy of bamboo charcoal in comparison with smectite to reduce the detrimental effect of aflatoxin B1 on in vitro rumen fermentation of a hay-rich feed mixture.

    PubMed

    Jiang, Ya-Hui; Wang, Ping; Yang, Hong-Jian; Chen, Ying

    2014-07-10

    Two commercial materials, a bamboo charcoal (BC) and a smectite clay (SC), were assessed in vitro with aflatoxin B1 (AFB1) in an equilibrium adsorption test. The adsorption capacity and proportion adsorbed (0.381 μg/mg, 0.955) for BC were greater than for SC (0.372 μg/mg, 0.931). The effects of in vitro ruminal fermentation of hay-rich feed incubated with 1.0 μg/mL AFB1 for 0-10 g/L doses of BC and SC were measured at 39 °C for 72 h. The BC and SC binders increased AFB1 loss at dosages ≥1.0 g/L (p < 0.0001). Average AFB1 loss (p < 0.0001) was greater for SC (0.904) than BC (0.881). Both SC and SC addition increased in vitro dry matter loss, and the average dry matter losses were similar. Asymptotic gas volume and volatile fatty acid production were greater for BC than for SC (p < 0.0001). Thus, BC may be as effective as SC in removing aflatoxin B1's detrimental effects on rumen degradability and fermentation under the occurrence of microbial aflatoxin degradation.

  7. The Efficacy of Bamboo Charcoal in Comparison with Smectite to Reduce the Detrimental Effect of Aflatoxin B1 on In Vitro Rumen Fermentation of a Hay-Rich Feed Mixture

    PubMed Central

    Jiang, Ya-Hui; Wang, Ping; Yang, Hong-Jian; Chen, Ying

    2014-01-01

    Two commercial materials, a bamboo charcoal (BC) and a smectite clay (SC), were assessed in vitro with aflatoxin B1 (AFB1) in an equilibrium adsorption test. The adsorption capacity and proportion adsorbed (0.381 μg/mg, 0.955) for BC were greater than for SC (0.372 μg/mg, 0.931). The effects of in vitro ruminal fermentation of hay-rich feed incubated with 1.0 μg/mL AFB1 for 0–10 g/L doses of BC and SC were measured at 39 °C for 72 h. The BC and SC binders increased AFB1 loss at dosages ≥1.0 g/L (p < 0.0001). Average AFB1 loss (p < 0.0001) was greater for SC (0.904) than BC (0.881). Both SC and SC addition increased in vitro dry matter loss, and the average dry matter losses were similar. Asymptotic gas volume and volatile fatty acid production were greater for BC than for SC (p < 0.0001). Thus, BC may be as effective as SC in removing aflatoxin B1’s detrimental effects on rumen degradability and fermentation under the occurrence of microbial aflatoxin degradation. PMID:25014194

  8. The Individual and Combined Effects of Deoxynivalenol and Aflatoxin B1 on Primary Hepatocytes of Cyprinus Carpio

    PubMed Central

    He, Cheng-Hua; Fan, Yan-Hong; Wang, Ying; Huang, Chao-Ying; Wang, Xi-Chun; Zhang, Hai-Bin

    2010-01-01

    Aflatoxin B1 (AFB1) and deoxynivalenol (DON) are important food-borne mycotoxins that have been implicated in animal and human health. In this study, individual and combinative effects of AFB1 and DON were tested in primary hepatocytes of Cyprinus carpio. The results indicated that the combinative effects of AFB1 and DON (0.01 μg/mL AFB1 and 0.25 μg/mL DON; 0.02 μg/mL AFB1 and 0.25 μg/mL DON; 0.02 μg/mL AFB1 and 0.5 μg/mL DON) were higher than that of individual mycotoxin (P < 0.05). The activity of AST, ALT and LDH in cell supernatant was higher than that of control group (P < 0.05) when the mycotoxins were exposed to primary hepatocytes for 4 h. The decreased cell number was observed in tested group by inverted light microscopy. The mitochondrial swelling, endoplasmic reticulum dilation and a lot of lipid droplets were observed in primary hepatocytes by transmission electron microscope. Therefore, this combination was classified as an additive response of the two mycotoxins. PMID:21152299

  9. Glycine N-methyltransferase affects the metabolism of aflatoxin B{sub 1} and blocks its carcinogenic effect

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yen, C.-H.; AIDS Prevention and Research Center, National Yang-Ming University, Taipei, Taiwan; Hung, J.-H.

    Previously, we reported that glycine N-methyltransferase (GNMT) knockout mice develop chronic hepatitis and hepatocellular carcinoma (HCC) spontaneously. For this study we used a phosphoenolpyruvate carboxykinase promoter to establish a GNMT transgenic (TG) mouse model. Animals were intraperitoneally inoculated with aflatoxin B{sub 1} (AFB{sub 1}) and monitored for 11 months, during which neither male nor female GNMT-TG mice developed HCC. In contrast, 4 of 6 (67%) male wild-type mice developed HCC. Immunofluorescent antibody test showed that GNMT was translocated into nuclei after AFB{sub 1} treatment. Competitive enzyme immunoassays indicated that after AFB{sub 1} treatment, the AFB{sub 1}-DNA adducts formed in stablemore » clones expressing GNMT reduced 51.4% compared to the vector control clones. Experiments using recombinant adenoviruses carrying GNMT cDNA (Ad-GNMT) further demonstrated that the GNMT-related inhibition of AFB{sub 1}-DNA adducts formation is dose-dependent. HPLC analysis of the metabolites of AFB{sub 1} in the cultural supernatants of cells exposed to AFB{sub 1} showed that the AFM{sub 1} level in the GNMT group was significantly higher than the control group, indicating the presence of GNMT can enhance the detoxification pathway of AFB{sub 1}. Cytotoxicity assay showed that the GNMT group had higher survival rate than the control group after they were treated with AFB{sub 1}. Automated docking experiments showed that AFB{sub 1} binds to the S-adenosylmethionine binding domain of GNMT. Affinity sensor assay demonstrated that the dissociation constant for GNMT-AFB{sub 1} interaction is 44.9 {mu}M. Therefore, GNMT is a tumor suppressor for HCC and it exerts protective effects in hepatocytes via direct interaction with AFB{sub 1}, resulting in reduced AFB{sub 1}-DNA adducts formation and cell death.« less

  10. Performance improvement of the one-dot lateral flow immunoassay for aflatoxin B1 by using a smartphone-based reading system.

    PubMed

    Lee, Sangdae; Kim, Giyoung; Moon, Jihea

    2013-04-18

    This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA) detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1) was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisition and data analysis. The detection limit of one-dot LFIA for AFB1 is 5 μg/kg. This method provided semi-quantitative analysis of AFB1 samples in the range of 5 to 1,000 μg/kg. Using combination of the one-dot LFIA and the Smartphone-based reading system, it is possible to conduct a more fast and accurate point-of-care diagnosis.

  11. A novel reduced graphene oxide/molybdenum disulfide/polyaniline nanocomposite-based electrochemical aptasensor for detection of aflatoxin B1.

    PubMed

    Geleta, Girma Selale; Zhao, Zhen; Wang, Zhenxin

    2018-03-26

    In this study, we developed a novel reduced graphene oxide/molybdenum disulfide/polyaniline@gold nanoparticles-based electrochemical aptasensor (termed as RGO/MoS2/PANI@AuNPs/Apt) for detection of aflatoxin B1 (AFB1). The RGO/MoS2/PANI nanocomposites were synthesized and characterized by multiple techniques, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), infrared spectroscopy (FTIR), UV-visible spectroscopy, and X-ray photoelectron spectroscopy (XPS). A glassy carbon electrode (GCE) was then modified by the RGO/MoS2/PANI nanocomposites, coated with a chitosan (Cs) film, and followed by AuNPs attachment for immobilizing the AFB1 aptamers. In the presence of AFB1, the AFB1 binding-induced conformation change of the immobilized aptamer on the electrode surface results in the reduction of the electron transfer from a [Fe(CN)6]3-/4- redox couple in the solution to the GCE surface. Therefore, the aptamer-AFB1 binding event can be easily monitored by the peak current change of the RGO/MoS2/PANI@AuNPs/Apt through differential pulse voltammetry (DPV) measurement. Under the optimized conditions, the as-developed RGO/MoS2/PANI@AuNPs/Apt exhibits a wide linear range from 0.01 fg mL-1 to 1.0 fg mL-1 and a remarkably low detection limit (3σ) of 0.002 fg mL-1. The aptasensor also has good reproducibility as well as shows high selectivity against other fungal toxins, such as OTA and FB1. Moreover, the practicability of the RGO/MoS2/PANI@AuNPs/Apt was demonstrated by the analysis of AFB1 in the spiked wine samples.

  12. Optical waveguide lightmode spectroscopy technique-based immunosensor development for aflatoxin B1 determination in spice paprika samples.

    PubMed

    Majer-Baranyi, Krisztina; Zalán, Zsolt; Mörtl, Mária; Juracsek, Judit; Szendrő, István; Székács, András; Adányi, Nóra

    2016-11-15

    Optical waveguide lightmode spectroscopy (OWLS) technique has been applied to label-free detection of aflatoxin B1 in a competitive immunoassay format, with the aim to compare the analytical goodness of the developed OWLS immunosenor with HPLC and enzyme-linked immunosorbent assay (ELISA) methods for the detection of aflatoxin in spice paprika matrix. We have also assessed applicability of the QuEChERS method prior to ELISA measurements, and the results were compared to those obtained by traditional solvent extraction followed by immunoaffinity clean-up. The AFB1 content of sixty commercial spice paprika samples from different countries were measured with the developed and optimized OWLS immunosensor. Comparing the results from the indirect immunosensor to that obtained by HPLC or ELISA provided excellent correlation (with regression coefficients above 0.94) indicating that the competitive OWLS immunosensor has a potential for quick determination of aflatoxin B1 in paprika samples. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. A SERS-active sensor based on heterogeneous gold nanostar core-silver nanoparticle satellite assemblies for ultrasensitive detection of aflatoxinB1

    NASA Astrophysics Data System (ADS)

    Li, Aike; Tang, Lijuan; Song, Dan; Song, Shanshan; Ma, Wei; Xu, Liguang; Kuang, Hua; Wu, Xiaoling; Liu, Liqiang; Chen, Xin; Xu, Chuanlai

    2016-01-01

    A surface-enhanced Raman scattering (SERS) sensor based on gold nanostar (Au NS) core-silver nanoparticle (Ag NP) satellites was fabricated for the first time to detect aflatoxinB1 (AFB1). We constructed the SERS sensor using AFB1 aptamer (DNA1)-modified Ag satellites and a complementary sequence (DNA2)-modified Au NS core. The Raman label (ATP) was modified on the surface of Ag satellites. The SERS signal was enhanced when the satellite NP was attached to the Au core NS. The AFB1 aptamer on the surface of Ag satellites would bind to the targets when AFB1 was present in the system, Ag satellites were then removed and the SERS signal decreased. This SERS sensor showed superior specificity for AFB1 and the linear detection range was from 1 to 1000 pg mL-1 with the limit of detection (LOD) of 0.48 pg mL-1. The excellent recovery experiment using peanut milk demonstrated that the sensor could be applied in food and environmental detection.A surface-enhanced Raman scattering (SERS) sensor based on gold nanostar (Au NS) core-silver nanoparticle (Ag NP) satellites was fabricated for the first time to detect aflatoxinB1 (AFB1). We constructed the SERS sensor using AFB1 aptamer (DNA1)-modified Ag satellites and a complementary sequence (DNA2)-modified Au NS core. The Raman label (ATP) was modified on the surface of Ag satellites. The SERS signal was enhanced when the satellite NP was attached to the Au core NS. The AFB1 aptamer on the surface of Ag satellites would bind to the targets when AFB1 was present in the system, Ag satellites were then removed and the SERS signal decreased. This SERS sensor showed superior specificity for AFB1 and the linear detection range was from 1 to 1000 pg mL-1 with the limit of detection (LOD) of 0.48 pg mL-1. The excellent recovery experiment using peanut milk demonstrated that the sensor could be applied in food and environmental detection. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr08372a

  14. Mechanisms of butylated hydroxytoluene chemoprevention of aflatoxicosis-inhibition of aflatoxin B{sub 1} metabolism

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Guarisco, John A.; Hall, Jeffery O.; Coulombe, Roger A.

    Chemoprevention of toxicoses and/or cancer through the use of nutrients or pharmacologic compounds is the subject of intense study. Among the many compounds examined, food additives such as antioxidants are being considered due to their ability to reduce disease formation by either induction or inhibition of key enzyme systems. One such compound, butylated hydroxytoluene (BHT), has been found to protect against cancer formation caused by exposure to aflatoxin B{sub 1} (AFB{sub 1}) in rodents. We have shown that dietary BHT protects against clinical signs of aflatoxicosis in turkeys, a species that is very susceptible to this mycotoxin. In this study,more » the effect of BHT on AFB{sub 1} metabolism and other cytochrome P450 (CYP)-related enzyme activities in turkey liver microsomes was examined to discern possible mechanisms of BHT-mediated protection against aflatoxicosis. Ethoxyresorufin O-deethylase (EROD), methoxyresorufin O-demethylase (MROD), prototype activities for CYP1A1 and 1A2, respectively, were decreased in the BHT fed (4000 ppm) animals, while oxidation of nifedipine, a prototype activity for CYP3A4, was increased. However, BHT added to microsomal incubations inhibited these CYP activities in a concentration-related manner. Importantly, BHT inhibited conversion of AFB{sub 1} to the reactive intermediate AFB{sub 1}-8,-9-epoxide (AFBO), exhibiting Michaelis-Menton competitive inhibition kinetics (Ki = 0.81 {mu}M). Likewise, microsomes prepared from turkeys fed BHT were significantly less active in AFBO formation compared to those from control birds. When turkeys were fed BHT for up to 40 days, residual BHT was present in liver, breast meat, thigh meat and abdominal fat in concentrations substantially below U.S. FDA guidelines for this antioxidant, but in concentrations greater than the Ki, likely sufficient to inhibit bioactivation of AFB{sub 1}in vivo. BHT-induced hydropic degeneration in the livers of BHT fed animals was significantly greater in

  15. Characterization of Lactic Acid Bacteria as Poultry Probiotic Candidates with Aflatoxin B1 Binding Activities

    NASA Astrophysics Data System (ADS)

    Damayanti, E.; Istiqomah, L.; Saragih, J. E.; Purwoko, T.; Sardjono

    2017-12-01

    Our previous studies have selected lactic acid bacteria (LAB) with antifungal activities from traditional fermented foods made from cassava (G7) and silage feed palm leaf (PDS5 and PDS3). In this study we evaluated their ability to bind aflatoxin B1 (AFB1) and probiotic characteristic. The probiotic characteristic assays of LAB consisted of resistance to acidic conditions (pH 3), gastric juice and bile salts 0.3%. We also carried out an in vitro evaluation of LAB aflatoxin binding ability in viable and non-viable cell for 24 and 48 hours of incubation. The measurement of aflatoxin content was performed by ELISA method using AgraQuant Total Aflatoxin Assay kit. The results showed that all isolates were potential as probiotics and the G7 isolate had the highest viability among other isolates in pH 3 (92.61 %) and the bile salts assay (97.71 %). The percentage of aflatoxin reduction between viable and non-viable cell from each LAB isolate were different. The highest aflatoxin reduction in viable cell assay was performed by G7 isolate (69.11 %) whereas in non-viable cell assay was performed by PDS3 isolate (73.75 %) during incubation time 48 hours. In this study, G7 isolate performed the best probiotic characteristics with the highest viability in acid pH assay, bile salt 0.3% assay and percentage of aflatoxin B1 reduction in viable cell condition. Molecular identification using 16S rRNA sequence analysis showed that G7 isolate had homology with Lactobacillus plantarum (99.9%). It was concluded that Lactobacillus plantarum G7 was potential as probiotic with aflatoxin binding activities.

  16. An attempt to model the probability of growth and aflatoxin B1 production of Aspergillus flavus under non-isothermal conditions in pistachio nuts.

    PubMed

    Aldars-García, Laila; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2015-10-01

    Human exposure to aflatoxins in foods is of great concern. The aim of this work was to use predictive mycology as a strategy to mitigate the aflatoxin burden in pistachio nuts postharvest. The probability of growth and aflatoxin B1 (AFB1) production of aflatoxigenic Aspergillus flavus, isolated from pistachio nuts, under static and non-isothermal conditions was studied. Four theoretical temperature scenarios, including temperature levels observed in pistachio nuts during shipping and storage, were used. Two types of inoculum were included: a cocktail of 25 A. flavus isolates and a single isolate inoculum. Initial water activity was adjusted to 0.87. Logistic models, with temperature and time as explanatory variables, were fitted to the probability of growth and AFB1 production under a constant temperature. Subsequently, they were used to predict probabilities under non-isothermal scenarios, with levels of concordance from 90 to 100% in most of the cases. Furthermore, the presence of AFB1 in pistachio nuts could be correctly predicted in 70-81 % of the cases from a growth model developed in pistachio nuts, and in 67-81% of the cases from an AFB1 model developed in pistachio agar. The information obtained in the present work could be used by producers and processors to predict the time for AFB1 production by A. flavus on pistachio nuts during transport and storage. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Performance Improvement of the One-Dot Lateral Flow Immunoassay for Aflatoxin B1 by Using a Smartphone-Based Reading System

    PubMed Central

    Lee, Sangdae; Kim, Giyoung; Moon, Jihea

    2013-01-01

    This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA) detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1) was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisition and data analysis. The detection limit of one-dot LFIA for AFB1 is 5 μg/kg. This method provided semi-quantitative analysis of AFB1 samples in the range of 5 to 1,000 μg/kg. Using combination of the one-dot LFIA and the Smartphone-based reading system, it is possible to conduct a more fast and accurate point-of-care diagnosis. PMID:23598499

  18. Aflatoxin B1 in betel nuts (Areca catechu L.) imported to Pakistan from different regions of South Asia.

    PubMed

    Asghar, Muhammad Asif; Iqbal, Javed; Ahmed, Aftab; Khan, Mobeen Ahmed; Shamsuddin, Zuzzer Ali

    2014-01-01

    Aflatoxin B1 (AFB1) levels were evaluated in betel nuts (Areca catechu L.) being imported to Pakistan during 2010-2011. In total, 278 betel nut samples (India = 21, Indonesia = 51, Sri-Lanka = 34 and Thailand = 172) were received from the Department of Customs and were analysed by thin layer chromatography (TLC). All Indian origin betel nuts showed AFB1 contamination ranging from 11.7-262.0 µg kg(-1) with a mean of 92.5 µg kg(-1). Among Indonesian and Sri Lankan shipments, 80.4% and 73.5% betel nuts were contaminated with AFB1 ranging between 3.3-39.2 and 6.5-103.4 µg kg(-1) with a mean of 11.6 and 35.0 µg kg(-1), respectively. However, only 30.2% of Thailand origin samples showed AFB1 contamination ranging 3.3-77.0 µg kg(-1) with a mean of 6.6 µg kg(-1). The widespread occurrence of AFB1 increases the hazard associated with betel nuts. Thus, strict control is a pre-requisite for the production and import/export of psychoactive substances as betel nuts.

  19. Crosstalk-eliminated quantitative determination of aflatoxin B1-induced hepatocellular cancer stem cells based on concurrent monitoring of CD133, CD44, and aldehyde dehydrogenase1.

    PubMed

    Ju, Hee; Shim, Yumi; Arumugam, Parthasarathy; Song, Joon Myong

    2016-01-22

    Cancer stem cells (CSCs), known as tumor initiating cells, have become a critically important issue for cancer therapy. Although much research has demonstrated the induction of hepato cellular carcinoma by aflatoxin B1, the formation of hepatocellular CSCs and their quantitative determination is hardly reported. In this work, it was found that hepatocellular CSCs were produced from HepG2 cells by aflatoxin B1-induced mutation, and their amount was quantitatively determined using crosstalk-eliminated multicolor cellular imaging based on quantum dot (Qdot) nanoprobes and an acousto-optical tunable filter (AOTF). Hepatocellular CSCs were acquired via magnetic bead-based sorting and observed using concurrent detection of three different markers: CD133, CD44, and aldehyde dehydrogenase1 (ALDH1). The DNA mutation of HepG2 cells caused by aflatoxin B1 was quantitatively observed via absorbance spectra of aflatoxin B1-8, 9-epoxide-DNA adducts. The percentages of hepatocellular CSCs formed in the entire HepG2 cells were determined to be 9.77±0.65%, 10.9±1.39%, 11.4±1.32%, and 12.8±0.7%, respectively, at 0 μM, 5 μM, 10 μM, and 20 μM of aflatoxin B1. The results matched well with those obtained utilizing flow cytometry. This study demonstrates that aflatoxin mediated mutation induced the conversion of hepatic cancer cell to hepatic CSCs by using a Qdot based constructed multicolor cellular imaging system. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  20. Application of SWIR hyperspectral imaging and chemometrics for identification of aflatoxin B1 contaminated maize kernels

    NASA Astrophysics Data System (ADS)

    Kimuli, Daniel; Wang, Wei; Wang, Wei; Jiang, Hongzhe; Zhao, Xin; Chu, Xuan

    2018-03-01

    A short-wave infrared (SWIR) hyperspectral imaging system (1000-2500 nm) combined with chemometric data analysis was used to detect aflatoxin B1 (AFB1) on surfaces of 600 kernels of four yellow maize varieties from different States of the USA (Georgia, Illinois, Indiana and Nebraska). For each variety, four AFB1 solutions (10, 20, 100 and 500 ppb) were artificially deposited on kernels and a control group was generated from kernels treated with methanol solution. Principal component analysis (PCA), partial least squares discriminant analysis (PLSDA) and factorial discriminant analysis (FDA) were applied to explore and classify maize kernels according to AFB1 contamination. PCA results revealed partial separation of control kernels from AFB1 contaminated kernels for each variety while no pattern of separation was observed among pooled samples. A combination of standard normal variate and first derivative pre-treatments produced the best PLSDA classification model with accuracy of 100% and 96% in calibration and validation, respectively, from Illinois variety. The best AFB1 classification results came from FDA on raw spectra with accuracy of 100% in calibration and validation for Illinois and Nebraska varieties. However, for both PLSDA and FDA models, poor AFB1 classification results were obtained for pooled samples relative to individual varieties. SWIR spectra combined with chemometrics and spectra pre-treatments showed the possibility of detecting maize kernels of different varieties coated with AFB1. The study further suggests that increase of maize kernel constituents like water, protein, starch and lipid in a pooled sample may have influence on detection accuracy of AFB1 contamination.

  1. Effect of UV irradiation on aflatoxin reduction: a cytotoxicity evaluation study using human hepatoma cell line.

    PubMed

    Patras, Ankit; Julakanti, Sharath; Yannam, Sudheer; Bansode, Rishipal R; Burns, Mallory; Vergne, Matthew J

    2017-11-01

    In this proof-of-concept study, the efficacy of a medium-pressure UV (MPUV) lamp source to reduce the concentrations of aflatoxin B 1 , aflatoxin B 2 , and aflatoxin G 1 (AFB 1, AFB 2 , and AFG 1 ) in pure water is investigated. Irradiation experiments were conducted using a collimated beam system operating between 200 to 360 nm. The optical absorbance of the solution and the irradiance of the lamp are considered in calculating the average fluence rate. Based on these factors, the UV dose was quantified as a product of average fluence rate and treatment time. Known concentrations of aflatoxins were spiked in water and irradiated at UV doses ranging from 0, 1.22, 2.44, 3.66, and 4.88 J cm -2 . The concentration of aflatoxins was determined by HPLC with fluorescence detection. LC-MS/MS product ion scans were used to identify and semi-quantify degraded products of AFB 1 , AFB 2 , and AFG 1 . It was observed that UV irradiation significantly reduced aflatoxins in pure water (p < 0.05). Irradiation doses of 4.88 J cm -2 reduced concentrations 67.22% for AFG 1 , 29.77% for AFB 2 , and 98.25% for AFB 1 (p < 0.05). Using this technique, an overall reduction of total aflatoxin content of ≈95% (p < 0.05) was achieved. We hypothesize that the formation of ˙OH radicals initiated by UV light may have caused photolysis of AFB 1 , AFB 2 , and AFG 1 molecules. In cell culture studies, our results demonstrated that the increase of UV dosage decreased the aflatoxin-induced cytotoxicity in HepG 2 cells. Therefore, our research finding suggests that UV irradiation can be used as an effective technique for the reduction of aflatoxins.

  2. Feasibility of detecting aflatoxin B1 on inoculated maize kernels surface using Vis/NIR hyperspectral imaging.

    PubMed

    Wang, Wei; Heitschmidt, Gerald W; Windham, William R; Feldner, Peggy; Ni, Xinzhi; Chu, Xuan

    2015-01-01

    The feasibility of using a visible/near-infrared hyperspectral imaging system with a wavelength range between 400 and 1000 nm to detect and differentiate different levels of aflatoxin B1 (AFB1 ) artificially titrated on maize kernel surface was examined. To reduce the color effects of maize kernels, image analysis was limited to a subset of original spectra (600 to 1000 nm). Residual staining from the AFB1 on the kernels surface was selected as regions of interest for analysis. Principal components analysis (PCA) was applied to reduce the dimensionality of hyperspectral image data, and then a stepwise factorial discriminant analysis (FDA) was performed on latent PCA variables. The results indicated that discriminant factors F2 can be used to separate control samples from all of the other groups of kernels with AFB1 inoculated, whereas the discriminant factors F1 can be used to identify maize kernels with levels of AFB1 as low as 10 ppb. An overall classification accuracy of 98% was achieved. Finally, the peaks of β coefficients of the discrimination factors F1 and F2 were analyzed and several key wavelengths identified for differentiating maize kernels with and without AFB1 , as well as those with differing levels of AFB1 inoculation. Results indicated that Vis/NIR hyperspectral imaging technology combined with the PCA-FDA was a practical method to detect and differentiate different levels of AFB1 artificially inoculated on the maize kernels surface. However, indicated the potential to detect and differentiate naturally occurring toxins in maize kernel. © 2014 Institute of Food Technologists®

  3. An enzyme-free catalytic DNA circuit for amplified detection of aflatoxin B1 using gold nanoparticles as colorimetric indicators

    NASA Astrophysics Data System (ADS)

    Chen, Junhua; Wen, Junlin; Zhuang, Li; Zhou, Shungui

    2016-05-01

    An enzyme-free biosensor for the amplified detection of aflatoxin B1 has been constructed based on a catalytic DNA circuit. Three biotinylated hairpin DNA probes (H1, H2, and H3) were designed as the assembly components to construct the sensing system (triplex H1-H2-H3 product). Cascaded signal amplification capability was obtained through toehold-mediated strand displacement reactions to open the hairpins and recycle the trigger DNA. By the use of streptavidin-functionalized gold nanoparticles as the signal indicators, the colorimetric readout can be observed by the naked eye. In the presence of a target, the individual nanoparticles (red) aggregate into a cross-linked network of nanoparticles (blue) via biotin-streptavidin coupling. The colorimetric assay is ultrasensitive, enabling the visual detection of trace levels of aflatoxin B1 (AFB1) as low as 10 pM without instrumentation. The calculated limit of detection (LOD) is 2 pM in terms of 3 times standard deviation over the blank response. The sensor is robust and works even when challenged with complex sample matrices such as rice samples. Our sensing platform is simple and convenient in operation, requiring only the mixing of several solutions at room temperature to achieve visible and intuitive results, and holds great promise for the point-of-use monitoring of AFB1 in environmental and food samples.An enzyme-free biosensor for the amplified detection of aflatoxin B1 has been constructed based on a catalytic DNA circuit. Three biotinylated hairpin DNA probes (H1, H2, and H3) were designed as the assembly components to construct the sensing system (triplex H1-H2-H3 product). Cascaded signal amplification capability was obtained through toehold-mediated strand displacement reactions to open the hairpins and recycle the trigger DNA. By the use of streptavidin-functionalized gold nanoparticles as the signal indicators, the colorimetric readout can be observed by the naked eye. In the presence of a target, the

  4. Aspergillus flavus aflatoxin occurrence and expression of aflatoxin biosynthesis genes in soil.

    PubMed

    Accinelli, Cesare; Abbas, H K; Zablotowicz, R M; Wilkinson, J R

    2008-05-01

    The carcinogen aflatoxin B1 (AFB1) produced by Aspergillus flavus is a major food safety concern in crops. However, information on AFB1 occurrence in soil and crop residue is scarce. A series of experiments investigated the occurrence of AFB1 in soil and corn residues and ascertained the ecology of A. flavus in a Dundee silt loam soil. Samples of untilled soil (0-2 cm) and residues were collected in March 2007 from plots previously planted with a corn isoline containing the Bacillus thuringiensis (Bt) endotoxin gene or the parental non-Bt isoline. AFB1 levels were significantly different in various corn residues. The highest AFB1 levels were observed in cobs containing grain, with 145 and 275 ng.g-1 in Bt and non-Bt, respectively (P > or = F = 0.001). Aflatoxin levels averaged 3.3 and 9.6 ng.g-1 in leaves and (or) stalks and cobs without grain, respectively. All soils had AFB1 ranging from 0.6 to 5.5 ng.g-1 with similar levels in plots from Bt and non-Bt corn. Based on cultural methods, soil contained from log10 3.1 to 4.5 A. flavus cfu.g-1 with about 60% of isolates producing aflatoxin. Laboratory experiments demonstrated that AFB1 is rapidly degraded in soil at 28 degrees C (half-life < or = 5 days). The potential of the soil A. flavus to produce aflatoxins was confirmed by molecular methods. Transcription of 5 aflatoxin biosynthesis genes, including aflD, aflG, aflP, aflR, and aflS, were detected by reverse transcription - polymerase chain reaction analysis in soil. Although AFB1 appears to be transient in soils, it is clear that AFB1 is produced in surface soil in the presence of corn residues, as indicated by A. flavus cfu levels, AFB1 detection, and expression of aflatoxin biosynthetic genes.

  5. Aflatoxin B1 Detoxification by Aspergillus oryzae from Meju, a Traditional Korean Fermented Soybean Starter.

    PubMed

    Lee, Kyu Ri; Yang, Sun Min; Cho, Sung Min; Kim, Myunghee; Hong, Sung-Yong; Chung, Soo Hyun

    2016-11-04

    Aflatoxins are classified as Group 1 (carcinogenic to humans) by the International Agency for Research on Cancer (IARC). In this study, a total of 134 fungal strains were isolated from 65 meju samples, and two fungal isolates were selected as potential aflatoxin B₁ (AFB₁)-biodetoxification fungi. These fungi were identified as Aspergillus oryzae MAO103 and A. oryzae MAO104 by sequencing the beta-tubulin gene. The two A. oryzae strains were able to degrade more than 90% of AFB1 (initial concentration: 40 µg/L) in a culture broth in 14 days. The mutagenic effects of AFB₁ treated with A. oryzae MAO103 and MAO104 significantly decreased to 5.7% and 6.4%, respectively, in the frame-shift mutation of Ames tests using Salmonella typhimurium TA 98. The base-substituting mutagenicity of AFB₁ was also decreased by the two fungi. Moreover, AFB1 production by A. flavus was significantly decreased by the two A. oryzae strains on soybean-based agar plates. Our data suggest that the two AFB1-detoxification A. oryzae strains have potential application to control AFB₁ in foods and feeds.

  6. Probability models for growth and aflatoxin B1 production as affected by intraspecies variability in Aspergillus flavus.

    PubMed

    Aldars-García, Laila; Berman, María; Ortiz, Jordi; Ramos, Antonio J; Marín, Sonia

    2018-06-01

    The probability of growth and aflatoxin B 1 (AFB 1 ) production of 20 isolates of Aspergillus flavus were studied using a full factorial design with eight water activity levels (0.84-0.98 a w ) and six temperature levels (15-40 °C). Binary data obtained from growth studies were modelled using linear logistic regression analysis as a function of temperature, water activity and time for each isolate. In parallel, AFB 1 was extracted at different times from newly formed colonies (up to 20 mm in diameter). Although a total of 950 AFB 1 values over time for all conditions studied were recorded, they were not considered to be enough to build probability models over time, and therefore, only models at 30 days were built. The confidence intervals of the regression coefficients of the probability of growth models showed some differences among the 20 growth models. Further, to assess the growth/no growth and AFB 1 /no- AFB 1 production boundaries, 0.05 and 0.5 probabilities were plotted at 30 days for all of the isolates. The boundaries for growth and AFB 1 showed that, in general, the conditions for growth were wider than those for AFB 1 production. The probability of growth and AFB 1 production seemed to be less variable among isolates than AFB 1 accumulation. Apart from the AFB 1 production probability models, using growth probability models for AFB 1 probability predictions could be, although conservative, a suitable alternative. Predictive mycology should include a number of isolates to generate data to build predictive models and take into account the genetic diversity of the species and thus make predictions as similar as possible to real fungal food contamination. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Effect of feed contamination with aflatoxin B1 and administration of exogenous corticosterone on Japanese quail biochemical and immunological parameters.

    PubMed

    Nazar, F N; Magnoli, A P; Dalcero, A M; Marin, R H

    2012-01-01

    Stress is the loss of homeostasis by external forces or stressors. Manipulation, transport, contamination, and other procedures involved in production could be considered stressors. Contamination is a problem commonly faced by producers in the poultry industry. Aflatoxicosis is one of the most common infections resulting from feed contaminated with Aspergillus flavus and Aspergillus parasiticus. This study evaluated the potential effects of the combined administration of aflatoxin B(1) (AFB(1)) and corticosterone on biochemical (concentration of globulins, proteins, and albumin) and immunological (inflammatory response and heterophil:lymphocyte ratio) parameters of Japanese quail. Potential sex effects on those parameters were also considered. The provision of corticosterone in drinking water is a method used for mimicking the effects of chronic stress in avian species. At 35 d of age, 24 mixed-sex groups of 4 animals (2 males and 2 females) were housed in cages and assigned to 1 of 4 treatments: plain drinking water and laying diet, corticosterone administration in drinking water, feed contamination with AFB(1) (100 μg/kg of feed), or corticosterone plus AFB(1) administration. There were 6 cages per treatment. No significant effect of sex in any of the parameters analyzed was detected. Hypoproteinemia, hypoalbuminemia, and hypoglobulinemia were observed in animals treated with corticosterone or contaminated feed. These responses were exacerbated when the factors were combined. The immunodepressive effect of corticosterone administration was confirmed, and a higher effect was noticed when combined with the aflatoxin contamination. Aflatoxin contamination affected birds' physiology similar to a chronic stressor stimulation because it elevates the heterophil:lymphocyte ratio. This study suggests that the effects of the AFB(1) contamination are further increased when overlapped with a chronic stressful stimulation and emphasizes the importance of controlling

  8. Analysis of fumonisin B1 removal by microorganisms in co-occurrence with aflatoxin B1 and the nature of the binding process.

    PubMed

    Pizzolitto, Romina P; Salvano, Mario A; Dalcero, Ana M

    2012-06-01

    The objectives of this investigation were to evaluate the ability of Saccharomyces cerevisiae CECT 1891 and Lactobacillus acidophilus 24 to remove fumonisin B(1) (FB(1)) from liquid medium; to determine the nature of the mechanism involved in FB(1)-microorganism interaction and to analyze whether the presence of aflatoxin B(1) (AFB(1)) interferes with the removal of FB(1) and vice versa. The results obtained indicated that: (i) both microorganisms were able to remove FB(1) from liquid medium; (ii) the removal was a fast and reversible process; (iii) cell viability was not necessary; (iv) the amount of FB(1) removed was both toxin- and microorganism concentration-dependent; (v) the process did not involve chemical modification of FB(1) molecules; and (vi) cell wall structural integrity of the microorganisms was required for FB(1) removal. Consequently, we propose that the mechanism involved in the removal of FB(1) is a physical adsorption (physisorption) of the toxin molecule to cell wall components of the microorganisms. It is highly probable that FB(1) and AFB(1) co-occur in contaminated foods, since the fungal genera Aspergillus and Fusarium frequently occur simultaneously. Therefore, we analyzed whether the presence of AFB(1) interferes with the removal of FB(1) by the microorganisms previously evaluated, and vice versa. Studies of co-occurrence of both mycotoxins clearly showed that they did not compete for binding sites on the microorganism cell wall and the presence of one toxin did not modify the efficiency of the organism in the removal of the other mycotoxin. These findings may be useful for optimization of mycotoxin binding and provide an important contribution to research on microorganisms with ability to remove these secondary metabolites. Copyright © 2012. Published by Elsevier B.V.

  9. Association between aflatoxin B1 occupational airway exposure and risk of hepatocellular carcinoma: a case-control study.

    PubMed

    Lai, Hao; Mo, Xianwei; Yang, Yang; He, Ke; Xiao, Jun; Liu, Chao; Chen, Jiansi; Lin, Yuan

    2014-10-01

    The aim of this study was to determine the airway exposure of sugar and papermaking factory workers to aflatoxin B1 (AFB1) and to explore the potential association between AFB1 airway exposure and the risk of hepatocellular carcinoma (HCC) in a case-control study. Dust samples were collected from the sugarcane bagasse warehouse, and presser and paper production workshops. Blood samples were collected from 181 workshop employees and 203 controls who worked outside the workshop. AFB1 albumin adducts were detected using a double antibody sandwich enzyme-linked immunosorbent assay (ELISA). To explore the association between AFB1 airway exposure and the risk of HCC, the medical records of 68 HCC patients who worked in a sugar and papermaking factory between January 1994 and December 2013 were analyzed. A questionnaire was used to collect information from 150 healthy controls who worked for the same company and lived near the factory. AFB1 was detected in the dust samples, but could not be detected in any of the rice samples. An analysis of serum samples revealed serum AFB1 albumin adducts in 102 (56.35 %) of the study participants. However, in the control group, only 12 (5.9 %) individuals had detectable levels of AFB1 albumin adducts. Those with airway exposure to Aspergillus flavus-contaminated dust had an elevated risk of HCC compared to those without exposure (odds ratio, 5.24; 95 % confidence interval, 2.77-9.88; P = 0.00). The findings of this study indicate that occupational AFB1 airway exposure might be associated with the risk of AFB1-related HCC among the population that was used in this study. Intervention programs aimed at reducing exposure to inhalational AFB1 are needed urgently. Additional suitably designed, multicenter, prospective studies using large samples are needed to further confirm the results.

  10. Understanding the sorption mechanisms of aflatoxin B1 to kaolinite, illite, and smectite clays via a comparative computational study.

    PubMed

    Kang, Fuxing; Ge, Yangyang; Hu, Xiaojie; Goikavi, Caspar; Waigi, Michael Gatheru; Gao, Yanzheng; Ling, Wanting

    2016-12-15

    In current adsorption studies of biotoxins to phyllosilicate clays, multiply weak bonding types regarding these adsorptions are not well known; the major attractive forces, especially for kaolinite and illite, are difficult to be identified as compared to smectite with exchangeable cations. Here, we discriminated the bonding types of aflatoxin B1 (AFB1) contaminant to these clays by combined batch experiment with model computation, expounded their bonding mechanisms which have been not quantitatively described by researchers. The observed adsorbent-to-solution distribution coefficients (K d ) of AFB1 presented in increasing order of 18.5-37.1, 141.6-158.3, and 354.6-484.7L/kg for kaolinite, illite, and smectite, respectively. Normalization of adsorbent-specific surface areas showed that adsorption affinity of AFB1 is mainly dependent on the outside surfaces of clay aggregates. The model computation and test of ionic effect further suggested that weakly electrostatic attractions ((Si/Al-OH) 2 ⋯(OC) 2 ) are responsible for AFB1-kaolinite adsorption (K d , 18.5-37.1L/kg); a moderate electron-donor-acceptor attraction ((CO) 2 ⋯K + ⋯(O-Al) 3 ) is related to AFB1-illite adsorption (K d , 141.6-158.3L/kg); a strong calcium-bridging linkage ((CO) 2 ⋯Ca 2+ ⋯(O-Si) 4 ) is involved in AFB1-smectite adsorption (K d , 354.6-484.7L/kg). Changes in Gibbs free energy (ΔG°) suggested that the computed result is reliable, providing a good reproduction of AFB1-clay interaction. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. RNA-Seq profiling reveals novel hepatic gene expression pattern in aflatoxin B1 treated rats.

    PubMed

    Merrick, B Alex; Phadke, Dhiral P; Auerbach, Scott S; Mav, Deepak; Stiegelmeyer, Suzy M; Shah, Ruchir R; Tice, Raymond R

    2013-01-01

    Deep sequencing was used to investigate the subchronic effects of 1 ppm aflatoxin B1 (AFB1), a potent hepatocarcinogen, on the male rat liver transcriptome prior to onset of histopathological lesions or tumors. We hypothesized RNA-Seq would reveal more differentially expressed genes (DEG) than microarray analysis, including low copy and novel transcripts related to AFB1's carcinogenic activity compared to feed controls (CTRL). Paired-end reads were mapped to the rat genome (Rn4) with TopHat and further analyzed by DESeq and Cufflinks-Cuffdiff pipelines to identify differentially expressed transcripts, new exons and unannotated transcripts. PCA and cluster analysis of DEGs showed clear separation between AFB1 and CTRL treatments and concordance among group replicates. qPCR of eight high and medium DEGs and three low DEGs showed good comparability among RNA-Seq and microarray transcripts. DESeq analysis identified 1,026 differentially expressed transcripts at greater than two-fold change (p<0.005) compared to 626 transcripts by microarray due to base pair resolution of transcripts by RNA-Seq, probe placement within transcripts or an absence of probes to detect novel transcripts, splice variants and exons. Pathway analysis among DEGs revealed signaling of Ahr, Nrf2, GSH, xenobiotic, cell cycle, extracellular matrix, and cell differentiation networks consistent with pathways leading to AFB1 carcinogenesis, including almost 200 upregulated transcripts controlled by E2f1-related pathways related to kinetochore structure, mitotic spindle assembly and tissue remodeling. We report 49 novel, differentially-expressed transcripts including confirmation by PCR-cloning of two unique, unannotated, hepatic AFB1-responsive transcripts (HAfT's) on chromosomes 1.q55 and 15.q11, overexpressed by 10 to 25-fold. Several potentially novel exons were found and exon refinements were made including AFB1 exon-specific induction of homologous family members, Ugt1a6 and Ugt1a7c. We find the

  12. Homogeneous electrochemical immunoassay of aflatoxin B1 in foodstuff using proximity-hybridization-induced omega-like DNA junctions and exonuclease III-triggered isothermal cycling signal amplification.

    PubMed

    Tang, Juan; Huang, Yapei; Liu, Huiqiong; Zhang, Cengceng; Tang, Dianping

    2016-12-01

    A new homogeneous electrochemical immunosensing platform was designed for sensitive detection of aflatoxin B 1 (AFB 1 ) in foodstuff. The system consisted of anti-AFB 1 antibody labeled DNA 1 (Ab-DNA 1 ), AFB 1 -bovine serum albumin (BSA)-conjugated DNA 2 (AFB 1 -DNA 2 ), and methylene blue functionalized hairpin DNA. Owing to a specific antigen-antibody reaction between anti-AFB 1 and AFB 1 -BSA, the immunocomplex formed assisted the proximity hybridization of DNA 1 with DNA 2 , thus resulting in the formation of an omega-like DNA junction. Thereafter, the junction opened the hairpin DNA to construct a new double-stranded DNA, which could be readily cleaved by exonuclease III to release the omega-like DNA junction and methylene blue. The dissociated DNA junction could repeatedly hybridize with residual hairpin DNA molecules with exonuclease III-based isothermal cycling amplification, thereby releasing numerous free methylene blue molecules into the detection solution. The as-produced free methylene blue molecules could be captured by a negatively charged indium tin oxide electrode, each of which could produce an electronic signal within the applied potentials. On introduction of target AFB 1 , the analyte competed with AFB 1 -DNA 2 for the conjugated anti-AFB 1 on the Ab-DNA 1 , subsequently decreasing the amount of omega-like DNA junctions formed, hence causing methylene blue labeled hairpin DNA to move far away from the electrode surface. Under optimal conditions the detectable electrochemical signal decreased with increasing amount of target AFB 1 in a dynamic working range of 0.01-30 ng mL -1 with a detection limit of 4.8 pg mL -1 . In addition, the precision and reproducibility of this system were acceptable. Finally, the method was further evaluated for analysis of naturally contaminated or AFB 1 -spiked peanut samples, giving results that matched well with those obtained with a commercial AFB 1 ELISA kit.

  13. Biomolecule-functionalized magnetic nanoparticles for flow-through quartz crystal microbalance immunoassay of aflatoxin B1.

    PubMed

    Wang, Li; Gan, Xian-Xue

    2009-01-01

    A flow-through quartz crystal microbalance (QCM) immunoassay method has been developed based on aflatoxin B(1) antibody (anti-AFB(1))-functionalized magnetic core-shell Fe(3)O(4)/SiO(2) composite nanoparticles (bionanoparticles) in this study. To construct such an assay protocol, anti-AFB(1), as a model protein, was initially covalently immobilized onto the Fe(3)O(4)/SiO(2) surface, and then the functionalized nanoparticles were attached to the surface of the QCM probe with an external magnet. The binding of target molecules onto the immobilized antibodies decreased the sensor's resonant frequency, and the frequency shift was proportional to the AFB(1) concentration in the range of 0.3-7.0 ng/ml. The regeneration of the developed immunosensor was carried out via attaching or detaching the external magnet from the detection cell. In addition, the selectivity, reproducibility, and stability of the proposed immunoassay system were acceptable. Compared with the conventional ELISAs, the proposed immunoassay system was simple and rapid without multiple labeling and separation steps. Importantly, the proposed immunoassay method could be further developed for the immobilization of other antigens or biocompounds.

  14. The effect of NovaSil dietary supplementation on the growth and health performance of Nile tilapia (Oreochromis niloticus) fed aflatoxin-B1 contaminated feed

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to evaluate the ability of NovaSil (NS) clay to sorb and mitigate the toxic effects of aflatoxin B1 (AFB1) in Nile tilapia (Oreochromis niloticus). Growth performance, specific innate immunological function, intestinal microbial community, and histology were evaluate...

  15. Hetero-enzyme-based two-round signal amplification strategy for trace detection of aflatoxin B1 using an electrochemical aptasensor.

    PubMed

    Zheng, Wanli; Teng, Jun; Cheng, Lin; Ye, Yingwang; Pan, Daodong; Wu, Jingjing; Xue, Feng; Liu, Guodong; Chen, Wei

    2016-06-15

    An electrochemical aptasensor for trace detection of aflatoxin B1 (AFB1) was developed by using an aptamer as the recognition unit while adopting the telomerase and EXO III based two-round signal amplification strategy as the signal enhancement units. The telomerase amplification was used to elongate the ssDNA probes on the surface of gold nanoparticles, by which the signal response range of the signal-off model electrochemical aptasensor could be correspondingly enlarged. Then, the EXO III amplification was used to hydrolyze the 3'-end of the dsDNA after the recognition of target AFB1, which caused the release of bounded AFB1 into the sensing system, where it participated in the next recognition-sensing cycle. With this two-round signal amplified electrochemical aptasensor, target AFB1 was successfully measured at trace concentrations with excellent detection limit of 0.6*10(-4)ppt and satisfied specificity due to the excellent affinity of the aptamer against AFB1. Based on this designed two-round signal amplification strategy, both the sensing range and detection limit were greatly improved. This proposed ultrasensitive electrochemical aptasensor method was also validated by comparison with the classic instrumental methods. Importantly, this hetero-enzyme based two-round signal amplified electrochemical aptasensor offers a great promising protocol for ultrasensitive detection of AFB1 and other mycotoxins by replacing the core recognition sequence of the aptamer. Copyright © 2016 Elsevier B.V. All rights reserved.

  16. Monoclonal IgA Antibodies for Aflatoxin Immunoassays

    PubMed Central

    Ertekin, Özlem; Pirinçci, Şerife Şeyda; Öztürk, Selma

    2016-01-01

    Antibody based techniques are widely used for the detection of aflatoxins which are potent toxins with a high rate of occurrence in many crops. We developed a murine monoclonal antibody of immunoglobulin A (IgA) isotype with a strong binding affinity to aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), aflatoxin G2 (AFG2) and aflatoxin M1 (AFM1). The antibody was effectively used in immunoaffinity column (IAC) and ELISA kit development. The performance of the IACs was compatible with AOAC performance standards for affinity columns (Test Method: AOAC 991.31). The total binding capacity of the IACs containing our antibody was 111 ng, 70 ng, 114 ng and 73 ng for AFB1, AFB2, and AFG1 andAFG2, respectively. Furthermore, the recovery rates of 5 ng of each AF derivative loaded to the IACs were determined as 104.9%, 82.4%, 85.5% and 70.7% for AFB1, AFB2, AFG1 and AFG2, respectively. As for the ELISA kit developed using non-oriented, purified IgA antibody, we observed a detection range of 2–50 µg/L with 40 min total test time. The monoclonal antibody developed in this research is hitherto the first presentation of quadruple antigen binding IgA monoclonal antibodies in mycotoxin analysis and also the first study of their utilization in ELISA and IACs. IgA antibodies are valuable alternatives for immunoassay development, in terms of both sensitivity and ease of preparation, since they do not require any orientation effort. PMID:27187470

  17. Low cost quantitative digital imaging as an alternative to qualitative in vivo bioassays for analysis of active aflatoxin B1.

    PubMed

    Rasooly, Reuven; Do, Paula M; Hernlem, Bradley J

    2016-06-15

    Aflatoxin B1 (AFB1) producing fungi contaminate food and feed and are a major health concern. To minimize the sources and incidence of AFB1 illness there is a need to develop affordable, sensitive mobile devices for detection of active AFB1. In the present study we used a low cost fluorescence detector and describe two quantitative assays for detection of detoxified and active AFB1 demonstrating that AFB1 concentration can be measured as intensity of fluorescence. When the assay plate containing increasing concentrations of AFB1 is illuminated with a 366 nm ultraviolet lamp, AFB1 molecules absorb photons and emit blue light with peak wavelength of 432 nm. The fluorescence intensity increased in dose dependent manner. However, this method cannot distinguish between active AFB1 which poses a threat to health, and the detoxified AFB1 which exhibits no toxicity. To measure the toxin activity, we used a cell based assay that makes quantification more robust and is capable of detecting multiple samples simultaneously. It is an alternative to the qualitative duckling bioassay which is the "gold-standard" assay currently being used for quantitative analysis of active AFB1. AFB1 was incubated with transduced Vero cells expressing the green fluorescence protein (GFP) gene. After excitation with blue light at 475 nm, cells emitted green light with emission peak at 509 nm. The result shows that AFB1 inhibits protein expression in a concentration dependent manner resulting in proportionately less GFP fluorescence in cells exposed to AFB1. The result also indicates strong positive linear relationship with R(2)=0.90 between the low cost CCD camera and a fluorometer, which costs 100 times more than a CCD camera. This new analytical method for measuring active AFB1 is low in cost and combined with in vitro assay, is quantitative. It also does not require the use of animals and may be useful especially for laboratories in regions with limited resources. Published by Elsevier B.V.

  18. Identification of O-methylsterigmatocystin as an aflatoxin B1 and G1 precursor in Aspergillus parasiticus.

    PubMed Central

    Bhatnagar, D; McCormick, S P; Lee, L S; Hill, R A

    1987-01-01

    An isolate of Aspergillus parasiticus CP461 (SRRC 2043) produced no detectable aflatoxins, but accumulated O-methylsterigmatocystin (OMST). When sterigmatocystin (ST) was fed to this isolate in a low-sugar medium, there was an increase in the accumulation of OMST, without aflatoxin synthesis. When radiolabeled [14C]OMST was fed to resting mycelia of a non-aflatoxin-, non-ST-, and non-OMST-producing mutant of A. parasiticus AVN-1 (SRRC 163), 14C-labeled aflatoxins B1 and G1 were produced; 10 nmol of OMST produced 7.8 nmol of B1 and 1.0 nmol of G1, while 10 nmol of ST produced 6.4 nmol of B1 and 0.6 nmol of G1. A time course study of aflatoxin synthesis in ST feeding experiments with AVN-1 revealed that OMST is synthesized by the mold during the onset of aflatoxin synthesis. The total amount of aflatoxins recovered from OMST feeding experiments was higher than from experiments in which ST was fed to the resting mycelia. These results suggest that OMST is a true metabolite in the aflatoxin biosynthetic pathway between sterigmatocystin and aflatoxins B1 and G1 and is not a shunt metabolite, as thought previously. PMID:3111363

  19. Inhibitory Effect of Cinnamaldehyde, Citral, and Eugenol on Aflatoxin Biosynthetic Gene Expression and Aflatoxin B1 Biosynthesis in Aspergillus flavus.

    PubMed

    Liang, Dandan; Xing, Fuguo; Selvaraj, Jonathan Nimal; Liu, Xiao; Wang, Limin; Hua, Huijuan; Zhou, Lu; Zhao, Yueju; Wang, Yan; Liu, Yang

    2015-12-01

    In order to reveal the inhibitory effects of cinnamaldehyde, citral, and eugenol on aflatoxin biosynthesis, the expression levels of 5 key aflatoxin biosynthetic genes were evaluated by real-time PCR. Aspergillus flavus growth and AFB1 production were completely inhibited by 0.80 mmol/L of cinnamaldehyde and 2.80 mmol/L of citral. However, at lower concentration, cinnamaldehyde (0.40 mmol/L), eugenol (0.80 mmol/L), and citral (0.56 mmol/L) significantly reduced AFB1 production with inhibition rate of 68.9%, 95.4%, and 41.8%, respectively, while no effect on fungal growth. Real-time PCR showed that the expressions of aflR, aflT, aflD, aflM, and aflP were down-regulated by cinnamaldehyde (0.40 mmol/L), eugenol (0.80 mmol/L), and citral (0.56 mmol/L). In the presence of cinnamaldehyde, AflM was highly down-regulated (average of 5963 folds), followed by aflP, aflR, aflD, and aflT with the average folds of 55, 18, 6.5, and 5.8, respectively. With 0.80 mmol/L of eugenol, aflP was highly down-regulated (average of 2061-folds), followed by aflM, aflR, aflD, and aflT with average of 138-, 15-, 5.2-, and 4.8-folds reduction, respectively. With 0.56 mmol/L of citral, aflT was completely inhibited, followed by aflM, aflP, aflR, and aflD with average of 257-, 29-, 3.5-, and 2.5-folds reduction, respectively. These results suggest that the reduction in AFB1 production by cinnamaldehyde, eugenol, and citral at low concentration may be due to the down-regulations of the transcription level of aflatoxin biosynthetic genes. Cinnamaldehyde and eugenol may be employed successfully as a good candidate in controlling of toxigenic fungi and subsequently contamination with aflatoxins in practice. © 2015 Institute of Food Technologists®

  20. Bypass of Aflatoxin B[subscript 1] Adducts by the Sulfolobus solfataricus DNA Polymerase IV

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Banerjee, Surajit; Brown, Kyle L.; Egli, Martin

    Aflatoxin B{sub 1} (AFB{sub 1}) is oxidized to an epoxide in vivo, which forms an N7-dG DNA adduct (AFB{sub 1}-N7-dG). The AFB{sub 1}-N7-dG can rearrange to a formamidopyrimidine (AFB{sub 1}-FAPY) derivative. Both AFB{sub 1}-N7-dG and the {beta}-anomer of the AFB{sub 1}-FAPY adduct yield G {yields} T transversions in Escherichia coli, but the latter is more mutagenic. We show that the Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) bypasses AFB{sub 1}-N7-dG in an error-free manner but conducts error-prone replication past the AFB{sub 1}-FAPY adduct, including misinsertion of dATP, consistent with the G {yields} T mutations observed in E. coli. Three ternarymore » (Dpo4-DNA-dNTP) structures with AFB{sub 1}-N7-dG adducted template:primers have been solved. These demonstrate insertion of dCTP opposite the AFB{sub 1}-N7-dG adduct, and correct vs incorrect insertion of dATP vs dTTP opposite the 5'-template neighbor dT from a primed AFB{sub 1}-N7-dG:dC pair. The insertion of dTTP reveals hydrogen bonding between the template N3 imino proton and the O{sup 2} oxygen of dTTP, and between the template T O{sup 4} oxygen and the N3 imino proton of dTTP, perhaps explaining why this polymerase does not efficiently catalyze phosphodiester bond formation from this mispair. The AFB{sub 1}-N7-dG maintains the 5'-intercalation of the AFB{sub 1} moiety observed in DNA. The bond between N7-dG and C8 of the AFB{sub 1} moiety remains in plane with the alkylated guanine, creating a 16{sup o} inclination of the AFB{sub 1} moiety with respect to the guanine. A binary (Dpo4-DNA) structure with an AFB{sub 1}-FAPY adducted template:primer also maintains 5'-intercalation of the AFB{sub 1} moiety. The {beta}-deoxyribose anomer is observed. Rotation about the FAPY C5-N{sup 5} bond orients the bond between N{sup 5} and C8 of the AFB{sub 1} moiety out of plane in the 5'-direction, with respect to the FAPY base. The formamide group extends in the 3'-direction. This improves stacking of the AFB

  1. A Comparative Study on Aflatoxin B1 Metabolism in Mice and Rats

    PubMed Central

    Steyn, M.; Pitout, M. J.; Purchase, I. F. H.

    1971-01-01

    In vivo metabolic studies on rats and mice revealed a marked difference in the fluorescent compounds produced after ingestion of aflatoxin B1. The mouse converted aflatoxin B1 to three unknown fluorescent compounds, designated x1, x2 and x3 and the known aflatoxin M1, while the rat was only capable of producing aflatoxin M1. The results suggested that metabolites x1, x2, x3 and aflatoxin M1 were not part of a major metabolic pathway, but produced independently. These unknown yellowish-green fluorescent compounds did not seem to be conjugated with sulphate or glucuronic acid. In vitro incubations of various mouse liver cell fractions with aflatoxin B1 showed that metabolites x1, x2, x3 and aflatoxin M1, could only be produced by the microsomal fraction and that NADPH was needed as a co-factor. The differences in aflatoxin metabolism by mice and rats are discussed in relation to the apparent resistance of the mouse to the carcinogenic effects of this toxin. PMID:4398926

  2. RNA-Seq Profiling Reveals Novel Hepatic Gene Expression Pattern in Aflatoxin B1 Treated Rats

    PubMed Central

    Merrick, B. Alex; Phadke, Dhiral P.; Auerbach, Scott S.; Mav, Deepak; Stiegelmeyer, Suzy M.; Shah, Ruchir R.; Tice, Raymond R.

    2013-01-01

    Deep sequencing was used to investigate the subchronic effects of 1 ppm aflatoxin B1 (AFB1), a potent hepatocarcinogen, on the male rat liver transcriptome prior to onset of histopathological lesions or tumors. We hypothesized RNA-Seq would reveal more differentially expressed genes (DEG) than microarray analysis, including low copy and novel transcripts related to AFB1’s carcinogenic activity compared to feed controls (CTRL). Paired-end reads were mapped to the rat genome (Rn4) with TopHat and further analyzed by DESeq and Cufflinks-Cuffdiff pipelines to identify differentially expressed transcripts, new exons and unannotated transcripts. PCA and cluster analysis of DEGs showed clear separation between AFB1 and CTRL treatments and concordance among group replicates. qPCR of eight high and medium DEGs and three low DEGs showed good comparability among RNA-Seq and microarray transcripts. DESeq analysis identified 1,026 differentially expressed transcripts at greater than two-fold change (p<0.005) compared to 626 transcripts by microarray due to base pair resolution of transcripts by RNA-Seq, probe placement within transcripts or an absence of probes to detect novel transcripts, splice variants and exons. Pathway analysis among DEGs revealed signaling of Ahr, Nrf2, GSH, xenobiotic, cell cycle, extracellular matrix, and cell differentiation networks consistent with pathways leading to AFB1 carcinogenesis, including almost 200 upregulated transcripts controlled by E2f1-related pathways related to kinetochore structure, mitotic spindle assembly and tissue remodeling. We report 49 novel, differentially-expressed transcripts including confirmation by PCR-cloning of two unique, unannotated, hepatic AFB1-responsive transcripts (HAfT’s) on chromosomes 1.q55 and 15.q11, overexpressed by 10 to 25-fold. Several potentially novel exons were found and exon refinements were made including AFB1 exon-specific induction of homologous family members, Ugt1a6 and Ugt1a7c. We find

  3. Determination of aflatoxins B1, B2, G1 and G2 in spices using a multifunctional column clean-up.

    PubMed

    Akiyama, H; Goda, Y; Tanaka, T; Toyoda, M

    2001-10-12

    A rapid and simple method using a multifunctional column, which contains lipophilic and charged active sites, was developed to analyse aflatoxins B1, B2, G1 and G2 in various spices, such as red pepper and nutmeg. After extraction by acetonitrile:water (9:1) and clean-up using MultiSep #228 column, the aflatoxins and aflatoxin-TFA derivatives are determined using LC with fluorescence detection. Recoveries of each aflatoxin B1, B2, G1 and G2 spiked to red pepper, white pepper, black pepper, nutmeg and tear grass at the level of 10 ng/g were over 80-85% in all instances. The minimum detectable concentration for aflatoxins in red pepper was 0.5 ng/g.

  4. Detection of serum AFB1-lysine adduct in Malaysia and its association with liver and kidney functions.

    PubMed

    Mohd Redzwan, S; Rosita, Jamaluddin; Mohd Sokhini, A M; Nurul 'Aqilah, A R; Wang, Jia-Sheng; Kang, Min-Su; Zuraini, Ahmad

    2014-01-01

    Aflatoxin is ubiquitously found in many foodstuffs and produced by Aspergillus species of fungi. Of many aflatoxin metabolites, AFB1 is classified by the International Agency for Research on Cancer (IARC) as group one carcinogen and linked to the development of hepatocellular carcinoma (HCC). The study on molecular biomarker of aflatoxin provides a better assessment on the extent of human exposure to aflatoxin. In Malaysia, the occurrences of aflatoxin-contaminated foods have been documented, but there is a lack of data on human exposure to aflatoxin. Hence, this study investigated the occurrence of AFB1-lysine adduct in serum samples and its association with liver and kidney functions. 5ml fasting blood samples were collected from seventy-one subjects (n=71) for the measurement of AFB1-lysine adduct, albumin, total bilirubin, AST (aspartate aminotransferase), ALT (alanine transaminase), ALP (alkaline phosphatase), GGT (gamma-glutamyl transpeptidase), creatinine and BUN (blood urea nitrogen). The AFB1-lysine adduct was detected in all serum samples (100% detection rate) with a mean of 6.85±3.20pg/mg albumin (range: 1.13-18.85pg/mg albumin). Male subjects (mean: 8.03±3.41pg/mg albumin) had significantly higher adduct levels than female subjects (mean: 5.64±2.46pg/mg albumin) (p<0.01). It was noteworthy that subjects with adduct levels greater than average (>6.85pg/mg albumin) had significantly elevated level of total bilirubin (p<0.01), GGT (p<0.05) and creatinine (p<0.01). Nevertheless, only the level of total bilirubin, (r=0.347, p-value=0.003) and creatinine (r=0.318, p-value=0.007) showed significant and positive correlation with the level of AFB1-lysine adduct. This study provides a valuable insight on human exposure to aflatoxin in Malaysia. Given that aflatoxin can pose serious problem to the health, intervention strategies should be implemented to limit/reduce human exposure to aflatoxin. Besides, a study with a big sample size should be warranted in

  5. Human health implications from co-exposure to aflatoxins and fumonisins in maize-based foods in Latin America: Guatemala as a case study

    USDA-ARS?s Scientific Manuscript database

    Co-occurence of fumonisin B1 (FB1) and aflatoxin B1 (AFB1) in maize has been demonstrated in many surveys. Combined-exposure to FB1 and AFB1 was of concern to the Joint FAO/WHO Expert Committee on Food Additives because of the known genotoxicity of AFB1 and the ability of FB1 to induce regenerative...

  6. Intercalation of aflatoxin B sub 1 in two oligodeoxynucleotide adducts: Comparative sup 1 H NMR analysis of d(ATC sup AFB GAT)ter dot d(ATCGAT) and d(AT sup ATB GCAT) sub 2

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gopalakrishnan, S.; Harris, T.M.; Stone, M.P.

    8,9-Dihydro-8-(N7-guanyl-(d(ATCGAT)))-9-hydroxyaflatoxin B{sub 1}{center dot}d(ATCGAT) and 8,9-dihydro-8-(N7-guanyl-(d(ATGCAT)))-9-hydroxyafltoxin B{sub 1}{center dot}8,9-dihydro-8-(N7-guanyl-(d(ATGCAT)))-9-hydroxyaflatoxin B{sub 1} were prepared by direct addition of aflatoxin B{sub 1} 8,9-expoxide to d(ATCGAT){sub 2} and d(ATGCAT){sub 2}, respectively. {sup 1}H NOE experiments, nonselective {sup 1}H T{sub 1} relaxation measurements, and {sup 1}H chemical shift perturbations demonstrate that in both modified oligodeoxynucleotides the aflatoxin moiety is intercalated above the 5{prime}-face of the modified guanine. The oligodeoxynucleotides remain right-handed, and perturbation of the B-DNA structure is localized adjacent to the adducted guanine. Aflatoxin-oligodeoxynucleotide {sup 1}H NOEs are observed between aflatoxin and the 5{prime}-neighbor base pair and include both the major groove andmore » the minor groove. The protons at C8 and C9 of the aflatoxin terminal furan ring exhibit slower spin-lattice relaxation as compared to other oligodeoxynucleotide protons, which supports the conclusion that they face into the major groove. Increased shielding is observed for aflatoxin protons. The difference in reaction stoichiometry is consistent with an intercalated transition-state complex between aflatoxin B{sub 1} 8,9-epoxide and B-DNA. Intercalation provides excellent positioning for nucleophilic attack by guanine N7 on aflatoxin B{sub 1} 8,9-epoxide, which probably accounts for the observed efficiency of adduct formation despite the relatively low DNA binding affinity observed for aflatoxin B{sub 1}.« less

  7. Assessing airborne aflatoxin B1 during on-farm grain handling activities.

    PubMed

    Selim, M I; Juchems, A M; Popendorf, W

    1998-04-01

    The presence of aflatoxin in corn and corn dust during relatively normal years and the increased risk of Aspergillus flavus infestation during drought conditions suggest that airborne agricultural exposures should be of considerable concern. Liquid extraction, thin layer chromatography, and high pressure liquid chromatography were used for the analysis of aflatoxin B1 in grain dust and bulk corn samples. A total of 24 samples of airborne dust were collected from 8 farms during harvest, 22 samples from 9 farms during animal feeding, and 14 sets of Andersen samples from 11 farms during bin cleaning. A total of 14 samples of settled dust and 18 samples of bulk corn were also collected and analyzed. The airborne concentration of aflatoxin B1 found in dust collected during harvest and grain unloading ranged from 0.04 to 92 ng/m3. Higher levels of aflatoxin B1 were found in the airborne dust samples collected from enclosed animal feeding buildings (5-421 ng/m3) and during bin cleaning (124-4849 ng/m3). Aflatoxin B1 up to 5100 ng/g were detected in settled dust collected from an enclosed animal feeding building; however, no apparent correlation was found between the airborne concentration of aflatoxin B1 and its concentration in settled dust or bulk corn. The data demonstrate that farmers and farm workers may be exposed to potentially hazardous concentrations of aflatoxin B1, particularly during bin cleaning and animal feeding in enclosed buildings.

  8. A large Raman scattering cross-section molecular embedded SERS aptasensor for ultrasensitive Aflatoxin B1 detection using CS-Fe3O4 for signal enrichment

    NASA Astrophysics Data System (ADS)

    Chen, Quansheng; Yang, Mingxiu; Yang, Xiaojing; Li, Huanhuan; Guo, Zhiming; Rahma, M. H.

    2018-01-01

    With growing concern on oil safety problems, developing a simple and sensitive method to detect Aflatoxin B1 (AFB1), a common mycotoxin in peanut oil, is very necessary. In this study, Surface-enhanced Raman Scattering (SERS) aptasensor was developed for ultrasensitive AFB1 detection using the amino-terminal AFB1 aptamer (NH2-DNA1); and thiol-terminal AFB1 complementary aptamer (SH-DNA2) conjugated magnetic-beads (CS-Fe3O4) as enrichment nanoprobe and AuNR@DNTB@Ag nanorods (ADANRs) as reporter nanoprobe respectively. 5,5‧-Dithiobis(2-nitrobenzoicacid) (DNTB) with large Raman scattering cross-section and no fluorescence interference was embedded in Au and Ag core/shell nanorods as Raman reporter molecules. CS-Fe3O4 possessed excellent biocompatibility and superparamagnetism for rapid signal enrichment. Therefore, NH2-DNA1-CS-Fe3O4 and SH-DNA2-ADANRs were fabricated via the hybrid reaction between aptamers and complementary aptamers. When there is AFB1, AFB1 would competitively combine with the NH2-DNA1-CS-Fe3O4 inducing the dissociation of SH-DNA2-ADANRs from CS-Fe3O4 and further decreasing the SERS signal. Based on this developed SERS aptasensor, a low limit of 0.0036 ng/mL and an effective linear detection range from 0.01 to 100 ng/mL with the correlation coefficient up to 0.986 for AFB1 detection were obtained. Moreover, the specificity of this SERS aptasensor was demonstrated by detecting other two mycotoxins and its accuracy for AFB1 detection in real peanut oil was further confirmed by standard addition recovery test.

  9. Aflatoxin B1-contaminated diet disrupts the blood-brain barrier and affects fish behavior: Involvement of neurotransmitters in brain synaptosomes.

    PubMed

    Baldissera, Matheus D; Souza, Carine F; Zeppenfeld, Carla Cristina; Descovi, Sharine N; Moreira, Karen Luise S; da Rocha, Maria Izabel U M; da Veiga, Marcelo L; da Silva, Aleksandro S; Baldisserotto, Bernardo

    2018-06-01

    It is known that the cytotoxic effects of aflatoxin B 1 (AFB 1 ) in endothelial cells of the blood-brain barrier (BBB) are associated with behavioral dysfunction. However, the effects of a diet contaminated with AFB 1 on the behavior of silver catfish remain unknown. Thus, the aim of this study was to evaluate whether an AFB 1 -contaminated diet (1177 ppb kg feed -1 ) impaired silver catfish behavior, as well as whether disruption of the BBB and alteration of neurotransmitters in brain synaptosomes are involved. Fish fed a diet contaminated with AFB 1 presented a behavioral impairment linked with hyperlocomotion on days 14 and 21 compared with the control group (basal diet). Neurotransmitter levels were also affected on days 14 and 21. The permeability of the BBB to Evans blue dye increased in the intoxicated animals compared with the control group, which suggests that the BBB was disrupted. Moreover, acetylcholinesterase (AChE) activity in brain synaptosomes was increased in fish fed a diet contaminated with AFB 1 , while activity of the sodium-potassium pump (Na + , K + -ATPase) was decreased. Based on this evidence, the present study shows that silver catfish fed a diet containing AFB 1 exhibit behavioral impairments related to hyperlocomotion. This diet caused a disruption of the BBB and brain lesions, which may contribute to the behavioral changes. Also, the alterations in the activities of AChE and Na + , K + -ATPase in brain synaptosomes may directly contribute to this behavior, since they may promote synapse dysfunction. In addition, the hyperlocomotion may be considered an important macroscopic marker indicating possible AFB 1 intoxication. Copyright © 2018 Elsevier B.V. All rights reserved.

  10. Organophilic treatments of bentonite increase the adsorption of aflatoxin B1 and protect stem cells against cellular damage.

    PubMed

    Nones, Janaína; Nones, Jader; Poli, Anicleto; Trentin, Andrea Gonçalves; Riella, Humberto Gracher; Kuhnen, Nivaldo Cabral

    2016-09-01

    Bentonite clays exhibit high adsorptive capacity for contaminants, including aflatoxin B1 (AFB1), a mycotoxin responsible for causing severe toxicity in several species including pigs, poultry and man. Organophilic treatments is known to increase the adsorption capacity of bentonites, and the primary aim of this study was to evaluate the ability of Brazilian bentonite and two organic salts - benzalkonium chloride (BAC) and cetyltrimethylammonium bromide (CTAB) to adsorb AFB1. For this end, 2(2) factorial designs were used in order to analyze if BAC or CTAB was able to increase AFB1 adsorption when submitted in different temperature and concentration. Both BAC and CTAB treatment (at 30°C and 2% of salt concentration) were found to increase the adsorption of AFB1 significantly compared with untreated bentonite. After organophilic bentonite treatments with BAC or CTAB, a vibration of CH stretch (2850 and 2920cm(-1)) were detected. A frequency of the SiO stretch (1020 and 1090cm(-1)) was changed by intercalation of organic cation. Furthermore, the interlayer spacing of bentonite increases to 1.23nm (d001 reflection at 2θ=7.16) and 1.22 (d001 reflection at 2θ=7.22) after the addition of BAC and CTAB, respectively. Another aim of the study was to observe the effects of these two bentonite salts in neural crest stem cell cultures. The two materials that were created by organophilic treatments were not found to be toxic to stem cells. Furthermore the results indicate that the two materials tested may protect the neural crest stem cells against damage caused by AFB1. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Rat liver endothelial and Kupffer cell-mediated mutagenicity and polycyclic aromatic hydrocarbons and aflatoxin B sub 1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Steinberg, P.; Schlemper, B.; Molitor, E.

    The ability of isolated rat liver endothelial and Kupffer cells to activate benzo(a)pyrene (BP), trans-7,8-dihydroxy-7,8-dihydrobenzo(a)pyrene (DDBP), trans-1,2-dihydroxy-1,2-dihydrochrysene (DDCH), and aflatoxin B{sub 1} (AFB{sub 1}) to mutagenic metabolites was assessed by means of a cell-mediated bacterial mutagenicity assay and compared with the ability of parenchymal cells to activate these compounds. Endothelial and Kupffer cells from untreated rats were able to activate AFB{sub 1} and DDBP; DDBP was activated even in the absence of an NADPH-generating system. Pretreating the animals with Aroclor 1254 strongly enhanced the mutagenicity of the dihydrodiol, whereas the mutagenicity of AFB{sub 1} showed a slight increase. BP andmore » DDCH were only activated by endothelial and Kupffer cells isolated from Aroclor 1254-pretreated rats. Parenchymal cells form untreated animals activated all four carcinogens tested; Aroclor 1254 enhanced the parenchymal cell-mediated mutagenicity of BP and DDCH but did not affect that of DDBP and clearly reduced that of AFB{sub 1}. The reduced mutagenicity of AFB{sub 1} correlates with the decrease in the amount of 2{alpha}-hydroxytestosterone formed when testosterone was incubated with parenchymal cell microsomes from Aroclor 1254-pretreated rats (compared with microsomes from untreated animals): the formation of 2{alpha}-hydroxytestosterone is specifically catalyzed by cytochrome P-450h, a hemoprotein thought to be involved in the activation of AFB{sub 1}. These results show that not only rat liver parenchymal cells, but also endothelial and Kupffer cells, activated several carcinogens to mutagenic metabolites.« less

  12. Control of aflatoxin production of Aspergillus flavus and Aspergillus parasiticus using RNA silencing technology by targeting aflD (nor-1) gene.

    PubMed

    Abdel-Hadi, Ahmed M; Caley, Daniel P; Carter, David R F; Magan, Naresh

    2011-06-01

    Aspergillus flavus and Aspergillus parasiticus are important pathogens of cotton, corn, peanuts and other oil-seed crops, producing toxins both in the field and during storage. We have designed three siRNA sequences (Nor-Ia, Nor-Ib, Nor-Ic) to target the mRNA sequence of the aflD gene to examine the potential for using RNA silencing technology to control aflatoxin production. Thus, the effect of siRNAs targeting of two key genes in the aflatoxin biosynthetic pathway, aflD (structural) and aflR (regulatory gene) and on aflatoxin B(1 )(AFB(1)), and aflatoxin G(1) (AFG(1)) production was examined. The study showed that Nor-Ib gave a significant decrease in aflD mRNA, aflR mRNA abundance, and AFB(1) production (98, 97 and 97% when compared to the controls) in A. flavus NRRL3357, respectively. Reduction in aflD and aflR mRNA abundance and AFB(1 )production increased with concentration of siRNA tested. There was a significant inhibition in aflD and AFB(1) production by A. flavus EGP9 and AFG(1 )production by A. parasiticus NRRL 13005. However, there was no significant decrease in AFG(1) production by A. parasiticus SSWT 2999. Changes in AFB(1) production in relation to mRNA levels of aflD showed a good correlation (R = 0.88; P = 0.00001); changes in aflR mRNA level in relation to mRNA level of aflD also showed good correlation (R = 0.82; P = 0.0001). The correlations between changes in aflR and aflD gene expression suggests a strong relationship between these structural and regulatory genes, and that aflD could be used as a target gene to develop efficient means for aflatoxin control using RNA silencing technology.

  13. New ion-exchanged zeolite derivatives: antifungal and antimycotoxin properties against Aspergillus flavus and aflatoxin B1

    NASA Astrophysics Data System (ADS)

    Savi, Geovana D.; Cardoso, Willian A.; Furtado, Bianca G.; Bortolotto, Tiago; Da Agostin, Luciana O. V.; Nones, Janaína; Torres Zanoni, Elton; Montedo, Oscar R. K.; Angioletto, Elidio

    2017-08-01

    Zeolites are microporous crystalline hydrated aluminosilicates with absorbent and catalytic properties. This material can be used in many applications in stored-pest management such as: pesticide and fertilizer carriers, animal feed additives, mycotoxin binders and food packaging materials. Herein, four 4A zeolite forms were prepared by ion-exchange and their antifungal effect against Aspergillus flavus was highlighted. Additionally, the antimycotoxin activity and the aflatoxin B1 (AFB1) adsorption capacity of these zeolites as well as their toxic effects on Artemia sp. were investigated. The ion-exchanged zeolites with Li+ and Cu2+ showed the best antifungal activity against A. flavus, including effects on conidia germination and hyphae morphological alterations. Regarding to antimycotoxin activity, all zeolite samples efficiently inhibited the AFB1 production by A. flavus. However, the ion-exchanged zeolites exhibited better results than the 4A zeolite. On the other hand, the AFB1 adsorption capacity was only observed by the 4A zeolite and zeolite-Li+. Lastly, our data showed that all zeolites samples used at effective concentrations for antifungal and antimycotoxin assays (2 mg ml-1) showed no toxic effects towards Artemia sp. Results suggest that some these ion-exchanged zeolites have great potential as an effective fungicide and antimycotoxin agent for agricultural and food safety applications.

  14. Survey of Deoxynivalenol and Aflatoxin B1 in Instant Noodles and Bread Consumed in Thailand by Using Liquid Chromatography-Tandem Mass Spectrometry.

    PubMed

    Pralatnet, Sasithorn; Poapolathep, Saranya; Giorgi, Mario; Imsilp, Kanjana; Kumagai, Susumu; Poapolathep, Amnart

    2016-07-01

    One hundred wheat product samples (50 instant noodle samples and 50 bread samples) were collected from supermarkets in Bangkok, Thailand. Deoxynivalenol (DON) and aflatoxin B1 (AFB1) contamination in these products was analyzed using a validated liquid chromatography-tandem mass spectrometry method. The limit of quantification values of DON and AFB1 in the instant noodles and bread were 2 and 1 ng g(-1), respectively. The survey found that DON was quantifiable in 40% of collected samples, in 2% of noodles (0.089 μg g(-1)), and in 78% of breads (0.004 to 0.331 μg g(-1)). AFB1 was below the limit of quantification of the method in all of the tested samples. The results suggest that the risk of DON exposure via noodles and breads is very low in urban areas of Thailand. No risk can be attributable to AFB1 exposure in the same food matrices, but further studies with a larger sample size are needed to confirm these data.

  15. Effects of airborne Aspergillus on serum aflatoxin B1 and liver enzymes in workers handling wheat flour.

    PubMed

    Saad-Hussein, A; Taha, M M; Fadl, N N; Awad, A-H; Mahdy-Abdallah, H; Moubarz, G; Aziz, H; El-Shamy, K A

    2016-01-01

    The present work aimed to investigate the relationship between occupational exposure to airborne molds, serum aflatoxin B1 (AFB1), and liver enzymes of workers handling wheat flour. The study included 90 bakers, 100 flour milling workers, and 100 controls with no exposure to flour dust. Workplace aspects such as temperature and relative humidity were measured. Airborne fungi were collected and identified. In all subjects included, the serum levels of AFB1, serum albumin (Alb), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) were measured. Air temperature and relative humidity were found to be higher in bakeries than in flour mill sections. Airborne Aspergillus species were isolated in dust particles <8 µm in size. The concentration of Aspergillus flavus and Aspergillus niger were higher in bakeries than in the flour mill sections. They were higher in the grinding section than in other mill sections. The serum AFB1-Alb adduct and ALP levels were significantly higher in bakers compared to milling workers (p < 0.0001, p = 0.05), respectively. The liver enzymes AST and ALT were significantly higher among milling workers and bakers than controls (p < 0.05, p < 0.0001), respectively. The duration of exposure was significantly correlated with serum AFB1 in bakers. Moreover, there was significant correlation between serum AFB1, each of ALT and AST levels in bakers. chronic occupational exposure to high concentrations of Aspergillus in workplaces may cause elevations in serum levels of AFB1 and liver enzymes in workers exposed to flour dust. Hence, worker protection measures should be consistently adopted and enforced at the workplace. © The Author(s) 2015.

  16. Reduction in the urinary aflatoxin M1 biomarker as an early indicator of the efficacy of dietary interventions to reduce exposure to aflatoxins.

    PubMed

    Mitchell, Nicole J; Kumi, Justice; Johnson, Natalie M; Dotse, Eunice; Marroquin-Cardona, Alicia; Wang, Jia-Sheng; Jolly, Pauline E; Ankrah, Nii-Ayi; Phillips, Timothy D

    2013-08-01

    Aflatoxin B1 is a persistent public health issue in Ghana. Assessment of AFB1 intervention efficacy is currently dependent on long-term biomarkers. This study was designed to determine whether daily AFM1 biomarker levels could be utilized as an early detection method for intervention efficacy. Participants were treated with a refined calcium montmorillonite clay (UPSN) or a placebo (calcium carbonate) in a crossover study. Urine samples were assessed for AFM1 levels daily. UPSN treatment reduced AFM1 biomarkers by 55% compared to the placebo. This is the first study to show that daily urinary AFM1 levels can be used as a biomarker of internal aflatoxin B1 exposure in short-term intervention trials to determine efficacy.

  17. Estimated exposure to zearalenone, ochratoxin A and aflatoxin B1 through the consume of bakery products and pasta considering effects of food processing.

    PubMed

    Bol, Emilli Keller; Araujo, Letícia; Veras, Flávio Fonseca; Welke, Juliane Elisa

    2016-03-01

    The objective of this research was to estimate the processing effect on mycotoxins levels and the exposure to zearalenone (ZEA), ochratoxin (OTA) and aflatoxin B1 (AFB1) through the consumption of pasta and bakery products. The higher reduction percentage of mycotoxins was observed in cake production (95, 90 and 70% for ZEA, OTA and AFB1, respectively). Bread and biscuit showed similar reduction in mycotoxins levels (89 and 90% for ZEA; 80 and 85% for OTA; 36 and 40% for AFB1, respectively). The lower reduction in the levels of mycotoxins has been observed for pasta (75, 65 and 10% for ZEA, OTA and AFB1, respectively). The consumption of these products could represent 12.6% of the maximum tolerable daily intake of ZEA and 30.5% of the tolerable weekly intake of OTA. The margin of exposure value related to the exposure to AFB1 was 24.6. The exposure to ZEA and OTA through the consumption of bakery products and pasta would not represent risk for consumer health, (although conjugated forms were not determined). However, the exposure to AFB1 represents a risk (even without considering the AFB1-conjugated forms). Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. Leaky Gut and Mycotoxins: Aflatoxin B1 Does Not Increase Gut Permeability in Broiler Chickens.

    PubMed

    Galarza-Seeber, Rosario; Latorre, Juan D; Bielke, Lisa R; Kuttappan, Vivek A; Wolfenden, Amanda D; Hernandez-Velasco, Xochitl; Merino-Guzman, Ruben; Vicente, Jose L; Donoghue, Annie; Cross, David; Hargis, Billy M; Tellez, Guillermo

    2016-01-01

    Previous studies conducted in our laboratory have demonstrated that intestinal barrier function can be adversely affected by diet ingredients or feed restriction, resulting in increased intestinal inflammation-associated permeability. Two experiments were conducted in broilers to evaluate the effect of three concentrations of Aflatoxin B1 (AFB1; 2, 1.5, or 1 ppm) on gastrointestinal leakage and liver bacterial translocation (BT). In experiment 1, 240 day-of-hatch male broilers were allocated in two groups, each group had six replicates of 20 chickens (n = 120/group): Control feed or feed + 2 ppm AFB1. In experiment 2, 240 day-of-hatch male broilers were allocated in three groups, each group had five replicates of 16 chickens (n = 80/group): Control feed; feed + 1 ppm AFB1; or feed + 1.5 ppm AFB1. In both experiments, chickens were fed starter (days 1-7) and grower diets (days 8-21) ad libitum and performance parameters were evaluated every week. At day 21, all chicks received an oral gavage dose of FITC-d (4.16 mg/kg) 2.5 h before collecting blood samples to evaluate gastrointestinal leakage of FITC-d. In experiment 2, a hematologic analysis was also performed. Liver sections were aseptically collected and cultured using TSA plates to determine BT. Cecal contents were collected to determine total colony-forming units per gram of Gram-negative bacteria, lactic acid bacteria (LAB), or anaerobes by plating on selective media. In experiment 2, liver, spleen, and bursa of Fabricius were removed to determine organ weight ratio, and also intestinal samples were obtained for morphometric analysis. Performance parameters, organ weight ratio, and morphometric measurements were significantly different between Control and AFB1 groups in both experiments. Gut leakage of FITC-d was not affected by the three concentrations of AFB1 evaluated (P > 0.05). Interestingly, a significant reduction in BT was observed in chickens that received 2 and 1

  19. DNA Sequence Modulates Geometrical Isomerism of the trans-8,9-Dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl-formamido)-9-hydroxy Aflatoxin B1 Adduct

    PubMed Central

    2016-01-01

    Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus flavus, is oxidized by cytochrome P450 enzymes to aflatoxin B1-8,9-epoxide, which alkylates DNA at N7-dG. Under basic conditions, this N7-dG adduct rearranges to yield the trans-8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl-formamido)-9-hydroxy aflatoxin B1 (AFB1–FAPY) adduct. The AFB1–FAPY adduct exhibits geometrical isomerism involving the formamide moiety. NMR analyses of duplex oligodeoxynucleotides containing the 5′-XA-3′, 5′-XC-3′, 5′-XT-3′, and 5′-XY-3′ sequences (X = AFB1–FAPY; Y = 7-deaza-dG) demonstrate that the equilibrium between E and Z isomers is controlled by major groove hydrogen bonding interactions. Structural analysis of the adduct in the 5′-XA-3′ sequence indicates the preference of the E isomer of the formamide group, attributed to formation of a hydrogen bond between the formyl oxygen and the N6 exocyclic amino group of the 3′-neighbor adenine. While the 5′-XA-3′ sequence exhibits the E isomer, the 5′-XC-3′ sequence exhibits a 7:3 E:Z ratio at equilibrium at 283 K. The E isomer is favored by a hydrogen bond between the formyl oxygen and the N4-dC exocyclic amino group of the 3′-neighbor cytosine. The 5′-XT-3′ and 5′-XY-3′ sequences cannot form such a hydrogen bond between the formyl oxygen and the 3′-neighbor T or Y, respectively, and in these sequence contexts the Z isomer is favored. Additional equilibria between α and β anomers and the potential to exhibit atropisomers about the C5–N5 bond do not depend upon sequence. In each of the four DNA sequences, the AFB1–FAPY adduct maintains the β deoxyribose configuration. Each of these four sequences feature the atropisomer of the AFB1 moiety that is intercalated above the 5′-face of the damaged guanine. This enforces the Ra axial conformation for the C5–N5 bond. PMID:25587868

  20. DNA Sequence Modulates Geometrical Isomerism of the trans-8,9- Dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl-formamido)- 9-hydroxy Aflatoxin B1 Adduct.

    PubMed

    Li, Liang; Brown, Kyle L; Ma, Ruidan; Stone, Michael P

    2015-02-16

    Aflatoxin B(1) (AFB(1)), a mycotoxin produced by Aspergillus flavus, is oxidized by cytochrome P450 enzymes to aflatoxin B(1)-8,9-epoxide, which alkylates DNA at N7-dG. Under basic conditions, this N7-dG adduct rearranges to yield the trans-8,9-dihydro-8-(2,6-diamino-4-oxo-3,4-dihydropyrimid-5-yl-formamido)-9-hydroxy aflatoxin B(1) (AFB(1)−FAPY) adduct. The AFB(1)−FAPY adduct exhibits geometrical isomerism involving the formamide moiety. NMR analyses of duplex oligodeoxynucleotides containing the 5′-XA-3′, 5′-XC-3′, 5′-XT-3′, and 5′-XY-3′ sequences (X = AFB(1)−FAPY; Y = 7-deaza-dG)demonstrate that the equilibrium between E and Z isomers is controlled by major groove hydrogen bonding interactions.Structural analysis of the adduct in the 5′-XA-3′ sequence indicates the preference of the E isomer of the formamide group,attributed to formation of a hydrogen bond between the formyl oxygen and the N(6) exocyclic amino group of the 3′-neighboradenine. While the 5′-XA-3′ sequence exhibits the E isomer, the 5′-XC-3′ sequence exhibits a 7:3 E:Z ratio at equilibrium at 283K. The E isomer is favored by a hydrogen bond between the formyl oxygen and the N(4)-dC exocyclic amino group of the 3′-neighbor cytosine. The 5′-XT-3′ and 5′-XY-3′ sequences cannot form such a hydrogen bond between the formyl oxygen and the 3′-neighbor T or Y, respectively, and in these sequence contexts the Z isomer is favored. Additional equilibria between α and β anomers and the potential to exhibit atropisomers about the C5−N(5) bond do not depend upon sequence. In each of the four DNA sequences, the AFB(1)−FAPY adduct maintains the β deoxyribose configuration. Each of these four sequences feature the atropisomer of the AFB(1) moiety that is intercalated above the 5′-face of the damaged guanine. This enforces the Ra axialc onformation for the C5−N(5) bond.

  1. Aflatoxin exposure during the first 1000 days of life in rural South Asia assessed by aflatoxin B1-lysine albumin biomarkers

    PubMed Central

    Groopman, John D.; Egner, Patricia A.; Schulze, Kerry J.; Wu, Lee S-F; Merrill, Rebecca; Mehra, Sucheta; Shamim, Abu A.; Ali, Hasmot; Shaikh, Saijuddin; Gernand, Alison; Khatry, Subarna K.; LeClerq, Steven C.; West, Keith P.; Christian, Parul

    2015-01-01

    Aflatoxin B1 is a potent carcinogen, occurring from mold growth that contaminates staple grains in hot, humid environments. In this investigation, aflatoxin B1-lysine albumin biomarkers were measured by mass spectrometry in rural South Asian women, during the first and third trimester of pregnancy, and their children at birth and at two years of age. These subjects participated in randomized community trials of antenatal micronutrient supplementation in Sarlahi District, southern Nepal and Gaibandha District in northwestern Bangladesh. Findings from the Nepal samples demonstrated exposure to aflatoxin, with 94% detectable samples ranging from 0.45 to 2939.30 pg aflatoxin B1-lysine/mg albumin during pregnancy. In the Bangladesh samples the range was 1.56 to 63.22 pg aflatoxin B1-lysine/mg albumin in the first trimester, 3.37 to 72.8 pg aflatoxin B1-lysine/mg albumin in the third trimester, 4.62 to 76.69 pg aflatoxin B1-lysine/mg albumin at birth and 3.88 to 81.44 pg aflatoxin B1-lysine/mg albumin at age two years. Aflatoxin B1-lysine adducts in cord blood samples demonstrated that the fetus had the capacity to convert aflatoxin into toxicologically active compounds and the detection in the same 2-year-old children illus trates exposure over the first 1000 days of life. PMID:25308602

  2. A conventional chemical reaction for use in an unconventional assay: A colorimetric immunoassay for aflatoxin B1 by using enzyme-responsive just-in-time generation of a MnO2 based nanocatalyst.

    PubMed

    Lai, Wenqiang; Zeng, Qiao; Tang, Juan; Zhang, Maosheng; Tang, Dianping

    2018-01-10

    The authors describe a colorimetric immunoassay for the model nalyte aflatoxin B 1 (AFB 1 ). It is based on the just-in-time generation of an MnO 2 nanocatalyst. Unlike previously developed immunoassay, the chromogenic reaction relies on the just-in-time formation of an oxidase mimic without the aid of the substrate. Potassium permanganate (KMnO 4 ) is converted into manganese dioxide (MnO 2 ) which acts as an oxidase mimic that catalyzes the oxidation 3,3',5,5'-tetramethylbenzidine (TMB) by oxygen to give a blue colored product. In the presence of ascorbic acid (AA), KMnO 4 is reduced to Mn(II) ions. This results in a decrease in the amount of MnO 2 nanocatalyst. Hence, the oxidation of TMB does not take place. By adding ascorbate oxidase, AA is converted into dehydroascorbic acid which cannot reduce KMnO 4 . Based on these observations, a colorimetric competitive enzyme immunoassay was developed where ascorbate oxidase and gold nanoparticle-labeled antibody against AFB 1 and magnetic beads carrying bovine serum albumin conjugated to AFB 1 are used for the determination of AFB 1 . In presence of AFB 1 , it will compete with the BSA-conjugated AFB 1 (on the magnetic beads) for the labeled antibody against AFB 1 on the gold nanoparticles. This makes the amount of ascorbate oxidase/anti-AFB 1 antibody-labeled gold nanoparticles, which conjugated on magnetic beads, reduce, and resulted in an increase of ascorbic acid. Under optimal conditions, the absorbance (measured at 652 nm) decreases with increasing AFB 1 concentrations in the range from 0.1 to 100 ng mL -1 , with a 0.1 ng mL -1 detection limit (at the 3S blank level). The accuracy of the assay was validated by analyzing spiked peanut samples. The results matched well with those obtained with a commercial ELISA kit. Conceivably, the method is not limited to aflatoxins but has a wide scope in that it may be applied to many other analytes for which respective antibodies are available. Graphical abstract

  3. The comparison of CHCA solvent compositions for improving LC-MALDI performance and its application to study the impact of aflatoxin B1 on the liver proteome of diabetes mellitus type 1 mice.

    PubMed

    Tsai, Fuu-Jen; Chen, Shih-Yin; Liu, Yu-Ching; Liao, Hsin-Yi; Chen, Chao-Jung

    2017-01-01

    In nanoflow liquid chromatography-matrix-assisted laser desorption/ionization tandem time-of-flight (nanoLC-MALDI-TOF/TOF) approaches, it is critical to directly apply small amounts of the sample elutes on the sample target using a nanoLC system due to its low flow rate of 200 ~ 300 nl/min. It is recommended to apply a sheath liquid containing a matrix with a several μL/min flow rate at the end of the nanoLC column to ensure a larger co-eluted droplet for more reproducible sample spotting and avoid the laborious task of post-manual matrix spotting. In this study, to achieve a better nanoLC-MALDI performance on sample spotting, we first compared α-Cyano-4-hydroxycinnamic acid (CHCA) solvent composition for efficiently concentrating nanoLC elutes on an anchor chip. The solvent composition of isopropanol (IPA): acetonitrile (ACN):acetone:0.1% Trifluoroacetic acid (TFA) (2:7:7:2) provided strong and homogeneous signals with higher peptide ion yields than the other solvent compositions. Then, nanoLC-MALDI-TOF/TOF was applied to study the impact of aflatoxin B1 on the liver proteome from diabetes mellitus type 1 mice. Aflatoxin B1 (AFB1), produced by Aspergillus flavus and Aspergillus parasiticus is a carcinogen and a known causative agent of liver cancer. To evaluate the effects of long-term exposure to AFB1 on type 1 diabetes mellitus (TIDM), the livers of T1DM control mice and mice treated with AFB1 were analyzed using isotope-coded protein labeling (ICPL)-based quantitative proteomics. Our results showed that gluconeogenesis, lipid, and oxidative phosphorylation mechanisms, normally elevated in T1DM, were disordered following AFB1 treatment. In addition, major urinary protein 1 (MUP1), an indicator of increased insulin sensitivity, was significantly decreased in the T1DM/AFB1 group and may have resulted in higher blood glucose levels compared to the T1DM group. These results indicate that T1DM patients should avoid the AFB1 intake, as they could lead to increased

  4. Fractionation of radioactivity in the milk of goats administered UC-aflatoxin B1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Goto, T.; Hsieh, D.P.

    A detailed fractionation of radioactivity in the milk of goats administered UC-aflatoxin B1 at low doses was performed. The milk collected in the first 24 h following dosing contained radioactivity equivalent to 0.45-1.1% of the dose given. The radioactivity in each sample was partitioned into 4 fractions: ether, protein, dichloromethane, and water-alcohol. Over 80% of the radioactivity was detected in the dichloromethane fraction, of which over 95% was attributable to aflatoxin M1. No aflatoxin B1 or other known aflatoxin metabolites were detected in any fraction. The results indicate that the major metabolite of aflatoxin B1 in goat milk is aflatoxinmore » M1 and that other metabolites, including conjugates, are of minor significance.« less

  5. Glutathione-S-transferase A3 knockout mice are sensitive to acute cytotoxic and genotoxic effects of aflatoxin B1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ilic, Zoran, E-mail: zxi01@health.state.ny.u; Crawford, Dana, E-mail: crawfod@mail.amc.ed; Egner, Patricia A., E-mail: pegner@jhsph.ed

    Aflatoxin B1 (AFB1) is a major risk factor for hepatocellular carcinoma (HCC) in humans. However, mice, a major animal model for the study of AFB1 carcinogenesis, are resistant, due to high constitutive expression, in the mouse liver, of glutathione S-transferase A3 subunit (mGSTA3) that is lacking in humans. Our objective was to establish that a mouse model for AFB1 toxicity could be used to study mechanisms of toxicity that are relevant for human disease, i.e., an mGSTA3 knockout (KO) mouse that responds to toxicants such as AFB1 in a manner similar to humans. Exons 3-6 of the mGSTA3 were replacedmore » with a neomycin cassette by homologous recombination. Southern blotting, RT-PCR, Western blotting, and measurement of AFB1-N{sup 7}-DNA adduct formation were used to evaluate the mGSTA3 KO mice. The KO mice have deletion of exons 3-6 of the mGSTA3 gene, as expected, as well as a lack of mGSTA3 expression at the mRNA and protein levels. Three hours after injection of 5 mg/kg AFB1, mGSTA3 KO mice have more than 100-fold more AFB1-N{sup 7}-DNA adducts in their livers than do similarly treated wild-type (WT) mice. In addition, the mGSTA3 KO mice die of massive hepatic necrosis, at AFB1 doses that have minimal toxic effects in WT mice. We conclude that mGSTA3 KO mice are sensitive to the acute cytotoxic and genotoxic effects of AFB1, confirming the crucial role of GSTA3 subunit in protection of normal mice against AFB1 toxicity. We propose the mGSTA3 KO mouse as a useful model with which to study the interplay of risk factors leading to HCC development in humans, as well as for testing of additional possible functions of mGSTA3.« less

  6. Spectra analysis of coating antigen: A possible explanation for difference in anti-AFB1 polyclonal antibody sensitivity

    NASA Astrophysics Data System (ADS)

    Ye, Yang; Liu, Aiping; Wang, Xiaohong; Chen, Fusheng

    2016-10-01

    For the detection of small hapten molecules, indirect competitive enzyme-linked immunosorbent assay (icELISA) is a preferred method. However, diverse coating antigen might bring different antiserum titer and sensitivity for the identical antiserum. In the present study, four AFB1-protein (aflatoxin B1-carrier protein) conjugates were prepared by activated ester method (AFB1O-BSA/AFB1O-OVA) and mannich method (AFB1-cBSA/AFB1-cOVA), and then applied as coating antigen for titer and sensitivity detection of the identical antiserum obtained from rabbit immunized by AFB1-KLH. Afterwards, the ultraviolet-visible, fluorescence and far-ultraviolet circular dichroism (far-UV CD) spectra were recorded for understanding the difference in titer and sensitivity obtained. Results revealed that AFB1O-BSA/AFB1O-OVA showed a strong intrinsic fluorescence band centered at 450 nm that originated from the emission of AFB1, which differed from AFB1-cBSA/AFB1-cOVA, while the decrease of α-helical and increase of β-sheet in AFB1-cBSA was the most remarkable. This indicated that the better sensitivity obtained by using AFB1O-BSA as coating antigen might be caused by its extended structure, because such structure affect the binding between AFB1 and antibody. The study might offer structural information for understanding the titer and sensitivity difference caused by coating antigen.

  7. A direct determination of AFBs in vinegar by aptamer-based surface plasmon resonance biosensor.

    PubMed

    Wu, Wenbo; Zhu, Zhiling; Li, Bingjie; Liu, Zhuqing; Jia, Lili; Zuo, Limin; Chen, Long; Zhu, Zhentai; Shan, Guangzhi; Luo, Shi-Zhong

    2018-05-01

    Aflatoxin (AFB) is one of the most toxic fungal metabolites produced by Aspergillus flavus, which may contaminate food and agricultural products. Herein, an aptamer-based surface plasmon resonance (SPR) biosensor was developed to detect AFBs. The chosen aptamer showed comparable interaction with the two AFBs, namely aflatoxin B1 (AFB1) and aflatoxin B2 (AFB2). Such phenomenon was rarely reported, and might lead to a simultaneous detection of both AFBs. In this study, AFB1 was used to systematically establish the detection method. In the SPR system, streptavidin proteins were immobilized on the surface of a CM5 sensor chip as a cross-linker and biotin-aptamers were captured through streptavidin-biotin interaction. After optimization, the assay showed a dynamic range between 0.09 and 200 ng mL -1 (linear range from 1.5 to 50 ng mL -1 and a LOD of 0.19 ng mL -1 ) of AFB1 in buffer. As expected, the aptasensor showed high specificity towards AFB1 and AFB2, but hardly bound to other toxins with similar structures, including ochratoxin A (OTA), ochratoxin B (OTB), Zeralenone (ZEA) and T-2 toxin (T-2). Determination of AFB1 in vinegar was further performed using the SPR biosensor. Recoveries of AFB1 from spiked samples ranged from 96.3 to 117.8%. The developed SPR assay is a simple, fast and sensitive approach for the detection of residual AFBs in agricultural products and foodstuffs like vinegar. Copyright © 2018 Elsevier Ltd. All rights reserved.

  8. Evaluation of particulate air samplers for airborne aflatoxin B1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Silas, J.C.; Harrison, M.A.; Carpenter, J.A.

    Five air samplers (Millipore, all-glass impinger, centrifugal, Andersen, and absorbent cotton) were evaluated for their ability to collect airborne grain particles contaminated with aflatoxin B1. Corn dust containing 100 micrograms aflatoxin B1/g was aerosolized within a containment system. Each device sampled 100 I air, thus exchanging the air in the chamber two times. Aflatoxin B1 was extracted from all sampling matrices and was detected and quantitated with thin-layer chromatography and scanning fluorodensitometry. The highest efficiency was obtained with the Millipore sampler, while the efficiencies of the centrifugal and the cotton samplers were almost identical. Efficiency of an Andersen was less,more » with no toxin recovered from an all-glass impinger. Measurement of particle size was accomplished with the Andersen sampler.« less

  9. Reduction of Aflatoxins in Apricot Kernels by Electronic and Manual Color Sorting

    PubMed Central

    Zivoli, Rosanna; Gambacorta, Lucia; Piemontese, Luca; Solfrizzo, Michele

    2016-01-01

    The efficacy of color sorting on reducing aflatoxin levels in shelled apricot kernels was assessed. Naturally-contaminated kernels were submitted to an electronic optical sorter or blanched, peeled, and manually sorted to visually identify and sort discolored kernels (dark and spotted) from healthy ones. The samples obtained from the two sorting approaches were ground, homogenized, and analysed by HPLC-FLD for their aflatoxin content. A mass balance approach was used to measure the distribution of aflatoxins in the collected fractions. Aflatoxin B1 and B2 were identified and quantitated in all collected fractions at levels ranging from 1.7 to 22,451.5 µg/kg of AFB1 + AFB2, whereas AFG1 and AFG2 were not detected. Excellent results were obtained by manual sorting of peeled kernels since the removal of discolored kernels (2.6%–19.9% of total peeled kernels) removed 97.3%–99.5% of total aflatoxins. The combination of peeling and visual/manual separation of discolored kernels is a feasible strategy to remove 97%–99% of aflatoxins accumulated in naturally-contaminated samples. Electronic optical sorter gave highly variable results since the amount of AFB1 + AFB2 measured in rejected fractions (15%–18% of total kernels) ranged from 13% to 59% of total aflatoxins. An improved immunoaffinity-based HPLC-FLD method having low limits of detection for the four aflatoxins (0.01–0.05 µg/kg) was developed and used to monitor the occurrence of aflatoxins in 47 commercial products containing apricot kernels and/or almonds commercialized in Italy. Low aflatoxin levels were found in 38% of the tested samples and ranged from 0.06 to 1.50 μg/kg for AFB1 and from 0.06 to 1.79 μg/kg for total aflatoxins. PMID:26797635

  10. Effects of temperature, water activity and incubation time on fungal growth and aflatoxin B1 production by toxinogenic Aspergillus flavus isolates on sorghum seeds.

    PubMed

    Lahouar, Amani; Marin, Sonia; Crespo-Sempere, Ana; Saïd, Salem; Sanchis, Vicente

    2016-01-01

    Sorghum, which is consumed in Tunisia as human food, suffers from severe colonization by several toxigenic fungi and contamination by mycotoxins. The Tunisian climate is characterized by high temperature and humidity that stimulates mold proliferation and mycotoxin accumulation in foodstuffs. This study investigated the effects of temperature (15, 25 and 37°C), water activity (aw, between 0.85 and 0.99) and incubation time (7, 14, 21 and 28 d) on fungal growth and aflatoxin B1 (AFB1) production by three Aspergillus flavus isolates (8, 10 and 14) inoculated on sorghum grains. The Baranyi model was applied to identify the limits of growth and mycotoxin production. Maximum diameter growth rates were observed at 0.99 a(w) at 37°C for two of the isolates. The minimum aw needed for mycelial growth was 0.91 at 25 and 37°C. At 15°C, only isolate 8 grew at 0.99 a(w). Aflatoxin B1 accumulation could be avoided by storing sorghum at low water activity levels (≤0.91 a(w)). Aflatoxin production was not observed at 15°C. This is the first work on the effects of water activity and temperature on A. flavus growth and AFB1 production by A. flavus isolates on sorghum grains. Copyright © 2015 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  11. Influence of Bacillus spp. isolated from maize agroecosystem on growth and aflatoxin B(1) production by Aspergillus section Flavi.

    PubMed

    Bluma, Romina V; Etcheverry, Miriam G

    2006-03-01

    A total of 59 bacteria of the Bacillus genus were isolated from different components of a maize agroecosystem and their antifungal activity against Aspergillus section Flavi was evaluated. Thirty-three and 46% of these bacteria were able to inhibit Aspergillus flavus Link and A. parasiticus Speare respectively at water activity (a(w)) 0.982; however, when a(w) was 0.955, these percentages were decreased and only three isolates were able to inhibit Aspergillus section Flavi. The majority of bacilli acted as contact antagonists, while a small number of isolates were able to form inhibition zones. In maize meal extract agar, Aspergillus section Flavi growth rate and aflatoxin B(1) (AFB(1)) production were significantly reduced when these strains were paired at a(w) 0.982 with bacilli at all inoculum levels studied. However, two bacilli isolated were able to reduce growth rate and aflatoxin production when a(w) was 0.955. Lag phase increase followed the same general pattern as growth rate reduction. When Aspergillus section Flavi was grown in sterile maize in the presence of three Bacillus strains at a(w) 0.982, the reduction in count (colony-forming units (cfu) g(-1) maize) was less than 30%, except when Aspergillus section Flavi grew with Bacillus amyloliquefaciens UNRCLR. However, levels of detectable AFB(1) were significantly reduced in these interactions at a(w) 0.982.

  12. Mutation of Chinese Hamster V79 cells and transformation and mutation of mouse fibroblast C3H/10T1/2 clone 8 cells by aflatoxin B1 and four other furocoumarins isolated from two Nigerian medicinal plants.

    PubMed

    Uwaifo, A O; Billings, P C; Heidelberger, C

    1983-03-01

    Mutation by aflatoxin B1 (AFB1), imperatorin, marmesin, chalepin, and 8-methoxypsoralen (MOP), with and without black light (BL; long-wavelength ultraviolet light) activation, was determined at the hypoxanthine-guanine phosphoribosyltransferase locus (8-azaguanine resistance) in Chinese hamster V79 cells and at the ouabain locus in mouse C3H/1OT1/2 cells. Transformation by these furocoumarins under the same activation conditions was also investigated in C3H/1OT1/2 cells. In V79 cells, AFB1 induced a 4-fold maximum mutation frequency over controls under BL activation at a concentration of 5 micrograms/ml; marmesin induced a 2-fold increased mutation frequency at 1.5 micrograms/ml; MOP induced a 19-fold increase at 10 micrograms/ml; chalepin induced a 3-fold increase at 5 micrograms/ml; and imperatorin induced a 20-fold increase at 10 micrograms/ml. Essentially no mutation was observed at the ouabain-resistant (Ouar) locus in C3H/1OT1/2 cells with any of these compounds. In the transformation assays, type II and type III foci were observed at a 1-microgram/ml addition of AFB1 with or without BL activation; while with MOP and imperatorin, these types of foci were observed only with BL activation. Marmesin, although relatively more cytotoxic than the other furocoumarins studied, with a 50% lethal dose of less than 0.5 micrograms/ml, was not as mutagenic or potentially carcinogenic as were AFB1, imperatorin, or MOP with BL activation. These furocoumarins are considered to be involved in the etiology of the high incidence of skin cancer in Nigeria. Our experiments reinforce that concept and suggest that exposure to these furocoumarins may constitute a real carcinogenic hazard.

  13. Risk assessment of exposure to aflatoxin B1 and ochratoxin A through consumption of different Pistachio (Pistacia vera L.) cultivars collected from four geographical regions of Iran.

    PubMed

    Taghizadeh, Seyedeh Faezeh; Rezaee, Ramin; Davarynejad, Gholamhossein; Asili, Javad; Nemati, Seyed Hossein; Goumenou, Marina; Tsakiris, Ioannis; Tsatsakis, Aristides M; Shirani, Kobra; Karimi, Gholamreza

    2018-07-01

    Iran is one of the main suppliers of pistachio for the European market accounting for over 90% of its demands; hence, efficient analytical methods are required for detection of mycotoxins contamination in pistachio kernels before exporting them. In this study, aflatoxin B1 (AFB1) and ochratoxin A (OTA) levels in five pistachio cultivars collected from four sites of Iran, were measured by HPLC. Based on the results, risk assessment for AFB1 and OTA residues was done. The highest mean concentrations of AFB1 and OTA were found in Ahmad-aghaei (4.33 and 2.19 ng/g, respectively) and Akbari (4.08 and 1.943 ng/g, respectively) cultivars from Rafsanjan, Iran. Even the highest concentrations of AFB1 and OTA in analyzed samples were lower than the corresponding maximum limits set by EU authorities. The hazard index (HI) value for consumers of Iranian pistachio is below one. It could be concluded that consumption of pistachio cultivated in these regions poses no health risk of mycotoxins exposure. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Does aflatoxin exposure in the United Kingdom constitute a cancer risk?

    PubMed Central

    Harrison, J C; Carvajal, M; Garner, R C

    1993-01-01

    Although the aflatoxins were discovered more than 30 years ago, there is still considerable controversy surrounding their human health effects. Most countries have introduced legislation to control the level of aflatoxins in food, but it is not known if these permitted levels still pose a significant cancer risk. Furthermore, it is unlikely that all the sources of human aflatoxin exposure have been discovered, nor if the liver is the only, or indeed, major target organ for aflatoxin-induced cancer in man. In our laboratory we have used both immunological and HPLC methods to examine human DNA from a variety of tissues and organs to identify and quantify aflatoxin DNA-adducts. We have already detected aflatoxin B1 (AFB1)-DNA adducts in formalin-fixed tissue from an acute poisoning incident in Southeast Asia. Here we have examined human colon and rectum DNA from normal and tumorous tissue obtained from cancer patients and colon, liver, pancreas, breast, and cervix DNA from autopsy specimens. AFB1-DNA adducts were detected in all tissue types examined and ranged from 0-60 adducts/10(6) nucleotides. Where sample size allowed, the adduct levels were confirmed by HPLC analysis. Tumor tissues tended to have higher adduct levels than normal tissue from the same individual, and levels generally increased with patient age. In samples analyzed by HPLC, the adducts present had the chromatographic properties of [8,9-dihydro-8-(N5-formyl)-2',5',6'-triamino-4'-oxo-(N5-pyramidyl) -9- hydroxy-aflatoxin B1, the ring-opened form of the AFB1-guanine adduct.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:8319666

  15. Biotransformation of aflatoxin B1 and its conjugated metabolites by rat gastrointestinal microfloras.

    PubMed Central

    Wei, C; Macy, J M; Hsieh, D P

    1981-01-01

    Rat cecal microflora from high- and low-fiber-fed animals hydrolyzed aflatoxin conjugates to metabolites indistinguishable from aflatoxin B1 and aflatoxin P1, but aflatoxicol was not a transformation product. PMID:6263185

  16. Impact of aflatoxin B1 on the pharmacokinetic disposition of enrofloxacin in broiler chickens.

    PubMed

    Kalpana, Starling; Srinivasa Rao, G; Malik, Jitendra K

    2015-09-01

    The potential impact of subchronic exposure of aflatoxin B1 was investigated on the pharmacokinetic disposition of enrofloxacin in broiler chickens. Broiler chickens given either normal or aflatoxin B1 (750μg/kg diet) supplemented diet for 6 weeks received a single oral dose of enrofloxacin (10mg/kg body wt). Blood samples were drawn from the brachial vein at predetermined time intervals after drug administration. Enrofloxacin plasma concentrations analyzed by RP-HPLC were significantly lower in aflatoxin B1-exposed broiler chickens at 0.167, 0.5 and 1.0h after drug administration. In aflatoxin B1-exposed broiler chickens, the absorption rate constant (ka) of enrofloxacin (0.20±0.05h(-1)) was significantly decreased as compared to the unexposed birds (0.98±0.31h(-1)). The values of [Formula: see text] , tmax and AUC0-∞ of enrofloxacin were nonsignificantly increased by 17%, 26% and 17% in aflatoxin-exposed broiler chickens, respectively. Subchronic aflatoxin B1 exposure markedly decreased the initial absorption of enrofloxacin without significantly influencing other pharmacokinetic parameters in broiler chickens. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Determination of Aflatoxin B1 in Smokeless Tobacco Products by use of UHPLC-MS/MS

    PubMed Central

    Zitomer, Nicholas; Rybak, Michael E.; Li, Zhong; Walters, Matthew J.; Holman, Matthew R.

    2017-01-01

    We have developed a UHPLC-MS/MS method for the detection and quantitation of aflatoxins in smokeless tobacco products and used it to determine aflatoxin B1 concentrations in 32 smokeless tobacco products commercially available in the US. Smokeless tobacco products were dried, milled and amended with 13C17-labelled internal standards, extracted in water/methanol solution in the presence of a surfactant, isolated through use of immunoaffinity column chromatography and reconstituted in mobile phase prior to UHPLC-MS/MS analysis. Our method was capable of baseline separation of aflatoxins B1, B2, G1 and G2 in a 2.5 min run by use of a fused core C18 column and a water/methanol gradient. MS/MS transition (m/z) 313.3>241.2 was used for aflatoxin B1 quantitation, with 313.3>285.1 used for confirmation. The limit of detection (LOD) for aflatoxin B1 was 0.007 parts per billion (ppb). Method imprecision for aflatoxin B1 (expressed as coefficient of variation) ranged from 5.5% to 9.4%. Spike recoveries were 105–111%. Aflatoxin B1 concentrations in the smokeless tobacco products analysed ranged from Aflatoxin B1 was most frequently detected in dry snuffs and chews, whereas all moist snuff products tested were below LOD. The amounts of aflatoxin B1 we detected were low relative to the 20 ppb regulatory limit established by the Food and Drug Administration for foods and feeds. PMID:26452144

  18. Rapid pretreatment and detection of trace aflatoxin B1 in traditional soybean sauce.

    PubMed

    Xie, Fang; Lai, WeiHua; Saini, Jasdeep; Shan, Shan; Cui, Xi; Liu, DaoFeng

    2014-05-01

    Soybean sauce, a traditional fermented food in China, has different levels of aflatoxin B1 pollution. Two kinds of direct and indirect immunomagnetic bead methods for the pretreatment of aflatoxin B1 were evaluated in this work. A method was established to detect aflatoxin B1 in soybean sauce using an immunomagnetic bead system for pretreatment and ELISA for quantification. The pretreatment method of immunomagnetic beads performed better compared with the conventional extraction and immunoaffinity column method. ELISA exhibited a good linear relationship at an aflatoxin B1 concentration of 0.05-0.3μg/kg (r(2)=0.9842). The average recoveries across spike levels varied from 0.5 to 7μg/kg were 83.6-104% with a relative standard deviation between 4.2% and 11.7%. With the advantages of rapid detection, easy operation, simple equipment, sensitivity, accuracy, and high recovery; this method can be well applied in the trace determination of aflatoxin B1 in soybean sauce samples. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Biosorption of B-aflatoxins Using Biomasses Obtained from Formosa Firethorn [Pyracantha koidzumii (Hayata) Rehder

    PubMed Central

    Ramales-Valderrama, Rosa Adriana; Vázquez-Durán, Alma; Méndez-Albores, Abraham

    2016-01-01

    Mycotoxin adsorption onto biomaterials is considered as a promising alternative for decontamination without harmful chemicals. In this research, the adsorption of B-aflatoxins (AFB1 and AFB2) using Pyracantha koidzumii biomasses (leaves, berries and the mixture of leaves/berries) from aqueous solutions was explored. The biosorbent was used at 0.5% (w/v) in samples spiked with 100 ng/mL of B-aflatoxin standards and incubated at 40 °C for up to 24 h. A standard biosorption methodology was employed and aflatoxins were quantified by an immunoaffinity column and UPLC methodologies. The biosorbent-aflatoxin interaction mechanism was investigated from a combination of zeta potential (ζ), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The highest aflatoxin uptakes were 86% and 82% at 6 h using leaves and the mixture of leaves/berries biomasses, respectively. A moderate biosorption of 46% was attained when using berries biomass. From kinetic studies, the biosorption process is described using the first order adsorption model. Evidence from FTIR spectra suggests the participation of hydroxyl, amine, carboxyl, amide, phosphate and ketone groups in the biosorption and the mechanism was proposed to be dominated by the electrostatic interaction between the negatively charged functional groups and the positively charged aflatoxin molecules. Biosorption by P. koidzumii biomasses has been demonstrated to be an alternative to conventional systems for B-aflatoxins removal. PMID:27420096

  20. Adsorption of aflatoxin B1, zearalenone and ochratoxin A by microorganisms isolated from Kefir grains.

    PubMed

    Taheur, Fadia Ben; Fedhila, Kais; Chaieb, Kamel; Kouidhi, Bochra; Bakhrouf, Amina; Abrunhosa, Luís

    2017-06-19

    A strategy to reduce the deleterious effects of mycotoxins is to use dietary supplements that contain microorganisms that bind mycotoxins and decrease their gastrointestinal absorption. Novel strains were isolated from a Kefir culture and assessed for their mycotoxin adsorption and biotransformation ability. The most active strains were identified using DNA sequencing, and the stability of microorganism/mycotoxin complexes was evaluated using buffer solutions to simulate the pH conditions in the gastrointestinal tract. Our results showed that the microorganism consortium of Kefir grains adsorbed 82 to 100% of aflatoxin B1 (AFB1), zearalenone (ZEA) and ochratoxin A (OTA) when cultivated in milk. The main strains that were capable of mycotoxin adsorption were identified as Lactobacillus kefiri, Kazachstania servazzii and Acetobacter syzygii. The strain L. kefiri KFLM3 was the most active, adsorbing 80 to 100% of the studied mycotoxins when cultivated in milk. Nonetheless, the strain K. servazzii KFGY7 retained more mycotoxin after the desorption experiments (65, 69 and 67% for AFB1, OTA and ZEA, respectively). These findings suggest that Kefir consumption may help to reduce gastrointestinal absorption of these mycotoxins and consequently reduce their toxic effects. The isolated strains may be of interest for the development of fermented dairy products for human consumption that have a new probiotic characteristic, the adsorption of mycotoxins. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Present and future directions of translational research on aflatoxin and hepatocellular carcinoma. A review.

    PubMed

    Wogan, Gerald N; Kensler, Thomas W; Groopman, John D

    2012-01-01

    The aflatoxins were discovered in toxic peanut meal causing "turkey X" disease, which killed large numbers of turkey poults, ducklings and chicks in the UK in the early 1960s. Extracts of toxic feed induced the symptoms in experimental animals, and purified metabolites with properties identical to aflatoxins B(1) and G(1) (AFB(1) and AFG(1)) were isolated from Aspergillus flavus cultures. Structure elucidation of aflatoxin B(1) was accomplished and confirmed by total synthesis in 1963. AFB(1) is a potent liver carcinogen in rodents, non-human primates, fish and birds, operating through a genotoxic mechanism involving metabolic activation to an epoxide, formation of DNA adducts and, in humans, modification of the p53 gene. Aflatoxins are unique among environmental carcinogens, in that elucidation of their mechanisms of action combined with molecular epidemiology provides a foundation for quantitative risk assessment; extensive evidence confirms that contamination of the food supply by AFB(1) puts an exposed population at increased risk of developing hepatocellular carcinoma (HCC). Molecular biomarkers to quantify aflatoxin exposure in individuals were essential to link aflatoxin exposure with liver cancer risk. Biomarkers were validated in populations with high HCC incidence in China and The Gambia, West Africa; urinary AFB(1)-N (7)-Guanine excretion was linearly related to aflatoxin intake, and levels of aflatoxin-serum albumin adducts also reflected aflatoxin intake. Two major cohort studies employing aflatoxin biomarkers identified their causative role in HCC etiology. Results of a study in Shanghai men strongly support a causal relationship between HCC risk and the presence of biomarkers for aflatoxin and HBV infection, and also show that the two risk factors act synergistically. Subsequent cohort studies in Taiwan confirm these results. IARC classified aflatoxin as a Group 1 human carcinogen in 1993, based on sufficient evidence in humans and experimental

  2. Fabrication of a novel nanocomposite based on sol-gel process for hollow fiber-solid phase microextraction of aflatoxins: B1 and B2, in cereals combined with high performane liquid chromatography-diode array detection.

    PubMed

    Es'haghi, Zarrin; Sorayaei, Hoda; Samadi, Fateme; Masrournia, Mahboubeh; Bakherad, Zohreh

    2011-10-15

    The new pre-concentration technique, hollow fiber-solid phase microextraction based on carbon nanotube reinforced sol-gel and liquid chromatography-photodiode array detection was applied to determination of aflatoxins B(1), B(2) (AFB(1), AFB(2)) in rice, peanut and wheat samples. This research provides an overview of trends related to synthesis of solid phase microextraction (SPME) sorbnents that improves the assay of aflatoxins as the semi-polar compounds in several real samples. It mainly includes summary and a list of the results for a simple carbon nanotube reinforced sol-gel in-fiber device. This device was used for extraction, pre-concentration and determination of aflatoxins B1, B2 in real samples. In this technique carbon nanotube reinforced sol was prepared by the sol-gel method via the reaction of phenyl trimethoxysilane (PTMS) with a basic catalyst (tris hydroxymethyl aminomethan). The influences of microextraction parameters such as pH, ageing time, carbon nanotube contents, desorption conditions, desorption solvent and agitation speed were investigated. Optimal HPLC conditions were: C(18) reversed phase column for separation, water-acetonitril-methanol (35:10:55) as the mobile phase and maximum wavelength for detection was 370 nm. The method was evaluated statistically and under optimized conditions, the detection limits for the analytes were 0.074 and 0.061 ng/mL for B1 and B2 respectively. Limit of quantification for B1 and B2 was 0.1 ng/mL too (n=7). The precisions were in the range of 2.829-2.976% (n=3), and linear ranges were within 0.1 and 400 ng/mL. The method was successfully applied to the analysis of cereals (peanut, wheat, rice) with the relative recoveries from 47.43% to 106.83%. Copyright © 2011 Elsevier B.V. All rights reserved.

  3. On the occurrence of aflatoxin M1 in milk and dairy products.

    PubMed

    Prandini, A; Tansini, G; Sigolo, S; Filippi, L; Laporta, M; Piva, G

    2009-05-01

    Aflatoxins are toxic fungal metabolites found in foods and feeds. When ruminants eat AFB(1)-feedstuffs, they metabolise the toxin and excrete AFM(1) in milk. To control AFM(1) in foods it is necessary to reduce AFB(1) contamination of feeds for dairy cattle by preventing fungal growth and AFB(1) formation in agricultural commodities intended for animal use. Corn and corn-based products are one of the most contaminated feedstuffs; therefore risk factor analysis of AFB(1) contamination in corn is necessary to evaluate risk of AFM(1) contamination in milk and milk products. During the corn silage production, the aflatoxins production is mostly influenced by: harvest time; fertilization; irrigation; pest control; silage moisture; and storage practices. Due to the lower moisture at harvest and to the conservation methods, the corn grain is mostly exposed to the contamination by Aspergillus species. Therefore, it is necessary to reduce the probability of this contaminant through choice of: hybrids; seeding time and density; suitable ploughing and fertirrigation; and chemical or biological control. Grains harvested with the lowest possible moisture and conservation moisture close to or less than 14% are necessary to reduce contamination risks, as is maintaining mass to homogeneous moisture. Kernel mechanical damage, grain cleaning practices and conservation temperature are also factors which need to be carefully controlled.

  4. Aflatoxins in spices marketed in Portugal.

    PubMed

    Martins, M L; Martins, H M; Bernardo, F

    2001-04-01

    Seventy-nine prepackaged samples of 12 different types of spice powders (five cardamom, five cayenne pepper, eight chilli, five cloves, seven cumin, five curry) powder, five ginger, five mustard, 10 nutmeg, 12 paprika, five saffron and seven white pepper) were selected from supermarkets and ethnic shops in Lisbon (Portugal) for estimation of aflatoxins by immunoaffinity column clean-up followed by HPLC. Aflatoxin B1 (AFB1) was detected in 34 samples of prepackaged spices (43.0%). All of the cayenne pepper samples were contaminated with levels ranging from 2 to 32 microg AFB1/kg. Three nutmeg samples contained levels ranging from 1 to 5 microg/kg, three samples had levels ranging from 6 to 20 microg/kg, and there were two with 54 microg/kg and 58 microg/ kg. Paprika contained levels of aflatoxin B1 ranging from 1 to 20 microg/kg. Chilli, cumin, curry powder, saffron and white pepper samples had levels ranging from 1 to 5 microg/kg. Aflotoxins were not detected in cardamon, cloves, ginger and mustard. None of the samples analysed contained aflatoxins B2, G1 and G2.

  5. Present and future directions of translational research on aflatoxin and hepatocellular carcinoma. A review

    PubMed Central

    Wogan, Gerald N.; Kensler, Thomas W.; Groopman, John D.

    2015-01-01

    The aflatoxins were discovered in toxic peanut meal causing “turkey X” disease, which killed large numbers of turkey poults, ducklings and chicks in the UK in the early 1960s. Extracts of toxic feed induced the symptoms in experimental animals, and purified metabolites with properties identical to aflatoxins B1 and G1 (AFB1 and AFG1) were isolated from Aspergillus flavus cultures. Structure elucidation of aflatoxin B1 was accomplished and confirmed by total synthesis in 1963. AFB1 is a potent liver carcinogen in rodents, non-human primates, fish and birds, operating through a genotoxic mechanism involving metabolic activation to an epoxide, formation of DNA adducts and, in humans, modification of the p53 gene. Aflatoxins are unique among environmental carcinogens, in that elucidation of their mechanisms of action combined with molecular epidemiology provides a foundation for quantitative risk assessment; extensive evidence confirms that contamination of the food supply by AFB1 puts an exposed population at increased risk of developing hepatocellular carcinoma (HCC). Molecular biomarkers to quantify aflatoxin exposure in individuals were essential to link aflatoxin exposure with liver cancer risk. Biomarkers were validated in populations with high HCC incidence in China and The Gambia, West Africa; urinary AFB1–N7-Guanine excretion was linearly related to aflatoxin intake, and levels of aflatoxin– serum albumin adducts also reflected aflatoxin intake. Two major cohort studies employing aflatoxin biomarkers identified their causative role in HCC etiology. Results of a study in Shanghai men strongly support a causal relationship between HCC risk and the presence of biomarkers for aflatoxin and HBV infection, and also show that the two risk factors act synergistically. Subsequent cohort studies in Taiwan confirm these results. IARC classified aflatoxin as a Group 1 human carcinogen in 1993, based on sufficient evidence in humans and experimental animals

  6. Optimization of chromatographic conditions for determination of aflatoxin B1, B2, G1 and G2 by using liquid chromatography-mass Spectrometry

    NASA Astrophysics Data System (ADS)

    Ramadhaningtyas, Dillani Putri; Aryana, Nurhani; Aristiawan, Yosi; Styarini, Dyah

    2017-11-01

    The optimization of instrument condition and chromatographic separation for analysis of aflatoxin B1, B2, G1 and G2 using liquid chromatography tandem with mass spectrometer detector was conducted in the aim to provide more accurate and reliable analysis results. The aflatoxin known to be serious threat for human health as it is classified as the carcinogenic compounds. The aflatoxin B1, B2, G1 and G2 were selected due to its extensive contamination in various agricultural commodities. The best chromatographic separation was obtained using C-18 column with gradient elution of solvent 5 mM ammonium acetate and 0.1% formic acid in methanol at 7 minutes runtime analysis. The linearity of the detector showed satisfied results as the coefficient determination found to be 0.9994, 0.9996, 0.9998 and 0.9987 for aflatoxin B1, G1, B2, and G2 respectively in the range concentration from 1 to 20 ng/g. The quantifier ion selected for the aflatoxin B1, B2, G1 and G2 was m/z 285.1, 259, 243 and 313 respectively. The instrument precision at these quantifier ions also showed satisfied result with %RSD was around 3.4 to 6.8%. The optimized method present in this study can be used for further sample analysis.

  7. Activated carbons as potentially useful non-nutritive additives to prevent the effect of fumonisin B1 on sodium bentonite activity against chronic aflatoxicosis.

    PubMed

    Monge, María Del Pilar; Magnoli, Alejandra Paola; Bergesio, Maria Virginia; Tancredi, Nestor; Magnoli, Carina E; Chiacchiera, Stella Maris

    2016-06-01

    Aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are mycotoxins that often co-occur in feedstuffs. The ingestion of AFB1 causes aflatoxicosis in humans and animals. Sodium bentonite (NaB), a cheap non-nutritive unselective sequestering agent incorporated in animal diets, can effectively prevent aflatoxicosis. Fumonisins are responsible for equine leukoencephalomalacia and porcine pulmonary oedema, and often have subclinical toxic effects in poultries. Fumonisin B1 and aflatoxin B1 are both strongly adsorbed in vitro on sodium bentonite. Co-adsorption studies, carried out with a weight ratio of FB1 to AFB1 that mimics the natural occurrence (200:1), showed that FB1 greatly decreases the in vitro ability of NaB to adsorb AFB1. The ability of two activated carbons to adsorb FB1 was also investigated. Both carbons showed high affinity for FB1. A complex behaviour of the FB1 adsorption isotherms with pH was observed. In vitro results suggest that under natural contamination levels of AFB1 and FB1, a mixture of activated carbon and sodium bentonite might be potentially useful for prevention of sub-acute aflatoxicosis.

  8. Infants’ Exposure to Aflatoxin M1 from Mother’s Breast Milk in Iran

    PubMed Central

    Ghiasian, SA; Maghsood, AH

    2012-01-01

    Background The occurrence of aflatoxin M1 (AFM1) in milk, especially breast milk, is a valuable biomarker for exposure determination to aflatoxin B1 (AFB1). In the present study, the risk of exposure to AFM1 in infants fed breast milk was investigated. Methods: An enzyme-linked immunosorbent assay (ELISA) was used for the analysis of AFM1 in breast milk samples from 132 lactating mothers referred to four urban Mothers and Babies Care Unit of Hamadan, western Iran. Results: AFM1 was detected in eight samples (6.06%) at mean concentration of 9.45 ng/L. The minimum and maximum of concentration was 7.1 to 10.8 ng/L, respectively. Although the concentration of AFM1 in none of the samples was higher than the maximum tolerance limit accepted by USA and European Union (25 ng/kg) however, 25% had a level of AFM1 above the allowable level of Australia and Switzerland legal limit (10 ng/L). Conclusions: Lactating mothers and infants in western parts of Iran could be at risk for AFB1 and AFM1 exposure, respectively. Considering all this information, the investigation of AFM1 in lactating mothers as a biomarker for post-natal exposure of infants to this carcinogen deserves further studies in various seasons and different parts of Iran. PMID:23113156

  9. Polymorphism in xeroderma pigmentosum complementation group C codon 939 and aflatoxin B1-related hepatocellular carcinoma in the Guangxi population.

    PubMed

    Long, Xi-Dai; Ma, Yun; Zhou, Yuan-Feng; Ma, Ai-Min; Fu, Guo-Hui

    2010-10-01

    Genetic polymorphisms in DNA repair genes may influence individual variations in DNA repair capacity, and this may be associated with the risk and outcome of hepatocellular carcinoma (HCC) related to aflatoxin B1 (AFB1) exposure. In this study, we focused on the polymorphism of xeroderma pigmentosum complementation group C (XPC) codon 939 (rs#2228001), which is involved in nucleotide excision repair. We conducted a case-control study including 1156 HCC cases and 1402 controls without any evidence of hepatic disease to evaluate the associations between this polymorphism and HCC risk and prognosis in the Guangxi population. AFB1 DNA adduct levels, XPC genotypes, and XPC protein levels were tested with a comparative enzyme-linked immunosorbent assay, TaqMan polymerase chain reaction for XPC genotypes, and immunohistochemistry, respectively. Higher AFB1 exposure was observed among HCC patients versus the control group [odds ratio (OR) = 9.88 for AFB1 exposure years and OR = 6.58 for AFB1 exposure levels]. The XPC codon 939 Gln alleles significantly increased HCC risk [OR = 1.25 (95% confidence interval = 1.03-1.52) for heterozygotes of the XPC codon 939 Lys and Gln alleles (XPC-LG) and OR = 1.81 (95% confidence interval = 1.36-2.40) for homozygotes of the XPC codon 939 Gln alleles (XPC-GG)]. Significant interactive effects between genotypes and AFB1 exposure status were also observed in the joint-effects analysis. This polymorphism, moreover, was correlated with XPC expression levels in cancerous tissues (r = -0.369, P < 0.001) and with the overall survival of HCC patients (the median survival times were 30, 25, and 19 months for patients with homozygotes of the XPC codon 939 Lys alleles, XPC-LG, and XPC-GG, respectively), especially under high AFB1 exposure conditions. Like AFB1 exposure, the XPC codon 939 polymorphism was an independent prognostic factor influencing the survival of HCC. Additionally, this polymorphism multiplicatively interacted with the xeroderma

  10. Long term administration of low doses of mycotoxins in poultry. 2. Residues of ochratoxin A and aflatoxins in broilers and laying hens after combined administration of ochratoxin A and aflatoxin B1.

    PubMed

    Micco, C; Miraglia, M; Benelli, L; Onori, R; Ioppolo, A; Mantovani, A

    1988-01-01

    The effects of combined administration of ochratoxin A (OA) and aflatoxin B1 (AFB1) on the occurrence and the levels of residues of mycotoxins in poultry have been investigated. Male broilers and laying hens were fed from 14 days old with standard diets contaminated with 50 micrograms/kg OA and 50 micrograms/kg AFB1. Two groups of broilers and hens were withdrawn from contaminated feed at 37 and 88 days, respectively. At the time of sacrifice no significant lesions were found. Residues were compared with those found after administration of either toxin alone in former trials. Combined treatment resulted in higher content of OA in broiler livers (40 versus 5.0 micrograms/kg) and, to a lesser extent, in kidneys and skin, and of AFB1 in broiler liver and kidney (0.15 versus 0.02 microgram/kg and 0.40 versus 0.05 microgram/kg respectively). Laying hens showed smaller differences (0.20 versus 0.10 microgram/kg in liver and 0.32 versus 0.08 in kidneys). Withdrawal from treatment led to the almost complete disappearance of OA residues in broilers and in hens. These results show a synergistic effect of OA and AFB1, particularly in broilers.

  11. Evaluation of the efficacy, acceptability and palatability of calcium montmorillonite clay used to reduce aflatoxin B1 dietary exposure in a crossover study in Kenya

    PubMed Central

    Awuor, Abigael O.; Yard, Ellen; Daniel, Johnni H.; Martin, Collen; Bii, Christine; Romoser, Amelia; Oyugi, Elvis; Elmore, Sarah; Amwayi, Samwel; Vulule, John; Zitomer, Nicholas C.; Rybak, Michael E.; Phillips, Timothy D.; Montgomery, Joel M.; Lewis, Lauren S.

    2017-01-01

    Acute aflatoxin exposure can cause death and disease (aflatoxicosis) in humans. Aflatoxicosis fatality rates have been documented to be as high as 40% in Kenya. The inclusion in the diet of calcium silicate 100 (ACCS100), a calcium montmorillonite clay, may reduce aflatoxin bioavailability, thus potentially decreasing the risk of aflatoxicosis. We investigated the efficacy, acceptability and palatability of ACCS100 in a population in Kenya with recurring aflatoxicosis outbreaks. Healthy adult participants were enrolled in this double-blinded, crossover clinical trial in 2014. Following informed consent, participants (n = 50) were randomised to receive either ACCS100 (3 g day−1) or placebo (3 g day−1) for 7 days. Treatments were switched following a 5-day washout period. Urine samples were collected daily and assessed for urinary aflatoxin M1 (AFM1). Blood samples were collected at the beginning and end of the trial and assessed for aflatoxin B1-lysine adducts from serum albumin (AFB1-lys). AFM1 concentrations in urine were significantly reduced while taking ACCS100 compared with calcium carbonate placebo (β = 0.49, 95% confidence limit = 0.32–0.75). The 20-day interval included both the placebo and ACCS100 treatments as well as a washout period. There were no statistically significant differences in reported taste, aftertaste, appearance, colour or texture by treatment. There were no statistically significant differences in self-reported adverse events by treatment. Most participants would be willing to take ACCS100 (98%) and give it to their children (98%). ACCS100 was effective, acceptable and palatable. More work is needed to test ACCS100 among vulnerable populations and to determine if it remains effective at the levels of aflatoxin exposure that induce aflatoxicosis. PMID:27603954

  12. The effect on performance and biochemical parameters when soil was added to aflatoxin-contaminated poultry rations.

    PubMed

    Madden, U A; Stahr, H M; Stino, F K

    1999-08-01

    The effects of silty clay loam soil on the performance and biochemical parameters of chicks were investigated when the soil was added to aflatoxin B1 (AFB1)-contaminated diets. One hundred 14-d-old White Leghorn chicks were fed a control ration (clean corn), a low aflatoxin-contaminated ration (120 ng AFB1/g), a high aflatoxin-contaminated ration (700 ng AFB1/g), or high aflatoxin-contaminated rations (700 ng AFB1/g) +10% or 25% soil. Body weight, feed consumption and blood samples were monitored weekly. Decreased feed consumption, body weight gain and efficiency of feed utilization, increased SGOT and LDH activities, and cholesterol and triglyceride concentrations, and decreased uric acid concentrations and ALP activity were observed in the chicks fed the high aflatoxin-contaminated ration without soil. Hepatomegaly was prominent in chicks fed the high aflatoxin-contaminated ration without soil, and some livers had extensive hepatocyte vacuolation, hepatocellular swelling, fatty change and hydropic degeneration, and stained positive for fat accumulation. Addition of soil reduced the detrimental effects of AFB1 for some parameters, although the reduction was less when 10% soil was fed compared with the 25% soil feeding.

  13. Evaluation of the bioactive extract of actinomyces isolated from the Egyptian environment against aflatoxin B1-induce cytotoxicity, genotoxicity and oxidative stress in the liver of rats.

    PubMed

    El-Nekeety, Aziza A; Salman, Asmaa S; Hathout, Amal S; Sabry, Bassem A; Abdel-Aziem, Sekena H; Hassan, Nabila S; Abdel-Wahhab, Mosaad A

    2017-07-01

    This study aimed to determine the bioactive compounds of actinomyces (ACT) isolated from the Egyptian environment (D-EGY) and to evaluate their protective activity against AFB 1 in female Sprague-Dawley rats. Six groups of animals were treated orally for 3 weeks included: C, the control group, T1, AFB 1 -treated group (80 μg/kg b.w), T2 and T3, the groups received ACT extract at low (25 mg/kg b.w) or high (50 mg/kg b.w) doses, T4 and T5, the groups received AFB 1 plus the low or high dose of ACT extract. Blood, bone marrow and tissue samples were collected for different analyses and histological examination. The results revealed the identification of 40 components, representing 99.98%. Treatment with AFB 1 disturbs liver function parameters, oxidative stress markers, antioxidant gene expressions, DNA fragmentation and induced severe histological changes. ACT extract at the low or high doses did not induce significant changes in all the tested parameters or histological picture of the liver. Moreover, ACT extract succeeded to induce a significant protection against the toxicity of AFB 1 . It could be concluded that the bioactive compounds in ACT are promise candidate for the development of food additive or drugs for the protection and treatment of liver disorders in the endemic area. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Dietary intake of aflatoxins in the adult Malaysian population - an assessment of risk.

    PubMed

    Chin, C K; Abdullah, A; Sugita-Konishi, Y

    2012-01-01

    Exposure to aflatoxins in the adult Malaysian diet was estimated by analysing aflatoxins in 236 food composites prepared as "ready for consumption". Dietary exposure to aflatoxin B1 (AFB1) ranged from 24.3 to 34.00 ng/kg b.w./day (lower to upper bound), with peanuts being the main contributor. Estimated liver cancer risk from this exposure was 0.61-0.85 cancers/100,000 population/year, contributing 12.4%-17.3% of the liver cancer cases. Excluding AFB1 occurrence data higher than 15 µg/kg reduced exposure by 65%-91% to 2.27-11.99 ng/kg b.w./day, reducing the cancer risk to 0.06-0.30 cancers/100,000 population/year (contributing 1.2%-6.1% liver cancer cases). Reducing further the ML of AFB1 from 15 to 5 µg/kg yielded 3%-7% greater drop in the exposure to 0.47-10.26 ng/kg b.w./day with an estimated risk of 0.01-0.26 cancers/100,000 population/year (0.2%-5.1% liver cancer cases attributed to dietary AFB1). These findings indicate that current MLs are adequate in protecting Malaysians' health.

  15. Determination of aflatoxins in medicinal plants by high-performance liquid chromatography-tandem mass spectrometry.

    PubMed

    Siddique, Nadeem A; Mujeeb, Mohd; Ahmad, Sayeed; Panda, Bibhu P; Makhmoor, Mohd

    2013-01-01

    The intention of the proposed work is to study the presence of the aflatoxins B1, B2, G1 and G2 in medicinal plants, namely Mucuna pruriens, Delphinium denudatum and Portulaca oleraceae. The aflatoxins were extracted, purified by immunoaffinity column chromatography and analysed by high-performance liquid chromatography-tandem quadrupole mass spectrometry with electrospray ionisation (HPLC-MS/MS). Fungal count was carried out in PDA media. A good linear relationship was found for AFB1, AFB2, AFG1 and AFG2 at 1-10 ppb (r>0.9995). The analyte accuracy under three different spiking levels was 86.7-108.1 %, with low per cent relative standard deviations in each case. The aflatoxins can be separated within 5 to7 min using an Agilent XDB C18-column. We found that AFB1 and AFB2 were in trace amounts below the detection limit in M. pruriens whilst they were not detected in D. denudatum. P. oleraceae was found to be contaminated with AFB1 and AFB2. AFG1 and AFG2 were not detected in M. pruriens, P. oleraceae and were below the detection limit in D. denudatum. This was consistent with very low numbers of fungal colonies observed after 6 hr of incubation. The analytical method developed is simple, precise, accurate, economical and can be effectively used to determine the aflatoxins in medicinal plants and therefore to control the quality of products. The aflatoxin levels in the plant extracts examined were related to the minimal fungal load in the medicinal plants examined.

  16. Inactivation of aflatoxin B1 by using the synergistic effect of hydrogen peroxide and gamma radiation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Patel, U.D.; Govindarajan, P.; Dave, P.J.

    Inactivation of aflatoxin B1 was studied by using gamma radiation and hydrogen peroxide. A 100-krad dose of gamma radiation was sufficient to inactivate 50 micrograms of aflatoxin B1 in the presence of 5% hydrogen peroxide, and 400 krad was required for total degradation of 100 micrograms of aflatoxin in the same system. Degradation of aflatoxin B1 was confirmed by high-pressure liquid chromatographic and thin-layer chromatographic analysis. Ames microsomal mutagenicity test showed loss of aflatoxin activity. This method of detoxification also reduces the toxin levels effectively in artificially contaminated groundnuts.

  17. Characterization of c-Ki-ras and N-ras oncogenes in aflatoxin B sub 1 -induced rat liver tumors

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    McMahon, G.; Davis, E.F.; Huber, L.J.

    c-Ki-ras and N-ras oncogenes have been characterized in aflatoxin B{sub 1}-induced hepatocellular carcinomas. Detection of different protooncogene and oncogene sequences and estimation of their frequency distribution were accomplished by polymerase chain reaction, cloning, and plaque screening methods. Two c-Ki-ras oncogene sequences were identified in DNA from liver tumors that contained nucleotide changes absent in DNA from livers of untreated control rats. Sequence changes involving G{center dot}C to T{center dot}A or G{center dot}C to A{center dot}T nucleotide substitutions in codon 12 were scored in three of eight tumor-bearing animals. Distributions of c-Ki-ras sequences in tumors and normal liver DNA indicated thatmore » the observed nucleotide changes were consistent with those expected to result from direct mutagenesis of the germ-line protooncogene by aflatoxin B{sub 1}. N-ras oncogene sequences were identified in DNA from two of eight tumors. Three N-ras gene regions were identified, one of which was shown to be associated with an oncogene containing a putative activating amino acid residing at codon 13. All three N-ras sequences, including the region detected in N-ras oncogenes, were present at similar frequencies in DNA samples from control livers as well as liver tumors. The presence of a potential germ-line oncogene may be related to the sensitivity of the Fischer rat strain to liver carcinogenesis by aflatoxin B{sub 1} and other chemical carcinogens.« less

  18. In vivo detoxification of aflatoxinB1 by magnetic carbon nanostructures prepared from bagasse.

    PubMed

    Khan, Farhat Ali; Zahoor, Muhammad

    2014-10-30

    Aflatoxins are serious hazard to poultry industry and human health. Broiler chickens fed on aflatoxin contaminated feed develop various abnormal signs and behavior including less attraction toward feed, abnormal faeces consistency, growth retardation, dirty and ruffled feather, abnormal organs size and weight and blood serum biochemistry. Therefore the study was aimed to detoxify aflatoxin B1 in poultry feed. In this study a novel adsorbent was prepared from bagasse, characterized in vitro and in vivo it was fed to different groups of poultry birds along with aflatoxin B1. The groups were given arbitrary names A, B, C, D, E and F. Group A was fed with normal decontaminated feed, group B was fed with aflatoxin contaminated (200 μg/kg feed) feed while the groups C, D, E and F were fed with aflatoxin contaminated (200 μg/kg feed) feed plus 0.2, 0.3, 0.4 and 0.5% adsorbent respectively. Clinical signs and behavior of the chicks; blood level of alanine transferase, alkaline phosphatase, serum albumen, serum total proteins and serum globulin; Mortality; Body and organ weights; Hemorrhages in organs etc. were monitored in order to study the efficacy of the adsorbent for binding of aflatoxin B1 in the gastrointestinal tract of chickens. Statistical approach was adopted to analyze the data. It was found that adsorbent amount 0.3%/kg feed was highly effective to adsorb and detoxify aflatoxin B1 in gastrointestinal tract of broiler chickens and pass safely leaving no harmful effects. However the results of groups E and F fed on 0.4% and 0.5% respectively showed slight variation in tested parameters from group A. The prepared adsorbent was efficient for the detoxification of aflatoxin B1 in gastrointestinal tract of chicks and no negative symptoms associated with the use of activated carbon as previously reported were observed for the adsorbent under study.

  19. New analytical techniques for mycotoxins in complex organic matrices. [Aflatoxins B1, B2, G1, and G2

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bicking, M.K.L.

    1982-07-01

    Air samples are collected for analysis from the Ames Solid Waste Recovery System. The high level of airborne fungi within the processing area is of concern due to the possible presence of toxic mycotoxins, and carcinogenic fungal metabolites. An analytical method has been developed to determine the concentration of aflatoxins B1, B2, G1, and G2 in the air of the plant which produces Refuse Derived Fuel (RDF). After extraction with methanol, some components in the matrix are precipitated by dissolving the sample in 30% acetonitrile/chloroform. An aliquot of this solution is injected onto a Styragel column where the sample componentsmore » undergo simultaneous size exclusion and reverse phase partitioning. Additional studies have provided a more thorough understanding of solvent related non-exclusion effects on size exclusion gels. The Styragel column appears to have a useable lifetime of more than six months. After elution from Styragel, the sample is diverted to a second column containing Florisil which has been modified with oxalic acid and deactivated with water. Aflatoxins are eluted with 5% water/acetone. After removal of this solvent, the sample is dissolved in 150 ..mu..L of a spotting solvent and the entire sample applied to a thin layer chromatography (TLC) plate using a unique sample applicator developed here. The aflatoxins on the TLC plate are analyzed by laser fluorescence. A detection limit of 10 pg is possible for aflatoxin standards using a nitrogen laser as the excitation source. Sample concentrations are determined by comparing with an internal standard, a specially synthesized aflatoxin derivative. In two separate RDF samples, aflatoxin B1 was found at levels of 6.5 and 17.0 ppB. The analytical method has also proven useful in the analysis of contaminated corn and peanut meal samples. 42 figures, 8 tables.« less

  20. Modulation of aflatoxin toxicity and biomarkers by lycopene in F344 rats

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tang, Lili; Southern Yangtze University, Wuxi; Guan Hongxia

    Modulation by lycopene of aflatoxin B{sub 1} (AFB{sub 1})-induced toxic effects, metabolism, and metabolic activations was studied in young F344 rats. Animals were pretreated orally with either corn oil (control group) or lycopene [100 mg/kg body weight (b.w.), intervention group] 5 days/week for 2 weeks. Control animals were then treated daily with AFB{sub 1} (250 {mu}g/kg b.w) alone. Intervention animals were administered lycopene (100 mg/kg b.w.) at 1 h following a daily treatment with AFB{sub 1} (250 {mu}g/kg b.w.). Pretreatment and intervention with lycopene significantly reduced the toxic effect caused by AFB{sub 1} and greatly modulated AFB{sub 1} metabolism andmore » metabolic activation. Urinary excretion of AFB{sub 1} phase 1 metabolites, AFM{sub 1}, AFQ{sub 1}, and AFP{sub 1}, was significantly decreased in lycopene-treated animals. Formation of serum AFB{sub 1}-albumin adducts was also significantly reduced. The rate of reduction was from approximately 30% on day 1 (p < 0.05) to 67.7% on day 15 (p < 0.001). Lycopene intervention also significantly reduced formation of AFB{sub 1}-DNA adducts in liver compared to control animals, with the highest reduction (52.7%) occurring on day 3 (p < 0.05). Levels of AFB{sub 1}-N {sup 7}-guanine excreted in urine were also significantly decreased. Urinary excretion of the phase 2 detoxification metabolite, AFB{sub 1}-mecapturic acid, was significantly increased in lycopene-intervened animals. AFB{sub 1}-induced urinary excretion of 8-hydroxydeoxyguanosine was also reduced to 50% on day 7 after lycopene intervention. Collectively, these results suggest that inhibition of phase 1 metabolism and metabolic activation, as well as induction of phase 2 detoxification enzyme activity are the potential mechanisms for the chemopreventive effects of lycopene.« less

  1. Aflatoxin metabolism in humans: detection of metabolites and nucleic acid adducts in urine by affinity chromatography

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Groopman, J.D.; Donahue, P.R.; Zhu, J.Q.

    A high-affinity IgM monoclonal antibody specific for aflatoxins was covalently bound to Sepharose 4B and used as a preparative column to isolate aflatoxin derivatives from the urine of people and experimental animals who had been exposed to the carcinogen environmentally or under laboratory conditions. Aflatoxin levels were quantified by radioimmunoassay and high-performance liquid chromatography after elution from the affinity column. In studies on rats injected with ( UC)aflatoxin B1, the authors identified the major aflatoxin-DNA adduct, 2,3-dihydro-2-(N7-guanyl)-3-hydroxy-aflatoxin B1 (AFB1-N7-Gua), and the oxidative metabolites M1 and P1 as the major aflatoxin species present in the urine. When this methodology was appliedmore » to human urine samples obtained from people from the Guangxi Province of China exposed to aflatoxin B1 through dietary contamination, the aflatoxin metabolites detected were also AFB1-N7-Gua and aflatoxins M1 and P1. Therefore, affinity chromatography using a monoclonal antibody represents a useful and rapid technique with which to isolate this carcinogen and its metabolites in biochemical epidemiology and for subsequent quantitative measurements, providing exposure information that can be used for risk assessment.« less

  2. Chemistry and Biology of Aflatoxin-DNA Adducts

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Stone, Michael P.; Banerjee, Surajit; Brown, Kyle L.

    Aspergillus flavus is a fungal contaminant of stored rice, wheat, corn, and other grainstuffs, and peanuts. This is of concern to human health because it produces the mycotoxin aflatoxin B{sub 1} (AFB{sub 1}), which is genotoxic and is implicated in the etiology of liver cancer. AFB{sub 1} is oxidized in vivo by cytochrome P450 to form aflatoxin B{sub 1} epoxide, which forms an N7-dG adduct (AFB{sub 1}-N7-dG) in DNA. The latter rearranges to a formamidopyrimidine (AFB{sub 1}-FAPY) derivative that equilibrates between {alpha} and {beta} anomers of the deoxyribose. In DNA, both the AFB{sub 1}-N7-dG and AFB{sub 1}-{beta}-FAPY adducts intercalate abovemore » the 5'-face of the damaged guanine. Each produces G {yields} T transversions in Escherichia coli, but the AFB{sub 1}-{beta}-FAPY adduct is more mutagenic. The Sulfolobus solfataricus P2 DNA polymerase IV (Dpo4) provides a model for understanding error-prone bypass of the AFB{sub 1}-N7-dG and AFB{sub 1}-{beta}-FAPY adducts. It bypasses the AFB{sub 1}-N7-dG adduct, but it conducts error-prone replication past the AFB{sub 1}-FAPY adduct, including mis-insertion of dATP, consistent with the G {yields} T mutations characteristic of AFB{sub 1} mutagenesis in E. coli. Crystallographic analyses of a series of binary and ternary complexes with the Dpo4 polymerase revealed differing orientations of the N7-C8 bond of the AFB{sub 1}-N7-dG adduct as compared to the N{sup 5}-C8 bond in the AFB{sub 1}-{beta}-FAPY adduct, and differential accommodation of the intercalated AFB{sub 1} moieties within the active site. These may modulate AFB{sub 1} lesion bypass by this polymerase.« less

  3. The potential effects of Zataria multiflora Boiss essential oil on growth, aflatoxin production and transcription of aflatoxin biosynthesis pathway genes of toxigenic Aspergillus parasiticus.

    PubMed

    Yahyaraeyat, R; Khosravi, A R; Shahbazzadeh, D; Khalaj, V

    2013-01-01

    This study aims at evaluating the effects of Zataria multiflora (Z. multiflora) essential oil (EO) on growth, aflatoxin production and transcription of aflatoxin biosynthesis pathway genes. Total RNAs of Aspergillus parasiticus (A.parasiticus) ATCC56775 grown in yeast extract sucrose (YES) broth medium treated with Z. multiflora EO were subjected to reverse transcription- polymerase chain reaction (RT-PCR). Specific primers of nor-1, ver-1, omt-A and aflR genes were used. In parallel mycelial dry weight of samples were measured and all the media were assayed by high-pressure liquid chromatography (HPLC) for aflatoxinB1 (AFB1), aflatoxinB2 (AFB2), aflatoxinG1 (AFG1), aflatoxinG2 (AFG2) and aflatoxin total (AFTotal) production. The results showed that mycelial dry weight and aflatoxin production reduce in the presence of Z. multiflora EO (100 ppm) on day 5 of growth. It was found that the expression of nor-1, ver-1, omt-A and aflR genes was correlated with the ability of fungus to produce aflatoxins on day 5 in YES medium. RT-PCR showed that in the presence of Z.multiflora EO (100 ppm) nor-1, ver-1 and omtA genes expression was reduced. It seems that toxin production inhibitory effects of Z. multiflora EO on day 5 may be at the transcription level and this herb may cause reduction in aflatoxin biosynthesis pathway genes activity.

  4. The potential effects of Zataria multiflora Boiss essential oil on growth, aflatoxin production and transcription of aflatoxin biosynthesis pathway genes of toxigenic Aspergillus parasiticus

    PubMed Central

    Yahyaraeyat, R.; Khosravi, A.R.; Shahbazzadeh, D.; Khalaj, V.

    2013-01-01

    This study aims at evaluating the effects of Zataria multiflora (Z. multiflora) essential oil (EO) on growth, aflatoxin production and transcription of aflatoxin biosynthesis pathway genes. Total RNAs of Aspergillus parasiticus (A.parasiticus) ATCC56775 grown in yeast extract sucrose (YES) broth medium treated with Z. multiflora EO were subjected to reverse transcription- polymerase chain reaction (RT-PCR). Specific primers of nor-1, ver-1, omt-A and aflR genes were used. In parallel mycelial dry weight of samples were measured and all the media were assayed by high-pressure liquid chromatography (HPLC) for aflatoxinB1 (AFB1), aflatoxinB2 (AFB2), aflatoxinG1 (AFG1), aflatoxinG2 (AFG2) and aflatoxin total (AFTotal) production. The results showed that mycelial dry weight and aflatoxin production reduce in the presence of Z. multiflora EO (100 ppm) on day 5 of growth. It was found that the expression of nor-1, ver-1, omt-A and aflR genes was correlated with the ability of fungus to produce aflatoxins on day 5 in YES medium. RT-PCR showed that in the presence of Z.multiflora EO (100 ppm) nor-1, ver-1 and omtA genes expression was reduced. It seems that toxin production inhibitory effects of Z. multiflora EO on day 5 may be at the transcription level and this herb may cause reduction in aflatoxin biosynthesis pathway genes activity. PMID:24294264

  5. Survey of aflatoxins in retail samples of whole and ground black and white peppercorns.

    PubMed

    Adzahan, N; Jalili, M; Jinap, S

    2009-01-01

    A total of 126 local and imported samples of commercial white and black pepper in Malaysia were analysed for aflatoxins B1, B2, G1 and G2 (AFB1, AFB2, AFG1, AFG2) content using high-performance liquid chromatography (HPLC) with a fluorescence detector (FD). An acetonitrile-methanol-water (17 : 29 : 54; v/v) mixture was used as a mobile phase and clean-up was using an immunoaffinity column (IAC). Seventy out of 126 (55.5%) samples were contaminated with total aflatoxins, although only low levels of aflatoxins were found ranging from 0.1 to 4.9 ng g(-1). Aflatoxin B1 showed the highest incidence of contamination and was found in all contaminated samples. There was a significant difference between type of samples and different brands (p < 0.05). The results showed black peppers were more contaminated than white peppers.

  6. Supercritical Fluid Extraction of Aflatoxin B 1 from Soil

    EPA Science Inventory

    This research describes the development of a Supercritical Fluid Extraction (SFE) method to recover aflatoxin B1 from fortified soil. The effects of temperature, pressure, modifier (identity and percentage), and extraction type were assessed. Using the optimized SFE conditions, ...

  7. Development of structure switching aptamer assay for detection of aflatoxin M1 in milk sample.

    PubMed

    Sharma, Atul; Catanante, Gaëlle; Hayat, Akhtar; Istamboulie, Georges; Ben Rejeb, Ines; Bhand, Sunil; Marty, Jean Louis

    2016-09-01

    The discovery of in-vitro systematic evolution of ligands by exponential enrichment (SELEX) process has considerably broaden the utility of aptamer as bio-recognition element, providing the high binding affinity and specificity against the target analytes. Recent research has focused on the development of structure switching signaling aptamer assay, transducing the aptamer- target recognition event into an easily detectable signal. In this paper, we demonstrate the development of structure switching aptamer assay for determination of aflatoxin M1 (AFM1) employing the quenching-dequenching mechanism. Hybridization of fluorescein labelled anti-AFM1 aptamer (F-aptamer) with TAMRA labelled complementary sequences (Q-aptamer) brings the fluorophore and the quencher into close proximity, which results in maximum fluorescence quenching. On addition of AFM1, the target induced conformational formation of antiparallel G-quadruplex aptamer-AFM1 complex results in fluorescence recovery. Under optimized experimental conditions, the developed method showed the good linearity with limit of detection (LOD) at 5.0ngkg(-1) for AFM1. The specificity of the sensing platform was carefully investigated against aflatoxin B1 (AFB1) and ochratoxin A (OTA). The developed assay platform showed the high specificity towards AFM1. The practical application of the developed aptamer assay was verified for detection of AFM1 in spiked milk samples. Good recoveries were obtained in the range from 94.40% to 95.28% (n=3) from AFM1 spiked milk sample. Copyright © 2016. Published by Elsevier B.V.

  8. The Hotspot for (Global) One Health in Primary Food Production: Aflatoxin M1 in Dairy Products.

    PubMed

    Frazzoli, Chiara; Gherardi, Paola; Saxena, Navneet; Belluzzi, Giancarlo; Mantovani, Alberto

    2016-01-01

    One Health involves the multifaceted environment-animal-human web: nevertheless, the role of toxicological issues has yet to be fully explored in this context. Aflatoxin B1 (AFB1) contamination of feeds is a risk for the health of several farm animals, including fishes; milk is the only food of animal origin where a significant feed-food carry over may occur. The main AFB1-related compound present in milk is the hydroxy-metabolite aflatoxin M1 (AFM1). Besides contamination of raw milk, AFM1 is of concern for the whole dairy chain; AFM1 may also contaminate the milk of several other ruminants used for milk/dairy production. In a One Health perspective, milk represents a sentinel matrix for AFB1 vulnerability of the agro-food system, that is crucial in a phase when food/nutritional security becomes a global issue and climatic changes may affect agricultural productions. In the global setting, food chain exposure to long-term toxicants, such as AFM1, is a growing concern for economically developing countries, whereas global trade and climatic change makes AFM1 an emerging hot issue in economically developed countries as well. We critically review the state of the art on AFM1 risk assessment and risk management using two scenarios as case studies: a European Union country where the health system aims at ensuring a high-level protection of food chain (Italy) and the world's largest (and economically developing) producer of dairy products by volume (India). The case studies are used to provide building blocks for a global One Health framework.

  9. Co-occurence of aflatoxins and fumonisins in maize: guatemala as a case study

    USDA-ARS?s Scientific Manuscript database

    Aflatoxin B1 (AFB1) and fumonisin B1 (FB1) are found in maize. AFB1 is a genotoxic carcinogen (IARC Group 1) and FB1 a liver cancer promoter in rodents and trout (IARC Group 2B). Therefore, the possibility of co-exposure is a health concern, most notably in areas where maize serves as a dietary st...

  10. Effects of lactic acid bacteria and smectite after aflatoxin B1 challenge on the growth performance, nutrient digestibility and blood parameters of broilers.

    PubMed

    Liu, N; Ding, K; Wang, J; Deng, Q; Gu, K; Wang, J

    2018-04-11

    This study aimed to investigate the effect of lactic acid bacteria (LAB) and smectite on the growth performance, nutrient digestibility and blood parameters of broilers that were fed diets contaminated with aflatoxin B 1 (AFB 1 ). A total of 480 newly hatched male Arbor Acres broilers were randomly allocated into four groups with six replicates of 20 chicks each. The broilers were fed diets with the AFB 1 (40 μg/kg) challenge or without (control) it and supplemented with smectite (3.0 g/kg) or LAB (4.0 × 10 10  CFU/kg) based on the AFB 1 diet. The trial lasted for 42 days. The results showed that during days 1-42 of AFB 1 challenge, the feed intake (FI) and body weight gain (BWG) were depressed (p < .05). The inclusion of LAB and smectite increased (p < .05) the BWG by 71.58 and 41.89 g/bird, respectively, which reached the level of the control diet (p ≥ .05), but there were no differences (p ≥ .05) in performance between LAB and smectite. LAB and smectite also increased (p < .05) the apparent total tract digestibility of the crude protein. Regarding the blood parameters, AFB 1 decreased (p < .05) the levels of red blood cell count, haematocrit, mean corpuscular volume, haemoglobin, albumin and total protein. In the meantime, the AFB 1 increased (p < .05) leucocyte counts, urea nitrogen, cholesterol, total bilirubin, creatinine, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase and alkaline phosphatase. By contrast, LAB and smectite affected (p < .05) these parameters in the opposite direction. It can be concluded that after the AFB 1 challenge, LAB and smectite have similar effects on the growth and health of the broilers, suggesting that LAB could be an alternative against AFB 1 in commercial animal feeds. © 2018 Blackwell Verlag GmbH.

  11. Vitamin E (α tocopherol) attenuates toxicity and oxidative stress induced by aflatoxin in rats.

    PubMed

    Yılmaz, Seval; Kaya, Emre; Comakli, Selim

    2017-09-01

    Aflatoxins are toxic metabolites produced by Aspergillus flavus and Aspergillus parasiticus and are classified as group I carcinogens by the International Agency for Research on Cancer (IARC). The purpose of this study was to investigate the possible preventive role of vitamin E (Vit E) on aflatoxin (AF) induced toxicity by using biochemical and histopathological approaches. Wistar-Albino rats were divided into 4 groups as follows: control group, Vit E group (Vit E was administered), AFB1 group (a single dose of AFB1 was administered), AF + Vit E group (AF and Vit E were administered). The effects of Vit E on AFB1 induced tissue toxicity were evaluated by using malondialdehyde (MDA), reduced glutathione (GSH) levels, antioxidant enzyme activities, and histopathological examination in tissues. AF caused the oxidative stress by the increased MDA level and the reduced GSH level, glutathioneS-transferase (GST), catalase (CAT), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and glucose-6-phosphate dehydrogenase (G6PD) activities in tissues. Plasma aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities, creatinine, and urea concentrations significantly increased; whereas, chloride, phosphorus, and magnesium concentrations were insignificantly affected. Plasma glucose, protein and sodium concentrations significantly decreased. Administration of AF caused hepatotoxicity, cardiotoxicity, and nephrotoxicity. As far as histopathological changes are concerned, a statistically significant difference was found in AFB1 group compared to the control group. Vit E considerably reduced plasma AST, ALT, ALP, LDH activities, and urea concentration and ameliorated the deleterious effects of AF on oxidative stress markers and pathological changes. This data indicated that the natural antioxidant Vit E might have a protective effect against AF-induced toxicity and oxidative stress.

  12. Enhancement and Analysis of Human Antiaflatoxin B1 (AFB1) scFv Antibody-Ligand Interaction Using Chain Shuffling.

    PubMed

    Rangnoi, Kuntalee; Choowongkomon, Kiattawee; O'Kennedy, Richard; Rüker, Florian; Yamabhai, Montarop

    2018-06-06

    A human antiaflatoxin B1 (AFB1) scFv antibody (yAFB1-c3), selected from a naı̈ve human phage-displayed scFv library, was used as a template for improving and analysis of antibody-ligand interactions using the chain-shuffling technique. The variable-heavy and variable-light (VH/VL)-shuffled library was constructed from the VH of 25 preselected clones recombined with the VL of yAFB1-c3 and vice versa. Affinity selection from these libraries demonstrated that the VH domain played an important role in the binding of scFv to free AFB1. Therefore, in the next step, VH-shuffled scFv library was constructed from variable-heavy (VH) chain repertoires, amplified from the naı̈ve library, recombined with the variable-light (VL) chain of the clone yAFB1-c3. This library was then used to select a specific scFv antibody against soluble AFB1 by a standard biopanning method. Three clones that showed improved binding properties were isolated. Amino acid sequence analysis indicated that the improved clones have amino acid mutations in framework 1 (FR1) and the complementarity determining region (CDR1) of the VH chain. One clone, designated sAFH-3e3, showed 7.5-fold improvement in sensitivity over the original scFv clone and was selected for molecular binding studies with AFB1. Homology modeling and molecular docking were used to compare the binding of this and the original clones. The results confirmed that VH is more important than VL for AFB1 binding.

  13. How does airway exposure of aflatoxin B1 affect serum albumin adduct concentrations? Evidence based on epidemiological study and animal experimentation.

    PubMed

    Mo, Xianwei; Lai, Hao; Yang, Yang; Xiao, Jun; He, Ke; Liu, Chao; Chen, Jiansi; Lin, Yuan

    2014-08-01

    Aflatoxin B1 (AFB1) airway inhalation represents an additional route of exposure to this toxin. However, the association between AFB1 inhalation and serum AFB1 albumin adducts remains unclear. The aim of this study was to explore the association between airway exposure to AFB1 and serum AFB1 albumin adduct concentrations via an epidemiological study, as well as in an AFB1 airway exposure animal model. Our epidemiological study was conducted in a sugar factory in the Guangxi Autonomous Region of China. In order to examine fungal contamination, air samples were obtained in the workshop and areas outside the workshop, such as the office and nearby store. Dust samples were also collected from the bagasse warehouse and presser workshop, and were analyzed using an indirect competitive enzyme-linked immunosorbent assay (ELISA). Additionally, blood samples were collected from a total of 121 workshop workers, and a control group (n = 80) was comprised of workers who undertook administrative tasks or other work outside the workshop. The animal experiment was conducted in the laboratory animal center of Guangxi Medical University, where a total of 60 adult male rabbits were involved in this study. By intubation, AFB1 was administered in three groups of rabbits daily, at dose rates of 0.075, 0.05 and 0.025 mg/kg/day for a period of 7 days. Blood samples were collected on day 1, day 3, day 7 and day 21, and the measurements of the AFB1 albumin adducts in the serum were performed by a double antibody sandwich ELISA. The epidemiological study showed that serum albumin adducts were detected in 67 workshop workers (55.37%), and the values ranged 6.4 pg/mg albumin to 212 pg/mg albumin (mean value: 51 ± 4.62 pg/mg albumin). In contrast, serum albumin adducts were detected in only 7 control group participants, with the values ranging from 9 pg AFB1/mg albumin to 59 pg/mg albumin (mean value: 20 ± 13.72 pg/mg albumin). The animal experiment revealed that the rabbits had detectable

  14. Natural occurrence of aflatoxins in Chinese peanut butter and sesame paste.

    PubMed

    Li, Feng-Qin; Li, Yu-Wei; Wang, Ye-Ru; Luo, Xue-Yun

    2009-05-13

    A study on the natural occurrence of aflatoxins in Chinese peanut butter and sesame paste samples was conducted. Aflatoxin B(1) (AFB(1)) was the predominant toxin detected abundantly and frequently at a level up to 68.51 microg/kg in 41 of 50 peanut butter samples and 20.45 microg/kg in 37 of 100 sesame paste samples analyzed by liquid chromatography (LC). Of the AFB(1)-positive samples, 15 (37%) and 1 (2%) peanut butter samples with AFB(1) exceed European Union (EU) and Chinese regulations, respectively, whereas 19 and 32% of sesame paste samples contained AFB(1) higher than Chinese and EU regulations, respectively. Fourteen and 1 peanut butter samples and 10 and 7 sesame paste samples, respectively, will be legally claimed as positive, rejected, and even banned with consideration of an uncertainty of 40% for AFB(1), based on EU and Chinese regulations. Seeking to balance health benefits with the potential trade disruptions that regulations can cause is the issue of concern.

  15. Identification of averantin as an aflatoxin B1 precursor: placement in the biosynthetic pathway.

    PubMed Central

    Bennett, J W; Lee, L S; Shoss, S M; Boudreaux, G H

    1980-01-01

    A new blocked mutant of Aspergillus parasiticus produces no detectable aflatoxin B1, but accumulates several polyhydroxyanthraquinones. One of these pigments was identified as averantin. This is the first report of its formation by A. parasiticus. Radiotracer studies with [14C]averantin showed that 15.3% of label from averantin was incorporated into aflatoxin B1. This incorporation was blocked by dichlorvos. With radiotracers and other mutants, averantin was placed after norsolorinic acid and before averufin in the biosynthetic pathway in which the general steps are norsolorinic acid leads to averantin leads to averufin leads to versiconal hemiacetal acetate leads to versicolorin A leads to sterigmatocystin leads to aflatoxin B1. PMID:7377778

  16. Aflatoxins contamination of maize in Serbia: the impact of weather conditions in 2015.

    PubMed

    Janić Hajnal, Elizabet; Kos, Jovana; Krulj, Jelena; Krstović, Saša; Jajić, Igor; Pezo, Lato; Šarić, Bojana; Nedeljković, Nataša

    2017-11-01

    In recent years climate changes recorded in temperate regions of Europe have led to aflatoxin (AF) contamination of maize. Thus, the aim of this study was to investigate the influence of weather conditions on levels of aflatoxin B 1 (AFB1), aflatoxin B 2 (AFB2), aflatoxin G 1 (AFG1) and aflatoxin G 2 (AFG2) in 180 maize samples collected from the main maize-growing regions (Western Bačka, North Banat, South Banat and Central Serbia) in Serbia after harvest in 2015. The concentrations of AFs were determined by a validated HPLC method with post-column derivatisation and fluorescence detection (HPLC-FLD). The presence of AFB1, AFB2, AFG1 and AFG2 was detected in 57.2%, 13.9%, 5.6% and 2.8% of maize samples in the concentration ranges of 1.3-88.8 µg kg - 1 , 0.60-2.8 µg kg - 1 , 1.8-28.5 µg kg - 1 and 2.1-7.5 µg kg - 1 respectively. The recorded smaller amount of precipitation and especially higher air temperatures during the summer of 2015 were favourable for AF production, which resulted in 32.2% and 21.1% of samples being unsuitable for human consumption, since AFB1 and the sum of AFs concentrations were above 5.0 and 10.0 µg kg - 1 respectively. Furthermore, the findings in this study indicate that the microclimate conditions in the investigated regions had a great influence on the contamination frequency of maize with AFs. The highest percentage of samples unsuitable for human consumption, considering both AFB1 and total AFs content were 72.5% and 51.5% respectively from Central Serbia, whilst the lowest percentages of 15.6% and 6.2% respectively were found in Western Bačka. These findings confirmed that maize should be continuously monitored in order to protect human and animal health from the harmful effects caused by AFs contamination.

  17. Pulmonary interstitial fibrosis with evidence of aflatoxin B1 in lung tissue

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dvorackova, I.; Pichova, V.

    Three cases of pulmonary interstitial fibrosis, two in agricultural workers and one in a textile worker, are reported. In lung samples of all three patients the presence of aflatoxin B1 was demonstrated by radioimmunoassay (RIA). A possible occupational risk of aflatoxin exposure via the respiratory tract is suggested.

  18. A novel signal amplification technology for ELISA based on catalyzed reporter deposition. Demonstration of its applicability for measuring aflatoxin B(1).

    PubMed

    Bhattacharya, D; Bhattacharya, R; Dhar, T K

    1999-11-19

    In an earlier communication we have described a novel signal amplification technology termed Super-CARD, which is able to significantly improve antigen detection sensitivity in conventional Dot-ELISA by approximately 10(5)-fold. The method utilizes hitherto unreported synthesized electron rich proteins containing multiple phenolic groups which, when immobilized over a solid phase as blocking agent, markedly increases the signal amplification capability of the existing CARD method (Bhattacharya, R., Bhattacharya, D., Dhar, T.K., 1999. A novel signal amplification technology based on catalyzed reporter deposition and its application in a Dot-ELISA with ultra high sensitivity. J. Immunol. Methods 227, 31.). In this paper we describe the utilization of this Super-CARD amplification technique in ELISA and its applicability for the rapid determination of aflatoxin B(1) (AFB(1)) in infected seeds. Using this method under identical conditions, the increase in absorbance over the CARD method was approximately 400%. The limit of detection of AFB(1) by this method was 0.1 pg/well, the sensitivity enhancement being 5-fold over the optimized CARD ELISA. Furthermore, the total incubation time was reduced to 16 min compared to 50 min for the CARD method. Assay specificity was not adversely affected and the amount of AFB(1) measured in seed extracts correlated well with the values obtained by conventional ELISA.

  19. Determination of aflatoxins in edible oil from markets in Hebei Province of China by liquid chromatography-tandem mass spectrometry.

    PubMed

    Yang, Li-xin; Liu, Yin-ping; Miao, Hong; Dong, Bin; Yang, Na-jing; Chang, Feng-qi; Yang, Li-xue; Sun, Jing-bo

    2011-01-01

    Analysis of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in 76 edible oil samples (peanut oil, soybean oil, corn embryo oil and blended oil) was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The oils were sampled from three areas (Shijiazhuang, Baoding and Tangshan) of Hebei Province of China. AFB1 was detected in 22 samples representing 28.9%, followed by AFB2 (7.89%) and AFG1 (3.95%), while no AFG2 contamination was detected in any samples. AFB1 levels in oil samples ranged 0.14-2.72 µg kg(-1) and AFB2 ranged 0.15-0.36 µg kg(-1), while lower levels of 0.01-0.02 µg kg(-1) for AFG1 were recorded. The paper is part of an on-going investigation of aflatoxin contamination in Chinese edible oils.

  20. Separation of aflatoxin B1 from synthetic physiological fluids using talc and diatomite: Kinetic and isotherm aspects.

    PubMed

    Sprynskyy, Myroslav; Krzemień-Konieczka, Iwona; Gadzała-Kopciuch, Renata; Buszewski, Bogusław

    2018-01-01

    The objective of the study was to examine adsorption of the aflatoxin B1 from synthetic gastric fluid and synthetic intestinal fluid by talc, raw and calcined diatomite. The kinetic and equilibrium adsorption processes were studied in the batch adsorption experiments applying high performance liquid chromatography for the aflatoxin B1 determination. The kinetic study showed a very fast adsorption of the aflatoxin B1 onto the selected adsorbents from the both physiological fluids with reaching equilibrium within 1-15min. The aflatoxin B1 was almost completely adsorbed in initial linear step of the kinetic process that can be described well by the zero-order kinetics model. The experimental data of the equilibrium adsorption were characterized using the Langmuir and Freundlich isotherm models. The high adsorption effectiveness was found in a range of 90%-100% and 60%-100% for the diatomite samples and the talc respectively at the initial concentrations of the aflatoxin B1 as 31-300ng/mL. The possible mechanisms of the aflatoxin adsorption onto the used mineral adsorbents are also discussed in the work. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Carryover of aflatoxins from feed to lambari fish (Astyanax altiparanae) tissues.

    PubMed

    Michelin, E C; Massocco, M M; Godoy, S H S; Baldin, J C; Yasui, G S; Lima, C G; Rottinghaus, G E; Sousa, R L M; Fernandes, A M

    2017-02-01

    The aim of this study was to verify the carryover of aflatoxin B 1 from feed to lambari fish. Aflatoxins (AF) were incorporated into feed, checking the levels by HPLC. Treatments were: Control, feed without toxin; A, feed + 10 µg AFB 1  kg - 1 ; B, feed + 20 µg AFB 1  kg - 1 ; and C, feed + 50 µg AFB 1  kg - 1 . Juveniles of lambari fish were placed in 12 aquariums at a density of 50 fish/m 2 . Fish were fed twice a day with extruded feed, at 5% of animal biomass. The unit sample was constituted by a pool of 10 fish. AFs B 1 , B 2 , G 1 , G 2 and M 1 were quantified by HPLC in fish muscle and liver after 30, 60, 90 and 120 days of experiment. There was accumulation of AFs is fish liver and muscle, mainly after 90 days. Fish from treatment C had higher levels of AFB 1 in muscle when compared with the others, and AFB 1 in muscle at 120 days was similar to the levels in feed. Therefore, when lambari fish is exposed on a daily and long-term basis to AFs in feed, the regulation limits for AFs in animal feed do not guarantee safety for consumers.

  2. The Hotspot for (Global) One Health in Primary Food Production: Aflatoxin M1 in Dairy Products

    PubMed Central

    Frazzoli, Chiara; Gherardi, Paola; Saxena, Navneet; Belluzzi, Giancarlo; Mantovani, Alberto

    2017-01-01

    One Health involves the multifaceted environment-animal-human web: nevertheless, the role of toxicological issues has yet to be fully explored in this context. Aflatoxin B1 (AFB1) contamination of feeds is a risk for the health of several farm animals, including fishes; milk is the only food of animal origin where a significant feed-food carry over may occur. The main AFB1-related compound present in milk is the hydroxy-metabolite aflatoxin M1 (AFM1). Besides contamination of raw milk, AFM1 is of concern for the whole dairy chain; AFM1 may also contaminate the milk of several other ruminants used for milk/dairy production. In a One Health perspective, milk represents a sentinel matrix for AFB1 vulnerability of the agro-food system, that is crucial in a phase when food/nutritional security becomes a global issue and climatic changes may affect agricultural productions. In the global setting, food chain exposure to long-term toxicants, such as AFM1, is a growing concern for economically developing countries, whereas global trade and climatic change makes AFM1 an emerging hot issue in economically developed countries as well. We critically review the state of the art on AFM1 risk assessment and risk management using two scenarios as case studies: a European Union country where the health system aims at ensuring a high-level protection of food chain (Italy) and the world’s largest (and economically developing) producer of dairy products by volume (India). The case studies are used to provide building blocks for a global One Health framework. PMID:28210616

  3. Individual and combined effects of Aflatoxin B1, Deoxynivalenol and Zearalenone on HepG2 and RAW 264.7 cell lines.

    PubMed

    Zhou, Hongyuan; George, Saji; Hay, Crystal; Lee, Joel; Qian, He; Sun, Xiulan

    2017-05-01

    To understand the combinatorial toxicity of mycotoxins, we measured the effects of individual, binary and tertiary combinations of Aflatoxin B 1 (AFB 1 ), Deoxynivalenol (DON) and Zearalenone (ZEN) on the cell viability and cellular perturbations of HepG2 and RAW 264.7 cells. The nature of mycotoxins interactions was assessed using mathematical modeling (Chou-Talalay). Mechanisms of cytotoxicity were studied using high content screening (HCS) that probed cytotoxicity responses, such as changes in intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP), intracellular calcium ([Ca 2+ ] i ) flux, and cell membrane damage. Our results showed that individual cytotoxicity of mycotoxins in a decreasing order was DON>AFB 1 >ZEN. Varying combinations of mycotoxins at differing concentrations showed different types of interactions. Most of the mixtures showed increasing toxic effects-synergism and/or addition while antagonistic effects were observed with combination of AFB 1 +ZEN. Generally, combination of mycotoxins showed significantly increased intracellular ROS production and [Ca 2+ ] i flux, and decreased MMP in both cell lines, showing that the synergistic and additive effects of mycotoxin combination originate from perturbations of multiple cellular functions. Additionally, this study demonstrated the applicability of HCS for gaining mechanistic understanding on the toxicity of individual as well as combinatorial mycotoxins, and also provided scientific bases for formulating regulatory policies. Copyright © 2017. Published by Elsevier Ltd.

  4. Rapid on-site sensing aflatoxin B1 in food and feed via a chromatographic time-resolved fluoroimmunoassay.

    PubMed

    Zhang, Zhaowei; Tang, Xiaoqian; Wang, Du; Zhang, Qi; Li, Peiwu; Ding, Xiaoxia

    2015-01-01

    Aflatoxin B1 poses grave threats to food and feed safety due to its strong carcinogenesis and toxicity, thus requiring ultrasensitive rapid on-site determination. Herein, a portable immunosensor based on chromatographic time-resolved fluoroimmunoassay was developed for sensitive and on-site determination of aflatoxin B1 in food and feed samples. Chromatographic time-resolved fluoroimmunoassay offered a magnified positive signal and low signal-to-noise ratio in time-resolved mode due to the absence of noise interference caused by excitation light sources. Compared with the immunosensing performance in previous studies, this platform demonstrated a wider dynamic range of 0.2-60 μg/kg, lower limit of detection from 0.06 to 0.12 µg/kg, and considerable recovery from 80.5% to 116.7% for different food and feed sample matrices. It was found to be little cross-reactivity with other aflatoxins (B2, G1, G2, and M1). In the case of determination of aflatoxin B1 in peanuts, corn, soy sauce, vegetable oil, and mouse feed, excellent agreement was found when compared with aflatoxin B1 determination via the conversational high-performance liquid chromatography method. The chromatographic time-resolved fluoroimmunoassay affords a powerful alternative for rapid on-site determination of aflatoxin B1 and holds a promise for food safety in consideration of practical food safety and environmental monitoring.

  5. Rapid On-Site Sensing Aflatoxin B1 in Food and Feed via a Chromatographic Time-Resolved Fluoroimmunoassay

    PubMed Central

    Wang, Du; Zhang, Qi; Li, Peiwu; Ding, Xiaoxia

    2015-01-01

    Aflatoxin B1 poses grave threats to food and feed safety due to its strong carcinogenesis and toxicity, thus requiring ultrasensitive rapid on-site determination. Herein, a portable immunosensor based on chromatographic time-resolved fluoroimmunoassay was developed for sensitive and on-site determination of aflatoxin B1 in food and feed samples. Chromatographic time-resolved fluoroimmunoassay offered a magnified positive signal and low signal-to-noise ratio in time-resolved mode due to the absence of noise interference caused by excitation light sources. Compared with the immunosensing performance in previous studies, this platform demonstrated a wider dynamic range of 0.2-60 μg/kg, lower limit of detection from 0.06 to 0.12 µg/kg, and considerable recovery from 80.5% to 116.7% for different food and feed sample matrices. It was found to be little cross-reactivity with other aflatoxins (B2, G1, G2, and M1). In the case of determination of aflatoxin B1 in peanuts, corn, soy sauce, vegetable oil, and mouse feed, excellent agreement was found when compared with aflatoxin B1 determination via the conversational high-performance liquid chromatography method. The chromatographic time-resolved fluoroimmunoassay affords a powerful alternative for rapid on-site determination of aflatoxin B1 and holds a promise for food safety in consideration of practical food safety and environmental monitoring. PMID:25874803

  6. Further evidence supporting the concurrent influence of aflatoxin and manganese

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Katzen, J.S.; Llewellyn, G.C.

    Trace elements, including manganese may afford protection from deleterious effects of aflatoxin. Young male Fischer rats received ip injections of aflatoxin B1 (AFB1) in dimethyl sulfoxide (DMSO), 1 mg/kg, 2 mg/kg or 4 mg/kg. Control groups received DMSO ip or no injection. All animals were intubated with 3 microCi of (/sup 54/Mn)-MnCl/sub 2/ 12 hr post-injection. Sacrifice occurred 72 hr after gavage of the radiolabel. All tested levels of AFB1 affected the loss of total body radioactivity. This response was observed within 12 hr when toxin-treated groups excreted almost 4 times more counts than controls. From 12-36 hr following radiolabelmore » administration, AFB1 appeared to enhance excretion; by 72 hr, toxin-treated animals (especially those receiving higher doses) appeared to conserve the metal. Aflatoxicosis manifested itself through reduced body weight gain. The data provide support evidence that Mn and AFB1 biointeract.« less

  7. The toxicity and decreased concentration of aflatoxin B in natural lactic acid fermented maize meal.

    PubMed

    Mokoena, M P; Chelule, P K; Gqaleni, N

    2006-04-01

    Aflatoxin B(1) (AFB(1)) is a mycotoxin which is known to frequently contaminate poorly stored food products destined for human consumption. This study was carried out to investigate the potential activity of lactic acid fermentation in reducing AFB(1) level in fermented maize meal products. Maize meal was spiked with 60 mug g(-1) AFB(1) and fermented, with or without starter culture, for 4 days at 25 degrees C. Unbound AFB(1) in solution and the pH of the media were monitored daily. A significant decrease (P < 0.05) in the level of unbound AFB(1) was observed (75% in the fourth day). Simultaneously, a progressive decrease in the pH of the media from 6.5 to 3.1 was also observed. AFB(1) was below the detection limit in commercial fermented porridge (amahewu) samples. Cytotoxicity tests on AFB(1)-spiked fermented extracts showed that those with a starter culture were comparatively less toxic (30-36%) than those with no added starter culture (24-30%). However, this difference was not significant (P > 0.05). These results indicate that lactic acid fermentation can significantly reduce the concentration of AFB(1) in maize to trace levels. However, the safety of fermented products has not been well studied, as the mechanism of AFB(1) removal is not well understood. Natural fermentation may potentially reduce exposure to natural toxins occurring in food.

  8. Effects of nanostructured zeolite and aflatoxin B1 in growth performance, immune parameters and pathological conditions of rainbow trout Oncorhynchus mykiss.

    PubMed

    Alinezhad, S; Faridi, M; Falahatkar, B; Nabizadeh, R; Davoodi, D

    2017-11-01

    The reduction of Aflatoxin B 1 (AF) in juvenile rainbow trout (Oncorhynchus mykiss) diet was analyzed after supplementing Nanostructured Zeolite (NZ) in a 56-day experiment. Two hundred and seventy juveniles with an average weight of 23 ± 3.7 g were placed in 6 different groups of C (control as a basal diet), NZ 0.5 (basal diet + 0.5% NZ), NZ 1 (basal diet + 1% NZ), AF 5 (basal diet + 5 mg AFB 1 ), AF 5 NZ 0.5 (basal diet + 5 mg AFB 1  + 0.5% NZ), AF 5 NZ 1 (basal diet + 5 mg AFB 1  + 1% NZ) with three replications and were fed four times a day based on their satiation. No significant differences were observed in terms of growth performance among the experimental groups (P > 0.05). However, hepatosomatic index in fish fed by AF 5 NZ 0.5 was reduced compared with NZ 0.5 group (P < 0.05). The carcass moisture content showed a higher amount in treatment AF 5 NZ 0.5 compared to the control group (P < 0.05). There was a decrease in fat content in treatment AF 5 compared to that of the control group (P < 0.05). Serum total protein, albumin and globulin levels in fish fed with aflatoxin were lower than in fish fed the diet without AF for all levels of NZ (P < 0.05); however, the interaction between AF and NZ was not significant (P > 0.05). Concentrations of C 3 , C 4 and immunoglobulin M together with serum lysozyme activity showed no significant differences among all treatments (P > 0.05). No considerable histopathological lesions were observed in liver, kidney and spleen for all treatments. Based on the results, NZ showed some effects on physiological functions in juvenile rainbow trout fed by 0.5% dietary NZ which could improve performance in this species. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Averufanin is an aflatoxin B1 precursor between averantin and averufin in the biosynthetic pathway.

    PubMed Central

    McCormick, S P; Bhatnagar, D; Lee, L S

    1987-01-01

    Wild-type Aspergillus parasiticus produces, in addition to the colorless aflatoxins, a number of pigmented secondary metabolites. Examination of these pigments demonstrated that a major component was an anthraquinone, averufanin. Radiolabeling studies with [14C]averufanin showed that 23% of the label was incorporated into aflatoxin B1 by the wild type and that 31% of the label was incorporated into O-methylsterigmatocystin by a non-aflatoxin-producing isolate. In similar studies with blocked mutants of A. parasiticus the 14C label from averufanin was accumulated in averufin (72%) and versicolorin A (54%) but not averantin. The results demonstrate that averufanin is a biosynthetic precursor of aflatoxin B1 between averantin and averufin. PMID:3103529

  10. Aflatoxin Toxicity Reduction in Feed by Enhanced Binding to Surface-Modified Clay Additives

    PubMed Central

    Jaynes, William F.; Zartman, Richard E.

    2011-01-01

    Animal feeding studies have demonstrated that clay additives, such as bentonites, can bind aflatoxins in ingested feed and reduce or eliminate the toxicity. Bentonite deposits are found throughout the world and mostly consist of expandable smectite minerals, such as montmorillonite. The surfaces of smectite minerals can be treated with organic compounds to create surface-modified clays that more readily bind some contaminants than the untreated clay. Montmorillonites treated with organic cations, such as hexadecyltrimethylammonium (HDTMA) and phenyltrimethylammonium (PTMA), more effectively remove organic contaminants, such as benzene and toluene, from water than untreated clay. Similarly, montmorillonite treated with PTMA (Kd = 24,100) retained more aflatoxin B1 (AfB1) from aqueous corn flour than untreated montmorillonite (Kd = 944). Feed additives that reduced aflatoxin toxicity in animal feeding studies adsorbed more AfB1 from aqueous corn flour than feed additives that were less effective. The organic cations HDTMA and PTMA are considered toxic and would not be suitable for clay additives used in feed or food, but other non-toxic or nutrient compounds can be used to prepare surface-modified clays. Montmorillonite (SWy) treated with choline (Kd = 13,800) and carnitine (Kd = 3960) adsorbed much more AfB1 from aqueous corn flour than the untreated clay (Kd = 944). A choline-treated clay prepared from a reduced-charge, high-charge montmorillonite (Kd = 20,100) adsorbed more AfB1 than the choline-treated high-charge montmorillonite (Kd = 1340) or the untreated montmorillonite (Kd = 293). Surface-modified clay additives prepared using low-charge smectites and nutrient or non-toxic organic compounds might be used to more effectively bind aflatoxins in contaminated feed or food and prevent toxicity. PMID:22069725

  11. Influence of gamma-irradiation and maize lipids on the production of aflatoxin B1 by Aspergillus flavus.

    PubMed

    Aziz, Nagy H; el-Zeany, Samia A; Moussa, Lotfy A A

    2002-10-01

    The effect of gamma-irradiation and maize lipids on aflatoxin B1 production by Aspergillus flavus artificially inoculated into sterilized maize at reduced water activity (aw 0.84) was investigated. By increasing the irradiation doses the total viable population of A. flavus decreased and the fungus was completely inhibited at 3.0 kGy. The amounts of aflatoxin B1 were enhanced at irradiation dose levels 1.0 and 1.5 kGy in both full-fat maize (FM) and defatted maize (DM) media and no aflatoxin B1 production at 3.0 kGy gamma-irradiation over 45 days of storage was observed. The level in free lipids of FM decreased gradually, whereas free fatty acid values and fungal lipase activity increased markedly by increasing the storage periods. The free fatty acid values decreased by increasing the irradiation dose levels and there was a significant enhancement of fungal lipase activity at doses of 1.0 and 1.50 kGy. The ability of A. flavus to grow at aw 0.84 and produce aflatoxin B1 is related to the lipid composition of maize. The enhancement of aflatoxin B1 at low doses was correlated to the enhancement of fungal lipase activity.

  12. Aflatoxin and ochratoxin A content of spices in Hungary.

    PubMed

    Fazekas, B; Tar, A; Kovács, M

    2005-09-01

    In October and November 2004, 91 spice samples (70 ground red pepper, six black pepper, five white pepper, five spice mix and five chilli samples), the majority of which originated from commercial outlets, were analysed for aflatoxins B1, B2, G1 and G2 (AFB1, AFB2, AFG1, AFG2) and ochratoxin A (OTA) content by high-performance liquid chromatography (HPLC) after immunoaffinity column clean-up. Eighteen of the 70 ground red pepper samples contained AFB1, seven of them in a concentration exceeding the 'maximum level' of 5 microg kg(-1) (range 6.1-15.7 microg kg(-1)). Of the other spices assayed, the AFB1 contamination of one chilli sample exceeded 5 microg kg(-1) (8.1 microg kg(-1)). Thirty-two of the 70 ground red pepper samples contained OTA, eight of them in a concentration exceeding the 10 microg kg(-1) 'maximum level' (range 10.6-66.2 microg kg(-1)). One chilli sample was contaminated with OTA at 2.1 microg kg(-1). The AFB1 and OTA contamination of ground red pepper exceeding the 'maximum level' (5 and 10 microg kg(-1), respectively) was obviously the consequence of mixing imported ground red pepper batches heavily contaminated with AFB1 and OTA with red pepper produced in Hungary. This case calls attention to the importance of consistently screening imported batches of ground red pepper for aflatoxin and ochratoxin A content and strictly prohibiting the use of batches containing mycotoxin concentrations exceeding the maximum permitted level.

  13. Surveys of rice sold in Canada for aflatoxins, ochratoxin A and fumonisins

    PubMed Central

    Bansal, J.; Pantazopoulos, P.; Tam, J.; Cavlovic, P.; Kwong, K.; Turcotte, A.-M.; Lau, B.P.-Y.; Scott, P.M.

    2011-01-01

    Approximately 200 samples of rice (including white, brown, red, black, basmati and jasmine, as well as wild rice) from several different countries, including the United States, Canada, Pakistan, India and Thailand, were analysed for aflatoxins, ochratoxin A (OTA) and fumonisins by separate liquid Chromatographic methods in two different years. The mean concentrations for aflatoxin B1 (AFB1) were 0.19 and 0.17 ng g−1 with respective positive incidences of 56% and 43% (≥ the limit of detection (LOD) of 0.002 ng g−1). Twenty-three samples analysed in the second year also contained aflatoxin B2 (AFB2) at levels ≥LOD of 0.002 ng g−1 The five most contaminated samples in each year contained 1.44–7.14 ng AFB1 g−1 (year 1) and 1.45–3.48 ng AFB1 g−1 (year 2); they were mostly basmati rice from India and Pakistan and black and red rice from Thailand. The average concentrations of ochratoxin A (OTA) were 0.05 and 0.005 ng g−1 in year 1 and year 2, respectively; incidences of samples containing ≥LOD of 0.05 ng g−1 were 43% and 1%, respectively, in the 2 years. All positive OTA results were confirmed by LC-MS/MS. For fumonisins, concentrations of fumonisin B1 (FB1) averaged 4.5 ng g−1 in 15 positive samples (≥0.7 ng g−1) from year 1 (n = 99); fumonisin B2 (FB2) and fumonisin B3 (FB3) were also present (≥1 ng g−1). In the second year there was only one positive sample (14 ng g−1 FB1) out of 100 analysed. All positive FB1 results were confirmed by LC-MS/MS. PMID:21623501

  14. Effect of γ irradiation on fungal load and aflatoxins reduction in red chillies

    NASA Astrophysics Data System (ADS)

    Iqbal, Shahzad Zafar; Bhatti, Ijaz Ahmad; Asi, Muhammad Rafique; Zuber, Mohammad; Shahid, Muhammad; Parveen, Ishrat

    2013-01-01

    Chillies are a very important cash crop of Pakistan. The effects of gamma irradiation on microbial load, aflatoxin B1 (AFB1) and total aflatoxins have been studied in chillies samples, collected from different districts of Punjab, Pakistan. Aflatoxins were analyzed using HPLC equipped with a fluorescence detector. The results revealed that among the Aspergillus species isolated, those belonging to section parasiticus were predominant. Gamma radiations of doses 2, 4 and 6 kGy were employed on fungi and chilli samples. The results have demonstrated that the dose of 6 kGy reduced the fungal load by 5 logs. Furthermore, 6 kGy reduced the level of AFB1 and total AFs in ground and whole chillies by 1-2 logs (α < 0.05).

  15. Detection of aflatoxin M1 in powdered milk and sweetened condensed milk products in several cities in Java with HPLC-fluorescence method

    NASA Astrophysics Data System (ADS)

    Wijaya, H.; Wardayanie, N. I.; Widjajanti, R.; Silitonga, R. F.

    2018-01-01

    Aflatoxin M1 (AFM1) is a hydroxylated metabolite of aflatoxin B1 (AFB1) produced by lactating animals due to consuming AFB1-contaminated feed. AFM1 can be found in dairy products because it is resistant to heat during processing. This study aimed to detect AFM1 in powdered milk and sweetened condensed milk sold in several cities in Java. The amount of powdered milk sample was 20, while the amount of sweetened condensed milk sample was 16. AFM1 detection in powdered milk and sweetened condensed milk was conducted by HPLC-fluorescence method. The results showed that the concentration of AFM1 in powdered milk ranged from undetectable to 0.549 μg/kg and the highest data (55%) was distributed in concentration range of >0.05 μg/kg - 0.2 μg/kg. On the other hand, AFM1 levels in sweetened condensed milk ranged from undetectable to 0.056 μg/kg and 43.75% data was distributed in concentration range of >0.025 μg/kg - 0.05 μg/kg. All powdered milk and sweetened condensed milk samples have met the maximum level of AFM1 according to Indonesian regulation.

  16. Assessment of aflatoxin exposure using serum and urinary biomarkers in São Paulo, Brazil: A pilot study.

    PubMed

    Jager, Alessandra V; Tonin, Fernando G; Baptista, Gabriela Z; Souto, Pollyana C M C; Oliveira, Carlos A F

    2016-05-01

    The aim of this study was to evaluate the human exposure of individuals from Pirassununga, Brazil, to dietary aflatoxins B1 (AFB1) and M1 (AFM1) by determination of serum AFB1-lysine and urinary aflatoxin biomarkers (AFM1 and AFB1-N(7)-guanine). The participants were recruited among employees from a Campus of the University of São Paulo, which provided food samples from their homes, as well as serum and urine samples four times every three months, from June 2011 until March 2012. The probable daily intake (PDI) of aflatoxin was estimated by using the results from analysis of food products collected by the time of samples collection, and data from a 24-hour dietary recall questionnaire. Analyses of AFB1 and AFM1 in food samples were conducted by high-performance liquid chromatography with fluorescence detection. Biomarkers in serum and urine were determined by tandem mass spectrometry. AFB1 and AFM1 were detected in 38 samples of cereals (28%, N=136) and 31 milk products (36%, N=86), respectively. AFB1-lysine and AFB1-N(7)-guanine and were not detected in serum or urine samples, respectively. However, AFM1 was found in 74 urine samples (65%), at mean levels in the 4 sampling times ranging from 0.37±0.23 to 1.70±2.88pg/mg creatinine. The mean PDI varied among different sampling times, ranging from 0.09±0.09 to 1.35±5.98ng/kg body weight/day. A modest though significant correlation (r=0.45; p=0.03; N=23) was found for the first time in Brazil between the AFM1 concentration in urine and the PDI for total aflatoxins (AFB1+AFM1) in sampling 1 (June 2011). Urinary AFM1 was confirmed as very sensitive for monitoring the human exposure to dietary aflatoxin. Further studies using serum and urinary biomarkers are needed to estimate the aflatoxin exposure of populations in higher risk areas in Brazil. Copyright © 2015 Elsevier GmbH. All rights reserved.

  17. Ameliorating Effects of Bacillus subtilis ANSB060 on Growth Performance, Antioxidant Functions, and Aflatoxin Residues in Ducks Fed Diets Contaminated with Aflatoxins

    PubMed Central

    Zhang, Liyuan; Ma, Qiugang; Ma, Shanshan; Zhang, Jianyun; Jia, Ru; Ji, Cheng; Zhao, Lihong

    2016-01-01

    Bacillus subtilis ANSB060 isolated from fish gut is very effective in detoxifying aflatoxins in feed and feed ingredients. The purpose of this research was to investigate the effects of B. subtilis ANSB060 on growth performance, body antioxidant functions, and aflatoxin residues in ducks fed moldy maize naturally contaminated with aflatoxins. A total of 1500 18-d-old male Cherry Valley ducks with similar body weight were randomly assigned to five treatments with six replicates of 50 ducks per repeat. The experiment design consisted of five dietary treatments labeled as C0 (basal diet containing 60% normal maize), M0 (basal diet containing 60% moldy maize contaminated with aflatoxins substituted for normal maize), M500, M1000, and M2000 (M0 +500, 1000 or 2000 g/t aflatoxin biodegradation preparation mainly consisted of B. subtilis ANSB060). The results showed that ducks fed 22.44 ± 2.46 μg/kg of AFB1 (M0) exhibited a decreasing tendency in average daily gain (ADG) and total superoxide dismutase (T-SOD) activity in serum, and T-SOD and glutathione peroxidase (GSH-Px) activities in the liver significantly decreased along with the appearance of AFB1 and AFM1 compared with those in Group C0. The supplementation of B. subtilis ANSB060 into aflatoxin-contaminated diets increased the ADG of ducks (p > 0.05), significantly improved antioxidant enzyme activities, and reduced aflatoxin accumulation in duck liver. In conclusion, Bacillus subtilis ANSB060 in diets showed an ameliorating effect to duck aflatoxicosis and may be a promising feed additive. PMID:28025501

  18. Aflatoxins, discolouration and insect damage in dried cowpea and pigeon pea in Malawi and the effectiveness of flotation/washing operation in eliminating the aflatoxins.

    PubMed

    Matumba, Limbikani; Singano, Lazarus; Pungulani, Lawrent; Mvula, Naomi; Matumba, Annie; Singano, Charles; Matita, Grey

    2017-05-01

    Aflatoxin contamination and biodeterioration were examined in 302 samples of dry cowpeas and pigeon peas that were randomly purchased from 9 districts of the Southern Region of Malawi during July and November 2015. Further, the impact of flotation/washing on aflatoxin levels on the pulses was elucidated. Aflatoxin analyses involved immunoaffinity column (IAC) clean-up and HPLC quantification with fluorescence detection (FLD) while legume biodeterioration assessments were done by visual inspection. Aflatoxins were frequently detected in cowpea (24%, max., 66 μg/kg) and pigeon pea (22%, max., 80 μg/kg) samples that were collected in the month of July. Lower aflatoxin incidence of 15% in cowpeas (max., 470 μg/kg) and 14% in pigeon peas (max., 377 μg/kg) was recorded in the November collection. Overall, aflatoxin levels were significantly higher in the pulses that were collected in November. However, there were no significant differences in the total aflatoxin (aflatoxin B 1 (AFB 1 ) + AFB 2 + AFG 1 + AFG 2 ) levels between the two types of pulses. Remarkably, in 76.2% of the aflatoxin positive cowpea and in 41.7% of the aflatoxin positive pigeon pea samples, aflatoxin G 1 concentration exceeded aflatoxin B 1. Insect damage percentage averaged at 18.1 ± 18.2% (mean ± SD) in the cowpeas and 16.1 ± 19.4% in pigeon peas. Mean discolouration percentage (number of pulses) of the cowpeas and pigeon peas was found to be at 6.7 ± 4.9 and 8.7 ± 6.2%, respectively. Washing and discarding the buoyant fraction was highly efficient in reducing aflatoxin levels; only 5.2 ± 11.1% of the initial aflatoxin level was found in the cleaned samples. In conclusion, cowpeas and pigeon peas sold on the local market in Malawi may constitute a hazard especially if floatation/washing step is skipped.

  19. Immunotoxicity of ochratoxin A and aflatoxin B1 in combination is associated with the nuclear factor kappa B signaling pathway in 3D4/21 cells.

    PubMed

    Hou, Lili; Gan, Fang; Zhou, Xuan; Zhou, Yajiao; Qian, Gang; Liu, Zixuan; Huang, Kehe

    2018-05-01

    The co-contamination of cereals, grains, crops, and animal feeds by mycotoxins is a universal problem. Humans and animals are exposed to several mycotoxins simultaneously as evidenced by extensive studies on this topic. Yet, most studies have addressed the effects of mycotoxins individually. Aflatoxin B1 and ochratoxin A can induce immunotoxicity. However, it remains unclear whether a combination of these mycotoxins aggravates immunotoxicity and the potential mechanism underlying this effect. In this study, we used the cell line 3D4/21, swine alveolus macrophages and innate immune cell. The results showed that the percentage of cell inhibition, annexin V/PI-positive rates, and the expression of pro-inflammatory cytokines (tumor necrosis factor alpha and interleukin-6) significantly increased and the release of lactate dehydrogenase and phagocytotic index were significantly decreased at different concentrations of aflatoxin B1 and ochratoxin A combination when compared with control. The combination of aflatoxin B1 and ochratoxin A significantly decreased the production of GSH and increased reactive oxygen species level. However, N-acetylcysteine suppressed the oxidative stress and alleviated the immunotoxicity induced by the combination. The combination of aflatoxin B1 and ochratoxin A markedly enhanced the degradation of IκBa, the phosphorylation of nuclear factor kappa B (p65), and the translocation of activated nuclear factor kappa B (NF-κB) into the nuclei as demonstrated by western blotting and confocal laser scanning microscopy. These effects could be reversed by BAY 11-7082, a specific inhibitor of NF-κB. Taken together, a combination of aflatoxin B1 and ochratoxin A could aggravate immunotoxicity by activating the NF-κB signaling pathway. Copyright © 2018 Elsevier Ltd. All rights reserved.

  20. Aflatoxin toxicity reduction in feed by enhanced binding to surface-modified clay additives.

    PubMed

    Jaynes, William F; Zartman, Richard E

    2011-06-01

    Animal feeding studies have demonstrated that clay additives, such as bentonites, can bind aflatoxins in ingested feed and reduce or eliminate the toxicity. Bentonite deposits are found throughout the world and mostly consist of expandable smectite minerals, such as montmorillonite. The surfaces of smectite minerals can be treated with organic compounds to create surface-modified clays that more readily bind some contaminants than the untreated clay. Montmorillonites treated with organic cations, such as hexadecyltrimethylammonium (HDTMA) and phenyltrimethylammonium (PTMA), more effectively remove organic contaminants, such as benzene and toluene, from water than untreated clay. Similarly, montmorillonite treated with PTMA (K(d) = 24,100) retained more aflatoxin B1 (AfB1) from aqueous corn flour than untreated montmorillonite (K(d) = 944). Feed additives that reduced aflatoxin toxicity in animal feeding studies adsorbed more AfB1 from aqueous corn flour than feed additives that were less effective. The organic cations HDTMA and PTMA are considered toxic and would not be suitable for clay additives used in feed or food, but other non-toxic or nutrient compounds can be used to prepare surface-modified clays. Montmorillonite (SWy) treated with choline (K(d) = 13,800) and carnitine (K(d) = 3960) adsorbed much more AfB1 from aqueous corn flour than the untreated clay (K(d) = 944). A choline-treated clay prepared from a reduced-charge, high-charge montmorillonite (K(d) = 20,100) adsorbed more AfB1 than the choline-treated high-charge montmorillonite (K(d) = 1340) or the untreated montmorillonite (K(d) = 293). Surface-modified clay additives prepared using low-charge smectites and nutrient or non-toxic organic compounds might be used to more effectively bind aflatoxins in contaminated feed or food and prevent toxicity.

  1. Development of Lateral Flow Immunochromatographic Strips for Micropollutant Screening Using Colorants of Aptamer-Functionalized Nanogold Particles, Part II: Experimental Verification with Aflatoxin B1 and Chloramphenicol.

    PubMed

    Zhang, Shan; Zhao, Shuai; Wang, Sai; Liu, Jiahui; Dong, Yiyang

    2018-05-09

    Lateral flow immunochromatographic strips based on colorants of aptamer-functionalized nanogold particles weredeveloped for the detection of micropollutants aflatoxin B1 (AFB1) and chloramphenicol (CAP). The lateral flow immunochromatographic strip was based on a competitive reaction of thiolated-aptamer between micropollutants and bio-DNA probe-streptavidin as capture material immobilized at the test line. General crucial parameters that might influence the sensitivity have been systematically investigated. To test the effectiveness and applicability of the optimized conditions, two structurally unrelated micropollutants, that is, AFB1 and CAP, were chosen for detection. In the present study, lateral flow immunochromatographic strips for AFB1 and CAP analysis by combining the high selectivity and affinity of aptamers with the unique optical properties of nanogold in municipal water samples were reported for the first time. With the optimized conditions, the immunochromatographic strip showed a visual LOD of 10 ppb and a quantitative LOD of 1.05 ppb using an immunochromatographic reader for AFB1 detection and a quantitative LOD of 63.4 ppb using an immunochromatographic reader for CAP detection. Furthermore, the sensitive strip provided a good linear detection range of approximately 0-50 ppm for AFB1 detection and a wider liner detection range of approximately 0-160 ppm for CAP detection. Moreover, the immunochromatographic strip provided recovery rates for water samples of 90-110% in the AFB1 analysis and 84-108% in the CAP analysis. The results demonstrated that the immunochromatographic strip has excellent potential for wide applicability and verified that the strip methods for the optimized conditions are applicable to a variety of micropollutants. The lateral flow immunochromatographic strip could be used as a simple, rapid, and efficient screening tool for rapid on-site detection of a variety of micropollutants.

  2. Reduction of aflatoxins (B₁, B₂, G₁, and G₂) in soybean-based model systems.

    PubMed

    Lee, Jongin; Her, Jae-Young; Lee, Kwang-Geun

    2015-12-15

    The effects of chemical, physical, and cooking treatments on the reduction of aflatoxin B1 (AFB1), B2, G1, and G2 in soybean matrix were investigated. A HPLC-FLD with a Kobra cell system was used for the quantitative analysis of aflatoxins (AFs). To decrease the level of AFs during the soaking process, the contaminated soybeans were submerged in organic acid solutions. The reduction rates of AFB1 in 1.0N citric acid, lactic acid, succinic acid, and tartaric acid for 18h were 94.1%, 92.7%, 62.0%, and 95.1%, respectively. In the case of pH and autoclave treatment, the level of AFB1 was significantly decreased during autoclaving process at pH 7.4, 9.0, and 11.1, compared with the non-autoclaved samples (p<0.05). In the case of physical treatment, the heating process at 100 and 150°C for 90min significantly decreased the level of AFB1 by 41.9% and 81.2%, respectively (p<0.05). The reduction rate of AFB1 after cooking was 97.9% for soybean milk and 33.6% for steamed soybeans. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Rapid detection of aflatoxin B1 in medicinal materials of radix and rhizome by gold immunochromatographic assay.

    PubMed

    Hu, Shurong; Dou, Xiaowen; Zhang, Lei; Xie, Yanjun; Yang, Shihai; Yang, Meihua

    2018-05-22

    A rapid screening of the most toxic aflatoxin B 1 (AFB 1 ) in medicinal materials of radix and rhizome was performed by an immune chromatography method for the first time. The colloidal gold immunochromatographic strip was prepared after optimization of the conjugation of gold particles with monoclonal antibody, the test line and the control line. Under optimized conditions, the detection limit of the constructed test strip was as low as 0.1 ng mL -1 and the total analysis was conducted within 15 min by naked eyes. Four kinds of medicinal materials (Gastrodia elata, Poria cocos, Bletilla striata and Radix Angelicae Dahuricae) were investigated by the strip. Various complex matrixes pay a significant influence on the feasibility and effectiveness of the strip screening in medicinal materials. Aiming to the characteristics of selected medicinal materials, the screening was successfully proceeded with extraction by 70% methanol-water as well as three-fold dilution in Gastrodia elata and Radix Angelicae Dahuricae, 70% methanol-PBS as well as four-fold dilution in Poria cocos., and 60% methanol-water as well as four-fold dilution in Bletilla striata. Among the collected 40 samples, one was found to be positive of AFB 1 with level above 5 μg kg -1 . The result was in a good agreement with those obtained from LC-MS/MS determination (6.12 μg kg -1 ). The gold immunochromatographic strip was demonstrated as a rapid, cost-effective, reliable and on-site screening technique for mycotoxins in starch and polysaccharides-rich herbal medicines. Copyright © 2018 Elsevier Ltd. All rights reserved.

  4. The effects of necrotic enteritis, aflatoxin B1, and virginiamycin on growth performance, necrotic enteritis lesion scores, and mortality in young broilers.

    PubMed

    Cravens, R L; Goss, G R; Chi, F; De Boer, E D; Davis, S W; Hendrix, S M; Richardson, J A; Johnston, S L

    2013-08-01

    The effects of increasing aflatoxin B1 concentration (0, 0.75, 1.5 mg/kg) on broilers with or without necrotic enteritis or virginiamycin were determined. In the 23-d study, 22 male Cobb 500 chicks per pen were allotted to 12 treatments (3 × 2 × 2 factorial arrangement) with 8 replications. Intestines of 5 birds per pen were examined for lesions on d 21. Birds were allowed to consume feed and water ad libitum. Aflatoxin was included in the diets from d 0. All birds received a 10× dose of coccidiosis vaccine on d 10. Pens of birds where necrotic enteritis was being induced were on Clostridium perfringens pathogen (CPP) contaminated litter from d 0. Aflatoxin decreased gain and feed intake and resulted in poorer feed:gain, increased mortality, and higher lesion scores. Inducing necrotic enteritis increased lesion scores and decreased feed intake and gain. Adding virginiamycin to the diets improved gain, feed intake, feed conversion, and decreased mortality. There was a 3-way interaction (aflatoxin × virginiamycin × CPP) on gain; increasing aflatoxin decreased gain and the effects of CPP and virginiamycin were dependent on aflatoxin concentration. In the absence of aflatoxin virginiamycin increased gain but was unable to prevent the growth suppression caused by CPP. At 0.75 mg/kg of aflatoxin virginiamycin no longer increased growth in non-CPP challenged birds but was able to increase growth in CPP-challenged birds. At the 1.5 mg/kg of aflatoxin concentration, virginiamycin increased gain in non-CPP-challenged birds but challenging birds with CPP had no effect on gain. Virginiamycin improved overall feed conversion with the greatest improvement at 1.5 mg/kg (aflatoxin × virginiamycin, P < 0.05). Aflatoxin increased lesion scores in unchallenged birds but not in challenged birds (aflatoxin × CPP, P < 0.001). Aflatoxin and necrotic enteritis decrease broiler performance and interact to decrease weight gain, virginiamycin helps improve gain in challenged birds at

  5. Aflatoxin M1 in Pasteurized Milk in Babol city, Mazandaran Province, Iran.

    PubMed

    Sefidgar, Saa; Mirzae, M; Assmar, M; Naddaf, Sr

    2011-01-01

    Aflatoxin M(1) (AFM(1)) is the metabolite of aflatoxin B1 (AFB(1)) and is found in milk when lactating animals are fed with contaminated feedstuff. The presence of AFM(1) in milk, pose a major risk for humans especially kids as it can have immunosuppressive, mutagenic, teratogenic and carcinogenic effects. The present study is aimed to investigate the occurrence of AFM(1) in subsidized pasteurized milk in Babol, Mazandaran Province, Iran. Some 72 pasteurized milk packages were collected from supermarkets in various districts of city during January to March 2006. Milk samples were centrifuged and amounts of 100 μl of skimmed milk were tested for AFM(1) contamination by competitive ELISA. All the samples (100%) exhibited contamination with AFM(1). The contamination levels means in January, February, and March were 227.85, 229.64, and 233.1ng/l, respectively. The amount of AFM(1) in all the samples were above 50ng/l, the threshold set by the European community regulations. Monitoring of AFM(1) level should be part of quality control procedures in dairy factories, particularly the ones providing infant's milk. Production of safer and healthier milk and other dairy products with minimum AFM(1) level can be achieved by adopting prophylactic measures including control of humidity and water content of feedstuff, which favors mould production.

  6. Aflatoxin M1 in Pasteurized Milk in Babol city, Mazandaran Province, Iran

    PubMed Central

    Sefidgar, SAA; Mirzae, M; Assmar, M; Naddaf, SR

    2011-01-01

    Background: Aflatoxin M1 (AFM1) is the metabolite of aflatoxin B1 (AFB1) and is found in milk when lactating animals are fed with contaminated feedstuff. The presence of AFM1 in milk, pose a major risk for humans especially kids as it can have immunosuppressive, mutagenic, teratogenic and carcinogenic effects. The present study is aimed to investigate the occurrence of AFM1 in subsidized pasteurized milk in Babol, Mazandaran Province, Iran. Methods: Some 72 pasteurized milk packages were collected from supermarkets in various districts of city during January to March 2006. Milk samples were centrifuged and amounts of 100 μl of skimmed milk were tested for AFM1 contamination by competitive ELISA. Results: All the samples (100%) exhibited contamination with AFM1. The contamination levels means in January, February, and March were 227.85, 229.64, and 233.1ng/l, respectively. The amount of AFM1 in all the samples were above 50ng/l, the threshold set by the European community regulations. Conclusion: Monitoring of AFM1 level should be part of quality control procedures in dairy factories, particularly the ones providing infant’s milk. Production of safer and healthier milk and other dairy products with minimum AFM1 level can be achieved by adopting prophylactic measures including control of humidity and water content of feedstuff, which favors mould production. PMID:23113064

  7. Model Averaging for Predicting the Exposure to Aflatoxin B1 Using DNA Methylation in White Blood Cells of Infants

    NASA Astrophysics Data System (ADS)

    Rahardiantoro, S.; Sartono, B.; Kurnia, A.

    2017-03-01

    In recent years, DNA methylation has been the special issue to reveal the pattern of a lot of human diseases. Huge amount of data would be the inescapable phenomenon in this case. In addition, some researchers interesting to take some predictions based on these huge data, especially using regression analysis. The classical approach would be failed to take the task. Model averaging by Ando and Li [1] could be an alternative approach to face this problem. This research applied the model averaging to get the best prediction in high dimension of data. In the practice, the case study by Vargas et al [3], data of exposure to aflatoxin B1 (AFB1) and DNA methylation in white blood cells of infants in The Gambia, take the implementation of model averaging. The best ensemble model selected based on the minimum of MAPE, MAE, and MSE of predictions. The result is ensemble model by model averaging with number of predictors in model candidate is 15.

  8. Aflatoxin and dimethyl sulfoxide influence on radiomanganese distribution and retention in neonate mice

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Thompson, J.S.; Llewellyn, G.C.

    The LD50 (7 d) for aflatoxin B/sub 1/ (AFB/sub 1/) in CD-1 neonate mice (3.1 g; 5 d of age) was determined to be 13.3 mg/kg. The vehicle was dimethyl sulfoxide (DMSO), given intraperitoneally, at 0.01 ml/animal (7 mg/kg). The solvent was nontoxic and caused no significant change in body weight in animals during an 11-d experimental period (17 d of age). Aflatoxin B/sub 1/ at 5.0 mg/kg and above caused reduced body weight gain. DMSO animals had a mean loss of more than 17% of the radiolabel over a 9-d period. Aflatoxin treatments reversed the DMSO loss of /supmore » 54/Mn in a concentration-related fashion, and generally, AFB/sub 1/ caused a conservation of the radioisotope. The radiolabel was redistributed into the following organs/tissues: liver > brain > bone > muscle = lungs > blood. Aflatoxin-treated animals showed a twofold increase of radiolabel in the liver as compared to controls. The DMSO itself failed to influence /sup 54/Mn influx into the liver. In general, control neonate mice, by 17 d of age, were retaining and redistributing the /sup 54/MnCl/sub 2/ and had not reached the time for sudden emergence of excretion common in rodents. DMSO was found not to be the most satisfactory solvent to use in the administration of aflatoxins, especially when manganese metabolism is being studied. Generally, both DMSO and AFB/sub 1/ influenced radiomanganese distribution, DMSO having a substantial influence. 27 references, 3 figures, 2 tables.« less

  9. Impact of maximum levels in European legislation on exposure of mycotoxins in dried products: case of aflatoxin B1 and ochratoxin A in nuts and dried fruits.

    PubMed

    Van de Perre, Evelien; Jacxsens, Liesbeth; Lachat, Carl; El Tahan, Fouad; De Meulenaer, Bruno

    2015-01-01

    In this study the impact of setting European criteria on exposure to aflatoxin B1 via nuts and figs and ochratoxin A via dried fruits is evaluated for the Belgian population, as an example of the European population. Two different scenarios were evaluated. In scenario 1 all collected literature data are considered, assuming that there is no border control nor legal limits in Europe. In the second scenario, contamination levels above the maximum limits are excluded. The results from scenario 1 demonstrated that if no regulation is in place, AFB1 and OTA concentrations reported in the analysed food can have potential health risk to the population. The estimated exposure of OTA for scenario 2 is below the TDI of 5 ng/kg BW⋅day, indicating that OTA concentrations accepted by EU legislation pose a low risk to the Belgian population. For AFB1, the MOE values of scenario 2 are above 10,000 and can be considered to be of low health concern, based on BDML10 for humans, except for figs (MOE = 5782). This means that for all matrices, with exception of figs, the maximum values of AFB1 in the European legislation are sufficient to be of a low health concern for consumers. Copyright © 2014. Published by Elsevier Ltd.

  10. Banana peel: an effective biosorbent for aflatoxins.

    PubMed

    Shar, Zahid Hussain; Fletcher, Mary T; Sumbal, Gul Amer; Sherazi, Syed Tufail Hussain; Giles, Cindy; Bhanger, Muhammad Iqbal; Nizamani, Shafi Muhammad

    2016-05-01

    This work reports the application of banana peel as a novel bioadsorbent for in vitro removal of five mycotoxins (aflatoxins (AFB1, AFB2, AFG1, AFG2) and ochratoxin A). The effect of operational parameters including initial pH, adsorbent dose, contact time and temperature were studied in batch adsorption experiments. Scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR) and point of zero charge (pHpzc) analysis were used to characterise the adsorbent material. Aflatoxins' adsorption equilibrium was achieved in 15 min, with highest adsorption at alkaline pH (6-8), while ochratoxin has not shown any significant adsorption due to surface charge repulsion. The experimental equilibrium data were tested by Langmuir, Freundlich and Hill isotherms. The Langmuir isotherm was found to be the best fitted model for aflatoxins, and the maximum monolayer coverage (Q0) was determined to be 8.4, 9.5, 0.4 and 1.1 ng mg(-1) for AFB1, AFB2, AFG1 and AFG2 respectively. Thermodynamic parameters including changes in free energy (ΔG), enthalpy (ΔH) and entropy (ΔS) were determined for the four aflatoxins. Free energy change and enthalpy change demonstrated that the adsorption process was exothermic and spontaneous. Adsorption and desorption study at different pH further demonstrated that the sorption of toxins was strong enough to sustain pH changes that would be experienced in the gastrointestinal tract. This study suggests that biosorption of aflatoxins by dried banana peel may be an effective low-cost decontamination method for incorporation in animal feed diets.

  11. High-performance liquid chromatography ultraviolet-photodiode array detection method for aflatoxin B1 in cattle feed supplements

    PubMed Central

    Mochamad, Lazuardi; Hermanto, Bambang

    2017-01-01

    Aim: The objective of the current study is to determine the concentration of aflatoxin B1 using high-performance liquid chromatography (HPLC) with a photodiode array (PDA) detector. Materials and Methods: Aflatoxin B1 certified reference grade from Trilogy Analytical Laboratory dissolved acetonitrile (ACN) at 10 µg/mL was using standard assessment. HPLC instruments such as ultraviolet-PDA detector used a Shimadzu LC-6AD pump with DGU-20A5 degasser, communication module-20A, and PDA detector SPD-M20A with FRC-10A fraction collector. The HPLC was set isocratic method at 354 nm with a reverse-phase ODS C18 column (LiChrospher® 100 RP-18; diameter, 5 µm) under a 20°C controlled column chamber. Rheodyne® sample loops were performed in 20 µL capacities. The mobile phase was performed at fraction 63:26:11 H2O: methanol:ACN at pH 6.8. A total of 1 kg of feed contained 10% bread crumbs and 30% concentrated, 40% forage, and 20% soybean dregs were using commercials samples. Samples were extracted by ACN and separated with solid phase extraction ODS 1 mL than elution with mobile phase to collect at drying samples performed. The samples were ready to use after added 1 mL mobile phase than injected into the system of HPLC. Results: We found that the retention time of aflatoxin B1 was approximately 10.858 min. Linearity of 0.01-0.08 µg/mL aflatoxin B1 dissolved in mobile phase was obtained at R2=0.9. These results demonstrate that these methods can be used to analyze aflatoxin B1 and gain 89-99% recovery. The limit of detection of this assay was obtained at 3.5 × 10−6 µg/mL. Conclusion: This method was easy to apply and suitable to analyzing at small concentrations of aflatoxin B1 in formulated product of feed cattle. PMID:28919686

  12. Five of 12 forms of vaccinia virus-expressed human hepatic cytochrome P450 metabolically activate aflatoxin B sub 1

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Aoyama, Toshifumi; Yamano, Shigeru; Gelboin, H.V.

    Twelve forms of human hepatic cytochrome P450 were expressed in hepatoma cells by means of recombinant vaccinia viruses. The expressed P450s were analyzed for their abilities to activate the potent hepatocarcinogen aflatoxin B{sub 1} to metabolites having mutagenic or DNA-binding properties. Five forms, P450s IA2, IIA3, IIB7, IIIA3, and IIIA4, activated aflatoxin B{sub 1} to mutagenic metabolites as assessed by the production of His revertants of Salmonella typhimurium in the Ames test. The same P450s catalyzed conversion of aflatoxin B{sub 1} to DNA-bound derivatives as judged by an in situ assay in which the radiolabeled carcinogen was incubated with cellsmore » expressing the individual P450 forms. Seven other human P450s, IIC8, IIC9, IID6, IIE1, IIF1, and IIIA5, and IVB1, did not significantly activate aflatoxin B{sub 1} as measured by both the Ames test and the DNA-binding assay. Moreover, polyclonal anti-rat liver P450 antibodies that crossreact with individual human P450s IA2, IIA3, IIIA3, and IIIA4 each inhibited aflatoxin B{sub 1} activation catalyzed by human liver S-9 extracts. Inhibition ranged from as low as 10% with antibody against IIA3 to as high as 65% with antibody against IIIA3 and IIIA4. These results establish that metabolic activation of aflatoxin B{sub 1} in human liver involves the contribution of multiple forms of P450.« less

  13. Aflatoxin B(1) in affecting broiler's performance, immunity, and gastrointestinal tract: a review of history and contemporary issues.

    PubMed

    Yunus, Agha W; Razzazi-Fazeli, E; Bohm, Josef

    2011-06-01

    Aflatoxin B(1) is a common contaminant of poultry feeds in tropical and subtropical climates. Research during the last five decades has well established the negative effects of the mycotoxin on health of poultry. However, the last ten years of relevant data have accentuated the potential of low levels of aflatoxin B(1) to deteriorate broiler performance. In this regard, any attempt to establish a dose-effect relationship between aflatoxin B(1) level and broiler performance is also complicated due to differences in types of broilers and length of exposure to the mycotoxin in different studies. Contrary to the prevalent notion regarding literature saturation with respect to aflatoxicosis of chicken, many areas of aflatoxicosis still need to be explored. Literature regarding effects of the mycotoxin on the gastrointestinal tract in this regard is particular scanty and non-conclusive. In addition to these issues, the metabolism of aflatoxin B(1) and recently proposed hypotheses regarding biphasic effects of the mycotoxin in broilers are briefly discussed.

  14. Enzymatic Formation of G-Group Aflatoxins and Biosynthetic Relationship between G- and B-Group Aflatoxins

    PubMed Central

    Yabe, Kimiko; Nakamura, Miki; Hamasaki, Takashi

    1999-01-01

    We detected biosynthetic activity for aflatoxins G1 and G2 in cell extracts of Aspergillus parasiticus NIAH-26. We found that in the presence of NADPH, aflatoxins G1 and G2 were produced from O-methylsterigmatocystin and dihydro-O-methylsterigmatocystin, respectively. No G-group aflatoxins were produced from aflatoxin B1, aflatoxin B2, 5-methoxysterigmatocystin, dimethoxysterigmatocystin, or sterigmatin, confirming that B-group aflatoxins are not the precursors of G-group aflatoxins and that G- and B-group aflatoxins are independently produced from the same substrates (O-methylsterigmatocystin and dihydro-O-methylsterigmatocystin). In competition experiments in which the cell-free system was used, formation of aflatoxin G2 from dihydro-O-methylsterigmatocystin was suppressed when O-methylsterigmatocystin was added to the reaction mixture, whereas aflatoxin G1 was newly formed. This result indicates that the same enzymes can catalyze the formation of aflatoxins G1 and G2. Inhibition of G-group aflatoxin formation by methyrapone, SKF-525A, or imidazole indicated that a cytochrome P-450 monooxygenase may be involved in the formation of G-group aflatoxins. Both the microsome fraction and a cytosol protein with a native mass of 220 kDa were necessary for the formation of G-group aflatoxins. Due to instability of the microsome fraction, G-group aflatoxin formation was less stable than B-group aflatoxin formation. The ordA gene product, which may catalyze the formation of B-group aflatoxins, also may be required for G-group aflatoxin biosynthesis. We concluded that at least three reactions, catalyzed by the ordA gene product, an unstable microsome enzyme, and a 220-kDa cytosol protein, are involved in the enzymatic formation of G-group aflatoxins from either O-methylsterigmatocystin or dihydro-O-methylsterigmatocystin. PMID:10473388

  15. Use of Cold Atmospheric Plasma to Detoxify Hazelnuts from Aflatoxins

    PubMed Central

    Siciliano, Ilenia; Spadaro, Davide; Prelle, Ambra; Vallauri, Dario; Cavallero, Maria Chiara; Garibaldi, Angelo; Gullino, Maria Lodovica

    2016-01-01

    Aflatoxins, produced by Aspergillus flavus and A. parasiticus, can contaminate different foodstuffs, such as nuts. Cold atmospheric pressure plasma has the potential to be used for mycotoxin detoxification. In this study, the operating parameters of cold atmospheric pressure plasma were optimized to reduce the presence of aflatoxins on dehulled hazelnuts. First, the effect of different gases was tested (N2, 0.1% O2 and 1% O2, 21% O2), then power (400, 700, 1000, 1150 W) and exposure time (1, 2, 4, and 12 min) were optimized. In preliminary tests on aflatoxin standard solutions, this method allowed to obtain a complete detoxification using a high power for a few minutes. On hazelnuts, in similar conditions (1000 W, 12 min), a reduction in the concentration of total aflatoxins and AFB1 of over 70% was obtained. Aflatoxins B1 and G1 were more sensitive to plasma treatments compared to aflatoxins B2 and G2, respectively. Under plasma treatment, aflatoxin B1 was more sensitive compared to aflatoxin G1. At the highest power, and for the longest time, the maximum temperature increment was 28.9 °C. Cold atmospheric plasma has the potential to be a promising method for aflatoxin detoxification on food, because it is effective and it could help to maintain the organoleptic characteristics. PMID:27128939

  16. Interaction of mammary bovine ABCG2 with AFB1 and its metabolites and regulation by PCB 126 in a MDCKII in vitro model.

    PubMed

    Manzini, L; Halwachs, S; Girolami, F; Badino, P; Honscha, W; Nebbia, C

    2017-12-01

    The ATP-binding cassette efflux transporter ABCG2 plays a key role in the mammary excretion of drugs and toxins in humans and animals. Aflatoxins (AF) are worldwide contaminants of food and feed commodities, while PCB 126 is a dioxin-like PCB which may contaminate milk and dairy products. Both compounds are known human carcinogens. The interactions between AF and bovine ABCG2 (bABCG2) as well as the effects of PCB 126 on its efflux activity have been investigated by means of the Hoechst H33342 transport assay in MDCKII cells stably expressing mammary bABCG2. Both AFB1 and its main milk metabolite AFM1 showed interaction with bABCG2 even at concentrations approaching the legal limits in feed and food commodities. Moreover, PCB 126 significantly enhanced bABCG2 functional activity. Specific inhibitors of either AhR (CH233191) or ABCG2 (Ko143) were able to reverse the PCB 126-induced increase in bABCG2 transport activity, showing the specific upregulation of the efflux protein by the AhR pathway. The incubation of PCB 126-pretreated cells with AFM1 was able to substantially reverse such effect, with still unknown mechanism(s). Overall, results from this study point to AFB1 and AFM1 as likely bABCG2 substrates. The PCB 126-dependent increased activity of the transporter could enhance the ABCG2-mediated excretion into dairy milk of chemicals (i.e., drugs and toxins) potentially harmful to neonates and consumers. © 2017 John Wiley & Sons Ltd.

  17. A Theoretical Study of 8-Chloro-9-Hydroxy-Aflatoxin B1, the Conversion Product of Aflatoxin B1 by Neutral Electrolyzed Water

    PubMed Central

    Escobedo-González, René; Méndez-Albores, Abraham; Villarreal-Barajas, Tania; Aceves-Hernández, Juan Manuel; Miranda-Ruvalcaba, René; Nicolás-Vázquez, Inés

    2016-01-01

    Theoretical studies of 8-chloro-9-hydroxy-aflatoxin B1 (2) were carried out by Density Functional Theory (DFT). This molecule is the reaction product of the treatment of aflatoxin B1 (1) with hypochlorous acid, from neutral electrolyzed water. Determination of the structural, electronic and spectroscopic properties of the reaction product allowed its theoretical characterization. In order to elucidate the formation process of 2, two reaction pathways were evaluated—the first one considering only ionic species (Cl+ and OH−) and the second one taking into account the entire hypochlorous acid molecule (HOCl). Both pathways were studied theoretically in gas and solution phases. In the first suggested pathway, the reaction involves the addition of chlorenium ion to 1 forming a non-classic carbocation assisted by anchimeric effect of the nearest aromatic system, and then a nucleophilic attack to the intermediate by the hydroxide ion. In the second studied pathway, as a first step, the attack of the double bond from the furanic moiety of 1 to the hypochlorous acid is considered, accomplishing the same non-classical carbocation, and again in the second step, a nucleophilic attack by the hydroxide ion. In order to validate both reaction pathways, the atomic charges, the highest occupied molecular orbital and the lowest unoccupied molecular orbital were obtained for both substrate and product. The corresponding data imply that the C9 atom is the more suitable site of the substrate to interact with the hydroxide ion. It was demonstrated by theoretical calculations that a vicinal and anti chlorohydrin is produced in the terminal furan ring. Data of the studied compound indicate an important reduction in the cytotoxic and genotoxic potential of the target molecule, as demonstrated previously by our research group using different in vitro assays. PMID:27455324

  18. Interlaboratory Validation of a Stable Isotope Dilution and Liquid Chromatography Tandem Mass Spectrometry Method for the Determination of Aflatoxins in Milk, Milk-Based Infant Formula, and Feed.

    PubMed

    Zhang, Kai; Liao, Chia-Ding; Prakash, Shristi; Conway, Michael; Cheng, Hwei-Fang

    2018-05-01

    An interlaboratory study was conducted to evaluate stable isotope dilution and LC tandem MS (MS/MS) for the determination of aflatoxins B1, B2, G1, G2, and M1 (AFB1, AFB2, AFG1, AFG2, and AFM1) in milk, milk-based infant formula (formula), and feed. Samples were first fortified with five 13C uniformly labeled aflatoxins {[13C]-internal standard (IS)} corresponding to the five native aflatoxins, which were subsequently extracted with acetonitrile-water (50 + 50, v/v), followed by centrifugation, filtration, and LC-MS/MS analysis. In addition to certified milk powder and animal feed, the three participating laboratories also analyzed milk, formula, and feed fortified with the five aflatoxins at concentrations ranging from 0.5 to 50 ng/g. The majority of recoveries ranged from 80 to 120%, with RSDs < 20%. Method LOQs were determined by the three laboratories using the three sample matrixes in replicates (n = 8), and the determined LOQs of AFB1, AFB2, AFG1, AFG2, and AFM1 ranged from 0.1 to 0.91, 0.24 to 0.64, 0.28 to 1.52, 0.19 to 3.80, and 0.12 to 0.45 ng/g, respectively. For detected aflatoxins in the certified materials, all measured concentrations were within ±25% of the certified values. Using [13C]-IS eliminated the need for matrix-matched calibration standards for quantitation, simplified sample preparation, and achieved simultaneous identification and quantitation of the aflatoxins in a simple LC-MS/MS procedure.

  19. Determination of aflatoxin B1 in food and feedstuffs in Cuba (1990 through 1996) using an immunoenzymatic reagent kit (Aflacen).

    PubMed

    Escobar, Arturo; Regueiro, Olga Sanchez

    2002-01-01

    The presence of aflatoxin B1 was analyzed in imported food and feedstuffs of national production in the period of 1990 through 1996, destined to animal and human consumption using an immunoenzymatic reagent kit (Aflacen, Ckure, la Habana, Cuba) with a detection limit of 0.3 microg/kg. It was found that the 17.04% of a total of 4,594 analyzed samples presented aflatoxin B1, and the biggest percentages were in sorghum and peanut with an 83.3 and 40.4%, respectively. The corn, oat, wheat, and soy are fundamental raw ingredients in the elaboration of concentrates. Percentages of contamination with aflatoxin B1 of 23.3, 10.7, 25, and 4.6 were found in corn, oat, wheat, and soy, respectively. Other analyzed foods like rice, beans, and peas presented percentages of contamination with aflatoxin B1 inferior to 5% of the analyzed samples. It was found that more than 455 samples surpassed the value of 10 microg/kg. Corn and peanut products present a high demand in population showing levels of contamination superior to 50 microg/kg. The 11.3% of the samples contaminated with aflatoxin B1 have values between 1 and 20 microg/kg, where peanut and concentrates show the highest percentages (21.9 and 18.7), respectively. These results show levels of aflatoxin B1 in the population that constitute a great risk for human and animal health.

  20. Effects of activated aflatoxin B/sub 1/ and caffeine on DNA replicon initiation in HeLa cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cramer, P.; Painter, R.B.

    1981-01-01

    Afatoxin B/sub 1/ (AFB/sub 1/) is activated by a rat microsomal extract (S-9) to form a product that inhibits DNA synthesis in HeLa cells. At 10/sup -7/ M, AFB/sub 1/ inhibited initiation of replicons, as shown in alkaline sucrose gradient profiles 30 min after incubation with the drug. Ninety minutes later, the profile of treated cells was similar to that of control, but 4 h later there was another effect on replicon initiation. At 10/sup -6/ M, the inhibition of initiation was greater than at 10/sup -7/ M and increased progressively. Four hours after removal of the drug, the gradientmore » profile showed low amounts of radioactivity in all size classes of DNA. When cells were incubated in medium containing caffeine (2 mM) even as late as 60 min after incubation with AFB/sub 1/, the inhibition of replicon initiation was prevented. If caffeine was later removed from the medium, replicon initiation was then inhibited. At 10/sup -7/ M or 10/sup -6/ M, AFB/sub 1/ had little immediate effect on chain elongation, but at 10/sup -5/ M, the gradient profiles showed an accumulation of low molecular weight DNA molecules, with no radioactivity in the region of high molecular weight DNA, owing to a block to chain elongation; this was not affected by caffeine. These results suggest that AFB/sub 1/ induces damage that changes the fonformation of chromatin so that initiation of new replicons cannot occur; in the presence of caffeine this change does not occur and DNA replication is not inhibited.« less

  1. Aflatoxin B1 in Affecting Broiler’s Performance, Immunity, and Gastrointestinal Tract: A Review of History and Contemporary Issues

    PubMed Central

    Yunus, Agha W.; Razzazi-Fazeli, E.; Bohm, Josef

    2011-01-01

    Aflatoxin B1 is a common contaminant of poultry feeds in tropical and subtropical climates. Research during the last five decades has well established the negative effects of the mycotoxin on health of poultry. However, the last ten years of relevant data have accentuated the potential of low levels of aflatoxin B1 to deteriorate broiler performance. In this regard, any attempt to establish a dose-effect relationship between aflatoxin B1 level and broiler performance is also complicated due to differences in types of broilers and length of exposure to the mycotoxin in different studies. Contrary to the prevalent notion regarding literature saturation with respect to aflatoxicosis of chicken, many areas of aflatoxicosis still need to be explored. Literature regarding effects of the mycotoxin on the gastrointestinal tract in this regard is particular scanty and non-conclusive. In addition to these issues, the metabolism of aflatoxin B1 and recently proposed hypotheses regarding biphasic effects of the mycotoxin in broilers are briefly discussed. PMID:22069726

  2. Near-infrared hyperspectral imaging for detecting Aflatoxin B1 of maize kernels

    USDA-ARS?s Scientific Manuscript database

    The feasibility of detecting the Aflatoxin B1 in maize kernels inoculated with Aspergillus flavus conidia in the field was assessed using near-infrared hyperspectral imaging technique. After pixel-level calibration, wavelength dependent offset, the masking method was adopted to reduce the noise and ...

  3. In vivo toxicity assessment of aflatoxin B1-contaminated corn after ozone degradation.

    PubMed

    Luo, Xiaohu; Li, Ke; Xing, Jiali; Qi, Lijun; Yang, Ming; Wang, Ren; Wang, Li; Li, Yanan; Chen, Zhengxing

    2018-02-01

    Corn is an important food and feedstuff in China and worldwide. The problems caused by aflatoxin B 1 -contaminated corn (ACC) are of great concern. Our previous studies have demonstrated that ozone can effectively degrade AFB 1 in corn, prompting us to investigate the in vivo toxicity of treated ACC. In this study, 35 Kunming mice were used to assess the in vivo toxicity of ozone treated ACC. Results indicated that compared to mice fed with basal feedstuff (provided by the Shanghai SLAC Laboratory), those fed with ACC have significantly decreased mean weight as well as total protein (TP), albumin (ALB), and globulin (GLB) contents (p < 0.05). On the other hand, the liver and kidney/body weight ratio as well as the serum alanine transaminase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) levels significantly increased (p < 0.05). Obvious histopathological changes were found in the liver and kidney. When mice were fed with the ozone-treated ACC, no significant differences were observed in the mean weight, the liver and kidney/body weight ratio and in the major serum indexes ALT, TP, ALB, and GLB (p > 0.05). However, AST and ALP significantly increased (p < 0.05), and slight histopathological changes were found in liver tissues. This study indicated that ACC may lead to significant changes in various physiological characteristics and biochemical indexes in liver and kidney tissues, but ozone treatment of ACC could significantly reduce these changes.

  4. Survey of aflatoxins in maize tortillas from Mexico City.

    PubMed

    Castillo-Urueta, Pável; Carvajal, Magda; Méndez, Ignacio; Meza, Florencia; Gálvez, Amanda

    2011-01-01

    In Mexico, maize tortillas are consumed on a daily basis, leading to possible aflatoxin exposure. In a survey of 396 2-kg samples, taken over four sampling days in 2006 and 2007 from tortilla shops and supermarkets in Mexico City, aflatoxin levels were quantified by HPLC. In Mexico, the regulatory limit is 12 µg kg⁻¹ total aflatoxins for maize tortillas. In this survey, 17% of tortillas contained aflatoxins at levels of 3-385 µg kg⁻¹ or values below the limit of quantification (12 µg kg⁻¹ and 87% were below the regulatory limit. Average aflatoxin concentrations in 56 contaminated samples were: AFB1 (12.1 µg kg⁻¹); AFB2 (2.7 µg kg⁻¹); AFG1 (64.1 µg kg⁻¹) and AFG2 (3.7 µg kg⁻¹), and total AF (20.3 µg kg⁻¹).

  5. Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis.

    PubMed

    Wang, Du; Zhang, Zhaowei; Li, Peiwu; Zhang, Qi; Zhang, Wen

    2016-07-14

    Rapid and quantitative sensing of aflatoxin B1 with high sensitivity and specificity has drawn increased attention of studies investigating soybean sauce. A sensitive and rapid quantitative immunochromatographic sensing method was developed for the detection of aflatoxin B1 based on time-resolved fluorescence. It combines the advantages of time-resolved fluorescent sensing and immunochromatography. The dynamic range of a competitive and portable immunoassay was 0.3-10.0 µg·kg(-1), with a limit of detection (LOD) of 0.1 µg·kg(-1) and recoveries of 87.2%-114.3%, within 10 min. The results showed good correlation (R² > 0.99) between time-resolved fluorescent immunochromatographic strip test and high performance liquid chromatography (HPLC). Soybean sauce samples analyzed using time-resolved fluorescent immunochromatographic strip test revealed that 64.2% of samples contained aflatoxin B1 at levels ranging from 0.31 to 12.5 µg·kg(-1). The strip test is a rapid, sensitive, quantitative, and cost-effective on-site screening technique in food safety analysis.

  6. The antioxidant effects of pumpkin seed oil on subacute aflatoxin poisoning in mice.

    PubMed

    Eraslan, Gökhan; Kanbur, Murat; Aslan, Öznur; Karabacak, Mürsel

    2013-12-01

    This study was aimed at the investigation of the antioxidant effect of pumpkin seed oil against the oxidative stress-inducing potential of aflatoxin. For this purpose, 48 male BALB/c mice were used. Four groups, each comprising 12 mice, were established. Group 1 was maintained as the control group. Group 2 was administered with pumpkin seed oil alone at a dose of 1.5 mL/kg.bw/day (∼1375mg/kg.bw/day). Group 3 received aflatoxin (82.45% AFB1 , 10.65% AFB2 , 4.13% AFG1, and 2.77% AFG2 ) alone at a dose of 625 μg/kg.bw/day. Finally, group 4 was given both 1.5 mL/kg.bw/day pumpkin seed oil and 625 μg/kg.bw/day aflatoxin. All administrations were oral, performed with the aid of a gastric tube and continued for a period of 21 days. At the end of day 21, the liver, lungs, kidneys, brain, heart, and spleen of the animals were excised, and the extirpated tissues were homogenized appropriately. Malondialdehyde (MDA) levels and catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities were determined in tissue homogenates. In conclusion, it was determined that aflatoxin exhibited adverse effects on most of the oxidative stress markers. The administration of pumpkin seed oil diminished aflatoxin-induced adverse effects. In other words, the values of the group, which was administered with both aflatoxin and pumpkin seed oil, were observed to have drawn closer to the values of the control group. Copyright © 2011 Wiley Periodicals, Inc.

  7. Development of a simple and convenient cell-based electrochemical biosensor for evaluating the individual and combined toxicity of DON, ZEN, and AFB1.

    PubMed

    Xia, Shuang; Zhu, Pei; Pi, Fuwei; Zhang, Yinzhi; Li, Yun; Wang, Jiasheng; Sun, Xiulan

    2017-11-15

    A simple and convenient cell-based electrochemical biosensor was developed to assess the individual and combined toxicity of deoxynivalenol (DON), zearalenone (ZEN), and Aflatoxin B 1 (AFB 1 ) on Hep G2 cells. The sensor was modified in succession with AuNPs (gold nanoparticles), cysteamine, and laminin. The cells interacting with laminin formed tight cell-to-electrode contacts, and collagen was used to maintain cell adhesion and viability. Electrochemical impedance spectroscopy (EIS) was developed to evaluate mycotoxin toxicity. Experimental results show that DON, ZEN, and AFB 1 caused a significant decrease in cell viability in a dose dependent manner. The EIS value decreased with concentrations of DON, ZEN, and AFB 1 in the range of 0.01-20, 0.1-50, and 0.1-3.5μg/mL, and IC 50 obtained using the developed method was 48.5, 59.0, and 3.10μg/mL, respectively. A synergistic effect was observed between DON and ZEN, an additive effect was observed between DON and AFB 1 , and an antagonism effect was found in the binary mixtures of ZEN and AFB 1 and ternary mixtures. These results were confirmed via CCK-8 assay. Utilizing SEM, we found that cells treated with mycotoxins caused significant changes in cell morphology, thus lessening cell adsorption and impedance reduction. Biological assay indicated that EIS patterns correlated with [Ca 2+ ] i concentrations and apoptosis and necrotic cells ratios, thus effecting electrochemical signals. This method is simpler, more convenient, sensitive, and has a quicker response rate than most conventional cytotoxicity evaluation methods. Copyright © 2017. Published by Elsevier B.V.

  8. Effects of aflatoxin and carotenoids on growth performance and immune response in mule ducklings.

    PubMed

    Cheng, Y H; Shen, T F; Pang, V F; Chen, B J

    2001-01-01

    The purpose of this study was to investigate if carotenoids could alleviate the adverse effects caused by aflatoxin with respect to growth performance and immune response. In two experiments, a total of 320 mule ducklings were assigned to 5 treatments, i.e. control, aflatoxin B(1) (AFB(1)) 200 ppb, AFB(1) +beta-carotene (BC) 200 ppm, AFB(1)+BC 400 ppm, and AFB(1)+astaxanthin (AS) 200 ppm. In experiment 1, the addition of beta-carotene or astaxanthin in the diet containing AFB(1) 200 ppb resulted in a significant decrease in average daily gain as compared with the control. AFB(1) 200 ppb alone and the addition of BC or AS on top of AFB(1) resulted in a significantly lower daily feed intake than for the control group. There were no significant differences in relative organ weights among treatment groups. Both treatments of BC 400 ppm and AS 200 ppm had significantly more macrophages harvested per duck than the control and AFB(1) 200 ppb treatments. However, there were no significant differences among treatments in percentages of phagocytotic macrophages and number of Candida albican phagocytized by phagocytotic macrophages. In experiment 2, blood biochemical parameters and antibody titers were evaluated. There were no significant differences among treatments in total bilirubin content and alkaline phosphatase activity in the serum or in antibody titers against fowl cholera. However, AFB(1) treatment had the highest activities of AST and ALT in the serum. The addition of BC 400 ppm on top of AFB(1) significantly reduced ALT activity as compared with the AFB(1) 200 ppb treatment. These results suggest that carotenoids could provide a slightly toxic alleviating effect on growth performance, enhance the chemotaxis ability of macrophages, and reduce ALT activity elevated by AFB(1).

  9. Effect of gamma-irradiation on aflatoxin B1 production by Aspergillus flavus and chemical composition of three crop seeds.

    PubMed

    Aziz, Nagy H; Mahrous, Souzan R

    2004-06-01

    The effect of gamma-irradiation on aflatoxin B1 production by Aspergillus flavus, and the chemical composition of some different crop seeds were investigated. A. flavus infected seeds behaved differently according to their principal constituents. A. flavus caused an increase in protein and decrease in lipids and carbohydrate contents of wheat, soyabean and fababean seeds. Growth of A. flavus and production of aflatoxin B1 was inhibited at a dose level of 5 kGy. A. flavus utilizes carbohydrates of seeds for its growth and aflatoxin production. Crops were arranged, in descending order, according to aflatoxin produced in seeds as wheat > soyabean > fababean. There were no changes in chemical constituents of irradiated seeds, such as protein, lipids, and carbohydrates.

  10. Development of hyperbranched polymers with non-covalent interactions for extraction and determination of aflatoxins in cereal samples.

    PubMed

    Liu, Xiaoyan; Li, Huihui; Xu, Zhigang; Peng, Jialin; Zhu, Shuqiang; Zhang, Haixia

    2013-10-03

    A novel approach for assembling homogeneous hyperbranched polymers based on non-covalent interactions with aflatoxins was developed; the polymers were used to evaluate the extraction of aflatoxins B1, B2, G1 and G2 (AFB1, AFB2, AFG1 and AFG2) in simulant solutions. The results showed that the extraction efficiencies of three kinds of synthesized polymers for the investigated analytes were not statistically different; as a consequence, one of the representative polymers (polymer I) was used as the solid-phase extraction (SPE) sorbent to evaluate the influences of various parameters, such as desorption conditions, pH, ionic strength, concentration of methanol in sample solutions, and the mass of the sorbent on the extraction efficiency. In addition, the extraction efficiencies for these aflatoxins were compared between the investigated polymer and the traditional sorbent C18. The results showed that the investigated polymer had superior extraction efficiencies. Subsequently, the proposed polymer for the SPE packing material was employed to enrich and analyze four aflatoxins in the cereal powder samples. The limits of detection (LODs) at a signal-to-noise (S/N) ratio of 3 were in the range of 0.012-0.120 ng g(-1) for four aflatoxins, and the limits of quantification (LOQs) calculated at S/N=10 were from 0.04 to 0.40 ng g(-1) for four aflatoxins. The recoveries of four aflatoxins from cereal powder samples were in the range of 82.7-103% with relative standard deviations (RSDs) lower than 10%. The results demonstrate the suitability of the SPE approach for the analysis of trace aflatoxins in cereal powder samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  11. Detoxification of aflatoxins on prospective approach: effect on structural, mechanical, and optical properties under pressures.

    PubMed

    Wei, Yong-Kai; Zhao, Xiao-Miao; Li, Meng-Meng; Yu, Jing-Xin; Gurudeeban, Selvaraj; Hu, Yan-Fei; Ji, Guang-Fu; Wei, Dong-Qing

    2018-06-01

    Aflatoxins are sequential of derivatives of coumarin and dihydrofuran with similar chemical structures and well-known carcinogenic agent. Many studies performed to detoxify aflatoxins, but the result is not ideal. Therefore, we studied structural, infrared spectrum, mechanical, and optical properties of these compounds in the aim of perspective physics. Mulliken charge distributions and infrared spectral analysis performed to understand the structural difference between the basic types of aflatoxins. In addition, the effect of pressure, different polarized, and incident directions on their structural changes was determined. It is found that AFB 1 is most stable structure among four basic types aflatoxins (AFB 1 , AFB 2 , AFG 1 , and AFG 2 ), and IR spectra are analyzed to exhibit the difference on structures of them. The mechanical properties of AFB 1 indicate that the structure of this toxin can be easily changed by pressure. The real [Formula: see text] and imaginary [Formula: see text] parts of the dielectric function, and the absorption coefficient [Formula: see text] and energy loss spectrum [Formula: see text] were also obtained under different polarized and incident directions. Furthermore, biological experiments needed to support the toxic level of AFB 1 using optical technologies.

  12. Occurrence of Aflatoxins in Selected Processed Foods from Pakistan

    PubMed Central

    Mushtaq, Muhammad; Sultana, Bushra; Anwar, Farooq; Khan, Muhammad Zargham; Ashrafuzzaman, Muhammad

    2012-01-01

    A total of 125 (ready to eat) processed food samples (70 intended for infant and 55 for adult intake) belonging to 20 different food categories were analyzed for aflatoxins contamination using Reverse Phase High Performance Liquid Chromatography (RP-HPLC) with fluorescent detection. A solvent mixture of acetonitrile-water was used for the extraction followed by immunoaffinity clean-up to enhance sensitivity of the method. The limit of detection (LOD) (0.01–0.02 ng·g−1) and limit of quantification (LOQ) (0.02 ng·g−1) was established for aflatoxins based on signal to noise ratio of 3:1 and 10:1, respectively. Of the processed food samples tested, 38% were contaminated with four types of aflatoxins, i.e., AFB1 (0.02–1.24 μg·kg−1), AFB2 (0.02–0.37 μg·kg−1), AFG1 (0.25–2.7 μg·kg−1) and AFG2 (0.21–1.3 μg·kg−1). In addition, the results showed that 21% of the processed foods intended for infants contained AFB1 levels higher than the European Union permissible limits (0.1 μg·kg−1), while all of those intended for adult consumption had aflatoxin contamination levels within the permitted limits. PMID:22942705

  13. Effect of ultra-superheated steam on aflatoxin reduction and roasted peanut properties.

    PubMed

    Pukkasorn, Parawee; Ratphitagsanti, Wannasawat; Haruthaitanasan, Vichai

    2018-06-01

    Aflatoxins are carcinogenic toxins produced by Aspergillus flavus and Aspergillus parasiticus that are found naturally in peanut. It requires extremely high temperatures to eliminate aflatoxins from the nuts. The aims of this study were to investigate the effect of ultra-superheated steam (USS) on the reduction of aflatoxin B 1 (AFB 1 ) accompanying the roasting process and to determine roasted peanut qualities that affected consumer acceptance. Whole peanut kernels were intentionally contaminated by AFB 1 standard solution at the level of 50 ± 10 µg kg -1 before subjecting to USS treatment at 300-400 °C between 10 and 80 s. The high temperature of USS could significantly decrease AFB 1 level to 9.83 ± 3.51, 15.33 ± 2.23 and 8.95 ± 2.32 µg kg -1 when 300 °C for 80 s, 350 °C for 40 s and 400 °C for 40 s were employed, respectively. AFB 1 was reduced as much as 83.86 ± 2.66% when 400 °C for 40 s was applied. The moisture content of treated peanuts was decreased to less than 3% and browning index was developed from 30.96 ± 1.59 to 95.76 ± 7.23. Higher roasting degree was obtained according to the increase in browning index. Oil quality showed that peroxide values and acid values were greatly below the allowance level. USS could effectively decrease AFB 1 and render expectable roasting qualities of peanut. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  14. Modified Rice Straw as Adsorbent Material to Remove Aflatoxin B1 from Aqueous Media and as a Fiber Source in Fino Bread

    PubMed Central

    Mohamed, Sherif R.; El-Desouky, Tarek A.; Hussein, Ahmed M. S.; Mohamed, Sherif S.; Naguib, Khayria M.

    2016-01-01

    The aims of the current work are in large part the benefit of rice straw to be used as adsorbent material and natural source of fiber in Fino bread. The rice straw was subjected to high temperature for modification process and the chemical composition was carried out and the native rice straw contained about 41.15% cellulose, 20.46% hemicellulose, and 3.91% lignin while modified rice straw has 42.10, 8.65, and 5.81%, respectively. The alkali number was tested and showed an increase in the alkali consumption due to the modification process. The different concentrations of modified rice straw, aflatoxin B1, and pH were tested for removal of aflatoxin B1 from aqueous media and the maximum best removal was at 5% modified rice straw, 5 ng/mL aflatoxin B1, and pH 7. The modified rice straw was added to Fino bread at a level of 5, 10, and 15% and the chemical, rheological, baking quality, staling, and sensory properties were studied. Modified rice straw induced an increase of the shelf life and the produced Fino bread has a better consistency. PMID:26989411

  15. Chronic aflatoxin exposure in children living in Bhaktapur, Nepal: Extension of the MAL-ED study

    USDA-ARS?s Scientific Manuscript database

    Fumonisin B1 (FB1) and aflatoxin B1 (AFB1) are toxic chemicals produced by molds. The molds that produce these two toxic chemicals are commonly found in corn and their co-occurence in corn has been demonstrated in many surveys. This study was conducted because it is suspected that exposure to eith...

  16. Aflatoxin B1 contamination in maize in Europe increases due to climate change

    NASA Astrophysics Data System (ADS)

    Battilani, P.; Toscano, P.; van der Fels-Klerx, H. J.; Moretti, A.; Camardo Leggieri, M.; Brera, C.; Rortais, A.; Goumperis, T.; Robinson, T.

    2016-04-01

    Climate change has been reported as a driver for emerging food and feed safety issues worldwide and its expected impact on the presence of mycotoxins in food and feed is of great concern. Aflatoxins have the highest acute and chronic toxicity of all mycotoxins; hence, the maximal concentration in agricultural food and feed products and their commodities is regulated worldwide. The possible change in patterns of aflatoxin occurrence in crops due to climate change is a matter of concern that may require anticipatory actions. The aim of this study was to predict aflatoxin contamination in maize and wheat crops, within the next 100 years, under a +2 °C and +5 °C climate change scenario, applying a modelling approach. Europe was virtually covered by a net, 50 × 50 km grids, identifying 2254 meshes with a central point each. Climate data were generated for each point, linked to predictive models and predictions were run consequently. Aflatoxin B1 is predicted to become a food safety issue in maize in Europe, especially in the +2 °C scenario, the most probable scenario of climate change expected for the next years. These results represent a supporting tool to reinforce aflatoxin management and to prevent human and animal exposure.

  17. Rapid analysis of aflatoxins B1, B2, and ochratoxin A in rice samples using dispersive liquid-liquid microextraction combined with HPLC.

    PubMed

    Lai, Xian-Wen; Sun, Dai-Li; Ruan, Chun-Qiang; Zhang, He; Liu, Cheng-Lan

    2014-01-01

    A novel, simple, and rapid method is presented for the analysis of aflatoxin B1, aflatoxin B2, and ochratoxin A in rice samples by dispersive liquid-liquid microextraction combined with LC and fluorescence detection. After extraction of the rice samples with a mixture of acetonitrile/water/acetic acid, mycotoxins were rapidly partitioned into a small volume of organic solvent (chloroform) by dispersive liquid-liquid microextraction. The three mycotoxins were simultaneously determined by LC with fluorescence detection after precolumn derivatization for aflatoxin B1 and B2. Parameters affecting both extraction and dispersive liquid-liquid microextraction procedures, including the extraction solvent, the type and volume of extractant, the volume of dispersive solvent, the addition of salt, the pH and the extraction time, were optimized. The optimized protocol provided an enrichment factor of approximately 1.25 and with detection of limits (0.06-0.5 μg/kg) below the maximum levels imposed by current regulations for aflatoxins and ochratoxin A. The mean recovery of three mycotoxins ranged from 82.9-112%, with a RSD less than 7.9% in all cases. The method was successfully applied to measure mycotoxins in commercial rice samples collected from local supermarkets in China. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  18. Risk Assessment on Dietary Exposure to Aflatoxin B1 in Post-Harvest Peanuts in the Yangtze River Ecological Region

    PubMed Central

    Ding, Xiaoxia; Wu, Linxia; Li, Peiwu; Zhang, Zhaowei; Zhou, Haiyan; Bai, Yizhen; Chen, Xiaomei; Jiang, Jun

    2015-01-01

    Based on the 2983 peanut samples from 122 counties in six provinces of China’s Yangtze River ecological region collected between 2009–2014, along with the dietary consumption data in Chinese resident nutrition and health survey reports from 2002 and 2004, dietary aflatoxin exposure and percentiles in the corresponding statistics were calculated by non-parametric probability assessment, Monte Carlo simulation and bootstrap sampling methods. Average climatic conditions in the Yangtze River ecological region were calculated based on the data from 118 weather stations via the Thiessen polygon method. The survey results found that the aflatoxin contamination of peanuts was significantly high in 2013. The determination coefficient (R2) of multiple regression reflected by the aflatoxin B1 content with average precipitation and mean temperature in different periods showed that climatic conditions one month before harvest had the strongest impact on aflatoxin B1 contamination, and that Hunan and Jiangxi provinces were greatly influenced. The simulated mean aflatoxin B1 intake from peanuts at the mean peanut consumption level was 0.777–0.790 and 0.343–0.349 ng/(kg·d) for children aged 2–6 and standard adults respectively. Moreover, the evaluated cancer risks were 0.024 and 0.011/(100,000 persons·year) respectively, generally less than China’s current liver cancer incidence of 24.6 cases/(100,000 persons·year). In general, the dietary risk caused by peanut production and harvest was low. Further studies would focus on the impacts of peanut circulation and storage on aflatoxin B1 contamination risk assessment in order to protect peanut consumers’ safety and boost international trade. PMID:26501322

  19. Surveys of aflatoxin B1 contamination of retail Turkish foods and of products intended for export between 2007 and 2009.

    PubMed

    Ulca, P; Evcimen, M K; Senyuva, H Z

    2010-01-01

    Surveys were carried out between 2007 and 2009 to determine the aflatoxin B1 content of 3345 commercial Turkish foodstuffs supplied by producers for testing for their own purposes or for export certification. To simplify the reporting of data, foods were categorized as: 1, high sugar products with nuts; 2, nuts and seeds; 3, spices; 4, grain; 5, cocoa products; 6, dried fruit and vegetables; 7, processed cereal products; 8, tea; and 9, baby food and infant formula. Aflatoxin analysis was carried out by high-performance liquid chromatography with fluorescence detection after immunoaffinity column clean-up, with a recoveries ranging from 91% to 99%, depending on the matrix. Of the 3345 samples analysed, 94% contained aflatoxin B1 below the European Union limit of 2 µg kg(-1), which applies to nuts, dried fruit, and cereals products. The 6% of the 206 contaminated samples were mainly nuts and spices. For pistachios, 24%, 38%, and 42% of the totals of 207, 182, and 24 samples tested for 2007, 2008 and 2009, respectively, were above 2 µg kg(-1), with 50 samples containing aflatoxin B1 at levels ranging from 10 to 477 µg kg(-1).

  20. Taxonomic comparison of three different groups of aflatoxin producers and a new efficient producer of aflatoxin B1, sterigmatocystin and 3-O-methylsterigmatocystin, Aspergillus rambellii sp. nov.

    PubMed

    Frisvad, Jens C; Skouboe, Pernille; Samson, Robert A

    2005-07-01

    Accumulation of the carcinogenic mycotoxin aflatoxin B, has been reported from members of three different groups of Aspergilli (4) Aspergillus flavus, A. flavus var. parvisclerotigenus, A. parasiticus, A. toxicarius, A. nomius, A. pseudotamarii, A. zhaoqingensis, A. bombycis and from the ascomycete genus Petromyces (Aspergillus section Flavi), (2) Emericella astellata and E. venezuelensis from the ascomycete genus Emericella (Aspergillus section Nidulantes) and (3) Aspergillus ochraceoroseus from a new section proposed here: Aspergillus section Ochraceorosei. We here describe a new species, A. rambellii referable to Ochraceorosei, that accumulates very large amounts of sterigmatocystin, 3-O-methylsterigmatocystin and aflatoxin B1, but not any of the other known extrolites produced by members of Aspergillus section Flavi or Nidulantes. G type aflatoxins were only found in some of the species in Aspergillus section Flavi, while the B type aflatoxins are common in all three groups. Based on the cladistic analysis of nucleotide sequences of ITS1 and 2 and 5.8S, it appears that type G aflatoxin producers are paraphyletic and that section Ochraceorosei is a sister group to the sections Flavi, Circumdati and Cervini, with Emericella species being an outgroup to these sister groups. All aflatoxin producing members of section Flavi produce kojic acid and most species, except A. bombycis and A. pseudotamarii, produce aspergillic acid. Species in Flavi, that produce B type aflatoxins, but not G type aflatoxins, often produced cyclopiazonic acid. No strain was found which produce both G type aflatoxins and cyclopiazonic acid. It was confirmed that some strains of A. flavus var. columnaris produce aflatoxin B2, but this extrolite was not detected in the ex type strain of that variety. A. flavus var. parvisclerotigenus is raised to species level based on the specific combination of small sclerotia, profile of extrolites and rDNA sequence differences. A. zhaoqingensis is regarded

  1. An outbreak of aflatoxin poisoning in dogs associated with aflatoxin B1-contaminated maize products.

    PubMed

    Wouters, Angelica Terezinha Barth; Casagrande, Renata Assis; Wouters, Flademir; Watanabe, Tatiane Terumi Negrão; Boabaid, Fabiana Marques; Cruz, Cláudio Estêvão Farias; Driemeier, David

    2013-03-01

    An aflatoxicosis outbreak affected 65 dogs from 9 different farms after they were fed diets with cooked corn meal as a common ingredient. Of the dogs, 60 died. Numerous dogs died on additional farms, but those dogs were not included in the study. The farmers acquired the contaminated maize products, in the form of whole corn grain or as corn meal, from the same supplier. The corn product was mixed with meat that was left over from home or commercial rations to form corn polenta, which was fed to the dogs. Necropsy was performed on 3 dogs. Two of the dogs died after a few days of refusing food, showing anorexia, polydipsia, icteric mucous membranes, hematemesis, hematochezia, or melena, and bleeding of the skin, eye, ear, and mouth. The primary necropsy findings included jaundice, hemorrhages in several organs, and yellowish enlarged liver with enhanced lobular pattern. The dog that experienced chronic ascites had a yellowish liver with reduced volume, irregular surface, and increased consistency. The main histological findings included hepatocyte fatty degeneration, biliary duct hyperplasia, cholestasis and, in the chronic case, hepatic fibrosis. High-performance liquid chromatography analysis of the corn meal from 2 affected farms revealed 1,640 ppb and 1,770 ppb of aflatoxin B1, respectively. The current study demonstrates an additional way that dogs can be exposed to, poisoned, and killed by aflatoxin.

  2. The effect of substrate, season, and agroecological zone on mycoflora and aflatoxin contamination of poultry feed from Khyber Pakhtunkhwa, Pakistan.

    PubMed

    Alam, Sahib; Shah, Hamid Ullah; Khan, Habibullah; Magan, Naresh

    2012-10-01

    To study the effects of and interactions among feed types, seasons, and agroecological zones on the total fungal viable count and aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1), and G2 (AFG2) production in poultry feed, an experiment was conducted using three-factorial design. A total of 216 samples of poultry feed ingredients, viz. maize, wheat, rice, cotton seed meal (CSM), and finished products, that is, starter and finisher broilers' rations, were collected from Peshawar, Swat, and D. I. Khan districts of Khyber Pakhtunkhwa, Pakistan, during the winter, spring, summer, and autumn seasons of the year 2007/2008. Analysis of variance showed that there was a complex interaction among all these factors and that this influenced the total fungal viable count and relative concentrations of the aflatoxins produced. Minimum total culturable fungi (6.43 × 10³ CFUs/g) were counted in CSM from D. I. Khan region in winter season while maximum (26.68 × 10³ CFUs/g) in starter ration from Peshawar region in summer. Maximum concentrations of AFB1 (191.65 ng/g), AFB2 (86.85 ng/g), and AFG2 (89.90 ng/g) were examined during the summer season whereas the concentration of AFG1 was maximum (167.82 ng/g) in autumn in finisher ration from Peshawar region. Minimum aflatoxins were produced in the winter season across all the three agroecological zones.

  3. Aflatoxins and ochratoxin a reduction in black and white pepper by gamma radiation

    NASA Astrophysics Data System (ADS)

    Jalili, M.; Jinap, S.; Noranizan, M. A.

    2012-11-01

    Irradiation is an important means of decontamination of food commodities, especially spices. The aim of the current study was to investigate the efficacy of gamma radiation (60Co) for decontaminating ochratoxin A (OTA) and aflatoxins B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) residues in artificially contaminated black and white pepper samples. The moisture content of the pepper samples was set at 12% or 18%, and the applied gamma dose ranged from 5 to 30 kGy. Mycotoxin levels were determined by high-performance liquid chromatography (HPLC) after immunoaffinity column (IAC) chromatography. Both the gamma irradiation dose and moisture content showed significant effects (P<0.05) on mycotoxin reduction. The maximum toxin reductions, found at 18% moisture content and 30 kGy, were 55.2%, 50.6%, 39.2%, 47.7% and 42.9% for OTA, AFB1, AFB2, AFG1 and AFG2, respectively.

  4. Characterization of the Critical Amino Acids of an Aspergillus parasiticus Cytochrome P-450 Monooxygenase Encoded by ordA That Is Involved in the Biosynthesis of Aflatoxins B1, G1, B2, and G2

    PubMed Central

    Yu, Jiujiang; Chang, Perng-Kuang; Ehrlich, Kenneth C.; Cary, Jeffrey W.; Montalbano, Beverly; Dyer, John M.; Bhatnagar, Deepak; Cleveland, Thomas E.

    1998-01-01

    The conversion of O-methylsterigmatocystin (OMST) and dihydro-O-methylsterigmatocystin to aflatoxins B1, G1, B2, and G2 requires a cytochrome P-450 type of oxidoreductase activity. ordA, a gene adjacent to the omtA gene, was identified in the aflatoxin-biosynthetic pathway gene cluster by chromosomal walking in Aspergillus parasiticus. The ordA gene was a homolog of the Aspergillus flavus ord1 gene, which is involved in the conversion of OMST to aflatoxin B1. Complementation of A. parasiticus SRRC 2043, an OMST-accumulating strain, with the ordA gene restored the ability to produce aflatoxins B1, G1, B2, and G2. The ordA gene placed under the control of the GAL1 promoter converted exogenously supplied OMST to aflatoxin B1 in Saccharomyces cerevisiae. In contrast, the ordA gene homolog in A. parasiticus SRRC 2043, ordA1, was not able to carry out the same conversion in the yeast system. Sequence analysis revealed that the ordA1 gene had three point mutations which resulted in three amino acid changes (His-400→Leu-400, Ala-143→Ser-143, and Ile-528→Tyr-528). Site-directed mutagenesis studies showed that the change of His-400 to Leu-400 resulted in a loss of the monooxygenase activity and that Ala-143 played a significant role in the catalytic conversion. In contrast, Ile-528 was not associated with the enzymatic activity. The involvement of the ordA gene in the synthesis of aflatoxins G1, and G2 in A. parasiticus suggests that enzymes required for the formation of aflatoxins G1 and G2 are not present in A. flavus. The results showed that in addition to the conserved heme-binding and redox reaction domains encoded by ordA, other seemingly domain-unrelated amino acid residues are critical for cytochrome P-450 catalytic activity. The ordA gene has been assigned to a new cytochrome P-450 gene family named CYP64 by The Cytochrome P450 Nomenclature Committee. PMID:9835571

  5. Aflatoxin M1 contamination of milk and ice cream in Abeokuta and Odeda local governments of Ogun State, Nigeria.

    PubMed

    Atanda, Olusegun; Oguntubo, Adenike; Adejumo, Oloyede; Ikeorah, John; Akpan, Iyang

    2007-07-01

    A survey was undertaken to determine the aflatoxin M(1) contamination of milk and some locally produced dairy products in Abeokuta and Odeda local governments of Ogun State, Nigeria. Samples of human and cow milk, yoghurt, "wara", ice cream and "nono" were collected randomly within the local governments and analysed for aflatoxin M(1) using the two-dimensional TLC. Aflatoxin M(1) contamination in the range of 2.04-4.00 microg l(-1) was noticed only in milk and ice cream. In particular, samples of human milk, cow milk and ice cream recorded high scores of 4.0 microg l(-1), 2.04 microg l(-1) and 2.23 microg l(-1), respectively in Abeokuta local governments and a score of 4.0 microg l(-1) for cow milk in Odeda local government. This indicates a high level contamination in the local governments since the weighted mean concentration of aflatoxin M1 in milk for African diet is 0.002 microg l(-1). Therefore the concentration of AFB1 in feeds which is transformed to AFM1 in milk should be reduced by good manufacturing and good storage practices. Furthermore, there is need for stringent quality control during processing and distribution of these products.

  6. A Survey of Aflatoxin-Producing Aspergillus sp. from Peanut Field Soils in Four Agroecological Zones of China

    PubMed Central

    Zhang, Chushu; Selvaraj, Jonathan Nimal; Yang, Qingli; Liu, Yang

    2017-01-01

    Peanut pods are easily infected by aflatoxin-producing Aspergillus sp.ecies from field soil. To assess the aflatoxin-producing Aspergillus sp. in different peanut field soils, 344 aflatoxin-producing Aspergillus strains were isolated from 600 soil samples of four agroecological zones in China (the Southeast coastal zone (SEC), the Yangtze River zone (YZR), the Yellow River zone (YR) and the Northeast zone (NE)). Nearly 94.2% (324/344) of strains were A. flavus and 5.8% (20/344) of strains were A. parasiticus. YZR had the highest population density of Aspergillus sp. and positive rate of aflatoxin production in isolated strains (1039.3 cfu·g−1, 80.7%), the second was SEC (191.5 cfu·g−1, 48.7%), the third was YR (26.5 cfu·g−1, 22.7%), and the last was NE (2.4 cfu·g−1, 6.6%). The highest risk of AFB1 contamination on peanut was in YZR which had the largest number of AFB1 producing isolates in 1g soil, followed by SEC and YR, and the lowest was NE. The potential risk of AFB1 contamination in peanuts can increase with increasing population density and a positive rate of aflatoxin-producing Aspergillus sp. in field soils, suggesting that reducing aflatoxigenic Aspergillus sp. in field soils could prevent AFB1 contamination in peanuts. PMID:28117685

  7. Highly Sensitive FRET-Based Fluorescence Immunoassay for Detecting of Aflatoxin B1 Using Magnetic/Silica Core-Shell as a Signal Intensifier.

    PubMed

    Kalarestaghi, Alireza; Bayat, Mansour; Hashemi, Seyed Jamal; Razavilar, Vadood

    2015-09-01

    Recently, some new nanobiosensors using different nanoparticles or microarray systems for detection of mycotoxins have been designed . However, rapid, sensitive and early detection of aflatoxicosis would be very helpful to distinguish high-risk persons. We report a highly sensitive competitive immunoassay using magnetic/silica core shell as a signal intensifier for the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET) from Cd/Te quantum dots (antiaflatoxin B1 antibody immobilized on the surface of Cd/Te quantum dots) to Rhodamine 123 (Rho 123-labeled aflatoxin B1 bound to albumin). The specific immune-reaction between the anti-aflatoxin B1 antibody on the QDs and the labeledaflatoxin B1 brings the Rho 123 fluorophore (acting as the acceptor) and the QDs (acting as the donor) in close spatial proximity and causes FRET to occur upon photo-excitation of the QDs. Using magnetic/silica core shell to intensify the obtained signal is the novelty of this study. Cd/Te QDs were synthesized by the simultaneous reduction of cadmium chloride and tellurium in the presence of sodium borohydride under nitrogen atmosphere. Magnetic nanoparticles were synthesized using FeSO 4 and FeCl 3 (1:2 molar ratio) and ammonia as an oxidizing agent under nitrogen atmosphere. The prepared magnetic nanoparticles shelled by silica using tetraethoxysilane in the presence of ammonia. Nanoparticles synthesis and monodispersity confirmed by TEM. Immobilization of Cd/Te QDs to antibodies and labeling of aflatoxin B1-albumin by Rho 123 were performed by EDC/NHS reaction in reaction mixture buffer, pH 6, at room temperature. By using the magnetic/silica core shell sensitivity of the system changed from 2×10 -11 in our previous study to 2×10 -12 in this work. The feasibility of the method established by the detection of aflatoxin B1 in spiked human serum. There is a linear relationship between the decreased fluorescence intensity of Rho 123 with increasing concentration

  8. Efficacy of adsorbents (bentonite and diatomaceous earth) and turmeric (Curcuma longa) in alleviating the toxic effects of aflatoxin in chicks.

    PubMed

    Dos Anjos, F R; Ledoux, D R; Rottinghaus, G E; Chimonyo, M

    2015-01-01

    A study was conducted to determine the efficacy of bentonite clay (BC), diatomaceous earth (DE) and turmeric powder (TUM) in alleviating the toxic effects of aflatoxin B1 (AFB1). A total of 250 Ross-308 d-old male broiler chicks were assigned to 10 dietary treatments (5 replicates of 5 chicks) from hatch to d 21. Dietary treatments were: basal diet; basal diet plus AFB1 (2 mg) or BC (0.75%), or DE (0.75%), or TUM (200 mg/kg curcuminoids) and different combinations of AFB1, BC, DE and TUM. Feed intake (FI), body weight gain (BWG) and feed gain (FG) of the birds fed on BC or DE separately were not different from control birds. Birds fed on TUM only had similar FI and FG but lower BWG than control chicks. Aflatoxin B1 reduced FI, BWG and serum concentrations of glucose, albumin, total protein calcium, but increased FG and relative liver and kidney weights. Chicks fed on the combination of AFB1 and BC had similar FI and FG to control chicks. Chicks fed on the combination of DE and AFB1 had lower FI (23.1%) and BWG (28.6%) compared with control chicks. Chicks fed on the combination of TUM and AFB1 also had decreased FI (26.2 %) and BWG (31%) compared with control chicks. Chicks fed on the combination of AFB1, BC and TUM consumed significantly higher amounts of feed compared with chicks fed on only AF, but gained less when compared with control diet chicks. Chicks fed on the combination of AFB1, DE and TUM diet had poorer growth performance than those fed on AFB1 alone. None of the combination diets reduced the severity of liver lesions.

  9. Common African cooking processes do not affect the aflatoxin binding efficacy of refined calcium montmorillonite clay

    PubMed Central

    Elmore, Sarah E.; Mitchell, Nicole; Mays, Travis; Brown, Kristal; Marroquin-Cardona, Alicia; Romoser, Amelia; Phillips, Timothy D.

    2013-01-01

    Aflatoxins are common contaminants of staple crops, such as corn and groundnuts, and a significant cause of concern for food safety and public health in developing countries. Aflatoxin B1 (AFB1) has been implicated in the etiology of acute and chronic disease in humans and animals, including growth stunting, liver cancer and death. Cost effective and culturally acceptable intervention strategies for the reduction of dietary AFB1 exposure are of critical need in populations at high risk for aflatoxicosis. Fermented gruels consisting of cornmeal are a common source for such exposure and are consumed by both children and adults in many countries with a history of frequent, high-level aflatoxin exposure. One proposed method to reduce aflatoxins in the diet is to include a selective enterosorbent, Uniform Particle Size NovaSil (UPSN), as a food additive in contaminated foods. For UPSN to be effective in this capacity, it must be stable in complex, acidic mixtures that are often exposed to heat during the process of fermented gruel preparation. Therefore, the objective of the present study was to test the ability of UPSN to sorb aflatoxin while common cooking conditions were applied. The influence of fermentation, heat treatment, acidity, and processing time were investigated with and without UPSN. Analyses were performed using the field-practical Vicam assay with HPLC verification of trends. Our findings demonstrated that UPSN significantly reduced aflatoxin levels (47-100%) in cornmeal, regardless of processing conditions. Upon comparison of each element tested, time appeared to be the primary factor influencing UPSN efficacy. The greatest decreases in AFB1 were reported in samples allowed to incubate (with or without fermentation) for 72 hrs. This data suggests that addition of UPSN to staple corn ingredients likely to contain aflatoxins would be a sustainable approach to reduce exposure. PMID:24311894

  10. Common African cooking processes do not affect the aflatoxin binding efficacy of refined calcium montmorillonite clay.

    PubMed

    Elmore, Sarah E; Mitchell, Nicole; Mays, Travis; Brown, Kristal; Marroquin-Cardona, Alicia; Romoser, Amelia; Phillips, Timothy D

    2014-03-01

    Aflatoxins are common contaminants of staple crops, such as corn and groundnuts, and a significant cause of concern for food safety and public health in developing countries. Aflatoxin B 1 (AFB 1 ) has been implicated in the etiology of acute and chronic disease in humans and animals, including growth stunting, liver cancer and death. Cost effective and culturally acceptable intervention strategies for the reduction of dietary AFB 1 exposure are of critical need in populations at high risk for aflatoxicosis. Fermented gruels consisting of cornmeal are a common source for such exposure and are consumed by both children and adults in many countries with a history of frequent, high-level aflatoxin exposure. One proposed method to reduce aflatoxins in the diet is to include a selective enterosorbent, Uniform Particle Size NovaSil (UPSN), as a food additive in contaminated foods. For UPSN to be effective in this capacity, it must be stable in complex, acidic mixtures that are often exposed to heat during the process of fermented gruel preparation. Therefore, the objective of the present study was to test the ability of UPSN to sorb aflatoxin while common cooking conditions were applied. The influence of fermentation, heat treatment, acidity, and processing time were investigated with and without UPSN. Analyses were performed using the field-practical Vicam assay with HPLC verification of trends. Our findings demonstrated that UPSN significantly reduced aflatoxin levels (47-100%) in cornmeal, regardless of processing conditions. Upon comparison of each element tested, time appeared to be the primary factor influencing UPSN efficacy. The greatest decreases in AFB 1 were reported in samples allowed to incubate (with or without fermentation) for 72 hrs. This data suggests that addition of UPSN to staple corn ingredients likely to contain aflatoxins would be a sustainable approach to reduce exposure.

  11. Co-exposure to fumonisins and aflatoxins in maize-based foods in central america: guatemala as a case study

    USDA-ARS?s Scientific Manuscript database

    Aflatoxin B1 (AFB1) is a human liver carcinogen having a genotoxic mechanism of action. The ceramide synthase inhibitor fumonisin B1 (FB1) is a liver cancer promoter in rats and trout. Both mycotoxins are found in maize so that the possibility of co-exposure is a health concern, especially in Centra...

  12. Antigenotoxic Studies of Different Substances to Reduce the DNA Damage Induced by Aflatoxin B1 and Ochratoxin A

    PubMed Central

    Madrigal-Santillán, Eduardo; Morales-González, José A.; Vargas-Mendoza, Nancy; Reyes-Ramírez, Patricia; Cruz-Jaime, Sandra; Sumaya-Martínez, Teresa; Pérez-Pastén, Ricardo; Madrigal-Bujaidar, Eduardo

    2010-01-01

    Mycotoxins are produced mainly by the mycelial structure of filamentous fungi, or more specifically, molds. These secondary metabolites are synthesized during the end of the exponential growth phase and appear to have no biochemical significance in fungal growth and development. The contamination of foods and feeds with mycotoxins is a significant problem for the adverse effects on humans, animals, and crops that result in illnesses and economic losses. The toxic effect of the ingestion of mycotoxins in humans and animals depends on a number of factors including intake levels, duration of exposure, toxin species, mechanisms of action, metabolism, and defense mechanisms. In general, the consumption of contaminated food and feed with mycotoxin induces to neurotoxic, immunosuppressive, teratogenic, mutagenic, and carcinogenic effect in humans and/or animals. The most significant mycotoxins in terms of public health and agronomic perspective include the aflatoxins, ochratoxin A (OTA), trichothecenes, fumonisins, patulin, and the ergot alkaloids. Due to the detrimental effects of these mycotoxins, several strategies have been developed in order to reduce the risk of exposure. These include the degradation, destruction, inactivation or removal of mycotoxins through chemical, physical and biological methods. However, the results obtained with these methods have not been optimal, because they may change the organoleptic characteristics and nutritional values of food. Another alternative strategy to prevent or reduce the toxic effects of mycotoxins is by applying antimutagenic agents. These substances act according to several extra- or intracellular mechanisms, their main goal being to avoid the interaction of mycotoxins with DNA; as a consequence of their action, these agents would inhibit mutagenesis and carcinogenesis. This article reviews the main strategies used to control AFB1 and ochratoxin A and contains an analysis of some antigenotoxic substances that reduce the

  13. Milk kefir: ultrastructure, antimicrobial activity and efficacy on aflatoxin B1 production by Aspergillus flavus.

    PubMed

    Ismaiel, Ahmed A; Ghaly, Mohamed F; El-Naggar, Ayman K

    2011-05-01

    The association of kefir microbiota was observed by electron microscopic examination. Scanning electron microscopic (SEM) observations revealed that kefir grain surface is very rough and the inner portions had scattered irregular holes on its surface. The interior of the grain comprised fibrillar materials which were interpreted as protein, lipid and a soluble polysaccharide, the kefiran complex that surrounds yeast and bacteria in the grain. Yeast was observed more clearly than bacteria on the outer portion of the grain. Transmission electron microscopic (TEM) observations of kefir revealed that the grain comprised a mixed culture of yeast and bacteria growing in close association with each other. Microbiota is dominated by budded and long-flattened yeast cells growing together with lactobacilli and lactococci bacteria. Bacterial cells with rounded ends were also observed in this mixed culture. Kefir grains, kefir suspensions, and kefiran were tested for antimicrobial activities against several bacterial and fungal species. The highest activity was obtained against Streptococcus faecalis KR6 and Fusarium graminearum CZ1. Growth of Aspergillus flavus AH3 producing for aflatoxin B1 for 10 days in broth medium supplemented with varying concentrations of kefir filtrate (%, v/v) showed that sporulation was completely inhibited at the higher concentrations of kefir filtrate (7-10%, v/v). The average values of both mycelial dry weights and aflatoxin B1 were completely inhibited at 10% (v/v). This is the first in vitro study about the antifungal characteristics of kefir against filamentous fungi which was manifested by applying its inhibitory effect on the productivity of aflatoxin B1 by A. flavus AH3.

  14. High sensitive and throughput screening of Aflatoxin using MALDI-TOF-TOF-PSD-MS/MS

    USDA-ARS?s Scientific Manuscript database

    We have achieved sensitive and efficient detection of aflatoxin B1(AFB1) through matrix-assisted laser desorption/ionization time-of-flight-time-of-flight mass spectrometry (MALDI-TOF-TOF) and post-source decay (PSD) tandem mass spectrometry (MS/MS) using an acetic acid – a-cyano-4-hydroxycinnamic a...

  15. Streptomyces-Aspergillus flavus interactions: impact on aflatoxin B accumulation.

    PubMed

    Verheecke, C; Liboz, T; Anson, P; Zhu, Y; Mathieu, F

    2015-01-01

    The aim of this work was to investigate the potential of Streptomyces sp. as biocontrol agents against aflatoxins in maize. As such, we assumed that Streptomyces sp. could provide a complementary approach to current biocontrol systems such as Afla-guard(®) and we focused on biocontrol that was able to have an antagonistic contact with A. flavus. A previous study showed that 27 (out of 38) Streptomyces sp. had mutual antagonism in contact with A. flavus. Among these, 16 Streptomyces sp. were able to reduce aflatoxin content to below 17% of the residual concentration. We selected six strains to understand the mechanisms involved in the prevention of aflatoxin accumulation. Thus, in interaction with A. flavus, we monitored by RT-qPCR the gene expression of aflD, aflM, aflP, aflR and aflS. All the Streptomyces sp. were able to reduce aflatoxin concentration (24.0-0.2% residual aflatoxin B1). They all impacted on gene expression, but only S35 and S38 were able to repress expression significantly. Indeed, S35 significantly repressed aflM expression and S38 significantly repressed aflR, aflM and aflP. S6 reduced aflatoxin concentrations (2.3% residual aflatoxin B1) and repressed aflS, aflM and enhanced aflR expression. In addition, the S6 strain (previously identified as the most reducing pure aflatoxin B1) was further tested to determine a potential adsorption mechanism. We did not observe any adsorption phenomenon. In conclusion, this study showed that Streptomyces sp. prevent the production of (aflatoxin gene expression) and decontamination of (aflatoxin B1 reduction) aflatoxins in vitro.

  16. Antifungal Activity and Aflatoxin Degradation of Bifidobacterium Bifidum and Lactobacillus Fermentum Against Toxigenic Aspergillus Parasiticus

    PubMed Central

    Ghazvini, Roshanak Daie; Kouhsari, Ebrahim; Zibafar, Ensieh; Hashemi, Seyed Jamal; Amini, Abolfazl; Niknejad, Farhad

    2016-01-01

    Food and feedstuff contamination with aflatoxins (AFTs) is a serious health problem for humans and animals, especially in developing countries. The present study evaluated antifungal activities of two lactic acid bacteria (LAB) against growth and aflatoxin production of toxigenic Aspergillus parasiticus. The mycelial growth inhibition rate of A. parasiticus PTCC 5286 was investigated in the presence of Bifidobacterium bifidum PTCC 1644 and Lactobacillus fermentum PTCC 1744 by the pour plate method. After seven days incubation in yeast extract sucrose broth at 30°C, the mycelial mass was weighed after drying. The inhibitory activity of LAB metabolites against aflatoxin production by A. parasiticus was evaluated using HPLC method. B. bifidum and L. fermentum significantly reduced aflatoxin production and growth rate of A. parasiticus in comparison with the controls (p≤0.05). LAB reduced total aflatoxins and B1, B2, G1 and G2 fractions by more than 99%. Moreover, LAB metabolites reduced the level of standard AFB1, B2, G1 and G2 from 88.8% to 99.8% (p≤0.05). Based on these findings, B. bifidum and L. fermentum are recommended as suitable biocontrol agents against the growth and aflatoxin production by aflatoxigenic Aspergillus species. PMID:28077976

  17. Antifungal Activity and Aflatoxin Degradation of Bifidobacterium Bifidum and Lactobacillus Fermentum Against Toxigenic Aspergillus Parasiticus.

    PubMed

    Ghazvini, Roshanak Daie; Kouhsari, Ebrahim; Zibafar, Ensieh; Hashemi, Seyed Jamal; Amini, Abolfazl; Niknejad, Farhad

    2016-01-01

    Food and feedstuff contamination with aflatoxins (AFTs) is a serious health problem for humans and animals, especially in developing countries. The present study evaluated antifungal activities of two lactic acid bacteria (LAB ) against growth and aflatoxin production of toxigenic Aspergillus parasiticus . The mycelial growth inhibition rate of A. parasiticus PTCC 5286 was investigated in the presence of Bifidobacterium bifidum PTCC 1644 and Lactobacillus fermentum PTCC 1744 by the pour plate method. After seven days incubation in yeast extract sucrose broth at 30°C, the mycelial mass was weighed after drying. The inhibitory activity of LAB metabolites against aflatoxin production by A. parasiticus was evaluated using HPLC method. B. bifidum and L. fermentum significantly reduced aflatoxin production and growth rate of A. parasiticus in comparison with the controls (p≤0.05). LAB reduced total aflatoxins and B 1 , B 2 , G 1 and G 2 fractions by more than 99%. Moreover, LAB metabolites reduced the level of standard AFB 1 , B 2 , G 1 and G 2 from 88.8% to 99.8% (p≤0.05). Based on these findings, B. bifidum and L. fermentum are recommended as suitable biocontrol agents against the growth and aflatoxin production by aflatoxigenic Aspergillus species.

  18. Intervention trial with calcium montmorillonite clay in a south Texas population exposed to aflatoxin.

    PubMed

    Pollock, Brad H; Elmore, Sarah; Romoser, Amelia; Tang, Lili; Kang, Min-Su; Xue, Kathy; Rodriguez, Marisa; Dierschke, Nicole A; Hayes, Holly G; Hansen, H Andrew; Guerra, Fernando; Wang, Jia-Sheng; Phillips, Timothy

    2016-08-01

    South Texas currently has the highest incidence of hepatocellular carcinoma (HCC) in the United States, a disease that disproportionately affects Latino populations in the region. Aflatoxin B1 (AFB1) is a potent liver carcinogen that has been shown to be present in a variety of foods in the United States, including corn and corn products. Importantly, it is a dietary risk factor contributing to a higher incidence of HCC in populations frequently consuming AFB1-contaminated diets. In a randomised double-blind placebo controlled trial, we evaluated the effects of a 3-month administration of ACCS100 (refined calcium montmorillonite clay) on serum AFB1-lysine adduct (AFB-Lys) level and serum biochemistry in 234 healthy men and women residing in Bexar and Medina counties, Texas. Participants recruited from 2012 to 2014 received either a placebo, 1.5 g or 3 g ACCS100 each day for 3 months, and no treatment during the fourth month. Adverse event rates were similar across treatment groups and no significant differences were observed for serum biochemistry and haematology parameters. Differences in levels of AFB-Lys at 1, 3 and 4 months were compared between placebo and active treatment groups. Although serum AFB-Lys levels were decreased by month 3 for both treatment groups, the low dose was the only treatment that was significant (p = 0.0005). In conclusion, the observed effect in the low-dose treatment group suggests that the use of ACCS100 may be a viable strategy to reduce dietary AFB1 bioavailability during aflatoxin outbreaks and potentially in populations chronically exposed to this carcinogen.

  19. Differences in response of glucuronide and glutathione conjugating enzymes to aflatoxin B/sub 1/ and N-acetylaminofluorene in underfed rats

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Rajpurohit, R.; Krishnaswamy, K.

    Changes in the hepatic drug/xenobiotic-metabolizing enzymes in underfed rats exposed to aflatoxin B/sub 1/ and N-acetylaminofluorene were investigated. Neither carcinogen, fed at the level of 10 ..mu..g and 0.667 mg per 100 g body weight, respectively, over a period of 3 wk, had any significant influence on cytochrome P-450 and aryl hydrocarbon hydroxylase in the undernourished rats. Significantly low activities of UDP-glucuronyltransferase and glutathione S-transferase were observed in food-restricted animals fed on aflatoxin B/sub 1/. N-acetylaminofluorene, on the other hand stimulated both the enzyme activities in the underfed group, to as much observed in the respective well-fed treated group. UDP-Glucuronyltransferasemore » and glutathione S-transferase in undernutrition seem to respond differently to aflatoxin B/sub 1/ and N-acetylaminofluorene. Further studies are needed to assess the possible consequences of such alterations.« less

  20. Natural postharvest aflatoxin occurrence in food legumes in the smallholder farming sector of Zimbabwe.

    PubMed

    Maringe, David Tinayeshe; Chidewe, Cathrine; Benhura, Mudadi Albert; Mvumi, Brighton Marimanzi; Murashiki, Tatenda Clive; Dembedza, Mavis Precious; Siziba, Lucia; Nyanga, Loveness Kuziwa

    2017-03-01

    Aflatoxins, mainly produced by Aspergillus flavus and Aspergillus parasiticus, are highly toxic and may lead to health problems such as liver cancer. Exposure to aflatoxins may result from ingestion of contaminated foods. Levels of AFB 1 , AFB 2 , AFG 1 and AFG 2 in samples of groundnuts (Arachis hypogaea), beans (Phaseolus vulgaris), cowpeas (Vigna unguiculata) and bambara nuts (Vigna subterranean) grown by smallholder farmers in Shamva and Makoni districts, Zimbabwe, were determined at harvesting, using high performance liquid chromatography after immunoaffinity clean-up. Aflatoxins were detected in 12.5% of groundnut samples with concentrations ranging up to 175.9 µg/kg. Aflatoxins were present in 4.3% of the cowpea samples with concentrations ranging from 1.4 to 103.4 µg/kg. Due to alarming levels of aflatoxins detected in legumes versus maximum permissible levels, there is a need to assist smallholder farmers to develop harvest control strategies to reduce contamination of aflatoxins in legumes.

  1. Tissue Distribution and Metabolism of Aflatoxin B1-14C in Broiler Chickens

    PubMed Central

    Mabee, Michael S.; Chipley, John R.

    1973-01-01

    The effects of administering low levels of aflatoxin B1-14C by crop intubation daily for 14 days to broiler chickens were determined. Studies on the distribution of 14C in the blood, selected organs, tissues, and excreta were conducted. No toxic effects were observed in broiler chickens during the 14 days of the experiment. The broiler chickens excreted 90.64% of the 14C administered. Of the 14C retained, 11.04, 9.83, 4.30, 12.52, 31.66, and 30.63% were detected in the blood, liver, heart, gizzard, breast, and leg, respectively. Chemical assay of those samples demonstrating radioactivity revealed that 81.2% of the radioactivity in these substrates was not extractable by classical extraction procedures while approximately 10% was extractable. Treatment of aqueous extracts for conjugated steroids by treatments with beta-glucuronidase revealed that 31.5% of the 14C detected in the aqueous extract was a liberated glucuronide conjugate of aflatoxin M1-14C. PMID:4715554

  2. Determination of the aflatoxin M1 (AFM1) from milk by direct analysis in real time - mass spectrometry (DART-MS)

    USDA-ARS?s Scientific Manuscript database

    Certain fungi that grow on crops can produce aflatoxins, which are highly carcinogenic. One of these, aflatoxin B1 can be metabolized by mammals to aflatoxin M1, a form that retains potent carcinogenicity and which can be excreted into milk. Direct analysis in real time (DART) ionization coupled to ...

  3. [Biological contamination by micromycetes in dried Boletus edulis: research of aflatoxin B1, B2 G1, G2 and ochratoxin A].

    PubMed

    Lorini, C; Rossetti, F; Palazzoni, S; Comodo, N; Bonaccorsi, G

    2008-01-01

    Aim of this survey is to identify those filamentous fungi which parasite Boletus edulis and its group and check the potential presence of secondary metabolites, specifically aflatoxin B1, total aflatoxins and ochratoxin A, in order to assess the risk to consumers' health. Forty samples of dried Boletus edulis, collected by two food industries which distribute the product in many Italian regions, have been analysed. The sampling plan has been conducted from November 2005 to March 2006, collecting 50 g from each commercial category of dried Boletus edulis available in the factory at the time of sampling. All the samples have been tested by visual macroscopic and stereoscopic assays; for some samples--those referred to commercial category presumably at higher risk--we have performed cultural assays as well, typization of isolated micromycetes, extraction and quantification of aflatoxins and ochratoxin A. Mycotoxin detection has been made by HPLC, using the UNI EN 14123 and UNI EN 14132 standard methods, respectively applied to aflatoxins determination in peanuts, pistachios, figs and paprika and to ochratoxin A in barley and coffee. Non pathogenic micromycetes, common in food products, have been frequently observed in cultural assays, while Aspergillus flavus and Aspergillus niger have been found in some samples. However the concentration of aflatoxins was always under the quantification limit. The survey confirm that, if the cold chain is kept throughout the process and the distribution, Boletus edulis and analogue mycetes are not a favourable substratum for the growth and the development of moulds.

  4. Time-course of germination, initiation of mycelium proliferation and probability of visible growth and detectable AFB1 production of an isolate of Aspergillus flavus on pistachio extract agar.

    PubMed

    Aldars-García, Laila; Sanchis, Vicente; Ramos, Antonio J; Marín, Sonia

    2017-06-01

    The aim of this work was to assess the temporal relationship among quantified germination, mycelial growth and aflatoxin B 1 (AFB1) production from colonies coming from single spores, in order to find the best way to predict as accurately as possible the presence of AFB1 at the early stages of contamination. Germination, mycelial growth, probability of growth and probability of AFB1 production of an isolate of Aspergillus flavus were determined at 25 °C and two water activities (0.85 and 0.87) on 3% Pistachio Extract Agar (PEA). The percentage of germinated spores versus time was fitted to the modified Gompertz equation for the estimation of the germination parameters (geometrical germination time and germination rate). The radial growth curve for each colony was fitted to a linear model for the estimation of the apparent lag time for growth and the growth rate, and besides the time to visible growth was estimated. Binary data obtained from growth and AFB1 studies were modeled using logistic regression analysis. Both water activities led to a similar fungal growth and AFB1 production. In this study, given the suboptimal set conditions, it has been observed that germination is a stage far from the AFB1 production process. Once the probability of growth started to increase it took 6 days to produce AFB1, and when probability of growth was 100%, only a 40-57% probability of detection of AFB1 production was predicted. Moreover, colony sizes with a radius of 1-2 mm could be a helpful indicator of the possible AFB1 contamination in the commodity. Despite growth models may overestimate the presence of AFB1, their use would be a helpful tool for producers and manufacturers; from our data 5% probability of AFB1 production (initiation of production) would occur when a minimum of 60% probability of growth is observed. Legal restrictions are quite severe for these toxins, thus their control from the early stages of contamination throughout the food chain is of paramount

  5. [Dietary exposure assessment of aflatoxin of foodstuff and edible oil from Shenzhen residents].

    PubMed

    Li, Ke; Qiu, Fen; Jiang, Lixin; Yang, Mei

    2014-07-01

    To assess the dietary exposure aflatoxin B1 and total aflatoxins of foodstuff and edible oil in Shenzhen residents. Aflatoxins in the samples were determined by the immuno-affinity column clean-up plus UPLC. The aflatoxin B1 and aflatoxins dietary exposure were calculated by the level of aflatoxins contamination in the food and consumption of dietary. The average diary aflatoxin B1 dietary exposure of the man of the 2 to 6, 7 to 14, 15 to 50 and > 50 age group in Shenzhen were 0.320, 0.385, 0.401 and 0.398 ng/(kg BW x d), the results of the woman were 0.282, 0.222, 0.367 and 0.470 ng/(kg BW x d) respectively. The total average daily dietary aflatoxin B1 exposure of the man were 0.012, 0.015, 0.016 and 0.016 ng/(kg BW x d) about each age group. The results of the woman were 78.4, 167, 113 and 103 ng/(kg BW d). According to the the average levels of consumption and the high levels of consumption, the risk of AFB, of the man were 0.012,0.015, 0.016, 0. 016 and 3.0, 8.2, 4.1, 4.4 cancer patient per one hundred thousand, respectively. The results of the woman were 0.010, 0.009, 0.014, 0.018 and 2.9, 6.7, 4.4, 4.0 cancer patient per one hundred thousand, respectively. 7 to 14 age group compared with adults age group face higher exposure levels. The rice and peanut oil are most primary aflatoxin dietary exposure sources in Shenzhen.

  6. Enzyme-linked immunosorbent assay for determination of aflatoxin M1 based on magnetic nanoparticles

    NASA Astrophysics Data System (ADS)

    Atanasova, M. K.; Ivanova, N. V.; Godjevargova, T. I.

    2017-02-01

    A sensitive enzyme immunoassay with magnetic nanoparticles (Method A) for the quantitative determination of aflatoxin M1 in milk was developed. This immunoassay was based on the immobilization of monoclonal antibody (mAb) on the modified magnetic nanoparticles (MNPs-NH2). It was observed that for each mg of the MNPs, 25 µg of antibody was immobilized. Both aflatoxin M1 in the sample and aflatoxin M1-BSA-peroxidase conjugate competed for the immobilized antibody. The proposed Method A was compared with other method (B). The Method B was based on the immobilization of aflatoxin M1-BSA conjugate on the MNPs-NH2, which competed with the aflatoxin M1 in the sample for binding to the added mAb. The binding of mAb to the aflatoxin M1-BSA-MNPs-NH2 was detected using a target secondary IgG-peroxidase antibody. The analytical characteristics of the two methods were compared. Real milk samples were investigated for present of aflatoxin M1. Two methods were based on the use of MNPs as a solid support for covalently immunoreagents immobilization. A comfortable separation of bound and free fraction of the tracer can be performed only through a simple collection of the MNPs by a permanent magnet. The application of MNPs helps to eliminate non-specific binding and to retain higher activity of bound biomolecules. The development of a MNPs-based ELISA for determination of aflatoxin M1 has a great potential to supersede the traditional ELISA for aflatoxin M1 diagnosis.

  7. Streptomyces sp. ASBV-1 reduces aflatoxin accumulation by Aspergillus parasiticus in peanut grains.

    PubMed

    Zucchi, T D; de Moraes, L A B; de Melo, I S

    2008-12-01

    To evaluate the ability of Streptomyces sp. (strain ASBV-1) to restrict aflatoxin accumulation in peanut grains. In the control of many phytopathogenic fungi the Streptomyces sp. ASBV-1 strain showed promise. An inhibitory test using this strain and A. parasiticus was conducted in peanut grains to evaluate the effects of this interaction on spore viability and aflatoxin accumulation. In some treatments the Streptomyces sp ASBV-1 strain reduced the viability of A. parasiticus spores by c. 85%, and inhibited aflatoxin accumulation in peanut grains. The values of these reductions ranged from 63 to 98% and from 67% to 96% for aflatoxins B(1) and G(1), respectively. It was demonstrated that Streptomyces sp. ASBV-1 is able to colonize peanut grains and thus inhibit the spore viability of A. parasiticus, as well as reducing aflatoxin production. The positive finding for aflatoxin accumulation reduction in peanut grains seems promising and suggests a wider use of this actinobacteria in biological control programmes.

  8. Aflatoxins, hydroxylated metabolites, and aflatoxicol from breast muscle of laying hens.

    PubMed

    Díaz-Zaragoza, M; Carvajal-Moreno, M; Méndez-Ramírez, I; Chilpa-Galván, N C; Avila-González, E; Flores-Ortiz, C M

    2014-12-01

    Aflatoxins (AF) are toxic fungal secondary metabolites that are pathological to animals and humans. This study identified and quantified AF (AFB(1), AFB(2), AFG(1), AFG(2)) and their hydroxylated metabolites (AFM(1), AFM(2), AFP(1)) and aflatoxicol (AFL) from laying hen breast muscles. Aflatoxins pass from cereal feed to the laying hen tissues, causing economic losses, and from there to humans. To detect the passage of AF from feed to hen breast muscle tissues, an experiment that included 25 Hy-Line W36 121-wk-old hens was performed for 8 d. Hens in individual cages were distributed into 3 groups: a control group, with feed free of AFB(1), and 2 experimental groups, with feed spiked with 2 AFB(1) dosages: 30 µg·kg(-1) (low) or 500 µg·kg(-1) (high). The daily feed consumption per hen was recorded and afterward hens were euthanized and breast muscles were collected, weighed, and dried individually. Aflatoxins were extracted by 2 chemical methods and quantified by HPLC. Both methods were validated by lineality (calibration curves), recovery percentage (>80%), limit of detection, and limit of quantification. The AF (µg·kg(-1)) averages recovered in control breast muscles were as follows: AFB(1) (18); AFG(1), AFM(2), and AFL (0); AFG(2) (1.3); AFM(1) (52), and AFP1 (79). Hens fed with feed spiked with 30 µg·kg(-1) of AFB(1) had AFG(1) (16); AFG(2) (72); AFM(1) (0); AFM(2) (18); AFP(1) (145); and AFL (5 µg·kg(-1)). Hens with feed spiked with 500 µg·kg(-1) of AFB(1) had AFG(1) (512); AFG(2) (7); AFM(1) (4,775); AFM(2) (0); AFP(1) (661); and AFL (21 µg·kg(-1)). The best AF extraction method was Qian and Yang's method, modified by adding additional AF from both Supelclean LC18 SPE columns; its limit of detection (0.5 ng·mL(-1)) was lower compared with that of Koeltzow and Tanner, which was 1 ng·mL(-1). ©2014 Poultry Science Association Inc.

  9. Characterization and competitive ability of non-aflatoxigenic Aspergillus flavus isolated from the maize agro-ecosystem in Argentina as potential aflatoxin biocontrol agents.

    PubMed

    Alaniz Zanon, María Silvina; Clemente, María Paz; Chulze, Sofía Noemí

    2018-07-20

    Aspergillus flavus is an opportunistic pathogen and may produce aflatoxins in maize, one of the most important crops in Argentina. A promising strategy to reduce aflatoxin accumulation is the biological control based on competitive exclusion. In order to select potential biocontrol agents among isolates from the maize growing region in Argentina, a total of 512 A. flavus strains were isolated from maize kernels and soil samples. Thirty-six per cent of the isolates from maize kernels did not produce detectable levels of aflatoxins, while 73% of the isolates from soil were characterized as non-aflatoxin producers. Forty percent and 49% of the isolates from maize kernels and soil samples, respectively, were not producers of cyclopiazonic acid (CPA). Sclerotia morphology was evaluated using Czapek Dox media. Eighty-six per cent of the isolates from maize kernels and 85% of the isolates from soil samples were L sclerotia morphotype (average diameter > 400 μm). The remaining isolates did not produce sclerotia. All isolates had MAT 1-1 idiomorph. The competitive ability of 9 non aflatoxigenic strains, 4 CPA(+) and 5 CPA(-), was evaluated in co-inoculations of maize kernels with an aflatoxigenic strain. All evaluated strains significantly (p < 0.05) reduced aflatoxin contamination in maize kernels. The aflatoxin B 1 (AFB 1 ) reduction ranged from 6 to 60%. The strain A. flavus ARG5/30 isolated from maize kernels would be a good candidate as a potential biocontrol agent to be used in maize, since it was characterized as neither aflatoxin nor CPA producer, morphotype L, MAT 1-1 idiomorph, and reduced AFB 1 content in maize kernels by 59%. This study showed the competitive ability of potential aflatoxin biocontrol agents to be evaluated under field trials in a maize agro-ecosystem in Argentina. Copyright © 2018 Elsevier B.V. All rights reserved.

  10. MYCOLOGICAL ANALYSIS AND AFLATOXIN B1 CONTAMINANT ESTIMATION OF HERBAL DRUG RAW MATERIALS

    PubMed Central

    Rajeshwari, Puttaswamy; Raveesha, KoteshwarAnandrao

    2016-01-01

    Background: The present study explores the fungal contamination of important herbal drug raw materials (HDRM), which are widely used in the preparation of many herbal drugs. Understanding of the microbial contamination status of HDRM is one of the important steps to ensure the safety and efficacy of herbal drugs. Materials and Methods: Eighteen samples of six herbal drug raw materials (HDRM) viz., Acorus calamus Linn., Cassia angustifolia Vahl., Centella asiatica (Linn.) Urban, Myristica fragrans Houtt., Tinospora cardifolia (Wild) Miers and Withania somnifera (Linn.) Dunal, were screened for fungal contamination, by employing serial dilution method. All the isolates of Aspergillus flavus were screened for their ability to produce aflatoxin B1 (AB1) and highly contaminated samples were subjected to AB1 estimation by using Thin Layer Chromatography (TLC), spectrophotometric method and occurrence of Aflatoxin B1 was confirmed by Liquid Chromatography-Mass Spectrometry analysis (LCMS). Results: A total of 302 isolates of 42 fungal species belonging to 17 genera were found in association with test the samples. More than 61% of A. flavus isolates tested positive for production of AB1 and highest yield recorded was 5008.20 ppb from the isolates of T. cordifolia. Amongthesix highly contaminated samples three samples tested positive for AB1. Highest AB1 was recorded from T. cordifolia (104.19 μg/kg), followed by A. calamus (13.73 μg/kg) and M. fragrans (12.02 μg/kg). Conclusion: Assessment of fungal and mycotoxin contamination should be a part of the quality check while selecting HDRM for manufacture of herbal products. Safe processing and storage practices are necessary. PMID:28487902

  11. Limited survey for the presence of aflatoxins in foods from local markets and supermarkets in Valencia, Spain.

    PubMed

    Blesa, J; Soriano, J M; Moltó, J C; Mañes, J

    2004-02-01

    Aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) were extracted by matrix solid-phase dispersion with C18 silica and acetonitrile as the eluting solvent, analysed by liquid chromatography with fluorescence detection and confirmed by liquid chromatography with mass spectrometry using an electrospray interface in 58 samples grouped as cereals, dried fruits, herbs and spices, pulses, snacks, and nuts and nut products collected from local markets and supermarkets in Valencia, Spain. All samples analysed by the proposed method were previously studied with an enzyme-linked immunosorbent assay as a screening protocol for the fast detection of mycotoxins. The samples containing residues (3/58) were hazelnut (0.42 and 0.52 microg kg(-1) for AFB1 and AFG1, respectively), nut cocktail (0.29 and 0.47 microg kg(-1) for AFB1 and AFG1, respectively) and pinhol (0.30 microg kg(-1) for AFG1). Such values were below the legislated maximum residue levels for the European Union.

  12. Transformation of adsorbed aflatoxin B1 on smectite at elevated temperatures

    USDA-ARS?s Scientific Manuscript database

    Aflatoxins cause liver damage and suppress immunity. Smectites can be used to reduce the bioavailability of aflatoxins through adsorption. To further reduce the toxicity of aflatoxins and to eliminate the treatments of aflatoxin-loaded smectites, degrading the adsorbed aflatoxin to nontoxic or less ...

  13. Effect of supplementation of fermented milk drink containing probiotic Lactobacillus casei Shirota on the concentrations of aflatoxin biomarkers among employees of Universiti Putra Malaysia: a randomised, double-blind, cross-over, placebo-controlled study.

    PubMed

    Mohd Redzwan, Sabran; Abd Mutalib, Mohd Sokhini; Wang, Jia-Sheng; Ahmad, Zuraini; Kang, Min-Su; Abdul Rahman, Nurul 'Aqilah; Nikbakht Nasrabadi, Elham; Jamaluddin, Rosita

    2016-01-14

    Human exposure to aflatoxin is through the diet, and probiotics are able to bind aflatoxin and prevent its absorption in the small intestine. This study aimed to determine the effectiveness of a fermented milk drink containing Lactobacillus casei Shirota (LcS) (probiotic drink) to prevent aflatoxin absorption and reduce serum aflatoxin B1-lysine adduct (AFB1-lys) and urinary aflatoxin M1 concentrations. The present study was a randomised, double-blind, cross-over, placebo-controlled study with two 4-week intervention phases. In all, seventy-one subjects recruited from the screening stage were divided into two groups--the Yellow group and the Blue group. In the 1st phase, one group received probiotic drinks twice a day and the other group received placebo drinks. Blood and urine samples were collected at baseline, 2nd and 4th week of the intervention. After a 2-week wash-out period, the treatments were switched between the groups, and blood and urine samples were collected at the 6th, 8th and 10th week (2nd phase) of the intervention. No significant differences in aflatoxin biomarker concentrations were observed during the intervention. A within-group analysis was further carried out. Aflatoxin biomarker concentrations were not significantly different in the Yellow group. Nevertheless, ANOVA for repeated measurements indicated that AFB1-lys concentrations were significantly different (P=0·035) with the probiotic intervention in the Blue group. The 2nd week AFB1-lys concentrations (5·14 (SD 2·15) pg/mg albumin (ALB)) were significantly reduced (P=0·048) compared with the baseline (6·24 (SD 3·42) pg/mg ALB). Besides, the 4th week AFB1-lys concentrations were significantly lower (P<0·05) with probiotic supplementation than with the placebo. Based on these findings, a longer intervention study is warranted to investigate the effects of continuous LcS consumption to prevent dietary aflatoxin exposure.

  14. Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element.

    PubMed

    Ertekin, Özlem; Öztürk, Selma; Öztürk, Zafer Ziya

    2016-08-11

    This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1.

  15. Label Free QCM Immunobiosensor for AFB1 Detection Using Monoclonal IgA Antibody as Recognition Element

    PubMed Central

    Ertekin, Özlem; Öztürk, Selma; Öztürk, Zafer Ziya

    2016-01-01

    This study introduces the use of an IgA isotype aflatoxin (AF) specific monoclonal antibody for the development of a highly sensitive Quartz Crystal Microbalance (QCM) immunobiosensor for the detection of AF in inhibitory immunoassay format. The higher molecular weight of IgA antibodies proved an advantage over commonly used IgG antibodies in label free immunobiosensor measurements. IgA and IgG antibodies with similar affinity for AF were used in the comparative studies. Sensor surface was prepared by covalent immobilization of AFB1, using self assembled monolayer (SAM) formed on gold coated Quartz Crystal, with 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy succinimide (EDC/NHS) method using a diamine linker. Nonspecific binding to the surface was decreased by minimizing the duration of EDC/NHS activation. Sensor surface was chemically blocked after AF immobilization without any need for protein blocking. This protein free sensor chip endured harsh solutions with strong ionic detergent at high pH, which is required for the regeneration of the high affinity antibody-antigen interaction. According to the obtained results, the detection range with IgA antibodies was higher than IgG antibodies in QCM immunosensor developed for AFB1. PMID:27529243

  16. Aflatoxin B1 and total fumonisin contamination and their producing fungi in fresh and stored sorghum grain in East Hararghe, Ethiopia.

    PubMed

    Taye, Wondimeneh; Ayalew, Amare; Chala, Alemayehu; Dejene, Mashilla

    2016-12-01

    Natural contamination of sorghum grains by aflatoxin B 1 and total fumonisin and their producing toxigenic fungi has been studied. A total of 90 sorghum grain samples were collected from small-scale farmers' threshing floors and 5-6 months later from underground pits during 2013 harvest from three districts of East Hararghe, Ethiopia. Mycotoxin analysis was done using enzyme-linked immunosorbent assay (ELISA). The limits of detection were in the range 0.01-0.03 μg kg -1 . The results revealed that all sorghum grain samples were contaminated with both Aspergillus and Fusarium species. Aflatoxin B 1 was detected at levels ranging from 1 grain. There were marked variations in aflatoxin B 1 concentrations between fresh and stored samples, with much higher levels in the latter. Total fumonisin levels varied between 907 and 2041 µg kg -1 grain across the samples. Lowest total fumonisin was recorded in freshly harvested sorghum grain samples. Sorghum is a main staple cereal in the studied districts and its consumption per day per person is high. Daily intake of low doses of mycotoxin-contaminated food stuff over a period of time could lead to chronic mycotoxicosis.

  17. Short communication: investigation of aflatoxin M1 levels in infant follow-on milks and infant formulas sold in the markets of Ankara, Turkey.

    PubMed

    Er, B; Demirhan, B; Yentür, G

    2014-01-01

    Aflatoxins are fungal toxins known to be carcinogenic and are classified as food contaminants. This study was performed to investigate aflatoxin (AF) M1 levels in baby foods sold in Ankara (Turkey) and to evaluate the obtained results according to the Turkish Food Codex (TFC). For this purpose, a total of 84 baby food samples (50 follow-on milks and 34 infant formulas) were obtained from different markets in Ankara and the presence of AFM1 in the samples was analyzed by ELISA. In 32 (38.1%) of 84 infant food samples, the presence of AFM1 was detected in concentrations ranging between 0.0055 and 0.0201 µg/kg. The mean level (± standard error) of AFM1 was found to be 0.0089 ± 0.0006 µg/kg in positive infant follow-on milks. Aflatoxin M1 was detected in only 1 infant formula sample (2.94%) at a concentration of 0.0061 µg/kg. The extrapolated levels of AFB1 contamination in feedstuffs were calculated based on levels of AFM1 in baby food samples. The data estimating AFB1 contamination in dairy cattle feedstuff indicate that contamination may range from 0.3410 to 1.2580 µg/kg, with the mean level (± standard error) being 0.5499 ± 0.0385 µg/kg, which is lower than the level set by the TFC and European Union regulations (5 µg/kg). According to the obtained results, the levels of AFM1 in analyzed samples were within the allowed limit (0.025 µg/kg) set in the TFC. Low levels of AFM1 in infant follow-on milks and infant formula samples obtained during the study do not pose a health risk to infants. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  18. Potential of essential oils for protection of grains contaminated by aflatoxin produced by Aspergillus flavus

    PubMed Central

    Esper, Renata H.; Gonçalez, Edlayne; Marques, Marcia O. M.; Felicio, Roberto C.; Felicio, Joana D.

    2014-01-01

    Aflatoxin B1 (AFB1) is a highly toxic and carcinogenic metabolite produced by Aspergillus species on food and agricultural commodities. Inhibitory effects of essential oils of Ageratum conyzoides (mentrasto) and Origanum vulgare (oregano) on the mycelial growth and aflatoxin B1 production by Aspergillus flavus have been studied previously in culture medium. The aim of this study was to evaluate aflatoxin B1 production by Aspergillus flavus in real food systems (corn and soybean) treated with Ageratum conyzoides (mentrasto) and Origanum vulgare (oregano) essential oils. Samples with 60 g of the grains were treated with different volumes of essential oils, 200, 100, 50, and 10 μL for oregano and 50, 30, 15, and 10 μL for mentrasto. Fungal growth was evaluated by disk diffusion method. Aflatoxin B1 production was evaluated inoculating suspensions of A. flavus containing 1.3 × 105 spores/mL in 60 g of grains (corn and soybeans) after adjusting the water activity at 0.94. Aflatoxin was quantified by photodensitometry. Fungal growth and aflatoxin production were inhibited by essential oils, but the mentrasto oil was more effective in soybeans than that of oregano. On the other hand, in corn samples, the oregano essential oil was more effective than that of mentrasto. Chemical compositions of the essential oils were also investigated. The GC/MS oils analysis showed that the main component of mentrasto essential oil is precocene I and of the main component of oregano essential oil is 4-terpineol. The results indicate that both essential oils can become an alternative for the control of aflatoxins in corn and soybeans. PMID:24926289

  19. Aflatoxin exposure during the first 36 months of life was not associated with impaired growth in Nepalese children: An extension of the MAL-ED study

    PubMed Central

    Hsu, Hui-Husan; Chandyo, Ram Krishna; Shrestha, Binob; Bodhidatta, Ladaporn; Tu, Yu-Kang; Gong, Yun-Yun; Egner, Patricia A.; Ulak, Manjeswori; Groopman, John D.; Wu, Felicia

    2017-01-01

    Exposure to aflatoxin, a mycotoxin common in many foods, has been associated with child growth impairment in sub-Saharan Africa. To improve our understanding of growth impairment in relation to aflatoxin and other risk factors, we assessed biospecimens collected in Nepalese children at 15, 24, and 36 months of age for aflatoxin exposure. Children (N = 85) enrolled in the Bhaktapur, Nepal MAL-ED study encompassed the cohort analysed in this study. Exposure was assessed through a plasma biomarker of aflatoxin exposure: the AFB1-lysine adduct. The aflatoxin exposures in the study participants were compared to anthropometrics at each time period (length-for-age [LAZ], weight-for-age [WAZ], and weight-for-length [WLZ] z-scores), growth trajectories over time, age, and breastfeeding status. Results demonstrated chronic aflatoxin exposure in this cohort of children, with a geometric mean of 3.62 pg AFB1-lysine/mg albumin. However, the chronic aflatoxin exposure in this cohort was not significantly associated with anthropometric z-scores, growth trajectories, age, or feeding status, based on the available time points to assess aflatoxin exposure. Low mean levels of aflatoxin exposure and infrequent occurrence of stunting, wasting, or underweight z-score values in this cohort are possible contributing factors to a lack of evidence for an association. Further research is needed to examine whether a threshold dose of aflatoxin exists that could induce child growth impairment. PMID:28212415

  20. Occurrence of aflatoxin M1 in UHT milk in Turkey.

    PubMed

    Unusan, Nurhan

    2006-11-01

    Aflatoxin M1 (AFM1) appears in milk as a direct result of the ingestion of food contaminated with aflatoxin B1 by cattle. The role of milk in human nutrition is well-known. The purpose of the study was to determine the levels of AFM1 in UHT milk samples in Central Anatolia, Turkey. The occurrence of AFM1 contamination in UHT milk samples was investigated by ELISA (Enzyme Linked Immunosorbent Assay) technique. A total of 129 samples of commercial UHT whole milk were analysed. The mean value was 108.17 ng/L. There was a high incidence rate of AFM1, with 75 (58.1%) milk samples being contaminated. Although 68 (53%) were below the limit, the remaining 61 (47%) were well above the limit permitted by the EU. Four of the samples exceeded the prescribed limit of US regulations. It can be concluded that AFM1 levels in the samples purchased in Central Anatolia Region, appear to be a serious public health problem at the moment. Dairy farmers must be educated by the government authorities on potential health consequences of aflatoxins.

  1. Impact of bacterial biocontrol agents on aflatoxin biosynthetic genes, aflD and aflR expression, and phenotypic aflatoxin B₁ production by Aspergillus flavus under different environmental and nutritional regimes.

    PubMed

    Al-Saad, Labeed A; Al-Badran, Adnan I; Al-Jumayli, Sami A; Magan, Naresh; Rodríguez, Alicia

    2016-01-18

    The objectives of this study were to examine the efficacy of four bacterial antagonists against Aspergillus flavus using 50:50 ratio of bacterial cells/conidia for the control of aflatoxin B1 (AFB1) production on two different nutritional matrices, nutrient and maize-based media at different water availabilities (0.98, 0.94 water activity (aw) on nutrient medium; 0.995, 0.98 aw on maize meal agar medium) at 35°C. The indicators of efficacy used were the relative expression of one structural and regulatory gene in the biosynthetic pathway (aflD and aflR respectively) and the production of AFB1. These studies showed that some of the bacterial species could significantly inhibit the relative expression of the aflD and aflR genes at both 0.98 and 0.94 aw on nutrient agar. On maize-based media some of the bacterial antagonists reduced the activity of both genes at 0.94 aw and some at 0.995 aw. However, the results for AFB1 production were not consistent with the effects on gene expression. Some bacterial species stimulated AFB1 production on both nutrient and maize-based media regardless of aw. However, some bacterial treatments did inhibit AFB1 production significantly when compared to the control. Overall, this study suggests that temporal studies are required on the biosynthetic genes under different environmental and nutritional conditions to evaluate the potential of antagonists to control AFB1. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Identification and quantification of aflatoxins and aflatoxicol from poultry feed and their recovery in poultry litter.

    PubMed

    Cortés, G; Carvajal, M; Méndez-Ramírez, I; Avila-González, E; Chilpa-Galván, N; Castillo-Urueta, P; Flores, C M

    2010-05-01

    Aflatoxins (AF) are toxic fungal secondary metabolites and are known mycotoxins pathological to animals and humans. Poultry litter is frequently used as a food supplement for ruminants, and when poultry feed contains AF, the litter becomes contaminated as well, thus having an effect on livestock health. This study identified and quantified AF (AFB(1), AFB(2), AFG(1), and AFG(2)) from poultry feed and their recovery, together with their metabolites (AFM(1), AFM(2), AFP(1), and aflatoxicol) in litter. An experiment with 25 Hy-Line W-36 hens, in their second production stage, 121 wk old, was carried out. Hens were distributed in 3 groups placed in individual cages and 1 ration of 250 g of feed was given to each hen daily. Nine hens of the control group were fed with clean feed, without AFB(1); the other 2 experimental groups, with 8 hens each, were fed with 2 AFB(1) concentrations: 30 and 500 microg.kg(-1). The feed was replaced and weighed daily throughout a 7-d period to register the amount of feed consumed by the hens. Litter from each hen was collected, weighed, and dried individually. The chemical analysis of 40 g of each one of the 200 feed and 200 litter samples was chemically extracted and concentrated with immunoaffinity columns for total AF. To quantify AF, calibration curves for each AF were done by HPLC. Feed samples of the 3 groups presented significant difference with AFB(2) and AFG(2), whereas in litter samples, there were significant differences for AFG(2) in the 500 microg.kg(-1) group. Poultry litter had traces of AFM(1), AFM(2), AFP(1), and AFL with no significant differences among treatments. Aflatoxin B(1) prevalence in litter samples can cause damages in livestock because this mycotoxin reduces the digestibility of ruminant feed up to 67%.

  3. Detoxification of Aflatoxin B1 by Antifungal Compounds from Lactobacillus brevis and Lactobacillus paracasei, Isolated from Dairy Products.

    PubMed

    Gomaa, Eman Zakaria; Abdelall, Manal Farouk; El-Mahdy, Omima Mohammed

    2018-06-01

    Aflatoxins are a large group of highly toxic, mutagenic, and carcinogenic mycotoxins produced by specific species of fungi. Potential contamination of food commodities by these compounds causes extensive damage that lead to great economic losses. This study explored the potential use of antifungal compounds, produced by Lactobacillus brevis and Lactobacillus paracasei, for growth inhibition and subsequent aflatoxin B1 production from select strains of Aspergillus flavus and Aspergillus parasiticus. Lactobacilli strains were isolated from traditional Egyptian dairy products, whereas fungal strains were isolated from infected cereal seeds. There were noticeable decreases in mycelium biomass and aflatoxin production as well. L. brevis exhibited the highest reduction of aflatoxin B1 production by A. flavus and A. parasiticus, 96.31 and 90.43%, respectively. The concentrations of amino acids of the antifungal compound produced by L. brevis were significantly higher than that produced by L. paracasei. Asparagine, glutamine, glycine, alanine, and leucine were the most concentrated amino acids for both strains. The antifungal compounds produced by L. brevis and L. paracasei were active in a wide range of pH, heat stable and inactivated by proteolytic enzymes (protease K and trypsin A). The expression of Omt-A gene that involved in the later step of aflatoxin production was evaluated by real-time PCR. There was a vigorous reduction at transcriptional level of Omt-A gene observed in A. flavus that is treated by L. brevis and L. paracasei (80 and 70%, respectively). However, the reduction of Omt-A gene observed in A. parasiticus that is treated by L. brevis and L. paracasei was 64.5 and 52%, respectively. Treating maize seeds with antifungal compounds exhibited great efficiency in controlling fungal infection and increasing seed germination. The results confirmed that lactic acid bacteria are a promising strategy to control food contamination of fermented food and dairy

  4. [Appearance of aflatoxin M1 during the manufacture of Camembert cheese].

    PubMed

    Frémy, J M; Roiland, J C

    1979-01-01

    Several classic cheese making of camembert are made from raw milk spiked with Aflatoxin M1. Three Aflatoxin levels 7.5 microgram/l, 3 microgram/l are used. In respective curds 35.6, 47.1 and 57.7% of Aflatoxin M1 are recovered and 64.4, 52.9 and 42.3% in respective whey. During the first 15 days of storage the Aflatoxin M1 content of different cheeses decrease respectively 25, 55, 75%. A similar experience is made with a milk contamined in Aflatoxin M1 C14 labelled. Same results are recovered, except about behaviour of Aflatoxin M1 in cheese: a same C14 activity is recovered during storage for 30 days.

  5. Behavior of 14C aflatoxin M1 during camembert cheese making.

    PubMed

    Fremy, J M; Roiland, J C; Gaymard, A

    1990-01-01

    Camembert cheeses are made from raw milk spiked with aflatoxin M1. Three aflatoxin M1 levels (7.5 micrograms/L, 3 micrograms/L, and 0.3 micrograms/L) are used. In curds 35.6, 47.1, and 57.7% of aflatoxin M1, respectively, are recovered, and in wheys 64.4, 52.9, and 42.3%, respectively, are recovered. During the first 15 days of storage, the aflatoxin M1 content of different cheeses decreases 25, 55, and 75%, respectively. A similar experiment is made with milk contaminated with 14C labeled aflatoxin M1. The same results are obtained, except for the behavior of aflatoxin M1 in cheese; the same 14C activity is recovered during storage for 30 days.

  6. Monoclonal Antibodies to Prevent Use of Mycotoxins as Biological Weapons

    DTIC Science & Technology

    2007-07-01

    conjugate to immunize rats. We found that vaccination by either the intraperitoneal or subcutaneous route induced very high aflatoxin B1-binding antibody...are small molecule secondary metabolites of the fungi Aspergillus flavus and Aspergillus parasiticus that are potent hepatic carcinogens and have a...their potential for use in the event of bioterrorism has been highlighted (1). Immunization with an aflatoxin B1 (AF-B1)-BSA conjugate previously

  7. Computational search for aflatoxin binding proteins

    NASA Astrophysics Data System (ADS)

    Wang, Ying; Liu, Jinfeng; Zhang, Lujia; He, Xiao; Zhang, John Z. H.

    2017-10-01

    Aflatoxin is one of the mycotoxins that contaminate various food products. Among various aflatoxin types (B1, B2, G1, G2 and M1), aflatoxin B1 is the most important and the most toxic one. In this study, through computational screening, we found that several proteins may bind specifically with different type of aflatoxins. Combination of theoretical methods including target fishing, molecular docking, molecular dynamics (MD) simulation, MM/PBSA calculation were utilized to search for new aflatoxin B1 binding proteins. A recently developed method for calculating entropic contribution to binding free energy called interaction entropy (IE) was employed to compute the binding free energy between the protein and aflatoxin B1. Through comprehensive comparison, three proteins, namely, trihydroxynaphthalene reductase, GSK-3b, and Pim-1 were eventually selected as potent aflatoxin B1 binding proteins. GSK-3b and Pim-1 are drug targets of cancers or neurological diseases. GSK-3b is the strongest binder for aflatoxin B1.

  8. Efficacy of Some Essential Oils Against Aspergillus flavus with Special Reference to Lippia alba Oil an Inhibitor of Fungal Proliferation and Aflatoxin B1 Production in Green Gram Seeds during Storage.

    PubMed

    Pandey, Abhay K; Sonker, Nivedita; Singh, Pooja

    2016-04-01

    During mycofloral analysis of green gram (Vigna radiata (L.) R. Wilczek) seed samples taken from different grocery stores by agar and standard blotter paper methods, 5 fungal species were identified, of which Aspergillus flavus exhibited higher relative frequency (75.20% to 80.60%) and was found to produce aflatoxin B1 . On screening of 11 plant essential oils against this mycotoxigenic fungi, Lippia alba essential oil was found to be most effective and showed absolute inhibition of mycelia growth at 0.28 μL/mL. The oil of L. alba was fungistatic and fungicidal at 0.14 and 0.28 μL/mL, respectively. Oil had broad range of fungitoxicity at its MIC value and was absolutely inhibited the AFB1 production level at 2.0 μL/mL. Chemical analysis of this oil revealed geranial (36.9%) and neral (29.3%) as major components followed by myrcene (18.6%). Application of a dose of 80 μL/0.25 L air of Lippia oil in the storage system significantly inhibited the fungal proliferation and aflatoxin production without affecting the seed germination rate. By the virtue of fungicidal, antiaflatoxigenic nature and potent efficacy in storage food system, L. alba oil can be commercialized as botanical fungicide for the protection of green gram seeds during storage. © 2016 Institute of Food Technologists®

  9. Socio-demographic determinants of aflatoxin B1-lysine adduct levels among pregnant women in Kumasi, Ghana.

    PubMed

    Shuaib, F M B; Jolly, P E; Ehiri, J E; Ellis, W O; Yatich, N J; Funkhouser, E; Person, S D; Williams, J H; Qian, G; Wang, J-S

    2012-12-01

    Aflatoxins are fungal metabolites that contaminate staple food crops in many developing countries. Although studies have linked these toxins to adverse birth outcomes and poor infant development, no study has investigated the socio-demographic and economic determinants of aflatoxin levels among pregnant women living in sub-Saharan Africa. A cross-sectional study was conducted among 785 pregnant women in Kumasi. Aflatoxin B1 lysine adduct levels (AF-ALB) were determined by High Performance Liquid Chromatography. Analysis of variance was used to determine mean log AF-ALB levels and significance of differences in these levels according to socio-demographic variables. Logistic regression was used to identify independent associations of socio-demographics with having AF-ALB levels (≥ 11.34 pg/mg; upper quartile). AF-ALB levels ranged from 0.44 pg/mg to 268.73 pg/mg albumin with a median level of 5.0 pg/mg. Bivariate analyses indicates that mean ln AF-ALB as well as the percent of women having high AF-ALB levels (≥ 11.34 pg/mg; upper quartile) were inversely associated with indices of higher socioeconomic status: higher education and income, being employed and having a flush toilet. Higher income, being employed, having one child (verses no children) and having a flush toilet (verses no toilet facilities) were each independently associated with a 30-40% reduced odds of high AF-ALB levels. Additional research is needed to investigate how socio-demographic and economic factors interact to influence aflatoxin ingestion by individuals in regions with high aflatoxin crop contamination. This knowledge can be used to formulate and implement policies that will reduce exposure of women and their unborn children to these toxins.

  10. Analysis of Aflatoxin M1 in Breast Milk and Its Association with Nutritional and Socioeconomic Status of Lactating Mothers in Lebanon.

    PubMed

    Elaridi, Jomana; Bassil, Maya; Kharma, Joelle Abi; Daou, Farah; Hassan, Hussein F

    2017-10-01

    Aflatoxin B 1 (AFB 1 ) is the most potent of the dietary aflatoxins, and its major metabolite, aflatoxin M 1 (AFM 1 ), is frequently found in the breast milk of lactating mothers. The aim of this study was to assess the occurrence and factors associated with AFM 1 contamination of breast milk collected from lactating mothers in Lebanon. A total of 111 breast milk samples were collected according to the guidelines set by the World Health Organization. Samples were analyzed with a competitive enzyme-linked immunosorbent assay between December 2015 and November 2016. A survey was used to determine the demographic and anthropometric characteristics of participating lactating mothers. Dietary habits were assessed using a semiquantitative food frequency questionnaire. Mean (±standard deviation) concentration of AFM 1 in the breast milk samples was 4.31 ± 1.8 ng/L, and 93.8% of samples contained AFM 1 at 0.2 to 7.9 ng/L. The mean concentration of AFM 1 was significantly lower (P < 0.05) in fall and winter (4.1 ± 1.9 ng/L) than in spring and summer (5.0 ± 1.7 ng/L). None of the samples exceeded the European Commission regulation limit (25 ng/L) for infant milk replacement formula. AFM 1 contamination was significantly associated (P < 0.05) with the daily consumption of white cheeses but not with the consumption of meat or cereal products. No significant association (P > 0.05) was observed between AFM 1 concentrations in breast milk and anthropometric sociodemographic factors (age and level of education) or the governorate of residence of the nursing mothers. The mean AFM 1 estimated daily intake was found to be 0.69 ng/day/kg of body weight. Although the incidence of AFM 1 contamination was low, our first-of-its-kind study highlights the importance of conducting investigations on mycotoxin contamination in breast milk and of developing protection strategies to tackle the exposure of infants to this potent chemical hazard.

  11. Phenolic extract of Dialium guineense pulp enhances reactive oxygen species detoxification in aflatoxin B₁ hepatocarcinogenesis.

    PubMed

    Adeleye, Abdulwasiu O; Ajiboye, Taofeek O; Iliasu, Ganiyat A; Abdussalam, Folakemi A; Balogun, Abdulazeez; Ojewuyi, Oluwayemisi B; Yakubu, Musa T

    2014-08-01

    This study investigated the effect of Dialium guineense pulp phenolic extract on aflatoxin B1 (AFB1)-induced oxidative imbalance in rat liver. Reactive oxygen species (ROS) scavenging potentials of free and bound phenolic extract of D. guineense (0.2-1.0 mg/mL) were investigated in vitro using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide ion (O2(-)), hydrogen peroxide (H2O2), hydroxyl radical, and ferric ion reducing system. In the in vivo study, 35 animals were randomized into seven groups of five rats each. Free and bound phenolic extract (1 mg/mL) produced 66.42% and 93.08%, 57.1% and 86.0%, 62.0% and 90.05%, and 60.11% and 72.37% scavenging effect on DPPH radical, O2(-) radical, H2O2, and hydroxyl radical, while ferric ion was significantly reduced. An AFB1-mediated decrease in the activities of ROS detoxifying enzymes (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glucose 6 phosphate dehydrogenase) was significantly attenuated (P<.05). AFB1-mediated elevation in the concentrations of oxidative stress biomarkers; malondialdehyde, conjugated dienes, lipid hydroperoxides, protein carbonyl, and percentage DNA fragmentation were significantly lowered by D. guineense phenolic extract (P<.05). Overall, the in vitro and in vivo effects suggest that D. guineense phenolic extract elicited ROS scavenging and detoxification potentials, as well as the capability of preventing lipid peroxidation, protein oxidation, and DNA fragmentation.

  12. Ionic-liquid-based dispersive liquid-liquid microextraction combined with magnetic solid-phase extraction for the determination of aflatoxins B1 , B2 , G1 , and G2 in animal feeds by high-performance liquid chromatography with fluorescence detection.

    PubMed

    Zhao, Jiao; Zhu, Yan; Jiao, Yang; Ning, Jinyan; Yang, Yaling

    2016-10-01

    A novel two-step extraction technique combining ionic-liquid-based dispersive liquid-liquid microextraction with magnetic solid-phase extraction was developed for the preconcentration and separation of aflatoxins in animal feedstuffs before high-performance liquid chromatography coupled with fluorescence detection. In this work, ionic liquid 1-octyl-3-methylimidazolium hexafluorophosphate was used as the extractant in dispersive liquid-liquid microextraction, and hydrophobic pelargonic acid modified Fe 3 O 4 magnetic nanoparticles as an efficient adsorbent were applied to retrieve the aflatoxins-containing ionic liquid. Notably, the target of magnetic nanoparticles was the ionic liquid rather than the aflatoxins. Because of the rapid mass transfer associated with the dispersive liquid-liquid microextraction and magnetic solid phase steps, fast extraction could be achieved. The main parameters affecting the extraction recoveries of aflatoxins were investigated and optimized. Under the optimum conditions, vortexing at 2500 rpm for 1 min in the dispersive liquid-liquid microextraction and magnetic solid-phase extraction and then desorption by sonication for 2 min with acetonitrile as eluent. The recoveries were 90.3-103.7% with relative standard deviations of 3.2-6.4%. Good linearity was observed with correlation coefficients ranged from 0.9986 to 0.9995. The detection limits were 0.632, 0.087, 0.422 and 0.146 ng/mL for aflatoxins B 1 , B2, G1, and G2, respectively. The results were also compared with the pretreatment method carried out by conventional immunoaffinity columns. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Behavior of sup 14 C aflatoxin M1 during camembert cheese making

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fremy, J.M.; Roiland, J.C.; Gaymard, A.

    Camembert cheeses are made from raw milk spiked with aflatoxin M1. Three aflatoxin M1 levels (7.5 micrograms/L, 3 micrograms/L, and 0.3 micrograms/L) are used. In curds 35.6, 47.1, and 57.7% of aflatoxin M1, respectively, are recovered, and in wheys 64.4, 52.9, and 42.3%, respectively, are recovered. During the first 15 days of storage, the aflatoxin M1 content of different cheeses decreases 25, 55, and 75%, respectively. A similar experiment is made with milk contaminated with {sup 14}C labeled aflatoxin M1. The same results are obtained, except for the behavior of aflatoxin M1 in cheese; the same 14C activity is recoveredmore » during storage for 30 days.« less

  14. Sensitive quantification of aflatoxin B1 in animal feeds, corn feed grain, and yellow corn meal using immunomagnetic bead-based recovery and real-time immunoquantitative-PCR.

    PubMed

    Babu, Dinesh; Muriana, Peter M

    2014-12-02

    Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1-10 μg/kg), addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B1, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup.

  15. Effects of dietary milk thistle on blood parameters, liver pathology, and hepatobiliary scintigraphy in white carneaux pigeons (Columba livia) challenged with B1 aflatoxin.

    PubMed

    Grizzle, Judith; Hadley, Tarah L; Rotstein, David S; Perrin, Shannon L; Gerhardt, Lillian E; Beam, James D; Saxton, Arnold M; Jones, Michael P; Daniel, Gregory B

    2009-06-01

    Milk thistle (Silybum marianum) has been used in humans for the treatment of liver disease because of its antioxidant properties and its ability to stabilize cell membranes and regulate cell permeability. To investigate possible hepatoprotective effects in birds, standardized extracts (80%) of silymarin from milk thistle were tested in white Carneaux pigeons (Columba livia). Pigeons were separated into 3 groups and fed diets formulated to provide milk thistle at a level of 0, 10, or 100 mg/kg body weight per day. After acclimation, the birds were challenged with B1 aflatoxin (3 mg/kg body weight for 2 consecutive days) by oral gavage. Liver function then was assessed by hematologic testing and plasma biochemical analysis, liver histopathology, and hepatobiliary scintigraphy. Results of histopathology and hepatobiliary scintigraphy showed no protective effects from milk-thistle administration. Aflatoxin challenge resulted in hepatic inflammation and necrosis, biliary-duct hyperplasia, and lymphocyte infiltration. All hepatobiliary scintigraphy elements increased significantly after aflatoxin challenge. Bile acid levels and plasma enzyme concentrations of aspartate aminotransferase, lactate dehydrogenase, alanine aminotransferase, and creatine phosphokinase all increased after aflatoxin exposure and were mostly unchanged with consumption of milk thistle. Only birds fed 10 mg/kg body weight milk thistle showed significant reductions in lactate dehydrogenase, alanine aminotransferase, and creatine phosphokinase concentrations after aflatoxin exposure. Our results show that consumption of milk thistle is not associated with hepatoprotective effects against acute B1 aflatoxin exposure in pigeons.

  16. Inhibition of aflatoxin B production of Aspergillus flavus, isolated from soybean seeds by certain natural plant products.

    PubMed

    Krishnamurthy, Y L; Shashikala, J

    2006-11-01

    The inhibitory effect of cowdung fumes, Captan, leaf powder of Withania somnifera, Hyptis suaveolens, Eucalyptus citriodora, peel powder of Citrus sinensis, Citrus medica and Punica granatum, neem cake and pongamia cake and spore suspension of Trichoderma harzianum and Aspergillus niger on aflatoxin B(1) production by toxigenic strain of Aspergillus flavus isolated from soybean seeds was investigated. Soybean seed was treated with different natural products and fungicide captan and was inoculated with toxigenic strain of A. flavus and incubated for different periods. The results showed that all the treatments were effective in controlling aflatoxin B(1) production. Captan, neem cake, spore suspension of T. harzianum, A. niger and combination of both reduced the level of aflatoxin B(1) to a great extent. Leaf powder of W. somnifera, H. suaveolens, peel powder of C. sinensis, C. medica and pongamia cake also controlled the aflatoxin B(1) production. All the natural product treatments applied were significantly effective in inhibiting aflatoxin B(1) production on soybean seeds by A. flavus. These natural plant products may successfully replace chemical fungicides and provide an alternative method to protect soybean and other agricultural commodities from aflatoxin B(1) production by A. flavus.

  17. Low-cost and highly sensitive immunosensing platform for aflatoxins using one-step competitive displacement reaction mode and portable glucometer-based detection.

    PubMed

    Tang, Dianping; Lin, Youxiu; Zhou, Qian; Lin, Yuping; Li, Peiwu; Niessner, Reinhard; Knopp, Dietmar

    2014-11-18

    Aflatoxins are highly toxic secondary metabolites produced by a number of different fungi and present in a wide range of food and feed commodities. Herein, we designed a simple and low-cost immunosensing platform for highly sensitive detection of mycotoxins (aflatoxin B1, AFB1, used as a model) on polyethylenimine (PEI)-coated mesoporous silica nanocontainers (PEI-MSN). The assay was carried out by using a portable personal glucometer (PGM) as the readout based on a competitive displacement reaction mode between target AFB1 and its pseudo-hapten (PEI-MSN) for monoclonal anti-AFB1 antibody (mAb). To construct such an assay protocol, two nanostructures including mAb-labeled gold nanoparticle (mAb-AuNP) and PEI-MSN were initially synthesized, and then numerous glucose molecules were gated into the pores based on the interaction between negatively charged mAb-AuNP and positively charged PEI-MSN. In the presence of target AFB1, a competitive-type displacement reaction was implemented between mAb-AuNP and PEI-MSN by target AFB1 through the specific antigen-antibody reaction. Accompanying the reaction, target AFB1 could displace the mAb-AuNP from the surface of PEI-MSN, resulting in the release of the loading glucose from the pores due to the gate opened. The released glucose molecules could be quantitatively determined by using a portable PGM. Under optimal conditions, the PGM signal increased with the increment of AFB1 concentration in the range from 0.01 to 15 μg/kg (ppb) with a detection limit (LOD) of 5 ng/kg (5 ppt) at the 3sblank criterion. The selectivity and precision were acceptable. Importantly, the methodology was further validated for assaying naturally contaminated or spiked blank peanut samples, and consistent results between the PGM-based immunoassay and the referenced enzyme-linked immunosorbent assay (ELISA) were obtained. Therefore, the developed immunoassay provides a promising approach for rapid screening of organic pollutants because it is simple

  18. Sensitive Quantification of Aflatoxin B1 in Animal Feeds, Corn Feed Grain, and Yellow Corn Meal Using Immunomagnetic Bead-Based Recovery and Real-Time Immunoquantitative-PCR

    PubMed Central

    Babu, Dinesh; Muriana, Peter M.

    2014-01-01

    Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be <20 μg/kg. The RT iq-PCR assay exhibited high antigen hook effect in samples containing aflatoxin levels higher than the quantification limits (0.1–10 μg/kg), addressed by comparing the quantification results of undiluted and diluted extracts. In testing the reliability of the immuno-PCR assay, samples were spiked with 200 μg/kg of aflatoxin B1, but the recovery of spiked aflatoxin was found to be poor. Considering the significance of determining trace levels of aflatoxins and their serious implications for animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup. PMID:25474493

  19. Analysis of the aflatoxin AFB1 from corn by direct analysis in real time - mass spectrometry (DART-MS)

    USDA-ARS?s Scientific Manuscript database

    Direct analysis in real time (DART) ionization coupled to a high resolution mass spectrometer (MS) was used for screening of aflatoxins from a variety of surfaces and the rapid quantitative analysis of aflatoxins extracted from corn. Sample preparation procedure and instrument parameter settings wer...

  20. Mycological and aflatoxin contamination of peanuts sold at markets in Kinshasa, Democratic Republic of Congo, and Pretoria, South Africa.

    PubMed

    Kamika, Ilunga; Mngqawa, Pamella; Rheeder, John P; Teffo, Snow L; Katerere, David R

    2014-01-01

    Peanut (Arachis hypogaea L.) is an important food crop in sub-Saharan Africa. In this survey, the mycological and aflatoxin contamination of peanuts collected from Kinshasa, Democratic Republic of Congo, and Pretoria, South Africa, was assessed. Twenty peanut samples were purchased randomly at informal markets in the two cities and analysed for mycoflora and aflatoxins (AFB1, AFB2, AFG1 and AFG2) using standard methods. The results indicated that 95% of the Kinshasa samples and 100% of the Pretoria samples were contaminated with aflatoxigenic fungi in the ranges 20-49,000 and 40-21,000 CFU/g, respectively. Seventy-five per cent of the Kinshasa samples and 35% of the Pretoria samples exceeded the maximum limits of AFB1 as set by The Joint FAO/WHO Expert Committee on Food Additives. Residents of both cities are at a high risk of aflatoxin exposure despite their apparent cultural, socio-economic, geographic and climatic differences. Further work needs to be done to understand the supply chains of peanut trade in informal markets of the two countries so that interventions are well targeted on a regional rather than a national level.

  1. Temporal Variation and Association of Aflatoxin B1 Albumin-Adduct Levels with Socio-Economic and Food Consumption Factors in HIV Positive Adults

    PubMed Central

    Jolly, Pauline E.; Akinyemiju, Tomi F.; Jha, Megha; Aban, Inmaculada; Gonzalez-Falero, Andrea; Joseph, Dnika

    2015-01-01

    The association between aflatoxin exposure and alteration in immune responses observed in humans suggest that aflatoxin could suppress the immune system and work synergistically with HIV to increase disease severity and progression to AIDS. No longitudinal study has been conducted to assess exposure to aflatoxin (AF) among HIV positive individuals. We examined temporal variation in AFB1 albumin adducts (AF-ALB) in HIV positive Ghanaians, and assessed the association with socioeconomic and food consumption factors. We collected socioeconomic and food consumption data for 307 HIV positive antiretroviral naive adults and examined AF-ALB levels at recruitment (baseline) and at six (follow-up 1) and 12 (follow-up 2) months post-recruitment, by age, gender, socioeconomic status (SES) and food consumption patterns. Generalized linear models were used to examine the influence of socioeconomic and food consumption factors on changes in AF-ALB levels over the study period, adjusting for other covariates. AF-ALB levels (pg/mg albumin) were lower at baseline (mean AF-ALB: 14.9, SD: 15.9), higher at six months (mean AF-ALB: 23.3, SD: 26.6), and lower at 12 months (mean AF-ALB: 15.3, SD: 15.4). Participants with the lowest SES had the highest AF-ALB levels at baseline and follow up-2 compared with those with higher SES. Participants who bought less than 20% of their food and who stored maize for less than two months had lower AF-ALB levels. In the adjusted models, there was a statistically significant association between follow up time and season (dry or rainy season) on AF-ALB levels over time (p = 0.04). Asymptomatic HIV-positive Ghanaians had high plasma AF-ALB levels that varied according to season, socioeconomic status, and food consumption patterns. Steps need to be taken to ensure the safety and security of the food supply for the population, but in particular for the most vulnerable groups such as HIV positive people. PMID:26633502

  2. Structure and Oxidation of Pyrrole Adducts Formed between Aflatoxin B2a and Biological Amines.

    PubMed

    Rushing, Blake R; Selim, Mustafa I

    2017-06-19

    Aflatoxin B 2a has been shown to bind to proteins through a dialdehyde intermediate under physiological conditions. The proposed structure of this adduct has been published showing a Schiff base interaction, but adequate verification using structural elucidation instrumental techniques has not been performed. In this work, we synthesized the aflatoxin B 2a amino acid adduct under alkaline conditions, and the formation of a new product was determined using high performance liquid chromatography-time-of-flight mass spectrometry. The resulting accurate mass was used to generate a novel proposed chemical structure of the adduct in which the dialdehyde forms a pyrrole ring with primary amines rather than the previously proposed Schiff base interaction. The pyrrole structure was confirmed using 1 H, 13 C, correlation spectroscopy, heteronuclear single quantum correlation, and heteronuclear multiple bond correlation NMR and tandem mass spectrometry. Reaction kinetics show that the reaction is overall second order and that the rate increases as pH increases. Additionally, this study shows for the first time that aflatoxin B 2a dialdehyde forms adducts with phosphatidylethanolamines and does so through pyrrole ring formation, which makes it the first aflatoxin-lipid adduct to be structurally identified. Furthermore, oxidation of the pyrrole adduct produced a product that was 16 m/z heavier. When the aflatoxin B 2a -lysine (ε) adduct was oxidized, it gave a product with an accurate mass, mass fragmentation pattern, and 1 H NMR spectrum that match aflatoxin B 1 -lysine, which suggest the transformation of the pyrrole ring to a pyrrolin-2-one ring. These data give new insight into the fate and chemical properties of biological adducts formed from aflatoxin B 2a as well as possible interferences with known aflatoxin B 1 exposure biomarkers.

  3. Control of Aspergillus section Flavi growth and aflatoxin accumulation by plant essential oils.

    PubMed

    Bluma, R; Amaiden, M R; Daghero, J; Etcheverry, M

    2008-07-01

    The antifungal effect of Pimpinella anisum (anise), Pëumus boldus (boldus), Mentha piperita (peppermint), Origanum vulgare (oregano) and Minthosthachys verticillata (peperina) essential oils against Aspergillus section Flavi (two isolates of Aspergillus parasiticus and two isolates of Aspergillus flavus) was evaluated in maize meal extract agar at 0.982 and 0.955 water activities, at 25 degrees C. The percentage of germination, germ-tube elongation rate, growth rate and aflatoxin B(1) (AFB(1)) accumulation at different essential oils concentrations were evaluated. Anise and boldus essential oils were the most inhibitory at 500 mg kg(-1) to all growth parameters of the fungus. These essential oils inhibited the percentage of germination, germ-tube elongation rate and fungal growth. AFB(1) accumulation was completely inhibited by anise, boldus and oregano essential oils. Peperina and peppermint essential oils inhibited AFB(1) production by 85-90% in all concentrations assayed. Anise and boldus essential oils could be considered as effective fungitoxicans for Aspergillus section flavi. Our results suggest that these phytochemical compounds could be used alone or in conjunction with other substances to control the presence of aflatoxigenic fungi in stored maize.

  4. Inhibition of aflatoxin biosynthesis in Aspergillus flavus by phenolic compounds extracted of Piper betle L.

    PubMed

    Yazdani, Darab; Mior Ahmad, Zainal Abidin; Yee How, Tan; Jaganath, Indu Bala; Shahnazi, Sahar

    2013-12-01

    Food contamination by aflatoxins is an important food safety concern for agricultural products. In order to identify and develop novel antifungal agents, several plant extracts and isolated compounds have been evaluated for their bioactivities. Anti-infectious activity of Piper betle used in traditional medicine of Malaysia has been reported previously. Crude methanol extract from P. betel powdered leaves was partitioned between chloroform and water. The fractions were tested against A. flavus UPMC 89, a strong aflatoxin producing strain. Inhibition of mycelial growth and aflatoxin biosynthesis were tested by disk diffusion and macrodillution techniques, respectively. The presence of aflatoxin was determined by thin-layer chromatography (TLC) and fluorescence spectroscopy techniques using AFB1 standard. The chloroform soluble compounds were identified using HPLC-Tandem mass spectrometry technique. The results, evaluated by measuring the mycelial growth and quantification of aflatoxin B1(AFLB1) production in broth medium revealed that chloroform soluble compounds extract from P. betle dried leaves was able to block the aflatoxin biosynthesis pathway at concentration of 500μg/ml without a significant effect on mycelium growth. In analyzing of this effective fractions using HPLC-MS(2) with ESI ionization technique, 11 phenolic compounds were identified. The results showed that the certain phenolic compounds are able to decline the aflatoxin production in A. flavus with no significant effect on the fungus mycelia growth. The result also suggested P. betle could be used as potential antitoxin product.

  5. Inhibitory effect of the essential oil of Curcuma longa L. and curcumin on aflatoxin production by Aspergillus flavus Link.

    PubMed

    Ferreira, Flavio Dias; Kemmelmeier, Carlos; Arrotéia, Carla Cristina; da Costa, Christiane Luciana; Mallmann, Carlos Augusto; Janeiro, Vanderly; Ferreira, Francine Maery Dias; Mossini, Simone Aparecida Galerani; Silva, Expedito Leite; Machinski, Miguel

    2013-01-15

    Aflatoxins are highly toxic, mutagenic, teratogenic and carcinogenic mycotoxins. Consumption of aflatoxin-contaminated food and commodities poses serious hazards to the health of humans and animals. Turmeric, Curcuma longa L., is a native plant of Southeast Asia and has antimicrobial, antioxidant and antifungal properties. This paper reports the antiaflatoxigenic activities of the essential oil of C. longa and curcumin. The medium tests were prepared with the oil of C. longa, and the curcumin standard at concentrations varied from 0.01% to 5.0%. All doses of the essential oil of the plant and the curcumin standard interfered with mycotoxin production. Both the essential oil and curcumin significantly inhibited the production of aflatoxins; the 0.5% level had a greater than 96% inhibitory effect. The levels of aflatoxin B(1) (AFB(1)) production were 1.0 and 42.7 μg/mL, respectively, for the samples treated with the essential oil of C. longa L. and curcumin at a concentration of 0.5%. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Natural occurrence of aflatoxins and ochratoxin A in processed spices marketed in Malaysia.

    PubMed

    Ali, Norhayati; Hashim, Noor Hasani; Shuib, Nor Shifa

    2015-01-01

    The analysis of aflatoxins (B1, B2, G1 and G2) and ochratoxin A (OTA) was performed in processed spices marketed in Penang, Malaysia, using immunoaffinity columns and HPLC equipped with fluorescence detector (HPLC-FD). The processed powdered spices analysed include dried chilli, fennel, cumin, turmeric, black and white pepper, poppy seed, coriander, 'garam masala', and mixed spices for fish, meat and chicken curry. Two different studies were carried out. The limit of detection (LOD) was 0.01 ng g(-1) for each aflatoxin (AF) and 0.10 ng g(-1) for OTA (signal-to-noise ratio = 3:1). In the first study, 34 commercial processed spices analysed with a mean level, range and incidence of positive samples for total AF were 1.61 ng g(-1), 0.01-9.34 ng g(-1) and 85%, respectively, and for AFB1 were 1.38 ng g(-1), 0.01-7.68 ng g(-1) and 85%, respectively. The mean level, range and incidence of positive samples for OTA were 2.21 ng g(-1), 0.14-20.40 ng g(-1) and 79%, respectively. Natural co-occurrence of AF and OTA was found in 25 (74%) samples. In the second study of 24 commercial processed spices, the mean level, range and incidence of positive samples for total AF were 8.38 ng g(-1), 0.32-31.17 ng g(-1) and 88%, respectively, and for AFB1 were 7.31 ng g(-1), 0.32-28.43 ng g(-1) and 83%, respectively. Fifteen positive samples for total AF and two positive samples for OTA exceeded the permissible Malaysian limit of 5 ng g(-1). Contamination of both mycotoxins in spices may represent another route of exposure to consumers due to their frequent and prolonged consumption, as spices are common ingredients in popular dishes among Asian countries.

  7. Aflatoxin and viral hepatitis exposures in Guatemala: Molecular biomarkers reveal a unique profile of risk factors in a region of high liver cancer incidence.

    PubMed

    Smith, Joshua W; Kroker-Lobos, Maria F; Lazo, Mariana; Rivera-Andrade, Alvaro; Egner, Patricia A; Wedemeyer, Heiner; Torres, Olga; Freedman, Neal D; McGlynn, Katherine A; Guallar, Eliseo; Groopman, John D; Ramirez-Zea, Manuel

    2017-01-01

    Liver cancer is an emerging global health issue, with rising incidence in both the United States and the economically developing world. Although Guatemala experiences the highest rates of this disease in the Western hemisphere and a unique 1:1 distribution in men and women, few studies have focused on this population. Thus, we determined the prevalence and correlates of aflatoxin B1 (AFB1) exposure and hepatitis virus infection in Guatemalan adults. Healthy men and women aged ≥40 years (n = 461), residing in five departments of Guatemala, were enrolled in a cross-sectional study from May-October of 2016. Serum AFB1-albumin adducts were quantified using isotope dilution mass spectrometry. Multivariate linear regression was used to assess relationships between AFB1-albumin adduct levels and demographic factors. Biomarkers of hepatitis B virus and hepatitis C virus infection were assessed by immunoassay and analyzed by Fisher's exact test. AFB1-albumin adducts were detected in 100% of participants, with a median of 8.4 pg/mg albumin (range, 0.2-814.8). Exposure was significantly higher (p<0.05) in male, rural, low-income, and less-educated participants than in female, urban, and higher socioeconomic status participants. Hepatitis B and C seropositivity was low (0.9% and 0.5%, respectively). Substantial AFB1 exposure exists in Guatemalan adults, concurrent with low prevalence of hepatitis virus seropositivity. Quantitatively, AFB1 exposures are similar to those previously found to increase risk for liver cancer in Asia and Africa. Mitigation of AFB1 exposure may reduce liver cancer incidence and mortality in Guatemala, warranting further investigation.

  8. Effects of harmane on growth and in vivo metabolism of aflatoxin B1 in male and female rats.

    PubMed

    Billaud, C

    1991-01-01

    The role of harmane, a beta-carboline formed during pyrolysis of tryptophan, on the metabolism of AFB1, growth and some parameters of the nutritional status was investigated in the rat. Male and female Wistar rats were fed a semi-synthetic diet containing AFB1 (2 ppm), harmane (250 ppm) or both compounds, for 33 days after weaning. Qualitative and quantitative differences in the urinary and faecal excretion of parental compound and metabolites were assessed by HPLC analysis. Harmane did not modify appreciably the growth and the other nutritional parameters studied. Similar excretion patterns of AFB1 metabolites were observed in males and females. Harmane caused a limited increase in the excretion of AFM1 in faeces but not in urine, without altering the growth process in rats of either sex.

  9. Efficacy of Lippia alba (Mill.) N.E. Brown essential oil and its monoterpene aldehyde constituents against fungi isolated from some edible legume seeds and aflatoxin B1 production.

    PubMed

    Shukla, Ravindra; Kumar, Ashok; Singh, Priyanka; Dubey, Nawal Kishore

    2009-10-31

    The present study deals with evaluation of antifungal properties of Lippia alba essential oil (EO) and two of its monoterpene aldehyde constituents against legume-contaminating fungi. Seventeen different fungal species were isolated from 11 varieties of legumes, and aflatoxigenic isolates of Aspergillus flavus were identified. Hydrodistillation method was used to extract the EO from fresh leaves. The GC and GC-MS analysis of EO revealed the monoterpene aldehydes viz. geranial (22.2%) and neral (14.2%) as the major components. The antifungal activity of EO, geranial and neral was evaluated by contact assay on Czapek's-dox agar. The EO (0.25-1 microL/mL) and its two constituents (1 microL/mL) showed remarkable antifungal effects against all the fungal isolates (growth inhibition range 32.1-100%). Their minimal inhibitory (MIC) and fungicidal (MFC) concentrations for A. flavus were lower than those of the systemic fungicide Bavistin. Aflatoxin B(1) (AFB(1)) production by three isolates of A. flavus was strongly inhibited even at the lower fungistatic concentration of EO and its constituents. There was no adverse effect of treatments on seed germination, and rather, there was enhanced seedling growth in the EO-treated seeds. It is concluded that L. alba EO and two of its constituents could be safely used as effective preservative for food legumes against fungal infections and mycotoxins.

  10. Effect of probiotic supplement on aflatoxicosis and gene expression in the liver of broiler chicken.

    PubMed

    Salem, Ramadan; El-Habashi, Nagwan; Fadl, Sabreen E; Sakr, Osama A; Elbialy, Zizy I

    2018-06-01

    The present study aimed to investigate the toxic effect of aflatoxin B1 (AFB1) and the effectiveness of a biological mycotoxin binder (Nutritox ® ) in detoxifying aflatoxicosis. Ninety one-day-old chicks were chosen and divided into 3 groups. The first group received standard basal diet only, the second group received a basal diet with AFB1 (0.25 mg/kg diet), and the third group received a basal diet with AFB1 (0.25 mg/kg diet) and Nutritox ® (1 kg/ton diet). AFB1 feed contamination significantly reduced growth performance and deteriorated FCR. Moreover, it significantly increased serum AST, ALT, and malondialdehyde and significantly decreased serum total protein, albumin, globulin, SOD, CAT activities and glutathione peroxidase gene expression as well. Aflatoxin residues were detected in the liver tissues. Furthermore, the liver and kidney of AFB1 treated group showed pathological changes. The supplementation of Nutritox ® significantly reduced aflatoxin levels in the liver and counteracted the negative effects of AFB1. Copyright © 2018 Elsevier B.V. All rights reserved.

  11. Aflatoxin exposure during the first 1000 days of life in rural South Asia assessed by aflatoxin B₁-lysine albumin biomarkers.

    PubMed

    Groopman, John D; Egner, Patricia A; Schulze, Kerry J; Wu, Lee S-F; Merrill, Rebecca; Mehra, Sucheta; Shamim, Abu A; Ali, Hasmot; Shaikh, Saijuddin; Gernand, Alison; Khatry, Subarna K; LeClerq, Steven C; West, Keith P; Christian, Parul

    2014-12-01

    Aflatoxin B1 is a potent carcinogen, occurring from mold growth that contaminates staple grains in hot, humid environments. In this investigation, aflatoxin B1-lysine albumin biomarkers were measured by mass spectrometry in rural South Asian women, during the first and third trimester of pregnancy, and their children at birth and at two years of age. These subjects participated in randomized community trials of antenatal micronutrient supplementation in Sarlahi District, southern Nepal and Gaibandha District in northwestern Bangladesh. Findings from the Nepal samples demonstrated exposure to aflatoxin, with 94% detectable samples ranging from 0.45 to 2939.30 pg aflatoxin B1-lysine/mg albumin during pregnancy. In the Bangladesh samples the range was 1.56 to 63.22 pg aflatoxin B1-lysine/mg albumin in the first trimester, 3.37 to 72.8 pg aflatoxin B1-lysine/mg albumin in the third trimester, 4.62 to 76.69 pg aflatoxin B1-lysine/mg albumin at birth and 3.88 to 81.44 pg aflatoxin B1-lysine/mg albumin at age two years. Aflatoxin B1-lysine adducts in cord blood samples demonstrated that the fetus had the capacity to convert aflatoxin into toxicologically active compounds and the detection in the same 2-year-old children illustrates exposure over the first 1000 days of life. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. RNA interference-based silencing of the alpha-amylase (amy1) gene in Aspergillus flavus decreases fungal growth and aflatoxin production in maize kernels.

    PubMed

    Gilbert, Matthew K; Majumdar, Rajtilak; Rajasekaran, Kanniah; Chen, Zhi-Yuan; Wei, Qijian; Sickler, Christine M; Lebar, Matthew D; Cary, Jeffrey W; Frame, Bronwyn R; Wang, Kan

    2018-06-01

    Expressing an RNAi construct in maize kernels that targets the gene for alpha-amylase in Aspergillus flavus resulted in suppression of alpha-amylase (amy1) gene expression and decreased fungal growth during in situ infection resulting in decreased aflatoxin production. Aspergillus flavus is a saprophytic fungus and pathogen to several important food and feed crops, including maize. Once the fungus colonizes lipid-rich seed tissues, it has the potential to produce toxic secondary metabolites, the most dangerous of which is aflatoxin. The pre-harvest control of A. flavus contamination and aflatoxin production is an area of intense research, which includes breeding strategies, biological control, and the use of genetically-modified crops. Host-induced gene silencing, whereby the host crop produces siRNA molecules targeting crucial genes in the invading fungus and targeting the gene for degradation, has shown to be promising in its ability to inhibit fungal growth and decrease aflatoxin contamination. Here, we demonstrate that maize inbred B104 expressing an RNAi construct targeting the A. flavus alpha-amylase gene amy1 effectively reduces amy1 gene expression resulting in decreased fungal colonization and aflatoxin accumulation in kernels. This work contributes to the development of a promising technology for reducing the negative economic and health impacts of A. flavus growth and aflatoxin contamination in food and feed crops.

  13. Aflatoxin (B1 , B2 , G1 , and G2 ) contamination in rice of Mexico and Spain, from local sources or imported.

    PubMed

    Suárez-Bonnet, Elena; Carvajal, Magda; Méndez-Ramírez, Ignacio; Castillo-Urueta, Pável; Cortés-Eslava, Josefina; Gómez-Arroyo, Sandra; Melero-Vara, José María

    2013-11-01

    Rice is an important cereal but it is often contaminated with aflatoxins (AFs). The purpose of this study was to identify and quantify AF (B1 , B2 , G1 , and G2 ) in 67 rice samples cultivated in Mexico and Spain, and from imported crops collected in 2008 and 2009. The methodology was validated, the rice samples were concentrated and purified with immunoaffinity columns and were quantified by high-pressure liquid chromatography (HPLC). The average total AF (AFt) in the Spanish rice was 37.3 μg/kg, the range was from 1.6 to 1383 μg/kg, the most contaminated samples being from San Juan de Aznalfarache, Sevilla (AFt = 138.6 μg/kg), from Tortosa, Tarragona (AFt = 104.6 μg/kg), and Calasparra, Murcia (AFt = 103.9 μg/kg). The rice imported from France to Spain had AFt of 26.6 μg/kg and from Pakistan AFt of 18.4 μg/kg, showing less AF contamination than the local one. The rice which originated from Mexico contained (AFt = 16.9 μg/kg), and those imported from the United States (AFt = 14.4 μg/kg) and Uruguay (AFt = 15.6 μg/kg). The imported rice had better quality in terms of the presence of AFs. © 2013 Institute of Food Technologists®

  14. Aflatoxin decomposition in various soils

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Angle, J.S.

    The persistence of aflatoxin in the soil environment could potentially result in a number of adverse environmental consequences. To determine the persistence of aflatoxin in soil, /sup 14/C-labeled aflatoxin B1, was added to silt loam, sandy loam, and silty clay loam soils and the subsequent release of /sup 14/CO/sub 2/ was determined. After 120 days of incubation, 8.1% of the original aflatoxin added to the silt loam soil was released as CO/sub 2/. Aflatoxin decomposition in the sandy loam soil proceeded more quickly than the other two soils for the first 20 days of incubation. After this time, the decompositionmore » rate declined and by the end of the study, 4.9% of the aflatoxin was released as CO/sub 2/. Aflatoxin decomposition proceeded most slowly in the silty clay loam soil. Only 1.4% of aflatoxin added to the soil was released as CO/sub 2/ after 120 days incubation. To determine whether aflatoxin was bound to the silty clay loam soil, aflatoxin B1 was added to this soil and incubated for 20 days. The soil was periodically extracted and the aflatoxin species present were determined using thin layer chromatographic (TLC) procedures. After one day of incubation, the degradation products, aflatoxins B2 and G2, were observed. It was also found that much of the aflatoxin extracted from the soil was not mobile with the TLC solvent system used. This indicated that a conjugate may have formed and thus may be responsible for the lack of aflatoxin decomposition.« less

  15. Assay for Aflatoxin Production by the Genera Aspergillus and Penicillium1

    PubMed Central

    Mislivec, Philip B.; Hunter, J. H.; Tuite, John

    1968-01-01

    A total of 260 isolates, including 43 species of Penicillium and 7 species of Aspergillus, were screened for their ability to produce aflatoxin on rice. Chloroform extracts were analyzed by thin-layer chromatography. None of the isolates produced aflatoxin. Certain species of Penicillium produced fluorescent substances that either were similar in RF or were of similar color to B and G aflatoxins. These substances were subsequently proved not to be aflatoxin by two-dimensional chromatography, by reaction with iodine fumes, or by both methods. PMID:5664121

  16. [Adsorption of aflatoxin on montmorillonite modified by low-molecular-weight humic acids].

    PubMed

    Yao, Jia-Jia; Kang, Fu-Xing; Gao, Yan-Zheng

    2012-03-01

    The adsorption of a typical biogenic toxin aflatoxin B1 on montmorillonite modified by low-molecular-weight humic acids (M(r) < 3 500) was investigated. The montmorillonite rapidly adsorbed the aflatoxin B1 until amounting to the maximal capacity, and then the adsorbed aflatoxin B1 slowly released into solution and reached the sorption equilibrium state after 12 h. The sorption isotherm of aflatoxin B1 by montmorillonite could be well described by Langmiur model, while the sorption isotherm by humic acid-modified montmorillonite was well fitted by using the Freundlich model. The modification of the montmorillonite with humic acids obviously enhanced its adsorption capacity for aflatoxin B1, and the amounts of aflatoxin adsorbed by modified montmorillonite were obviously higher than those by montmorillonite. The sorption enhancement by humic acid modification was attributed to (1) the enlarged adsorption sites which owed to the surface collapse of crystal layers induced by organic acids, and (2) the binding of aflatoxin with the humic acid sorbed on mineral surface. In addition, the adsorption amounts of aflatoxin by montmorillonite and modified montmorillonite increased with the increase of pH values in solution, and more significant enhancement was observed for the latter than the former, which attributed to the release of humic acids from the modified montmorillonite with the high pH values in solution. This indicates that increasing the pH values resulted in the enhanced hydrophilic property and the release of the organic acids presented in modified montmorillonite, and more sorption sites were available for aflatoxin on the modified montmorillonite. Results of this work would strengthen our understanding of the behavior and fate of biological contaminants in the environment.

  17. Determination of Aflatoxin M1 Contamination and Integrity as well as Credibility

    PubMed Central

    Ataee, R; Tavana, A Mehrabi; Ataee, MH

    2012-01-01

    Aspergillums can produce and secrete directly aflatoxin M1. During the previous decade several papers pertaining to aflatoxin M1 have been published in different journals. Not mention of their more or less scientific aspects, they have fundamentally some problems in different features. In this paper we are going to have a bird’s eye view on some articles published on this topic. It is suggested that complete research must be performed in order to find out the source of aflatoxin M1 contamination. PMID:23304667

  18. Effect of Carum copticum essential oil on growth and aflatoxin formation by Aspergillus strains.

    PubMed

    Kazemi, M

    2015-01-01

    The objectives of this study were to determine the antiaflatoxin B1 activity in vitro of the essential oil (EO) extracted from the seeds of Carum copticum and to evaluate its antifungal activity in vivo as a potential food preservative. The C. copticum EO exhibited noticeable inhibition on dry mycelium and synthesis of aflatoxin B1 (AFB1) by Aspergillus flavus, completely inhibiting AFB1 production at 4 μL/mL. C. copticum EOs showed the lowest percentages of decayed cherry tomatoes for all fungi compared with the control at 100 μL/mL with values of 5.01 ± 67% for A. flavus and 5.98 ± 54% for Aspergillus niger. The results indicated that the percentage of infected fruits is significantly (p < 0.01) reduced by the EO at 16°C for 30 days. In this case, the oil at 100 μL/mL concentration showed the highest inhibition of fungal infection with a value of 80.45% compared with the control. Thus, the EO of dill could be used to control food spoilage and as a potential source of food preservative.

  19. Nanoencapsulated Illicium verum Hook.f. essential oil as an effective novel plant-based preservative against aflatoxin B1 production and free radical generation.

    PubMed

    Dwivedy, Abhishek Kumar; Singh, Vipin Kumar; Prakash, Bhanu; Dubey, Nawal Kishore

    2018-01-01

    The study reports efficacy of Illicium verum essential oil (IvEO) against food borne moudls and its nanoencapsulation for enhancing antifungal and antiaflatoxigenic potency. Chemical characterization of the IvEO showed anethole (89.12%) as major compound followed by estragole (4.859%). The IvEO showed broad fungitoxic spectrum against common food borne moulds. It's minimum inhibitory concentration (MIC) and minimum aflatoxin B 1 inhibitory concentration (MAIC) against aflatoxigenic strain Aspergillus flavus LHP-PV-1 were 0.7, and 0.5 μL/mL respectively. Morphological observations of treatment sets by SEM and TEM along with decrease in ergosterol content and enhanced leakage of Ca 2+ , K + and Mg 2+ ions denoted fungal cell membrane as site of action. The IvEO showed promising free radical scavenging activity and favourable safety profile with high LD 50 value on mice. The IvEO also exhibited considerable protection of Pistacia vera from fungal contamination and complete protection from aflatoxin B 1 contamination in storage containers. Nanoencapsulated IvEO in gel form and lyophilized form exhibited enhanced efficacy as fungal inhibitor and aflatoxin suppressor. The chemically characterised IvEO may be recommended as plant based preservative having favourable safety and its nanocapsules may be of industrial significance as shelf life enhancer of food items. This is the first report on in situ antiaflatoxigenic efficacy and nanoencapsulation of IvEO. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Antigenotoxic Effect of Piperine in Broiler Chickens Intoxicated with Aflatoxin B1.

    PubMed

    da Silva Cardoso, Verônica; Vermelho, Alane Beatriz; Ribeiro de Lima, Cristina Amorim; Mendes de Oliveira, Jéssica; Freire de Lima, Marco Edilson; Pinto da Silva, Lúcia Helena; Direito, Glória Maria; Miranda Danelli, Maria das Graças

    2016-10-31

    Piperine is an abundant amide extracted from black pepper seeds which has been shown to have protective effects against cytotoxic and genotoxic carcinogenesis induced by certain chemical carcinogens and aflatoxin B₁ (AFB₁) in vitro. The aim of this work was to study, in vivo, the antigenotoxic potential of feed-added piperine on broiler chickens experimentally intoxicated with AFB₁, using micronucleus and comet assays. The antigenotoxicity assessment of 9-day-old chicks was performed on a total of 60 chickens divided into four groups of 15 broilers each: (C) control, (P) 60 mg·piperine kg -1 feed, (A) 0.5 mg·AFB₁·kg -1 body weight, (daily by oral route), and (P + A) co-treatment with piperine and AFB₁. The experiment was conducted for 26 days. Chicks intoxicated with AFB₁ showed significant genotoxic effects in the first 24 h post intoxication, and the effects remained in the other periods analyzed (48, 72, and 96 h and 26 days of treatment). The DNA damage in peripheral blood cells, the number of erythrocytes with micronuclei, and polychromatic-to-normochromatic erythrocyte ratio were significantly reduced or absent in the piperine/AFB₁ group. No significant differences were observed between the group piperine/AFB₁ and the control and piperine-alone groups. The addition 60 mg·kg -1 of piperine to the diet of the broiler chicks was safe, promoting beneficial effects in poultry health with respect to the toxic effects 0.5 mg·AFB₁·kg -1 body weight.

  1. Effect of aflatoxin B₁ on IgA⁺ cell number and immunoglobulin mRNA expression in the intestine of broilers.

    PubMed

    Jiang, Min; Fang, Jing; Peng, Xi; Cui, Hengmin; Yu, Zhengqiang

    2015-01-01

    Aflatoxin B1 (AFB1) is the most toxic group of mycotoxins produced by two species of the Aspergillus, common contaminants of food and animal feed. The purpose of our study was to determine the effect of AFB1 on the number of IgA(+) cell and immunoglobulin mRNA expression in the intestine of broilers. One hundred and fifty six one-day-old healthy Cobb broilers were randomly divided into the control group (the dosage of 0 mg/kg AFB1) and AFB1 group (the dosage of 0.6 mg/kg AFB1) with three replicates per group and 26 birds per replicate for 21 days, respectively. After necropsy at 7, 14 and 21 days of age, duodenum, jejunum and ileum samples were taken for analyzing IgA(+) cell by immunohistochemistry and IgA, pIgR, IgM and IgG mRNA expression by qRT-PCR. IgA(+) cells were mainly distributed in the lamina propria of small intestinal mucosa in both groups at 14 and 21 days of age. A significant decrease in the number of IgA(+) cells in the duodenum, jejunum and ileum was revealed in the AFB1 group compared with that of the control group. The expression levels of IgA, pIgR, IgM and IgG mRNA in the intestinal mucosa were lower in the AFB1 group than those in the control group at 14 and 21 days of age. Our data demonstrated that the dosage of 0.6 mg/kg AFB1 in broiler diet reduced the number of IgA(+) cell and the expression of IgA, pIgR, IgM and IgG mRNA in the small intestine.

  2. Effects of feeding corn naturally contaminated with aflatoxin on growth performance, apparent ileal digestibility, serum hormones levels and gene expression of Na+, K+-ATPase in ducklings.

    PubMed

    Abbasi, Farzana; Liu, Jingbo; Zhang, Hongfu; Shen, Xiaoyun; Luo, Xuegang

    2018-01-01

    A 14-d trial was conducted to determine the effects of feeding corn naturally contaminated with aflatoxin B 1 (AFB 1 ) on growth performance, apparent ileal digestibility, serum hormones levels and gene expression of Na + , K + -ATPase in ducklings. A total of 704 ducklings were blocked on the basis of sex and body weight (BW), and then allocated randomly to one of the following two treatments: i) CON, basal diet and ii) AFB 1 , diets with 100% of normal corn replaced with AFB 1 contaminated corn. There were 22 pens per treatment and 16 birds per pen. The concentration of AFB 1 was 195.4 and 124.35 μg/kg in the contaminated corn and AFB 1 diet, respectively. The AFB 1 decreased average daily gain, average daily feed intake, d 7 BW, final BW in the whole trial, and feed conversion ratio (FCR) during d 8 to 14 and d 1 to 14 by 10% to 47% (p<0.05), while FCR during d 1 to 7 was increased (p<0.05). AFB 1 did not affect mortality to 7 d of age, and then increased to 5.8% from 8 to 14 d of age (p<0.01). Apparent ileal gross energy digestibility was reduced by AFB 1 , whereas apparent ileal digestibility of dry matter, nitrogen, and amino acid was improved (p<0.01). Feeding AFB 1 diets increased serum concentration of leptin and insulin-like growth factors-1 (IGF-1) (p<0.05), but had no effect on neuropeptide Y, ghrelin, cholecystokinin-8 or insulin (p>0.05). Dietary treatments did not influence relative expression of jejunal Na + , K + -ATPase gene (p>0.05). Taken together, feeding corn naturally contaminated with AFB 1 reduced growth performance, improved apparent ileal digestibility, and affected serum leptin and IGF-1 in ducklings from d 1 to 14.

  3. Development of an Ultrasonication-Assisted Extraction Based HPLC With a Fluorescence Method for Sensitive Determination of Aflatoxins in Highly Acidic Hibiscus sabdariffa.

    PubMed

    Liu, Xiaofei; Ying, Guangyao; Sun, Chaonan; Yang, Meihua; Zhang, Lei; Zhang, Shanshan; Xing, Xiaoyan; Li, Qian; Kong, Weijun

    2018-01-01

    The high acidity and complex components of Hibiscus sabdariffa have provided major challenges for sensitive determination of trace aflatoxins. In this study, sample pretreatment of H. sabdariffa was systematically developed for sensitive high performance liquid chromatography-fluorescence detection (HPLC-FLD) after ultrasonication-assisted extraction, immunoaffinity column (IAC) clean-up and on-line post-column photochemical derivatization (PCD). Aflatoxins B 1 , B 2 , G 1 , G 2 were extracted from samples by using methanol/water (70:30, v/v ) with the addition of NaCl. The solutions were diluted 1:8 with 0.1 M phosphate buffer (pH 8.0) to negate the issues of high acidity and matrix interferences. The established method was validated with satisfactory linearity ( R > 0.999), sensitivity (limits of detection (LODs) and limits of quantitation (LOQs) of 0.15-0.65 and 0.53-2.18 μg/kg, respectively), precision (RSD <11%), stability (RSD of 0.2-3.6%), and accuracy (recovery rates of 86.0-102.3%), which all met the stipulated analytical requirements. Analysis of 28 H. sabdariffa samples indicated that one sample incubated with Aspergillus flavus was positive with aflatoxin B 1 (AFB 1 ) at 3.11 μg/kg. The strategy developed in this study also has the potential to reliably extract and sensitively detect more mycotoxins in other complex acidic matrices, such as traditional Chinese medicines, foodstuffs, etc.

  4. Efficacy of chemically characterized Piper betle L. essential oil against fungal and aflatoxin contamination of some edible commodities and its antioxidant activity.

    PubMed

    Prakash, Bhanu; Shukla, Ravindra; Singh, Priyanka; Kumar, Ashok; Mishra, Prashant Kumar; Dubey, Nawal Kishore

    2010-08-15

    The study investigates fungal contamination in some dry fruits, spices and areca nut and evaluation of the essential oil (EO) of Piper betle var. magahi for its antifungal, antiaflatoxigenic and antioxidant properties. A total of 1651 fungal isolates belonging to 14 species were isolated from the samples and Aspergillus was recorded as the dominant genus with 6 species. Eleven aflatoxin B(1) (AFB(1)) producing strains of A. flavus were recorded from the samples. Eugenol (63.39%) and acetyleugenol (14.05%) were the major components of 32 constituents identified from the Piper betle EO through GC and GC-MS analysis. The minimum inhibitory concentration (MIC) of P. betle EO was found 0.7 microl/ml against A.flavus. The EO reduced AFB(1) production in a dose dependent manner and completely inhibited at 0.6 microl/ml. This is the first report on efficacy of P. betle EO as aflatoxin suppressor. EO also exhibited strong antioxidant potential as its IC(50) value (3.6 microg/ml) was close to that of ascorbic acid (3.2 microg/ml) and lower than that of the synthetic antioxidants such as butylated hydroxytouene (BHT) (7.4 microg/ml) and butylated hydroxyanisole (BHA) (4.5 microg/ml). P. betle EO thus exhibited special merits possessing antifungal, aflatoxin suppressive and antioxidant characters which are desirable for an ideal preservative. Hence, its application as a plant based food additive in protection and enhancement of shelf life of edible commodities during storage and processing is strongly recommended in view of the toxicological implications by synthetic preservatives. Copyright 2010 Elsevier B.V. All rights reserved.

  5. Aflatoxin-induced biochemical changes in liver of mice and its mitigation by black tea extract.

    PubMed

    Jha, Anamika; Shah, Komal; Verma, Ramtej J

    2012-01-01

    Aflatoxin belongs to the class of naturally occurring mycotoxins, food contaminants having potent carcinogenicity. We have evaluated the ameliorative role of black tea extract on aflatoxin-induced biochemical changes in the liver of albino male mice. Adult male mice were orally administered with 750 and 1500 pg of aflatoxin in 0.2 mL olive oil/kg b.w./day for 30 days. Oral administration of aflatoxin caused, as compared with controls, significant, dose-dependent reduction in DNA, RNA, protein and glycogen contents; however, cholesterol content and phsphorylase activity were significantly increased. Black tea is one of the most potent antioxidants containing numerous bioactive phytonurtients having therapeutic applications. Aflatoxin-induced changes in the liver of mice were significantly ameliorated on co-treatment of black tea extract (2% infusion in water).

  6. Aflatoxins and ochratoxin A in different cocoa clones (Theobroma cacao L.) developed in the southern region of Bahia, Brazil.

    PubMed

    Maciel, Leonardo Fonseca; Felício, Ana Lúcia de Souza Madureira; Miranda, Lucas Caldeirão Rodrigues; Pires, Tassia Cavalcante; Bispo, Eliete da Silva; Hirooka, Elisa Yoko

    2018-01-01

    Brazil is the sixth largest producer of cocoa beans in the world, after Côte d'Ivoire, Ghana, Indonesia, Nigeria and Cameroon. The southern region of Bahia stands out as the country's largest producer, accounting for approximately 60% of production. Due to damage caused by infestation of the cocoa crop with the fungus Moniliophthora perniciosa, which causes 'witch's broom disease', research in cocoa beans has led to the cloning of species that are resistant to the disease; however, there is little information about the development of other fungal genera in these clones, such as Aspergillus, which do not represent a phytopathogenicity problem but can grow during the pre-processing of cocoa beans and produce mycotoxins. Thus, the aim of this work was to determine the presence of aflatoxin (AF) and ochratoxin A (OTA) in cocoa clones developed in Brazil. Aflatoxin and ochratoxin A contamination were determined in 130 samples from 13 cocoa clones grown in the south of Bahia by ultra-performance liquid chromatography with a fluorescence detector. The method was evaluated for limit of detection (LOD) (0.05-0.90 μg kg -1 ), limit of quantification (0.10-2.50 μg kg -1 ) and recovery (RSD) (89.40-95.80%) for AFB 1 , AFB 2 , AFG 1 , AFG 2 and OTA. Aflatoxin contamination was detected in 38% of the samples in the range of 1 , with AFB 1 in 25% of the total samples, whereas ochratoxin A was positive in 18% of the samples in the range of 1 .

  7. The master transcription factor mtfA governs aflatoxin production, morphological development, and pathogenicity in the fungus Aspergillus flavus

    USDA-ARS?s Scientific Manuscript database

    Aspergillus flavus produces a variety of toxic secondary metabolites, among them the aflatoxins (AFs) are the most well-known. These compounds are highly mutagenic and carcinogenic, particularly AFB1. A. flavus is capable of colonizing economically important crops contaminating them with AFs. Molecu...

  8. High Aflatoxin Production on a Chemically Defined Medium 1

    PubMed Central

    Reddy, T. V.; Viswanathan, L.; Venkitasubramanian, T. A.

    1971-01-01

    Aspergillus parasiticus ATCC 15517 produced 28 to 30 mg of aflatoxin per 100 ml of a medium containing sucrose, asparagine, and salts in stationary and shaken cultures. In the absence of asparagine in the medium, the toxin yields fell drastically, and the thin-layer chromatograms of the chloroform extracts of the cultures indicated the total absence of aflatoxin G1 and the presence of new intense blue and green fluorescent bands having RF values lower than aflatoxins. Initial pH was critical and had to be around 4.5 for good growth and high toxin production on this medium. Optimum concentrations of KH2PO4 and MgSO4·7H2O in the medium were much lower than those normally used in fungal growth media. PMID:5119206

  9. Distribution and stability of aflatoxin M1 during processing, ripening and storage of Telemes cheese.

    PubMed

    Govaris, A; Roussi, V; Koidis, P A; Botsoglou, N A

    2001-05-01

    Telemes cheeses were produced using milk that was artificially-contaminated with aflatoxin M1 at the levels of 0.050 and 0.100 microg/l. The cheeses produced in the two cheese-making trials were allowed to ripen for 2 months and stored for an additional 4 months to simulate commercial production of Telemes cheese. Concentrations of aflatoxin M1 in whey, curd, brine, and the produced cheeses were determined at intervals by liquid chromatography and fluorometric detection coupled with immunoaffinity column extraction. Concentrations of aflatoxin M1 in the produced curds were found to be 3.9 and 4.4 times higher than those in milk, whereas concentrations in whey were lower than those in curd and milk. Aflatoxin M1 was present in cheese at higher concentrations at the beginning than at the end of the ripening/storage period, and it declined to concentrations 2.7 and 3.4 times higher than those initially present in milk by the end of the sixth month of storage. Concentrations of aflatoxin M1 in brine started low and increased by the end of the ripening/storage period but only a portion of the amounts of aflatoxin M1 lost from cheese was found in the brine. Results showed that Telemes cheeses produced from milk containing aflatoxin M1 at a concentration close to either the maximum acceptable level of 0.05 microg/l set by the European union (EU) or at double this value, will contain the toxin at a level that is much lower or slightly higher, respectively, than the maximum acceptable level of 0.250 microg of aflatoxin M1/kg cheese set by some countries.

  10. Rapid, economical qualitative method for separation of aflatoxins B-1, B-2 & G-1, G-2 by dry column chromatography.

    PubMed

    Megalla, S E

    1983-12-01

    A good correlation of four components of aflatoxins was accomplished by using the dry column chromatography method. The decolorization process of interfering substances, by 0.01 N KOH and defatting the extract with petroleum ether yields a clean residue for DCC separation. It is clear that the dry column chromatography is a very simple and time-saving procedure for separation of aflatoxins. DCC columns are more economical than precoated 'thick layer' preparative plates and, in DCC, no large developing tanks need to be used. Hazards associated with the use of large volumes of flammable solvents are greatly reduced.

  11. Reduction of aflatoxins by Rhizopus oryzae and Trichoderma reesei.

    PubMed

    Hackbart, H C S; Machado, A R; Christ-Ribeiro, A; Prietto, L; Badiale-Furlong, E

    2014-08-01

    This study evaluated the ability of the microorganisms Rhizopus oryzae (CCT7560) and Trichoderma reesei (QM9414), producers of generally recognized as safe (GRAS) enzymes, to reduce the level of aflatoxins B1, B2, G1, G2, and M1. The variables considered to the screening were the initial number of spores in the inoculum and the culture time. The culture was conducted in contaminated 4 % potato dextrose agar (PDA) medium, and the residual mycotoxins were determined every 24 h by HPLC-FL. The fungus R. oryzae has reduced aflatoxins B1, B2, and G1 in the 96 h and aflatoxins M1 and G2 in the range of 120 h of culture by approximately 100 %. The fungus T. reesei has reduced aflatoxins B1, B2, and M1 in the 96 h and aflatoxin G1 in the range of 120 h of culture by approximately 100 %. The highest reduction occurred in the middle of R. oryzae culture.

  12. A systems approach to model the relationship between aflatoxin gene cluster expression, environmental factors, growth and toxin production by Aspergillus flavus

    PubMed Central

    Abdel-Hadi, Ahmed; Schmidt-Heydt, Markus; Parra, Roberto; Geisen, Rolf; Magan, Naresh

    2012-01-01

    A microarray analysis was used to examine the effect of combinations of water activity (aw, 0.995–0.90) and temperature (20–42°C) on the activation of aflatoxin biosynthetic genes (30 genes) in Aspergillus flavus grown on a conducive YES (20 g yeast extract, 150 g sucrose, 1 g MgSO4·7H2O) medium. The relative expression of 10 key genes (aflF, aflD, aflE, aflM, aflO, aflP, aflQ, aflX, aflR and aflS) in the biosynthetic pathway was examined in relation to different environmental factors and phenotypic aflatoxin B1 (AFB1) production. These data, plus data on relative growth rates and AFB1 production under different aw × temperature conditions were used to develop a mixed-growth-associated product formation model. The gene expression data were normalized and then used as a linear combination of the data for all 10 genes and combined with the physical model. This was used to relate gene expression to aw and temperature conditions to predict AFB1 production. The relationship between the observed AFB1 production provided a good linear regression fit to the predicted production based in the model. The model was then validated by examining datasets outside the model fitting conditions used (37°C, 40°C and different aw levels). The relationship between structural genes (aflD, aflM) in the biosynthetic pathway and the regulatory genes (aflS, aflJ) was examined in relation to aw and temperature by developing ternary diagrams of relative expression. These findings are important in developing a more integrated systems approach by combining gene expression, ecophysiological influences and growth data to predict mycotoxin production. This could help in developing a more targeted approach to develop prevention strategies to control such carcinogenic natural metabolites that are prevalent in many staple food products. The model could also be used to predict the impact of climate change on toxin production. PMID:21880616

  13. Molecularly imprinted polymer nanoparticle-based assay (MINA): application for fumonisin B1 determination.

    PubMed

    Munawar, Hasim; Smolinska-Kempisty, Katarzyna; Cruz, Alvaro Garcia; Canfarotta, Francesco; Piletska, Elena; Karim, Khalku; Piletsky, Sergey A

    2018-06-20

    The enzyme-linked immunosorbent assay (ELISA) has been used as a standard tool for monitoring food and animal feed contamination from the carcinogenic fumonisin B1 (FB1). Unfortunately, ELISA is not always efficient due to the instability of the antibody and enzyme components in the immunoassay, the presence of natural enzyme inhibitors in the samples and the high levels of non-specific protein binding. Additionally, the production of antibodies for ELISA can be time-consuming and costly, due to the involvement of animals in the manufacturing process. To overcome these limiting factors, a molecularly imprinted nanoparticle based assay (MINA) has been developed, where the molecularly imprinted nanoparticles (nanoMIPs) replace the primary antibody used in a competitive ELISA. Herein, computational modelling was used to design the nanoMIPs by selecting monomers that specifically interact with FB1. The affinity of the monomers to FB1 was verified by measuring their binding in affinity chromatography experiments. The nanoMIPs were produced by solid phase synthesis and the results showed that nanoMIPs had a hydrodynamic diameter of around 249 ± 29 nm. The assay tested in model samples is highly selective and does not show cross-reactivity with other mycotoxins such as fumonisin B2 (FB2), aflatoxin B1 (AFB1), citrinin (CTT), zearalenone (ZEA), and deoxynivalenol (DON). The MINA allows the detection of FB1 in the concentration range of 10 pM-10 nM with a detection limit of 1.9 pM and a recovery of 108.13-113.76%.

  14. [Morphological characteristics and physiological properties of aflatoxin B1 producing and non-producing Aspergillus flavus strains].

    PubMed

    Kogbo, W; Lemarinier, S; Boutibonnes, P

    1985-09-01

    Comparison between about 80 strains of Aspergillus flavus, belonging to the series flavus and oryzae, obtained from international collections but also isolated from French or African substrates revealed the following observations: 1. Cultural and morphological characteristics of toxicogenic and atoxicogenic strains of A. flavus are similar. However, the former produce a diffusible yellow pigment in 83% of isolates. 2. The two groups of conidiospores have the same resistance to UV irradiation (254 nm, 5 and 10 min). All the strains are equally sensitive to 4 antifungal antibiotics: nystatine, ketoconazole, clotrimazole and amphotericine. 3. A difference was seen in the capacity to produce enzymes as alpha-galactosidase, beta-galactosidase and beta-glucosidase, implicated in the glucid metabolism. The specific hydrolytic activity has been confirmed by the characterization of a large amount of beta-galactosidase and by a diauxic growth on glucose medium supplemented by lactose. Possible relationship between these characters and aflatoxin B1 production by A. flavus strains is discussed.

  15. Development of an Ultrasonication-Assisted Extraction Based HPLC With a Fluorescence Method for Sensitive Determination of Aflatoxins in Highly Acidic Hibiscus sabdariffa

    PubMed Central

    Liu, Xiaofei; Ying, Guangyao; Sun, Chaonan; Yang, Meihua; Zhang, Lei; Zhang, Shanshan; Xing, Xiaoyan; Li, Qian; Kong, Weijun

    2018-01-01

    The high acidity and complex components of Hibiscus sabdariffa have provided major challenges for sensitive determination of trace aflatoxins. In this study, sample pretreatment of H. sabdariffa was systematically developed for sensitive high performance liquid chromatography-fluorescence detection (HPLC-FLD) after ultrasonication-assisted extraction, immunoaffinity column (IAC) clean-up and on-line post-column photochemical derivatization (PCD). Aflatoxins B1, B2, G1, G2 were extracted from samples by using methanol/water (70:30, v/v) with the addition of NaCl. The solutions were diluted 1:8 with 0.1 M phosphate buffer (pH 8.0) to negate the issues of high acidity and matrix interferences. The established method was validated with satisfactory linearity (R > 0.999), sensitivity (limits of detection (LODs) and limits of quantitation (LOQs) of 0.15–0.65 and 0.53–2.18 μg/kg, respectively), precision (RSD <11%), stability (RSD of 0.2–3.6%), and accuracy (recovery rates of 86.0–102.3%), which all met the stipulated analytical requirements. Analysis of 28 H. sabdariffa samples indicated that one sample incubated with Aspergillus flavus was positive with aflatoxin B1 (AFB1) at 3.11 μg/kg. The strategy developed in this study also has the potential to reliably extract and sensitively detect more mycotoxins in other complex acidic matrices, such as traditional Chinese medicines, foodstuffs, etc. PMID:29681848

  16. Detection of aflatoxin B₁ with immunochromatographic test strips: Enhanced signal sensitivity using gold nanoflowers.

    PubMed

    Ji, Yanwei; Ren, Meiling; Li, Yanping; Huang, Zhibing; Shu, Mei; Yang, Hongwei; Xiong, Yonghua; Xu, Yang

    2015-09-01

    Immunochromatographic test strips (ICTS) are commonly limited to higher concentrations of analytes. This limitation stems from the relatively low sensitivity of conventional gold nanospheres (AuNSs with a diameter of 20 nm) to emit detectable brightness values. The larger multi-branched gold nanoflowers (AuNFs) with a higher optical brightness as well as good colloidal stability exhibit significant improvements over conventional AuNSs for enhanced sensitivity of ICTS. In this study, blue AuNFs with an average diameter of 75±5 nm were synthetized and employed as a signal amplification probe for ultrasensitive and quantitative detection of aflatoxin B1 (AFB1) in rice. A portable optical strip reader was used to record the optical densities of test and control lines of the strip. Under the optimal conditions, the AuNF based ICTS system accurately detected AFB1 linearly and dynamically over the range of 0.5-25 pg/mL with a half maximal inhibitory concentration at 4.17 pg/mL. The inhibitory concentration was achieved 10 times lower than that of the traditional AuNS based ICTS systems (41.25 pg/mL). The limit of detection for AFB1 in rice extract was achieved at 0.32 pg/mL. In summary, AuNFs are a novel probe that exhibited excellent sensitivity in the ICTS system and could be used for ultrasensitive detection of other analytes in food safety monitoring, and even medical diagnostics. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. Dietary exposure to aflatoxin B1, ochratoxin A and fuminisins of adults in Lao Cai province, Viet Nam: A total dietary study approach.

    PubMed

    Huong, Bui Thi Mai; Tuyen, Le Danh; Tuan, Do Huu; Brimer, Leon; Dalsgaard, Anders

    2016-12-01

    Aflatoxins, fumonisins and ochratoxin A that contaminate various agricultural commodities are considered of significant toxicity and potent human carcinogens. This study took a total dietary study approach and estimated the dietary exposure of these mycotoxins for adults living in Lao Cai province, Vietnam. A total of 42 composite food samples representing 1134 individual food samples were prepared according to normal household practices and analysed for the three mycotoxins. Results showed that the dietary exposure to aflatoxin B1 (39.4 ng/kg bw/day) and ochratoxin A (18.7 ng/kg bw/day) were much higher than recommended provisional tolerable daily intake (PTDI) values mainly due to contaminated cereals and meat. The exposure to total fumonisins (1400 ng/kg bw/day) was typically lower than the PTDI value (2000 ng/kg bw/day). The estimated risk of liver cancer associated with exposure to aflatoxin B1 was 2.7 cases/100,000 person/year. Margin of exposure (MOE) of renal cancer linked to ochratoxin A and liver cancer associated with fumonisins were 1124 and 1954, respectively indicating risk levels of public health concern. Further studies are needed to evaluate the efficiency of technical solutions which could reduce mycotoxin contamination as well as to determine the health effects of the co-exposure to different types of mycotoxins. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. [Survey of aflatoxins contamination of foodstuffs and edible oil in Shenzhen].

    PubMed

    Li, Ke; Qiu, Fen; Yang, Mei; Liang, Zhaohai; Zhou, Haitao

    2013-07-01

    To identify the aflatoxins contamination of foodstuffs and edible oil sold in Shenzhen. As research subjects stratified random sampling of 238 foodstuffs and edible oil, and applied with immuno-affinity column clean-up plus UPLC to determine the content of aflatoxin B1, B2, G1, and G2. Positive ratio of aflatoxin in rice, rice products, wheat flour, corn flour, edible oil were 35.3%, 33.8%, 13.9%, 46.7% and 24.5%,respectively. There were statistical differences between the positive ratio of aflatoxin in stereotypes packaged rice (26.5%) and bulk rice (56.3%) (chi2 = 11.6, P < 0.05). There were statistical differences between the positive ratio of aflatoxin in the rice produced in the area north of the Yangtze River (27.3%) and in the rice (41.4%) produced in the area south of the Yangtze River (chi2 = 7.257, P < 0.05). Aflatoxin B1 and B2 were detected in rice products, wheat flour, corn flour. Positive ratio of aflatoxin B1, B2, G1, and G2 were 24.5%, 24.5%, 11.3% and 3.8% in the edible oil,respectively. The over standard rate of aflatoxin B1 was 5.66%, excessive samples were producted bulk and self-pressed peanut oil from unlicensed workshop. All the four kinds of aflatoxin were detected, while subtype B1 and B2 dominated aflatoxin contamination in the rice and edible oil samples. There are differences between in the northern and southern rice, and the same as in the stereotypes packaged and bulk rice sold at Shenzhen.

  19. Effects of clay after an aflatoxin challenge on aflatoxin clearance, milk production, and metabolism of Holstein cows.

    PubMed

    Sulzberger, S A; Melnichenko, S; Cardoso, F C

    2017-03-01

    Oral supplementation of clay to dairy cattle has been reported to reduce toxicity of aflatoxin (AF) in contaminated feed. The objective of this study was to determine the effects of 3 concentrations of dietary clay supplementation in response to an AF challenge. Ten multiparous rumen-cannulated Holstein cows [body weight (mean ± SD) = 669 ± 20 kg and 146 ± 69 d in milk], were assigned to 1 of 5 treatments in a randomized replicated 5 × 5 Latin square design balanced to measure carryover effects. Periods (21 d) were divided in an adaptation phase (d 1 to 14) and a measurement phase (d 15 to 21). From d 15 to 17, cows received an AF challenge. The challenge consisted of 100 μg of aflatoxin B 1 (AFB 1 )/kg of dietary dry matter intake (DMI). The material was fitted into 10-mL gelatin capsules and administered into the rumen through a rumen-cannula based on the average DMI obtained on d 12 to 14. Treatments were no clay plus an AF challenge (POS); 3 different concentrations of clay (0.5, 1, or 2% of dietary DMI) plus an AF challenge; and a control consisting of no clay and no AF challenge (C). Statistical analysis was performed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Two contrasts, CONT1 (POS vs. C) and CONT2 (POS vs. the average of 0.5, 1, and 2% clay), were compared along with the linear and quadratic treatment effects (POS, 0.5%, 1%, 2%). Cows supplemented with clay had lower AF excretion in milk as aflatoxin M 1 (AFM 1 ; 0.5% = 20.83 μg/d, 1% = 22.82 μg/d, and 2% = 16.51 μg/d) and AF transfer from rumen fluid to milk (AFM 1; 0.5% = 1.01%, 1% = 0.98%, and 2% = 0.74%) compared with cows in POS (AFM 1 = 27.81 μg/d and AF transfer = 1.37%, CONT2). Similarly, concentrations of AFM 1 in milk (0.5% = 0.35 μg/kg, 1% = 0.30 μg/kg, 2% = 0.25 μg/kg), AFB 1 in feces (0.5% = 1.79 μg/g, 1% = 1.52 μg/kg, 2% = 1.48 μg/kg), and AFB 1 in rumen fluid (0.5% = 0.05 μg/kg, 1% = 0.02 μg/kg, 2% = 0.02 μg/kg) were reduced in cows fed clay compared

  20. Aflatoxicosis chemoprevention by probiotic Lactobacillius and lack of effect on the major histocompatibility complex.

    PubMed

    Rawal, Sumit; Bauer, Miranda M; Mendoza, Kristelle M; El-Nezami, Hani; Hall, Jeffery R; Kim, Ji Eun; Stevens, John R; Reed, Kent M; Coulombe, Roger A

    2014-10-01

    Turkeys are extremely sensitive to aflatoxin B1 (AFB1) which causes decreased growth, immunosuppression and liver necrosis. The purpose of this study was to determine whether probiotic Lactobacillus, shown to be protective in animal and clinical studies, would likewise confer protection in turkeys, which were treated for 11 days with either AFB1 (AFB; 1 ppm in diet), probiotic (PB; 1 × 10(11) CFU/ml; oral, daily), probiotic + AFB1 (PBAFB), or PBS control (CNTL). The AFB1 induced drop in body and liver weights were restored to normal in CNTL and PBAFB groups. Hepatotoxicity markers were not significantly reduced by probiotic treatment. Major histocompatibility complex (MHC) genes BG1 and BG4, which are differentially expressed in liver and spleens, were not significantly affected by treatments. These data indicate modest protection, but the relatively high dietary AFB1 treatment, and the extreme sensitivity of this species may reveal limits of probiotic-based protection strategies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. GST-M1 is transcribed moreso than AKR7A2 in AFB₁-exposed human monocytes and lymphocytes.

    PubMed

    Bahari, Abbas; Mehrzad, Jalil; Mahmoudi, Mahmoud; Bassami, Mohammad Reza; Dehghani, Hesam

    2015-01-01

    Glutathione-S-transferases (GST) and aldo-keto reductases (AKR) are key aflatoxin (AF)-detoxifying enzymes. In this study, the expression of GST-M1, GST-T1, AKR-7A2, and AKR-7A3 genes in human monocytes and lymphocytes was analyzed after in vitro exposure to 10 or 100 ng AFB1/ml for 2 h. Unlike in pilot studies that showed that all four examined genes were present in HepG2 cells, in lymphocytes and monocytes, only GST-M1 and AKR-7A2 were detected. In fact, the induced expression of both GST-M1 and AKR-7A2 genes in human monocytes was moreso than that seen in AFB1-exposed lymphocytes. In addition, analyses of the effects of the exposures on cell cycle status were performed as, in cells lacking adequate detoxification capacities, it would be expected the cells would arrest at checkpoints in the cell cycle or progress to apoptotic/necrotic states. The results here indicated that only the high dose of AFB1 led to a change in cell cycle profiles and only in the monocytes (i.e. cells in S phase were significantly reduced). In general, the results here strongly suggest that human immune cell lineages appear to be able to increase their expression of AFB1-detoxifying enzymes (albeit to differing degrees) and, as a result, are able to counter potential toxicities from AFB1 and (likely) its metabolites.

  2. Installation Restoration Program. Phase 1. Records Search, Hazardous Materials Disposal Sites, Eglin AFB, Florida

    DTIC Science & Technology

    1981-10-01

    Okaloosa Co. 3-12 Crestview South Quadrangle, Florida-Okaloosa Co. -13 Mossy ead Quadrangle, Florida-Walton Co 3-14 E-1 EGUN AFB 4,h aa V, WRAMATIO...I) Um AYPM hIN1DS SP * So". AM T 0 Tou A is a O I. h E-2 ES ENGINEERING -SCIENCE V%777-7NN7 .7-7 EGUN AFB 0.4 .m ... (1) WMg LAN DOSAMU *Y* CO u...r Oumm *p S(S Ann~ Il~~O 0*aAMa 2-3 ES ENGINEER ING - SCIENCE EGUN AFB %Tt MAR ESTHER * : . . - . - ~ - ~ r W R G H T L A N D F I L L 3 SITE 0296

  3. Identification and reduction of urinary aflatoxin metabolites in dogs.

    PubMed

    Bingham, A K; Huebner, H J; Phillips, T D; Bauer, J E

    2004-11-01

    Hydrated sodium calcium aluminosilicate (HSCAS) is a phyllosilicate clay commonly used as an anticaking agent in animal feeds. HSCAS tightly and selectively adsorbs aflatoxin. In 1998, 55 dogs died in Texas after eating dog food containing aflatoxin (150-300 ppb). The corn in the diets was contaminated with aflatoxin. Six dogs were given a low-level, sub-clinical dose of aflatoxin B(1). On average, 71.5% of aflatoxin M(1) cleared within 6 h after dosing, increasing to 90.4% after 12 h. Aflatoxin M(1) was no longer detectable in urine after 48 h. Aflatoxin P(1) was not found in urine compared to large amounts of M(1) and trace amounts of Q(1). In a crossover study, six dogs randomly fed a commercial dog food (no-clay control) or coated with HSCAS (0.5% by weight) were subsequently administered a sub-clinical dose of aflatoxin B(1). Diets were switched and the process repeated. The HSCAS-coated diet significantly reduced urinary aflatoxin M(1) by 48.4%+/-16.6 SD versus the control diet. In conclusion, HSCAS protects dogs fed diets with even minimal aflatoxin contamination. Despite regular and careful ingredient screening for aflatoxin, low concentrations may reach the final product undetected. Therefore, HSCAS may provide the pet food industry further assurance of canine diet safety.

  4. Some high-performance liquid-chromatographic studies of the metabolism of aflatoxins by rat liver microsomal preparations.

    PubMed Central

    Neal, G E; Colley, P J

    1978-01-01

    The metabolism of aflatoxin B1 in vitro was examined in rat liver microsomal preparations. 2. H.p.l.c. (high-performance liquid-chromatographic) systems were used. A silica column was used to separate non-polar metabolites. A system utilizing a reversed-phase column which separates both poar and non-polar metabolites was also developed. 3. The principal metabolites of aflatoxin B1 found were aflatoxin M1, aflatoxin Q1 and a compound which co-chromatographed with a degradation product of aflatoxin B1 2,3-dihydrodiol. 4. The time course of metabolism of aflatoxin B1 by microsomal preparations isolated from control and phenobarbitone-pretreated rats was examined. The rate and extent of metabolism was greater with microsomal preparations from the latter. The formation of aflatoxin Q1 was enhanced 4--5-fold by phenobarbitone pretreatment, whereas the production of aflatoxin M1 was only increased 1--2-fold. The formation of the degradation product of aflatoxin B1 2,3-dihydrodiol was increased 4--5-fold by the pretreatment with phenobarbitone. 5. The microsomal metabolism of aflatoxins M1, P1 and Q1 was examined. Aflatoxin M1 apparently underwent very limited microsomal metabolism to more polar compounds. Aflatoxin P1 was not metabolized. The situation with aflatoxin Q1 was complicated in that it was metabolized in the absence of NADPH to an unidentified metabolite. Aflatoxin B1 appeared as a metabolite of aflatoxin Q1 only when NADPH was present, and the formation of more polar metabolites was also then observed. PMID:728090

  5. Development and validation of a high-performance liquid chromatography method with post-column derivatization for the detection of aflatoxins in cereals and grains.

    PubMed

    Asghar, Muhammad Asif; Iqbal, Javed; Ahmed, Aftab; Khan, Mobeen Ahmed; Shamsuddin, Zuzzer Ali; Jamil, Khalid

    2016-06-01

    A novel, reliable and rapid high-performance liquid chromatography (HPLC) method with post-column derivatization was developed and validated. The HPLC method was used for the simultaneous determination of aflatoxin B1 (AFB1), B2 (AFB2), G1 (AFG1) and G2 (AFG2) in various cereals and grains. Samples were extracted with 80:20 (v/v) methanol:water and purified using C18 (40-63 μm) solid-phase extraction cartridges. AFs were separated using a LiChroCART-RP-18 (5 μm, 250 × 4.0 mm(2)) column. The mobile phase consisted of methanol:acetonitrile:buffer (17.5:17.5:65 v/v) (pH 7.4) delivered at the flow rate of 1.0 mL min(-1) The fluorescence of each AF was detected at λex = 365 nm and λem = 435 nm. All four AFs were properly resolved within the total run time of 20 min. The established method was extensively validated as a final verification of the method development by the evaluation of selectivity (AFB1, AFB2, AFG1 and AFG2), linearity (R(2) ≥ 0.9994), precision (average SD ≤ 2.79), accuracy (relative mean error ≤ -5.51), robustness (p < 0.0080), ruggedness (p < 0.0100) and average recoveries (89.2-97.8%). The limits of quantification of AFB1, AFB2, AFG1 and AFG2 were 0.080, 0.073, 0.062 and 0.066 ng g(-1), respectively. Finally, the developed method was applied for the analysis of AFs in 45 samples comprising rice (n = 20), wheat (n = 15) and maize (n = 10). The results showed that 65% of rice, 20% of wheat and 80% of maize samples were found contaminated with AFs. Thus, according to the achieved results, it is suggested that the newly developed HPLC method could be effectively applied for the routine analysis of the AFs in different cereals and grains. © The Author(s) 2014.

  6. Step of Dichlorvos Inhibition in the Pathway of Aflatoxin Biosynthesis

    PubMed Central

    Yao, Raymond C.; Hsieh, Dennis P. H.

    1974-01-01

    Dichlorvos (dimethyl 2,2-dichlorovinyl phosphate) inhibits the biosynthesis of aflatoxin by Aspergillus parasiticus. Cultures treated with dichlorvos excrete an orange pigment which can be converted into aflatoxin B1 by the untreated mycelia. The orange pigment was partially identified as an acetyl derivative of versiconol-type compound. In the presence of dichlorvos, sterigmatocystin is converted into aflatoxin B1 without being interfered, but averufin is converted into the orange pigment instead of aflatoxin B1. Therefore, dichlorvos appears to block an enzymatic step in the aflatoxin biosynthetic pathway, which lies beyond averufin but before sterigmatocystin, at the formation of the orange pigment. PMID:4844267

  7. Target-induced displacement reaction accompanying cargo release from magnetic mesoporous silica nanocontainers for fluorescence immunoassay.

    PubMed

    Tang, Dianping; Liu, Bingqian; Niessner, Reinhard; Li, Peiwu; Knopp, Dietmar

    2013-11-05

    A new fluorescence immunoassay strategy based on a target-induced displacement reaction with cargo release from protein-gated carbohydrate-functionalized magnetic mesoporous silica nanoparticles (MMSN) was developed for sensitive detection of small molecular mycotoxins (aflatoxin B1, AFB1 used in this case). To construct such an assay system, MMSN was initially functionalized with mannose-terminated silanes, then capped with biotinylated concanavalin A (Con A) entrapped rhodamine B (RB) within the pores through the carbohydrate-protein interaction, and then biotinylated monoclonal anti-AFB1 capture antibody was conjugated to Con A-functionalized MMSN by the streptavidin-biotin chemistry. Gold nanoparticles (AuNP) heavily functionalized with invertase and bovine serum albumin-AFB1 conjugate were utilized as the trace tag. With AFB1 introduction, a competitive immunoreaction for the immobilized anti-AFB1 antibody on the MMSN was started between target analyte and the labeled AFB1 on the AuNP. Accompanied by AuNP, the carried invertase hydrolyzed sucrose in glucose and fructose. The generated glucose competed with the mannose for Con A and displaced the Con A-antibody complex from the MMSN, resulting in the opening of molecular gates owing to the uncapping of MMSN, thereby the entrapped RB could release from the pores. The released RB could be quantitatively determined by a fluorometer. Under optimal conditions, the fluorescence intensity decreased with the increasing AFB1 concentration in the range from 0.01 to 5 ng mL(-1) with a detection limit (LOD) of 8 pg mL(-1) at the 3sblank criterion. Intra- and interbatch assay precisions were lower than 9 and 9.5% (CV), respectively. The method featured unbiased identification of negative (blank) and positive samples. No significant differences at the 0.05 significance level were encountered in the analysis of naturally contaminated peanut samples between the fluorescence immunoassay and a commercialized enzyme

  8. Definition of metabolism-dependent xenobiotic toxicity with co-cultures of human hepatocytes and mouse 3T3 fibroblasts in the novel integrated discrete multiple organ co-culture (IdMOC) experimental system: results with model toxicants aflatoxin B1, cyclophosphamide and tamoxifen.

    PubMed

    Li, Albert P; Uzgare, Aarti; LaForge, Yumiko S

    2012-07-30

    The integrated discrete multiple organ co-culture system (IdMOC) allows the co-culturing of multiple cell types as physically separated cells interconnected by a common overlying medium. We report here the application of IdMOC with two cell types: the metabolically competent primary human hepatocytes, and a metabolically incompetent cell line, mouse 3T3 fibroblasts, in the definition of the role of hepatic metabolism on the cytotoxicity of three model toxicants: cyclophosphamide (CPA), aflatoxin B1 (AFB) and tamoxifen (TMX). The presence of hepatic metabolism in IdMOC with human hepatocytes was demonstrated by the metabolism of the P450 isoform 3A4 substrate, luciferin-IPA. The three model toxicants showed three distinct patterns of cytotoxic profile: TMX was cytotoxic to 3T3 cells in the absence of hepatocytes, with slightly lower cytotoxicity towards both 3T3 cells and hepatocytes in the IdMOC. AFB was selective toxic towards the human hepatocytes and relatively noncytotoxic towards 3T3 cells both in the presence and absence of the hepatocytes. CPA cytotoxicity to the 3T3 cells was found to be significantly enhanced by the presence of the hepatocytes, with the cytotoxicity dependent of the number of hepatocytes, and with the cytotoxicity attenuated by the presence of a non-specific P450 inhibitor, 1-aminobenzotriazole. We propose here the following classification of toxicants based on the role of hepatic metabolism as defined by the human hepatocyte-3T3 cell IdMOC assay: type I: direct-acting cytotoxicants represented by TMX as indicated by cytotoxicity in 3T3 cells in the absence of hepatocytes; type II: metabolism-dependent cytotoxicity represented by AFB1 with effects localized within the site of metabolic activation (i. e. hepatocytes); and type III: metabolism-dependent cytotoxicity with metabolites that can diffuse out of the hepatocytes to cause toxicity in cells distal from the site of metabolism, as exemplified by CPA. Copyright © 2012 Elsevier Ireland

  9. High pressure liquid chromatographic determination of aflatoxins in spices.

    PubMed

    Awe, M J; Schranz, J L

    1981-11-01

    High pressure liquid chromatography with fluorescence detection is used to determine aflatoxin in 5 common spices. A 10 micrometer microparticulate silica gel column is used with a dichloromethane-cyclohexane-acetonitrile solvent system to resolve aflatoxins B1, G1, B2, and G2. The fluorescence detector contained a silica gel-packed flowcell. Samples of black, white, and red pepper, ginger, and nutmeg were extracted according to a previously published method. Recoveries from aflatoxin-free samples of white pepper, ginger, and red pepper spiked with 1-50 micrograms aflatoxin/kg ranged from 64 to 92%.

  10. Cobalt-Porphyrin-Platinum-Functionalized Reduced Graphene Oxide Hybrid Nanostructures: A Novel Peroxidase Mimetic System For Improved Electrochemical Immunoassay

    NASA Astrophysics Data System (ADS)

    Shu, Jian; Qiu, Zhenli; Wei, Qiaohua; Zhuang, Junyang; Tang, Dianping

    2015-10-01

    5,10,15,20-Tetraphenyl-21H,23H-porphine cobalt flat stacking on the reduced graphene oxide with platinum nanoparticles (PtNPs/CoTPP/rGO) were first synthesized and functionalized with monoclonal rabbit anti-aflatoxin B1 antibody (anti-AFB1) for highly efficient electrochemical immunoassay of aflatoxin B1 (AFB1) in this work. Transmission electron microscopy (TEM), atomic force microscope (AFM) and spectral techniques were employed to characterize the PtNPs/CoTPP/rGO hybrids. Using anti-AFB1-conjugated PtNPs/CoTPP/rGO as the signal-transduction tag, a novel non-enzymatic electrochemical immunosensing system was designed for detection of target AFB1 on the AFB1-bovine serum albumin-functionalized sensing interface. Experimental results revealed that the designed immunoassay could exhibit good electrochemical responses for target analyte and allowed the detection of AFB1 at a concentration as low as 5.0 pg mL-1 (5.0 ppt). Intra- and inter-assay coefficients of variation were below 10%. Importantly, the methodology was further validated for analyzing naturally contaminated or spiked blank peanut samples with consistent results obtained by AFB1 ELISA kit, thus providing a promising approach for quantitative monitoring of organic pollutants.

  11. Effect of temperature on aflatoxin production in Mucuna pruriens seeds.

    PubMed Central

    Roy, A K; Chourasia, H K

    1989-01-01

    This paper describes the effect of temperature on the level of aflatoxin production in Mucuna pruriens seeds. The highest level of aflatoxin B1 (1.75 micrograms/g) was detected in the samples incubated at 25 degrees C for three weeks. At 20, 30, and 35 degrees C, aflatoxin levels were 0.30 to 0.56, 0.37 to 1.20, and 0.26 to 0.65 micrograms/g, respectively. The lowest concentration of aflatoxin B1 (0.10 to 0.29 microgram/g) was produced at 15 degrees C. PMID:2719482

  12. Occurrence of aflatoxin M1 in human milk samples in Vojvodina, Serbia: Estimation of average daily intake by babies.

    PubMed

    Radonić, Jelena R; Kocić Tanackov, Sunčica D; Mihajlović, Ivana J; Grujić, Zorica S; Vojinović Miloradov, Mirjana B; Škrinjar, Marija M; Turk Sekulić, Maja M

    2017-01-02

    The objectives of the study were to determine the aflatoxin M1 content in human milk samples in Vojvodina, Serbia, and to assess the risk of infants' exposure to aflatoxins food contamination. The growth of Aspergillus flavus and production of aflatoxin B1 in corn samples resulted in higher concentrations of AFM1 in milk and dairy products in 2013, indicating higher concentrations of AFM1 in human milk samples in 2013 and 2014 in Serbia. A total number of 60 samples of human milk (colostrum and breast milk collected 4-8 months after delivery) were analyzed for the presence of AFM1 using the Enzyme Linked Immunosorbent Assay method. The estimated daily intake of AFM1 through breastfeeding was calculated for the colostrum samples using an average intake of 60 mL/kg body weight (b.w.)/day on the third day of lactation. All breast milk collected 4-8 months after delivery and 36.4% of colostrum samples were contaminated with AFM1. The greatest percentage of contaminated colostrum (85%) and all samples of breast milk collected 4-8 months after delivery had AFM1 concentration above maximum allowable concentration according to the Regulation on health safety of dietetic products. The mean daily intake of AFM1 in colostrum was 2.65 ng/kg bw/day. Results of our study indicate the high risk of infants' exposure, who are at the early stage of development and vulnerable to toxic contaminants.

  13. Occurrence of Aflatoxins and Aflatoxin-Producing Strains of Aspergillus spp. in Soybeans 1

    PubMed Central

    Bean, George A.; Schillinger, John A.; Klarman, William L.

    1972-01-01

    Above average rainfall in Maryland during August, September, and October 1971 resulted in heavy mold growth in soybeans while still in the field. Of 28 samples of soybean seed, aflatoxins were found in 14, 2 of which had been used in poultry feed. Aflatoxins were identified by thin-layer chromatography, spectrophotometry, and chicken embryo bioassay. Aspergillus spp. were isolated from 11 samples, and 5 of these isolates produced aflatoxins when grown in liquid culture. PMID:4673021

  14. Aflatoxin M1 in Tarhana chips.

    PubMed

    Özçam, Mustafa; Obuz, Ersel; Tosun, Halil

    2014-01-01

    Tarhana chips are a popular traditional fermented food consumed widely in the Kahramanmaraş region of Turkey. Tarhana chips are different from many other types of fermented food in that they are produced in the form of tortilla chips. Cereal and yoghurt are the main ingredients in Tarhana chips. Aflatoxin M1 (AFM1) levels in dairy and dairy-based products are of concern for human health. To investigate AFM1 contamination, a total of 40 samples were collected from Kahramanmaraş region and AFM1 levels were determined by competitive enzyme-linked immunosorbent assay (ELISA). Furthermore, physicochemical characteristics of Tarhana chips were investigated and compared with classic fried chips in terms of nutritional value. Based on data obtained from enzyme-linked immunosorbent assay, 21 (52.5%) out of 40 samples contained AFM1 in the range 0.5-36.6 ng/kg, so AFM1 levels of all samples were below the legal limit.

  15. Identification of aflatoxin biosynthesis genes by genetic complementation in an Aspergillus flavus mutant lacking the aflatoxin gene cluster.

    PubMed Central

    Prieto, R; Yousibova, G L; Woloshuk, C P

    1996-01-01

    Aspergillus flavus mutant strain 649, which has a genomic DNA deletion of at least 120 kb covering the aflatoxin biosynthesis cluster, was transformed with a series of overlapping cosmids that contained DNA harboring the cluster of genes. The mutant phenotype of strain 649 was rescued by transformation with a combination of cosmid clones 5E6, 8B9, and 13B9, indicating that the cluster of genes involved in aflatoxin biosynthesis resides in the 90 kb of A. flavus genomic DNA carried by these clones. Transformants 5E6 and 20B11 and transformants 5E6 and 8B9 accumulated intermediate metabolites of the aflatoxin pathway, which were identified as averufanin and/or averufin, respectively.These data suggest that avf1, which is involved in the conversion of averufin to versiconal hemiacetal acetate, was present in the cosmid 13B9. Deletion analysis of 13B9 located the gene on a 7-kb DNA fragment of the cosmid. Transformants containing cosmid 8B9 converted exogenously supplied O-methylsterigmatocystin to aflatoxin, indicating that the oxidoreductase gene (ord1), which mediates the conversion of O-methylsterigmatocystin to aflatoxin, is carried by this cosmid. The analysis of transformants containing deletions of 8B9 led to the localization of ord1 on a 3.3-kb A. flavus genomic DNA fragment of the cosmid. PMID:8967772

  16. Multiclonal plastic antibodies for selective aflatoxin extraction from food samples.

    PubMed

    Bayram, Engin; Yılmaz, Erkut; Uzun, Lokman; Say, Rıdvan; Denizli, Adil

    2017-04-15

    Herein, we focused on developing a new generation of monolithic columns for extracting aflatoxin from real food samples by combining the superior features of molecularly imprinted polymers and cryogels. To accomplish this, we designed multiclonal plastic antibodies through simultaneous imprinting of aflatoxin subtypes B1, B2, G1, and G2. We applied Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and spectrofluorimetry to characterize the materials, and conducted selectivity studies using ochratoxin A and aflatoxin M1 (a metabolite of aflatoxin B1), as well as other aflatoxins, under competitive conditions. We determined optimal aflatoxin extraction conditions in terms of concentration, flow rate, temperature, and embedded particle amount as up to 25ng/mL for each species, 0.43mL/min, 7.0, 30°C, and 200mg, respectively. These multiclonal plastic antibodies showed imprinting efficiencies against ochratoxin A and aflatoxin M1 of 1.84 and 26.39, respectively, even under competitive conditions. Finally, we tested reusability, repeatability, reproducibility, and robustness of columns throughout inter- and intra-column variation studies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Aflatoxin M1 in human breast milk in southeastern Turkey.

    PubMed

    Kılıç Altun, Serap; Gürbüz, Semra; Ayağ, Emin

    2017-05-01

    This study was performed to determine aflatoxin M 1 (AFM 1 ) in human breast milk samples collected in Şanlıurfa, located in Southeastern region of Turkey, and to investigate a possible correlation between AFM 1 occurrence (frequency and levels) and sampling seasons. Human breast milk samples collected in December 2014 and in June 2015 from a total of 74 nursing women, both outpatient and inpatient volunteers in hospitals located in Şanlıurfa, Turkey, were analyzed using competitive enzyme-linked immunosorbent assay (ELISA) for the presence of AFM 1 . AFM 1 was detected in 66 (89.2%) out of 74 samples at an average concentration of 19.0 ± 13.0 ng/l (min.-max., 9.6-80 ng/l). There was a statistically significant difference between December and June concerning AFM 1 levels (p < 0.05). Further detailed studies will be needed to determine the main sources of aflatoxins in food, to establish protection strategies against maternal and infant exposure to these mycotoxins.

  18. Occurrence of aflatoxin M₁ in commercial pasteurized milk samples in Sari, Mazandaran province, Iran.

    PubMed

    Mohammadi, Hamidreza; Shokrzadeh, Mohammad; Aliabadi, Zahra; Riahi-Zanjani, Bamdad

    2016-05-01

    The frequency and levels of aflatoxin M1 (AFM1) in pasteurized milk samples in Sari, located in Mazandaran province, Iran, were determined by enzyme immunoassay. Seventy-six samples of pasteurized milk from different retail stores were randomly collected over four seasons during the year 2015. AFM1 contamination was detected in all milk samples. The mean concentration of aflatoxin M1 was 65.8 ng/l, with a range of 11.7-106.6 ng/l. The highest AFM1 level was detected in milk samples collected during spring. Forty-six (60.53 %) samples had AFM1 levels that exceeded the maximum acceptable levels (50 ng/l) recommended by the European Union (EU). Comparison of these results with previously published data for AFM1 in milk in Iran shows that the percentage of samples exceeding the EU maximum level is consistently high over the years, indicating a general problem related to AFB1 contamination in dairy feedingstuff.

  19. Coxiella burnetii Avirulent Nine Mile Phase II Induces Caspase-1-Dependent Pyroptosis in Murine Peritoneal B1a B Cells.

    PubMed

    Schoenlaub, Laura; Cherla, Rama; Zhang, Yan; Zhang, Guoquan

    2016-12-01

    Our recent study demonstrated that virulent Coxiella burnetii Nine Mile phase I (NMI) is capable of infecting and replicating within peritoneal B1a cells and that B1a cells play an important role in host defense against C. burnetii infection in mice. However, it remains unknown if avirulent Nine Mile phase II (NMII) can infect and replicate in B1a cells and whether NMI and NMII can differentially interact with B1a cells. In this study, we examined if NMI and NMII can differentially modulate host cell apoptotic signaling in B1a cells. The results showed that NMII induced dose-dependent cell death in murine peritoneal B1a cells but NMI did not, suggesting that NMI and NMII may differentially activate host cell apoptotic signaling in B1a cells. Western blotting indicated that NMII-induced B1a cell death was not dependent on either caspase-3 or PARP-1 cleavage, but cleavage of caspase-1 was detected in NMII-infected B1a cells. In addition, inhibition or deficiency of caspase-1 activity blocked NMII-induced B1a cell death. These results suggest that NMII induces a caspase-1-dependent pyroptosis in murine peritoneal B1a cells. We also found that heat-killed NMII and type 4 secretion system (T4SS) mutant NMII were unable to induce B1a cell death and that NMII infection did not induce cell death in peritoneal B1a cells from Toll-like receptor 2 (TLR-2)- or NLRP3 inflammasome-deficient mice. These data suggest that NMII-induced caspase-1-dependent pyroptosis may require its T4SS and activation of the TLR-2 and NLRP3 signaling pathways. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  20. Mycotoxins in commercial dry pet food in China.

    PubMed

    Shao, Manyu; Li, Li; Gu, Zuli; Yao, Ming; Xu, Danning; Fan, Wentao; Yan, Liping; Song, Suquan

    2018-05-10

    The aim of this study was to investigate the occurrence of the most common mycotoxins in commercial dry dog food using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Aflatoxin B1 (AFB1), aflatoxin G1 (AFG1), zearalenone (ZEN), deoxynivalenol (DON), T-2, beauvericin (BEA), citrinin (CIT), ochratoxin A (OTA), fumonisin B1 (FB1) were included in this study. The results showed that all these analytes could be found in the samples. Furthermore, only one sample was found free of mycotoxins contamination. All other samples (96.9%) were contaminated by at least three different types of mycotoxins. Among these mycotoxins, DON, ZEN, AFB1, FB1, CIT and BEA exhibited relatively high incidence, with occurrence rates of 78.1%, 62.5%, 87.5%, 93.8%, 68.8 and 96.9%. Furthermore, it is worth noting that AFB1 concentration in all AFB1 positive samples exceeded the maximum limits set by the EU, with concentrations ranging from 30.3 µg/kg to 242.7 µg/kg.

  1. Benzo[a]pyrene, Aflatoxine B1 and Acetaldehyde Mutational Patterns in TP53 Gene Using a Functional Assay: Relevance to Human Cancer Aetiology

    PubMed Central

    Paget, Vincent; Lechevrel, Mathilde; André, Véronique; Le Goff, Jérémie; Pottier, Didier; Billet, Sylvain; Garçon, Guillaume; Shirali, Pirouz; Sichel, François

    2012-01-01

    Mutations in the TP53 gene are the most common alterations in human tumours. TP53 mutational patterns have sometimes been linked to carcinogen exposure. In hepatocellular carcinoma, a specific G>T transversion on codon 249 is classically described as a fingerprint of aflatoxin B1 exposure. Likewise G>T transversions in codons 157 and 158 have been related to tobacco exposure in human lung cancers. However, controversies remain about the interpretation of TP53 mutational pattern in tumours as the fingerprint of genotoxin exposure. By using a functional assay, the Functional Analysis of Separated Alleles in Yeast (FASAY), the present study depicts the mutational pattern of TP53 in normal human fibroblasts after in vitro exposure to well-known carcinogens: benzo[a]pyrene, aflatoxin B1 and acetaldehyde. These in vitro patterns of mutations were then compared to those found in human tumours by using the IARC database of TP53 mutations. The results show that the TP53 mutational patterns found in human tumours can be only partly ascribed to genotoxin exposure. A complex interplay between the functional impact of the mutations on p53 phenotype and the cancer natural history may affect these patterns. However, our results strongly support that genotoxins exposure plays a major role in the aetiology of the considered cancers. PMID:22319594

  2. Electrophoretically deposited reduced graphene oxide platform for food toxin detection

    NASA Astrophysics Data System (ADS)

    Srivastava, Saurabh; Kumar, Vinod; Ali, Md Azahar; Solanki, Pratima R.; Srivastava, Anchal; Sumana, Gajjala; Saxena, Preeti Suman; Joshi, Amish G.; Malhotra, B. D.

    2013-03-01

    Reduced graphene oxide (RGO) due to its excellent electrochemical properties and large surface area, has recently aroused much interest for electrochemical biosensing application. Here, the chemically active RGO has been synthesized and deposited onto an indium tin oxide (ITO) coated glass substrate by the electrophoretic deposition technique. This novel platform has been utilized for covalent attachment of the monoclonal antibodies of aflatoxin B1 (anti-AFB1) for food toxin (AFB1) detection. The electron microscopy, X-ray diffraction, and UV-visible studies reveal successful synthesis of reduced graphene oxide while the XPS and FTIR studies suggest its carboxylic functionalized nature. The electrochemical sensing results of the anti-AFB1/RGO/ITO based immunoelectrode obtained as a function of aflatoxin concentration show high sensitivity (68 μA ng-1 mL cm-2) and improved detection limit (0.12 ng mL-1). The association constant (ka) for antigen-antibody interaction obtained as 5 × 10-4 ng mL-1 indicates high affinity of antibodies toward the antigen (AFB1).Reduced graphene oxide (RGO) due to its excellent electrochemical properties and large surface area, has recently aroused much interest for electrochemical biosensing application. Here, the chemically active RGO has been synthesized and deposited onto an indium tin oxide (ITO) coated glass substrate by the electrophoretic deposition technique. This novel platform has been utilized for covalent attachment of the monoclonal antibodies of aflatoxin B1 (anti-AFB1) for food toxin (AFB1) detection. The electron microscopy, X-ray diffraction, and UV-visible studies reveal successful synthesis of reduced graphene oxide while the XPS and FTIR studies suggest its carboxylic functionalized nature. The electrochemical sensing results of the anti-AFB1/RGO/ITO based immunoelectrode obtained as a function of aflatoxin concentration show high sensitivity (68 μA ng-1 mL cm-2) and improved detection limit (0.12 ng mL-1). The

  3. Physicochemical and immunological characterization of chitosan-coated bacteriophage nanoparticles for in vivo mycotoxin modeling.

    PubMed

    de Andrade, Carla Yoko Tanikawa; Yamanaka, Isabel; Schlichta, Laís S; Silva, Sabrina Karim; Picheth, Guilherme F; Caron, Luiz Felipe; de Moura, Juliana; de Freitas, Rilton Alves; Alvarenga, Larissa Magalhães

    2018-04-01

    To propose a novel modeling of aflatoxin immunization and surrogate toxin conjugate from AFB1 vaccines, an immunogen based on the mimotope, (i.e. a peptide-displayed phage that mimics aflatoxins epitope without toxin hazards) was designed. The recombinant phage 3P30 was identified by phage display technology and exhibited the ability to bind, dose dependent, specifically to its cognate target - anti-AFB1 antibody. In immunization assay, the phage-displayed mimotope and its peptide chemically synthesized were able to induce specific anti-AFB1 antibodies, indicating the proof of concept for aflatoxin mimicry. Furthermore, the phage 3P30 was homogeneously coated with chitosan, which also provided a tridimensional matrix network for mucosal delivery. After intranasal immunization, chitosan coated phages improved specific immunogenicity compared to the free antigen. It can be concluded that affinity-selected phage may contribute to the rational design of epitope-based vaccines in a prospectus for the control of aflatoxins and possibly other mycotoxins, and that chitosan coating improved the vectorization of the vaccine by the mucosal route. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Deciphering the Anti-Aflatoxinogenic Properties of Eugenol Using a Large-Scale q-PCR Approach

    PubMed Central

    Caceres, Isaura; El Khoury, Rhoda; Medina, Ángel; Lippi, Yannick; Naylies, Claire; Atoui, Ali; El Khoury, André; Oswald, Isabelle P.; Bailly, Jean-Denis; Puel, Olivier

    2016-01-01

    Produced by several species of Aspergillus, Aflatoxin B1 (AFB1) is a carcinogenic mycotoxin contaminating many crops worldwide. The utilization of fungicides is currently one of the most common methods; nevertheless, their use is not environmentally or economically sound. Thus, the use of natural compounds able to block aflatoxinogenesis could represent an alternative strategy to limit food and feed contamination. For instance, eugenol, a 4-allyl-2-methoxyphenol present in many essential oils, has been identified as an anti-aflatoxin molecule. However, its precise mechanism of action has yet to be clarified. The production of AFB1 is associated with the expression of a 70 kB cluster, and not less than 21 enzymatic reactions are necessary for its production. Based on former empirical data, a molecular tool composed of 60 genes targeting 27 genes of aflatoxin B1 cluster and 33 genes encoding the main regulatory factors potentially involved in its production, was developed. We showed that AFB1 inhibition in Aspergillus flavus following eugenol addition at 0.5 mM in a Malt Extract Agar (MEA) medium resulted in a complete inhibition of the expression of all but one gene of the AFB1 biosynthesis cluster. This transcriptomic effect followed a down-regulation of the complex composed by the two internal regulatory factors, AflR and AflS. This phenomenon was also influenced by an over-expression of veA and mtfA, two genes that are directly linked to AFB1 cluster regulation. PMID:27128940

  5. Degradation kinetics of aflatoxin B1 and B2 in filter paper and rough rice by using pulsed light irradiation

    USDA-ARS?s Scientific Manuscript database

    Rough rice is susceptible to contamination by aflatoxins, which are highly toxic, mutagenic and carcinogenic compounds. To develop aflatoxin degradation technology for rice with the use of pulsed light (PL) treatment, the objective of this study was to investigate the degradation characters of aflat...

  6. Stable expression of rat cytochrome P-450IIB1 cDNA in Chinese hamster cells (V79) and metabolic activation of aflatoxin B1.

    PubMed Central

    Doehmer, J; Dogra, S; Friedberg, T; Monier, S; Adesnik, M; Glatt, H; Oesch, F

    1988-01-01

    V79 Chinese hamster fibroblasts are widely used for mutagenicity testing but have the serious limitation that they do not express cytochromes P-450, which are needed for the activation of many promutagens to mutagenic metabolites. A full-length cDNA clone encoding the monooxygenase cytochrome P-450IIB1 under control of the simian virus 40 early promoter was constructed and cointroduced with the selection marker neomycin phosphotransferase (conferring resistance to G418) into V79 Chinese hamster cells. G418-resistant cells were selected, established as cell lines, and tested for cytochrome P-450IIB1 expression and enzymatic activity. Two cell lines (SD1 and SD3) were found that stably produce cytochrome P-450IIB1. Although purified cytochromes P-450 possess monooxygenase activity only after reconstitution with cytochrome P-450 reductase and phospholipid, the gene product of the construct exhibited this activity. This implies that the gene product is intracellularly localized in a way that allows access to the required components. If compared with V79 cells, the mutation rate for the hypoxanthine phosphoribosyltransferase (HPRT) locus in SD1 cells is markedly increased when exposed to aflatoxin B1, which is activated by this enzyme. Images PMID:3137560

  7. Efficacy of chemically characterized Foeniculum vulgare Mill seed essential oil in protection of raw tobacco leaves during storage against fungal and aflatoxin contamination.

    PubMed

    Kedia, A; Dwivedy, A K; Pandey, A K; Kumar, R R; Regmi, P; Dubey, N K

    2015-10-01

    To report fungal and aflatoxin contamination in stored tobacco leaves and the potential of Foeniculum vulgare (fennel) seed essential oil (EO) as a plant-based preservative in protection of tobacco during storage. Mycological analysis of tobacco samples was done by surface sterilization and serial dilution tests. The Aspergillus flavus isolates were screened for their toxigenicity. Both in vivo and in vitro tests were done to evaluate antifungal and antiaflatoxigenic efficacy of chemically characterized EO. The mycoflora analysis revealed 108 fungal colonies belonging to five genera and nine species. All A. flavus isolates were found aflatoxigenic during screening. Gas chromatography and mass spectrometry analysis of EO identified 19 components (99·66%); estragole being the major component (47·49%). The EO showed broad fungitoxicity at 1·25 μl ml(-1) and 100% inhibition to AFB1 production as well as ergosterol synthesis at 1·0 μl ml(-1) concentration. EO showed 100% protection of stored tobacco samples from aflatoxin B1 contamination. The fennel EO can thus be formulated as a plant-based preservative for food items. The present investigation comprises the first report on antiaflatoxin efficacy of fennel oil and its potency in the protection of tobacco leaves from fungal and aflatoxin contamination during storage. © 2015 The Society for Applied Microbiology.

  8. Mouse models to study the interaction of risk factors for human liver cancer.

    PubMed

    Sell, Stewart

    2003-11-15

    Each of the risk factors for human liver cancer (aflatoxin exposure, hepatitis B virus-associated liver injury, p53 loss, p53ser249 mutation, and male sex) also increases the incidence of hepatocellular carcinoma (HCC) in mouse models of hepatocarcinogenesis. Neonatal mice, partially hepatectomized adult mice, and p53-deficient mice each have a higher hepatocyte proliferation rate, are less able to detoxify AFB1, and form more DNA adducts than do normal wild-type controls. However, transgenic hepatitis B surface antigen mice, expressing hepatitis B surface antigen under control of the albumin promoter (alb/psx), are able to detoxify AFB1 at the same level as do wild-type mice. Thus, AFB1-induced HCC development in neonatal mice and p53+/- mice may be due to "immature" carcinogen metabolism, whereas increased HCC in transgenic hepatitis B virus mice may be due to promotion effects of increased proliferation. Future studies will explore the effects of modifying factors on the development of HCC.

  9. Evaluation of Saccharomyces cerevisiae as an antiaflatoxicogenic agent in broiler feedstuffs.

    PubMed

    Pizzolitto, R P; Armando, M R; Salvano, M A; Dalcero, A M; Rosa, C A

    2013-06-01

    Aflatoxins (AF) are the most important mycotoxins produced by toxigenic strains of various Aspergillus spp. Biological decontamination of mycotoxins using microorganisms is a well-known strategy for the management of mycotoxins in feeds. Saccharomyces cerevisiae strains have been reported to bind aflatoxin B1 (AFB1). The aim of this study was to evaluate the ability of S. cerevisiae CECT 1891 in counteracting the deleterious effects of AFB1 in broiler chicks. Experimental aflatoxicosis was induced in 6-d-old broilers by feeding them 1.2 mg of AFB1/kg of feed for 3 wk, and the yeast strain was administrated in feed (10(10) cells/kg), in the drinking water (5 × 10(9) cells/L), or a combination of both treatments. A total of 160 chicks were randomly divided into 8 treatments (4 repetitions per treatment). Growth performance was measured weekly from d 7 to 28, and serum biochemical parameters, weights, and histopathological examination of livers were determined at d 28. The AFB1 significantly decreased the BW gain, feed intake, and impaired feed conversion rate. Moreover, AFB1 treatment decreased serum protein concentration and increased liver damage. The addition of S. cerevisiae strain to drinking water, to diets contaminated with AFB1, showed a positive protection effect on the relative weight of the liver, histopathology, and biochemical parameters. Furthermore, dietary addition of the yeast strain to drinking water alleviated the negative effects of AFB1 on growth performance parameters. In conclusion, this study suggests that in feed contaminated with AFB1, the use of S. cerevisiae is an alternative method to reduce the adverse effects of aflatoxicosis. Thus, apart from its excellent nutritional value, yeast can also be used as a mycotoxin adsorbent.

  10. Identification of the Anti-Aflatoxinogenic Activity of Micromeria graeca and Elucidation of Its Molecular Mechanism in Aspergillus flavus

    PubMed Central

    El Khoury, Rhoda; Caceres, Isaura; Puel, Olivier; Bailly, Sylviane; Atoui, Ali; Oswald, Isabelle P.; El Khoury, André; Bailly, Jean-Denis

    2017-01-01

    Of all the food-contaminating mycotoxins, aflatoxins, and most notably aflatoxin B1 (AFB1), are found to be the most toxic and economically costly. Green farming is striving to replace fungicides and develop natural preventive strategies to minimize crop contamination by these toxic fungal metabolites. In this study, we demonstrated that an aqueous extract of the medicinal plant Micromeria graeca—known as hyssop—completely inhibits aflatoxin production by Aspergillus flavus without reducing fungal growth. The molecular inhibitory mechanism was explored by analyzing the expression of 61 genes, including 27 aflatoxin biosynthesis cluster genes and 34 secondary metabolism regulatory genes. This analysis revealed a three-fold down-regulation of aflR and aflS encoding the two internal cluster co-activators, resulting in a drastic repression of all aflatoxin biosynthesis genes. Hyssop also targeted fifteen regulatory genes, including veA and mtfA, two major global-regulating transcription factors. The effect of this extract is also linked to a transcriptomic variation of several genes required for the response to oxidative stress such as msnA, srrA, catA, cat2, sod1, mnsod, and stuA. In conclusion, hyssop inhibits AFB1 synthesis at the transcriptomic level. This aqueous extract is a promising natural-based solution to control AFB1 contamination. PMID:28257049

  11. Aflatoxin contamination of red chili pepper from Bolivia and Peru, countries with high gallbladder cancer incidence rates.

    PubMed

    Asai, Takao; Tsuchiya, Yasuo; Okano, Kiyoshi; Piscoya, Alejandro; Nishi, Carlos Yoshito; Ikoma, Toshikazu; Oyama, Tomizo; Ikegami, Kikuo; Yamamoto, Masaharu

    2012-01-01

    Chilean red chili peppers contaminated with aflatoxins were reported in a previous study. If the development of gallbladder cancer (GBC) in Chile is associated with a high level of consumption of aflatoxin-contaminated red chili peppers, such peppers from other countries having a high GBC incidence rate may also be contaminated with aflatoxins. We aimed to determine whether this might be the case for red chili peppers from Bolivia and Peru. A total of 7 samples (3 from Bolivia, 4 from Peru) and 3 controls (2 from China, 1 from Japan) were evaluated. Aflatoxins were extracted with acetonitrile:water (9:1, v/v) and eluted through an immuno-affinity column. The concentrations of aflatoxins B1, B2, G1, and G2 were measured using high-performance liquid chromatography (HPLC), and then the detected aflatoxins were identified using HPLC-mass spectrometry. In some but not all of the samples from Bolivia and Peru, aflatoxin B1 or aflatoxins B1 and B2 were detected. In particular, aflatoxin B1 or total aflatoxin concentrations in a Bolivian samples were above the maximum levels for aflatoxins in spices proposed by the European Commission. Red chili peppers from Bolivia and Peru consumed by populations having high GBC incidence rates would appear to be contaminated with aflatoxins. These data suggest the possibility that a high level of consumption of aflatoxin-contaminated red chili peppers is related to the development of GBC, and the association between the two should be confirmed by a case-control study.

  12. Aflatoxin Control in Maize by Trametes versicolor

    PubMed Central

    Scarpari, Marzia; Bello, Cristiano; Pietricola, Chiara; Zaccaria, Marco; Bertocchi, Luigi; Angelucci, Alessandra; Ricciardi, Maria Rosaria; Scala, Valeria; Parroni, Alessia; Fabbri, Anna A.; Reverberi, Massimo; Zjalic, Slaven; Fanelli, Corrado

    2014-01-01

    Aspergillus flavus is a well-known ubiquitous fungus able to contaminate both in pre- and postharvest period different feed and food commodities. During their growth, these fungi can synthesise aflatoxins, secondary metabolites highly hazardous for animal and human health. The requirement of products with low impact on the environment and on human health, able to control aflatoxin production, has increased. In this work the effect of the basidiomycete Trametes versicolor on the aflatoxin production by A. flavus both in vitro and in maize, was investigated. The goal was to propose an environmental loyal tool for a significant control of aflatoxin production, in order to obtain feedstuffs and feed with a high standard of quality and safety to enhance the wellbeing of dairy cows. The presence of T. versicolor, grown on sugar beet pulp, inhibited the production of aflatoxin B1 in maize by A. flavus. Furthermore, treatment of contaminated maize with culture filtrates of T. versicolor containing ligninolytic enzymes, showed a significant reduction of the content of aflatoxin B1. PMID:25525683

  13. Aflatoxin control in maize by Trametes versicolor.

    PubMed

    Scarpari, Marzia; Bello, Cristiano; Pietricola, Chiara; Zaccaria, Marco; Bertocchi, Luigi; Angelucci, Alessandra; Ricciardi, Maria Rosaria; Scala, Valeria; Parroni, Alessia; Fabbri, Anna A; Reverberi, Massimo; Zjalic, Slaven; Fanelli, Corrado

    2014-12-17

    Aspergillus flavus is a well-known ubiquitous fungus able to contaminate both in pre- and postharvest period different feed and food commodities. During their growth, these fungi can synthesise aflatoxins, secondary metabolites highly hazardous for animal and human health. The requirement of products with low impact on the environment and on human health, able to control aflatoxin production, has increased. In this work the effect of the basidiomycete Trametes versicolor on the aflatoxin production by A. flavus both in vitro and in maize, was investigated. The goal was to propose an environmental loyal tool for a significant control of aflatoxin production, in order to obtain feedstuffs and feed with a high standard of quality and safety to enhance the wellbeing of dairy cows. The presence of T. versicolor, grown on sugar beet pulp, inhibited the production of aflatoxin B1 in maize by A. flavus. Furthermore, treatment of contaminated maize with culture filtrates of T. versicolor containing ligninolytic enzymes, showed a significant reduction of the content of aflatoxin B1.

  14. The Aspergillus flavus Homeobox Gene, hbx1, is Required for Development and Aflatoxin Production.

    PubMed

    Cary, Jeffrey W; Harris-Coward, Pamela; Scharfenstein, Leslie; Mack, Brian M; Chang, Perng-Kuang; Wei, Qijian; Lebar, Matthew; Carter-Wientjes, Carol; Majumdar, Rajtilak; Mitra, Chandrani; Banerjee, Sourav; Chanda, Anindya

    2017-10-12

    Homeobox proteins, a class of well conserved transcription factors, regulate the expression of targeted genes, especially those involved in development. In filamentous fungi, homeobox genes are required for normal conidiogenesis and fruiting body formation. In the present study, we identified eight homeobox ( hbx ) genes in the aflatoxin-producing ascomycete, Aspergillus flavus , and determined their respective role in growth, conidiation and sclerotial production. Disruption of seven of the eight genes had little to no effect on fungal growth and development. However, disruption of the homeobox gene AFLA_069100, designated as hbx1 , in two morphologically different A. flavus strains, CA14 and AF70, resulted in complete loss of production of conidia and sclerotia as well as aflatoxins B₁ and B₂, cyclopiazonic acid and aflatrem. Microscopic examination showed that the Δ hbx1 mutants did not produce conidiophores. The inability of Δ hbx1 mutants to produce conidia was related to downregulation of brlA (bristle) and abaA (abacus), regulatory genes for conidiophore development. These mutants also had significant downregulation of the aflatoxin pathway biosynthetic genes aflC , aflD , aflM and the cluster-specific regulatory gene, aflR . Our results demonstrate that hbx1 not only plays a significant role in controlling A. flavus development but is also critical for the production of secondary metabolites, such as aflatoxins.

  15. Effect of ultraviolet radiation A and B on growth and mycotoxin production by Aspergillus carbonarius and Aspergillus parasiticus in grape and pistachio media.

    PubMed

    García-Cela, Esther; Marin, Sonia; Sanchis, Vicente; Crespo-Sempere, Ana; Ramos, Antonio J

    2015-01-01

    The effects of two exposure times per day (6 and 16 h) of UV-A or UV-B radiation, combined with dark and dark plus light incubation periods during 7-21 d on fungal growth and mycotoxins production of Aspergillus species were studied. Aspergillus carbonarius and Aspergillus parasiticus were inoculated on grape and pistachio media under diurnal and nocturnal temperatures choosing light photoperiod according to harvest conditions of these crops in Spain. Ultraviolet irradiation had a significant effect on A. carbonarius and A. parasiticus colony size (diameter, biomass dry weight, and colony density) and mycotoxin accumulation, although intraspecies differences were observed. Inhibition of A. carbonarius fungal growth decreased when exposure time was reduced from 16 h to 6 h, but this was not always true for ochratoxin A (OTA) production. OTA reduction was higher under UV-A than UV-B radiation and the reduction increased along time conversely to the aflatoxins (AFs). Aflatoxin B1 (AFB1) was the main toxin produced by A. parasiticus except in the UV-B light irradiated colonies which showed a higher percentage of AFG than AFB. Morphological changes were observed in colonies grown under UV-B light. Copyright © 2014 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  16. Association between Urinary Aflatoxin (AFM1) and Dietary Intake among Adults in Hulu Langat District, Selangor, Malaysia

    PubMed Central

    Sulaiman, Siti Husna

    2018-01-01

    Aflatoxin is a food contaminant and its exposure through the diet is frequent and ubiquitous. A long-term dietary aflatoxin exposure has been linked to the development of liver cancer in populations with high prevalence of aflatoxin contamination in foods. Therefore, this study was conducted to identify the association between urinary aflatoxin M1 (AFM1), a biomarker of aflatoxin exposure, with the dietary intake among adults in Hulu Langat district, Selangor, Malaysia. Certain food products have higher potential for aflatoxin contamination and these were listed in a Food Frequency Questionnaire, which was given to all study participants. This allowed us to record consumption rates for each food product listed. Concomitantly, urine samples were collected, from adults in selected areas in Hulu Langat district, for the measurement of AFM1 levels using an ELISA kit. Of the 444 urine samples collected and tested, 199 were positive for AFM1, with 37 of them exceeding the limit of detection (LOD) of 0.64 ng/mL. Cereal products showed the highest consumption level among all food groups, with an average intake of 512.54 g per day. Chi-square analysis showed that consumption of eggs (X2 = 4.77, p = 0.03) and dairy products (X2 = 19.36, p < 0.01) had significant associations with urinary AFM1 but both food groups were having a phi and Cramer’s V value that less than 0.3, which indicated that the association between these food groups’ consumption and AFM1 level in urine was weak. PMID:29642443

  17. Role of metabolism and viruses in aflatoxin-induced liver cancer

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Groopman, John D.; Kensler, Thomas W.

    The use of biomarkers in molecular epidemiology studies for identifying stages in the progression of development of the health effects of environmental agents has the potential for providing important information for critical regulatory, clinical and public health problems. Investigations of aflatoxins probably represent one of the most extensive data sets in the field and this work may serve as a template for future studies of other environmental agents. The aflatoxins are naturally occurring mycotoxins found on foods such as corn, peanuts, various other nuts and cottonseed and they have been demonstrated to be carcinogenic in many experimental models. As amore » result of nearly 30 years of study, experimental data and epidemiological studies in human populations, aflatoxin B{sub 1} was classified as carcinogenic to humans by the International Agency for Research on Cancer. The long-term goal of the research described herein is the application of biomarkers to the development of preventative interventions for use in human populations at high-risk for cancer. Several of the aflatoxin-specific biomarkers have been validated in epidemiological studies and are now being used as intermediate biomarkers in prevention studies. The development of these aflatoxin biomarkers has been based upon the knowledge of the biochemistry and toxicology of aflatoxins gleaned from both experimental and human studies. These biomarkers have subsequently been utilized in experimental models to provide data on the modulation of these markers under different situations of disease risk. This systematic approach provides encouragement for preventive interventions and should serve as a template for the development, validation and application of other chemical-specific biomarkers to cancer or other chronic diseases.« less

  18. Occurrence of mycotoxins in refrigerated pizza dough and risk assessment of exposure for the Spanish population.

    PubMed

    Quiles, Juan Manuel; Saladino, Federica; Mañes, Jordi; Fernández-Franzón, Mónica; Meca, Giuseppe

    2016-08-01

    Mycotoxins are toxic metabolites produced by filamentous fungi, as Aspergillus, Penicillium and Fusarium. The first objective of this research was to study the presence of mycotoxins in 60 samples of refrigerated pizza dough, by extraction with methanol and determination by liquid chromatography associated with tandem mass spectrometry (LC-MS/MS). Then, the estimated dietary intakes (EDIs) of these mycotoxins, among the Spanish population, was calculated and the health risk assessment was performed, comparing the EDIs data with the tolerable daily intake values (TDIs). The mycotoxins detected in the analyzed samples were aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1), zearalenone (ZEA), enniatin A (ENA), enniatin A1 (ENA1), enniatin (ENB), enniatin B1 (ENB1) and BEA (beauvericin) with average concentration of the positive samples of 4.09 μg/kg, 0.50 μg/kg, 0.79 μg/kg, 77.78 μg/kg, 14.96 μg/kg, 4.54 μg/kg, 3.37 μg/kg, 1.69 μg/kg and 22.39 μg/kg, respectively. The presence of ZEA, ENA1, ENB and ENB1 was detected in 100% of the samples, AFB2 in 32%, AFB1 in 23%, ENA in 8% and BEA in 3%. Twelve percent of the samples contaminated with AFB1 and 12% of the doughs contaminated with ZEA exceeded the EU legislated maximum limits. The dietary intakes were estimated considering three different age groups of population, and the EDIs calculated for the mycotoxins detected in the samples were all below the established TDI. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Comparison of homogenization techniques and incidence of aflatoxin contamination in dried figs for export.

    PubMed

    Bircan, Cavit

    2009-01-01

    To determine differences in mean aflatoxin contamination and subsample variance from dry and slurry homogenizations, 10 kg of six different, naturally contaminated dried fig samples were collected from various exporting companies in accordance with the EU Commission Directive. The samples were first dry-mixed for 5 min using a blender and sub-sampled seven times; the remainder was slurry homogenized (1 : 1, v/v) and sub-sampled seven times. Aflatoxin B1 and total aflatoxin levels were recorded and coefficient of variations (CV) computed for all sub-samples. Only a small reduction in sub-sample variations, indicated by the lower CV values, and slight differences in mean aflatoxin B1 and total aflatoxin levels were observed when slurry homogenization was applied. Therefore, 7326 dried figs, destined for export from Turkey to the EU and collected during the 2008 crop year, were dry-homogenized and tested for aflatoxins (B1, B2, G1 and G2) by immunoaffinity column clean-up using RP-HPLC. While 34% of the samples contained detectable levels of total aflatoxins (0.20-208.75 µg kg(-1)), only 9% of them exceeded the EU limit of 4 µg kg(-1) in the range 2.0-208.75 µg kg(-1), respectively. A substantial increase in the incidence of aflatoxins was observed in 2008, most likely due to the drought stress experienced in Aydin province as occurred in 2007.

  20. Development and application of solvent-free extraction for the detection of aflatoxin M1 in dairy products by enzyme immunoassay.

    PubMed

    Anfossi, Laura; Calderara, Marianna; Baggiani, Claudio; Giovannoli, Cristina; Arletti, Enrico; Giraudi, Gianfranco

    2008-03-26

    The official methods for the quantification of aflatoxin M1 in dairy products (cheese and yogurt) include extraction into dichloromethane or chloroform, evaporation of the solvent, partitioning of the reconstituted residue with hexane, and subsequent analysis. To secure a rapid and inexpensive screen for aflatoxin M1 contamination, a sensitive competitive ELISA, using a rabbit polyclonal antibody, was developed for measuring aflatoxin M1 in milk and used in a comparative study for measuring the extraction efficiency of aflatoxin M1 in aqueous or organic solvent buffers using yogurt samples. An aqueous sodium citrate solution was found to be suitable for extracting aflatoxin M1, thus eliminating the need for organic solvents. The citrate extraction proved to be efficient (recovery ranged from 70 to 124%) in fortified samples of very different kinds of dairy products, including yogurt and six types of cheese. Fourteen yogurt and cheese samples were extracted with citrate solution and analyzed by ELISA. A good correlation was observed (y=0.95x-0.59, r2=0.98) when the data were compared with those obtained through the official method, across a wide range of aflatoxin M1 contaminations (10-200 ng/kg).

  1. Salt-Inducible Kinase 1 (SIK1) is Induced by Alcohol and Suppresses Microglia Inflammation via NF-κB Signaling.

    PubMed

    Zhang, Yu; Gao, Weida; Yang, Kongbin; Tao, Haiquan; Yang, Haicheng

    2018-06-19

    Alcohol consumption has been shown to cause neuroinflammation and increase a variety of immune-related signaling processes. Microglia are a crucial part of alcohol-induced neuroinflammation and undergo apoptosis. Even though the importance of these inflammatory processes in the effects of alcohol-related neurodegeneration have been established, the mechanism of alcohol-induced microglia apoptosis is unknown. In prior research, we discovered that alcohol increases expression of salt-inducible kinase 1 (SIK1) in rodent brain tissue. In this study, we sought to determine what role SIK1 expression plays in alcohol-induced neuroinflammation as well as whether and by what mechanism it regulates microglia apoptosis. Adult C57BL/6 mice were divided into four groups and for 3 weeks treated with either 0%, 5%, 10%, or 15% alcohol during 3 hour periods. The mice were sacrificed and their brains excised for analysis. Additionally, primary microglia were isolated from neonatal mice. SIK1 expression in alcohol-treated brain tissue and microglia was analyzed via RT-PCR and western blotting. TUNEL staining, caspase-3, and caspase-9 activity assays were performed to evaluate microglial apoptosis. Cell fluorescence staining and NF-κB luciferase activity assays were used to evaluate the effects of SIK1 expression on the NF-κB signaling pathway. SIK1 expression was increased in the brains of mice that consumed alcohol, and this effect was seen in mouse primary microglia. SIK1 knockdown in microglia increased alcohol-induced apoptosis in these cells. Furthermore, SIK1 reduced NF-κB signaling pathway factors, and SIK1 knockdown in microglia promoted alcohol-induced NF-κB activity. TUNEL staining, caspase-3, and caspase-9 activity assays consistently revealed that alcohol-induced microglial apoptosis was inhibited by depletion of p65. Finally, we determined that NF-κB signaling is required for alcohol-induced, SIK1-mediated apoptosis in microglia. This study establishes for the

  2. Occurrence of aflatoxins in oilseeds providing cocoa-butter substitutes.

    PubMed Central

    Kershaw, S J

    1982-01-01

    Four oilseeds providing cocoa-butter substitutes--shea, pentadecima, illipe, and salseed--when tested as substrates for aflatoxin production by two strains of Aspergillus parasiticus, gave varying levels of aflatoxin. Aflatoxins were found at low levels occurring naturally in moldy shea-nuts, but none of 21 commercial shea-nut samples contained greater than 20 micrograms of aflatoxin B1 per kg. PMID:6808919

  3. Occurrence of aflatoxins in oilseeds providing cocoa-butter substitutes.

    PubMed

    Kershaw, S J

    1982-05-01

    Four oilseeds providing cocoa-butter substitutes--shea, pentadecima, illipe, and salseed--when tested as substrates for aflatoxin production by two strains of Aspergillus parasiticus, gave varying levels of aflatoxin. Aflatoxins were found at low levels occurring naturally in moldy shea-nuts, but none of 21 commercial shea-nut samples contained greater than 20 micrograms of aflatoxin B1 per kg.

  4. Human aflatoxin exposure in Kenya, 2007: a cross-sectional study

    PubMed Central

    Yard, Ellen E.; Daniel, Johnni H.; Lewis, Lauren S.; Rybak, Michael E.; Paliakov, Ekaterina M.; Kim, Andrea A.; Montgomery, Joel M.; Bunnell, Rebecca; Abudo, Mamo Umuro; Akhwale, Willis; Breiman, Robert F.; Sharif, Shahnaaz K.

    2013-01-01

    Aflatoxins contaminate approximately 25% of agricultural products worldwide. They can cause liver failure and liver cancer. Kenya has experienced multiple aflatoxicosis outbreaks in recent years, often resulting in fatalities. However, the full extent of aflatoxin exposure in Kenya has been unknown. Our objective was to quantify aflatoxin exposure across Kenya. We analysed aflatoxin levels in serum specimens from the 2007 Kenya AIDS Indicator Survey – a nationally representative, cross-sectional serosurvey. KAIS collected 15,853 blood specimens. Of the 3180 human immunodeficiency virus-negative specimens with ≥1 mL sera, we randomly selected 600 specimens stratified by province and sex. We analysed serum specimens for aflatoxin albumin adducts by using isotope dilution MS/MS to quantify aflatoxin B1-lysine, and normalised with serum albumin. Aflatoxin concentrations were then compared by demographic, socioeconomic and geographic characteristics. We detected serum aflatoxin B1-lysine in 78% of serum specimens (range = 1.78 pg/mg albumin). Aflatoxin exposure did not vary by sex, age group, marital status, religion or socioeconomic characteristics. Aflatoxin exposure varied by province (p < 0.05); it was highest in Eastern (median = 7.87 pg/mg albumin) and Coast (median = 3.70 pg/mg albumin) provinces and lowest in Nyanza (median = aflatoxin exposure is a public health problem throughout Kenya, and it could be substantially impacting human health. Wide-scale, evidence-based interventions are urgently needed to decrease exposure and subsequent health effects. PMID:23767939

  5. Aspergillus and aflatoxin in groundnut (Arachis hypogaea L.) and groundnut cake in Eastern Ethiopia.

    PubMed

    Mohammed, Abdi; Chala, Alemayehu; Dejene, Mashilla; Fininsa, Chemeda; Hoisington, David A; Sobolev, Victor S; Arias, Renee S

    2016-12-01

    This study was conducted to assess major Aspergillus species and aflatoxins associated with groundnut seeds and cake in Eastern Ethiopia and evaluate growers' management practices. A total of 160 groundnut seed samples from farmers' stores and 50 groundnut cake samples from cafe and restaurants were collected. Fungal isolation was done from groundnut seed samples. Aspergillus flavus was the dominant species followed by Aspergillus parasiticus. Aflatoxin analyses of groundnut seed samples were performed using ultra performance liquid chromatography; 22.5% and 41.3% of samples were positive, with total aflatoxin concentrations of 786 and 3135 ng g -1 from 2013/2014 and 2014/2015 samples, respectively. The level of specific aflatoxin concentration varied between 0.1 and 2526 ng g -1 for B 2 and B 1 , respectively. Among contaminated samples of groundnut cake, 68% exhibited aflatoxin concentration below 20 ng g -1 , while as high as 158 ng g -1 aflatoxin B 1 was recorded. The study confirms high contamination of groundnut products in East Ethiopia.

  6. Reduced Graphene Oxide-Gold Nanoparticle Nanoframework as a Highly Selective Separation Material for Aflatoxins.

    PubMed

    Guo, Wenbo; Wu, Lidong; Fan, Kai; Nie, Dongxia; He, Weijing; Yang, Junhua; Zhao, Zhihui; Han, Zheng

    2017-11-03

    Graphene-based materials have been studied in many applications, owing to the excellent electrical, mechanical, and thermal properties of graphene. In the current study, an environmentally friendly approach to the preparation of a reduced graphene oxide-gold nanoparticle (rGO-AuNP) nanocomposite was developed by using L-cysteine and vitamin C as reductants under mild reaction conditions. The rGO-AuNP material showed a highly selective separation ability for 6 naturally occurring aflatoxins, which are easily adsorbed onto traditional graphene materials but are difficult to be desorbed. The specificity of the nanocomposite was evaluated in the separation of 6 aflatoxin congeners (aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, aflatoxin M1 and aflatoxin M2) from 23 other biotoxins (including, ochratoxin A, citrinin, and deoxynivalenol). The results indicated that this material was specific for separating aflatoxin congeners. The synthesized material was further validated by determining the recovery (77.6-105.0%), sensitivity (limit of detection in the range of 0.05-0.21 μg kg -1 ), and precision (1.5-11.8%), and was then successfully applied to the separation of aflatoxins from real-world maize, wheat and rice samples.

  7. Fungi, aflatoxins, and cyclopiazonic acid associated with peanut retailing in Botswana.

    PubMed

    Mphande, Fingani A; Siame, Bupe A; Taylor, Joanne E

    2004-01-01

    Peanuts are important food commodities, but they are susceptible to fungal infestation and mycotoxin contamination. Raw peanuts were purchased from retail outlets in Botswana and examined for fungi and mycotoxin (aflatoxins and cyclopiazonic acid) contamination. Zygomycetes were the most common fungi isolated; they accounted for 41% of all the isolates and were found on 98% of the peanut samples. Among the Zygomycetes, Absidia corymbifera and Rhizopus stolonifer were the most common. Aspergillus spp. accounted for 35% of all the isolates, with Aspergillus niger being the most prevalent (20.4%). Aspergillus flavus/parasiticus were also present and accounted for 8.5% of all the isolates, with A. flavus accounting for the majority of the A. flavus/parasiticus identified. Of the 32 isolates of A. flavus screened for mycotoxin production, 11 did not produce detectable aflatoxins, 8 produced only aflatoxins B1 and B2, and 13 produced all four aflatoxins (B1, B2, G1, and G2) in varying amounts. Only 6 of the A. flavus isolates produced cyclopiazonic acid at concentrations ranging from 1 to 55 microg/kg. The one A. parasiticus isolate screened also produced all the four aflatoxins (1,200 microg/kg) but did not produce cyclopiazonic acid. When the raw peanut samples (n = 120) were analyzed for total aflatoxins, 78% contained aflatoxins at concentrations ranging from 12 to 329 microg/kg. Many of the samples (49%) contained total aflatoxins at concentrations above the 20 microg/kg limit set by the World Health Organization. Only 21% (n = 83) of the samples contained cyclopiazonic acid with concentrations ranging from 1 to 10 microg/kg. The results show that mycotoxins and toxigenic fungi are common contaminants of peanuts sold at retail in Botswana.

  8. Toxigenic potentiality of Aspergillus flavus and Aspergillus parasiticus strains isolated from black pepper assessed by an LC-MS/MS based multi-mycotoxin method.

    PubMed

    Yogendrarajah, Pratheeba; Devlieghere, Frank; Njumbe Ediage, Emmanuel; Jacxsens, Liesbeth; De Meulenaer, Bruno; De Saeger, Sarah

    2015-12-01

    A liquid chromatography triple quadrupole tandem mass spectrometry method was developed and validated to determine mycotoxins, produced by fungal isolates grown on malt extract agar (MEA). All twenty metabolites produced by different fungal species were extracted using acetonitrile/1% formic acid. The developed method was applied to assess the toxigenic potentiality of Aspergillus flavus (n = 11) and Aspergillus parasiticus (n = 6) strains isolated from black peppers (Piper nigrum L.) following their growth at 22, 30 and 37 °C. Highest mean radial colony growth rates were observed at 30 °C for A. flavus (5.21 ± 0.68 mm/day) and A. parasiticus (4.97 ± 0.33 mm/day). All of the A. flavus isolates produced aflatoxin B1 and O-methyl sterigmatocystin (OMST) while 91% produced aflatoxin B2 (AFB2) and 82% of them produced sterigmatocystin (STERIG) at 30 °C. Except one, all the A. parasiticus isolates produced all the four aflatoxins, STERIG and OMST at 30 °C. Remarkably high AFB1 was produced by some A. flavus isolates at 22 °C (max 16-40 mg/kg). Production of mycotoxins followed a different trend than that of growth rate of both species. Notable correlations were found between different secondary metabolites of both species; R(2) 0.87 between AFB1 and AFB2 production. Occurrence of OMST could be used as a predictor for AFB1 production. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Evaluation of atoxigenic isolates of Aspergillus flavus as potential biocontrol agents for aflatoxin in maize.

    PubMed

    Atehnkeng, J; Ojiambo, P S; Ikotun, T; Sikora, R A; Cotty, P J; Bandyopadhyay, R

    2008-10-01

    Aflatoxin contamination resulting from maize infection by Aspergillus flavus is both an economic and a public health concern. Therefore, strategies for controlling aflatoxin contamination in maize are being investigated. The abilities of eleven naturally occurring atoxigenic isolates in Nigeria to reduce aflatoxin contamination in maize were evaluated in grain competition experiments and in field studies during the 2005 and 2006 growing seasons. Treatments consisted of inoculation of either grains in vials or ears at mid-silking stage in field plots, with the toxigenic isolate (La3228) or atoxigenic isolate alone and co-inoculation of each atoxigenic isolate and La3328. Aflatoxin B(1) + B(2) concentrations were significantly (p < 0.05) lower in the co-inoculation treatments compared with the treatment in which the aflatoxin-producing isolate La3228 was inoculated alone. Relative levels of aflatoxin B(1) + B(2) reduction ranged from 70.1% to 99.9%. Among the atoxigenics, two isolates from Lafia, La3279 and La3303, were most effective at reducing aflatoxin B(1) + B(2) concentrations in both laboratory and field trials. These two isolates have potential value as agents for the biocontrol of aflatoxin contamination in maize. Because these isolates are endemic to West Africa, they are both more likely than introduced isolates to be well adapted to West African environments and to meet regulatory concerns over their use throughout that region.

  10. An evaluation of aflatoxin and cyclopiazonic acid production in Aspergillus oryzae.

    PubMed

    Kim, Nam Yeun; Lee, Jin Hee; Lee, Inhyung; Ji, Geun Eog

    2014-06-01

    To date, edible fungi such as Aspergillus flavus var. oryzae (A. oryzae) has been considered as safe. However, some strains can produce mycotoxins. Thus, the biosynthetic ability to produce mycotoxins should be reevaluated to determine the safety of edible fungi. We analyzed the production of aflatoxins and cyclopiazonic acid (CPA) from edible fungi such as A. oryzae isolated from various Korean foods using multiplex PCR, enzyme-linked immunosorbent assay, and high-performance liquid chromatography (HPLC). In the multiplex PCR analysis of aflatoxin biosynthetic genes omtB, aflR, ver-1, and omtA, 5 of 19 Aspergillus strains produced all PCR products. Among them, aflatoxin B1 and aflatoxin B2 were detected from only A. flavus KACC 41403 by HPLC. Aflatoxins were not detected from the other four strains that produced all positive PCR bands. Aflatoxin also was not detected from 12 strains that had PCR patterns without aflR or ver-1 and from 2 strains that did not produce any of the expected PCR products. Only the seven A. oryzae strains that produced all of the positive PCR bands including the CPA biosynthetic genes maoA, dmaT, and pks-nrps produced CPA. CPA and aflatoxin production must be evaluated before A. oryzae strains are used for the development of fermented foods.

  11. Effectiveness of pulsed light treatment for degradation and detoxification of aflatoxin B1 and B2 in rough rice and rice bran

    USDA-ARS?s Scientific Manuscript database

    Aflatoxins primarily accumulate in the hull and bran layers of rough rice making these by-products of rice milling unsuitable for animal feed or human consumption. Contaminated rough rice is also a potential source of aflatoxin exposure to workers handling the grain during post-harvest storage and p...

  12. Insulin induces alpha1B-adrenergic receptor phosphorylation and desensitization.

    PubMed

    García-Sáinz, J Adolfo; Romero-Avila, M Teresa; Molina-Muñoz, Tzindilú; Medina, Luz del Carmen

    2004-09-03

    The ability of insulin to induce alpha1B-adrenoceptor phosphorylation and desensitization was tested in two model systems: rat-1 cells that stably express alpha1B-adrenoceptors, through transfection, and endogenously express insulin receptors and DDT1 MF2 cells that endogenously express both receptors. Insulin induced concentration-dependent increases in the phosphorylation state of the adrenergic receptors in the two models with similar EC50 values (0.5-2 nM). The effect was rapid in the two systems but it was sustained in rat-1 cells and transient in DDT1 MF2 cells. In both cell lines, the insulin-mediated phosphorylation of alpha1B-adrenoceptors was blocked by wortmannin and LY 294002, and by staurosporine and bisindolylmaleimide I, indicating that the effect involved phosphoinositide 3-kinase and protein kinase C activities. The adrenoceptor phosphorylation induced by insulin was associated to desensitization as evidences by a diminished elevation of intracellular calcium in response to noradrenaline. Inhibitors of phosphoinositide 3-kinase and protein kinase C blocked the functional desensitization induced by insulin.

  13. Biosorption of B-aflatoxins Using Biomasses Obtained from Formosa Firethorn [Pyracantha koidzumii (Hayata) Rehder].

    PubMed

    Ramales-Valderrama, Rosa Adriana; Vázquez-Durán, Alma; Méndez-Albores, Abraham

    2016-07-13

    Mycotoxin adsorption onto biomaterials is considered as a promising alternative for decontamination without harmful chemicals. In this research, the adsorption of B-aflatoxins (AFB₁ and AFB₂) using Pyracantha koidzumii biomasses (leaves, berries and the mixture of leaves/berries) from aqueous solutions was explored. The biosorbent was used at 0.5% (w/v) in samples spiked with 100 ng/mL of B-aflatoxin standards and incubated at 40 °C for up to 24 h. A standard biosorption methodology was employed and aflatoxins were quantified by an immunoaffinity column and UPLC methodologies. The biosorbent-aflatoxin interaction mechanism was investigated from a combination of zeta potential (ζ), Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The highest aflatoxin uptakes were 86% and 82% at 6 h using leaves and the mixture of leaves/berries biomasses, respectively. A moderate biosorption of 46% was attained when using berries biomass. From kinetic studies, the biosorption process is described using the first order adsorption model. Evidence from FTIR spectra suggests the participation of hydroxyl, amine, carboxyl, amide, phosphate and ketone groups in the biosorption and the mechanism was proposed to be dominated by the electrostatic interaction between the negatively charged functional groups and the positively charged aflatoxin molecules. Biosorption by P. koidzumii biomasses has been demonstrated to be an alternative to conventional systems for B-aflatoxins removal.

  14. Quantitative Scrutinization of Aflatoxins in Different Spices from Pakistan

    PubMed Central

    Kashif, Aiza; Kanwal, Kinza; Khan, Abdul Muqeet; Abbas, Mateen

    2016-01-01

    The current research work aimed to access the contamination level of aflatoxins B1, B2, G1, and G2 in the household spices that are widely consumed in huge amounts. 200 different spice samples, 100 packed and 100 unpacked, were analyzed for the aflatoxins profile by HPLC with an incidence of 61.5% contamination out of which 53.66% samples exceed the EU limit. The results disclosed that the unpacked samples are more contaminated as compared to the packed samples except for white cumin seeds. Among packed and unpacked samples of spices, the maximum value of aflatoxins was detected in fennel, that is, 27.93 μg/kg and 67.04 μg/kg, respectively. The lowest concentration of aflatoxin was detected in cinnamon in packed form (0.79 μg/kg) and in the unpacked samples of white cumin seeds which is 1.75 μg/kg. Caraway seeds and coriander in its unpacked form showed positive results whereas black pepper (packed and unpacked) was found free from aflatoxins. This is the first report on the occurrence of aflatoxins in packed and unpacked samples of spices from Pakistan. To ensure safe consumption of spices, there should be constant monitoring of aflatoxin and more studies need to be executed with the intention of preventing mycotoxin accretion in this commodity. PMID:27781067

  15. Annexin A1, Annexin A2, and Dyrk 1B are upregulated during GAS1-induced cell cycle arrest.

    PubMed

    Pérez-Sánchez, Gilberto; Jiménez, Adriana; Quezada-Ramírez, Marco A; Estudillo, Enrique; Ayala-Sarmiento, Alberto E; Mendoza-Hernández, Guillermo; Hernández-Soto, Justino; Hernández-Hernández, Fidel C; Cázares-Raga, Febe E; Segovia, Jose

    2018-05-01

    GAS1 is a pleiotropic protein that has been investigated because of its ability to induce cell proliferation, cell arrest, and apoptosis, depending on the cellular or the physiological context in which it is expressed. At this point, we have information about the molecular mechanisms by which GAS1 induces proliferation and apoptosis; but very few studies have been focused on elucidating the mechanisms by which GAS1 induces cell arrest. With the aim of expanding our knowledge on this subject, we first focused our research on finding proteins that were preferentially expressed in cells arrested by serum deprivation. By using a proteomics approach and mass spectrometry analysis, we identified 17 proteins in the 2-DE protein profile of serum deprived NIH3T3 cells. Among them, Annexin A1 (Anxa1), Annexin A2 (Anxa2), dual specificity tyrosine-phosphorylation-regulated kinase 1B (Dyrk1B), and Eukaryotic translation initiation factor 3, F (eIf3f) were upregulated at transcriptional the level in proliferative NIH3T3 cells. Moreover, we demonstrated that Anxa1, Anxa2, and Dyrk1b are upregulated at both the transcriptional and translational levels by the overexpression of GAS1. Thus, our results suggest that the upregulation of Anxa1, Anxa2, and Dyrk1b could be related to the ability of GAS1 to induce cell arrest and maintain cell viability. Finally, we provided further evidence showing that GAS1 through Dyrk 1B leads not only to the arrest of NIH3T3 cells but also maintains cell viability. © 2017 Wiley Periodicals, Inc.

  16. Oral administration of aflatoxin G₁ induces chronic alveolar inflammation associated with lung tumorigenesis.

    PubMed

    Liu, Chunping; Shen, Haitao; Yi, Li; Shao, Peilu; Soulika, Athena M; Meng, Xinxing; Xing, Lingxiao; Yan, Xia; Zhang, Xianghong

    2015-02-03

    Our previous studies showed oral gavage of aflatoxin G₁ (AFG₁) induced lung adenocarcinoma in NIH mice. We recently found that a single intratracheal administration of AFG₁ caused chronic inflammatory changes in rat alveolar septum. Here, we examine whether oral gavage of AFG₁ induces chronic lung inflammation and how it contributes to carcinogenesis. We evaluated chronic lung inflammatory responses in Balb/c mice after oral gavage of AFG₁ for 1, 3 and 6 months. Inflammatory responses were heightened in the lung alveolar septum, 3 and 6 months after AFG₁ treatment, evidenced by increased macrophages and lymphocytes infiltration, up-regulation of NF-κB and p-STAT3, and cytokines production. High expression levels of superoxide dismutase (SOD-2) and hemoxygenase-1 (HO-1), two established markers of oxidative stress, were detected in alveolar epithelium of AFG₁-treated mice. Promoted alveolar type II cell (AT-II) proliferation in alveolar epithelium and angiogenesis, as well as increased COX-2 expression were also observed in lung tissues of AFG₁-treated mice. Furthermore, we prolonged survival of the mice in the above model for another 6 months to examine the contribution of AFG₁-induced chronic inflammation to lung tumorigenesis. Twelve months later, we observed that AFG₁ induced alveolar epithelial hyperplasia and adenocarcinoma in Balb/c mice. Up-regulation of NF-κB, p-STAT3, and COX-2 was also induced in lung adenocarcinoma, thus establishing a link between AFG₁-induced chronic inflammation and lung tumorigenesis. This is the first study to show that oral administration of AFG₁ could induce chronic lung inflammation, which may provide a pro-tumor microenvironment to contribute to lung tumorigenesis. Copyright © 2014. Published by Elsevier Ireland Ltd.

  17. Cytochrome P450 1B1 contributes to angiotensin II-induced hypertension and associated pathophysiology.

    PubMed

    Jennings, Brett L; Sahan-Firat, Seyhan; Estes, Anne M; Das, Kanak; Farjana, Nasreen; Fang, Xiao R; Gonzalez, Frank J; Malik, Kafait U

    2010-10-01

    Hypertension is the leading cause of cardiovascular diseases, and angiotensin II is one of the major components of the mechanisms that contribute to the development of hypertension. However, the precise mechanisms for the development of hypertension are unknown. Our recent study showing that angiotensin II-induced vascular smooth muscle cell growth depends on cytochrome P450 1B1 led us to investigate its contribution to hypertension caused by this peptide. Angiotensin II was infused via miniosmotic pump into rats (150 ng/kg per minute) or mice (1000 μg/kg per day) for 13 days resulting in increased blood pressure, increased cardiac and vascular hypertrophy, increased vascular reactivity to vasoconstrictor agents, increased vascular reactive oxygen species production, and endothelial dysfunction in both species. The increase in blood pressure and associated pathophysiological changes were minimized by the cytochrome P450 1B1 inhibitor 2,3',4,5'-tetramethoxystilbene in both species and was markedly reduced in Cyp1b1(-/-) mice. These data suggest that cytochrome P450 1B1 contributes to angiotensin II-induced hypertension and associated pathophysiological changes. Moreover, 2,3',4,5'-tetramethoxystilbene, which prevents both cytochrome P450 1B1-dependent and -independent components of angiotensin II-induced hypertension and inhibits associated pathophysiological changes could be clinically useful in the treatment of hypertension and associated cardiovascular and inflammatory diseases.

  18. CYTOCHROME P450 1B1 CONTRIBUTES TO ANGIOTENSIN II-INDUCED HYPERTENSION AND ASSOCIATED PATHOPHYSIOLOGY

    PubMed Central

    Jennings, Brett L.; Sahan-Firat, Seyhan; Estes, Anne M.; Das, Kanak; Farjana, Nasreen; Fang, Xiao R.; Gonzalez, Frank J.; Malik, Kafait U.

    2010-01-01

    Hypertension is the leading cause of cardiovascular diseases, and angiotensin II is one of the major components of the mechanisms that contribute to the development of hypertension. However, the precise mechanisms for the development of hypertension are unknown. Our recent study that angiotensin II-induced vascular smooth muscle cell growth is dependent on cytochrome P450 1B1 led us to investigate its contribution to hypertension caused by this peptide. Angiotensin II was infused via miniosmotic pump into rats (150 ng/kg/min) or mice (1000 μg/kg/day) for 13 days resulting in increased blood pressure, increased cardiac and vascular hypertrophy, increased vascular reactivity to vasoconstrictor agents, increased reactive oxygen species production, and endothelial dysfunction in both species. The increase in blood pressure and associated pathophysiological changes were minimized by the cytochrome P450 1B1 inhibitor, 2,3′,4,5′-tetramethoxystilbene in both species and was markedly reduced in Cyp1b1-/- mice. These data suggest that cytochrome P450 1B1 contributes to angiotensin II-induced hypertension and associated pathophysiological changes. Moreover, 2,3′,4,5′-tetramethoxystilbene which prevents both cytochrome P450 1B1-dependent and independent components of angiotensin II-induced hypertension and inhibits associated pathophysiological changes could be clinically useful in the treatment of hypertension and associated cardiovascular and inflammatory diseases. PMID:20805442

  19. The first report of A. novoparasiticus, A. arachidicola and A. pseudocaelatus in Brazilian corn kernels.

    PubMed

    Viaro, Helena Paula; da Silva, Josué José; de Souza Ferranti, Larissa; Bordini, Jaqueline Gozzi; Massi, Fernanda Pelisson; Fungaro, Maria Helena Pelegrinelli

    2017-02-21

    Maize is one of the most important commercial crops cultivated throughout the world, mostly in tropical and subtropical countries. It is highly susceptible to mycotoxins, toxic secondary metabolites produced by fungi. In this study, we assessed freshly harvested corn produced in Brazil for aflatoxin contamination and the presence of Aspergillus. B type aflatoxins (AFB 1 +AFB 2 ) were detected in 56% of 16 grain samples, while G type aflatoxins (AFG 1 +AFG 2 ) were detected in 25%. Of the total number of grains (n=1920) evaluated for the presence of fungi species, 4.7% were infected with Aspergillus species, 74.5% and 16.7% respectively with Fusarium and Penicillium species and 4.1% with other fungi genera. In total, 89 Aspergillus isolates were identified, most (86 isolates) characterized as belonging to Aspergillus section Flavi, and the remainder to Aspergillus section Cremei (2 isolates) and Aspergillus section Terrei (1 isolate). All the isolates of section Flavi were subjected to molecular analysis. They were found to belong to six species, including Aspergillus novoparasiticus, Aspergillus arachidicola and Aspergillus pseudocaelatus, all aflatoxins B and G producing species, which are herein described for the first time infecting corn kernels. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. Detection of mycotoxins using imaging surface plasmon resonance (iSPR)

    USDA-ARS?s Scientific Manuscript database

    Significant progress has been made in the development of biosensors that can be used to detect mycotoxins. One technology that has been extensively tested is surface plasmon resonance (SPR). In 2003 a multi-toxin method was reported that detected aflatoxin B1 (AFB1), zearalenone (ZEA), fumonisin B1 ...

  1. Occurrence of Ochratoxins, Fumonisin B2 , Aflatoxins (B1 and B2 ), and Other Secondary Fungal Metabolites in Dried Date Palm Fruits from Egypt: A Mini-Survey.

    PubMed

    Abdallah, Mohamed F; Krska, Rudolf; Sulyok, Michael

    2018-02-01

    This study was conducted to investigate the natural co-occurrence of 295 fungal and bacterial metabolites in 28 samples of dried date palm fruits collected from different shops distributed in Assiut Governorate, Upper Egypt in 2016. Extraction and quantification of the target analytes were done using the "dilute and shoot" approach followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. In total, 30 toxic fungal metabolites were detected. Among these metabolites, 4 types of ochratoxins including ochratoxin type A and B were quantified in 3 samples (11%) with a contamination range from 1.48 to 6070 μg/kg for ochratoxin A and from 0.28 to 692 μg/kg for ochratoxin B. In addition, fumonisin B 2 was observed in 2 (7%) samples with contamination levels ranging from 4.99 to 16.2 μg/kg. The simultaneous detection of fumonisin B 2 in the same contaminated samples with ochratoxins indicates the fungal attack by Aspergillus niger species during storage. Only 1 sample was contaminated with aflatoxin B 1 (14.4 μg/kg) and B 2 (2.44 μg/kg). The highest maximum concentration (90400 μg/kg) was for kojic acid that contaminated 43% of the samples. To the best of the authors' knowledge, this is the first report of the natural co-occurrence of fumonisin B 2 and ochratoxin A and B in addition to a wide range of other fungal metabolites in date palm fruits. Mycotoxins are secondary metabolites produced by different fungi. These metabolites pose a potential risk on human health since they contaminate many food commodities. Among these, date palm fruits which are an integral part of diet in several countries. Therefore, detection of mycotoxins is a prerequisite to insure the safety of food. Here, different types of mycotoxins have been detected in levels that may have health hazard. © 2018 Institute of Food Technologists®.

  2. Operable Unit B1 Interim Record of Decision. Installation Restoration Program (IRP)

    DTIC Science & Technology

    1993-07-28

    Geology ..................................... I-1 2.0 SITE HISTORY AND ENFORCEMENT ACTIVITIES ........... 11-5 2.1 Background on Contamination Problems at...McClellan AFB OU BI . 11-5 2.2 Previous Studies ................................ 11-5 2.3 Regulatory and Enforcement History .................... 11-7...1-4 2-1 History of OU BI ...................................... 11-6 4-1 Cross-Section A-A’ of OU B.I

  3. Survey of fungal counts and natural occurrence of aflatoxins in Malaysian starch-based foods.

    PubMed

    Abdullah, N; Nawawi, A; Othman, I

    1998-01-01

    In a survey of starch-based foods sampled from retail outlets in Malaysia, fungal colonies were mostly detected in wheat flour (100%), followed by rice flour (74%), glutinous rice grains (72%), ordinary rice grains (60%), glutinous rice flour (48%) and corn flour (26%). All positive samples of ordinary rice and glutinous rice grains had total fungal counts below 10(3) cfu/g sample, while among the positive rice flour, glutinous rice flour and corn flour samples, the highest total fungal count was more than 10(3) but less than 10(4) cfu/g sample respectively. However, in wheat flour samples total fungal count ranged from 10(2) cfu/g sample to slightly more than 10(4) cfu/g sample. Aflatoxigenic colonies were mostly detected in wheat flour (20%), followed by ordinary rice grains (4%), glutinous rice grains (4%) and glutinous rice flour (2%). No aflatoxigenic colonies were isolated from rice flour and corn flour samples. Screening of aflatoxin B1, aflatoxin B2, aflatoxin G1 and aflatoxin G2 using reversed-phase HPLC were carried out on 84 samples of ordinary rice grains and 83 samples of wheat flour. Two point four percent (2.4%) of ordinary rice grains were positive for aflatoxin G1 and 3.6% were positive for aflatoxin G2. All the positive samples were collected from private homes at concentrations ranging from 3.69-77.50 micrograms/kg. One point two percent (1.2%) of wheat flour samples were positive for aflatoxin B1 at a concentration of 25.62 micrograms/kg, 4.8% were positive for aflatoxin B2 at concentrations ranging from 11.25-252.50 micrograms/kg, 3.6% were positive for aflatoxin G1 at concentrations ranging from 25.00-289.38 micrograms/kg and 13.25% were positive for aflatoxin G2 at concentrations ranging from 16.25-436.25 micrograms/kg. Similarly, positive wheat flour samples were mostly collected from private homes.

  4. Ibuprofen Inhibits Colitis-Induced Overexpression of Tumor-Related Rac1b1

    PubMed Central

    Matos, Paulo; Kotelevets, Larissa; Goncalves, Vania; Henriques, Andreia; Zerbib, Philippe; Moyer, Mary Pat; Chastre, Eric; Jordan, Peter

    2013-01-01

    The serrated pathway to colorectal tumor formation involves oncogenic mutations in the BRAF gene, which are sufficient for initiation of hyperplastic growth but not for tumor progression. A previous analysis of colorectal tumors revealed that overexpression of splice variant Rac1b occurs in around 80% of tumors with mutant BRAF and both events proved to cooperate in tumor cell survival. Here, we provide evidence for increased expression of Rac1b in patients with inflamed human colonic mucosa as well as following experimentally induced colitis in mice. The increase of Rac1b in the mouse model was specifically prevented by the nonsteroidal anti-inflammatory drug ibuprofen, which also inhibited Rac1b expression in cultured HT29 colorectal tumor cells through a cyclooxygenase inhibition.independent mechanism. Accordingly, the presence of ibuprofen led to a reduction of HT29 cell survival in vitro and inhibited Rac1b-dependent tumor growth of HT29 xenografts. Together, our results suggest that stromal cues, namely, inflammation, can trigger changes in Rac1b expression in the colon and identify ibuprofen as a highly specific and efficient inhibitor of Rac1b overexpression in colorectal tumors. Our data suggest that the use of ibuprofen may be beneficial in the treatment of patients with serrated colorectal tumors or with inflammatory colon syndromes. PMID:23359345

  5. In Situ Soil Venting - Full Scale Test Hill AFB, Guidance Document, Literature Review. Volume 1

    DTIC Science & Technology

    1991-08-01

    AD-A254 924 1’) VOL I IN SITU SOIL VENTING - FULL SCALE TEST HILL AFB, GUIDANCE DOCUMENT, LITERATURE REVIEW D. W. DEPAO, S. E. HERBES, J. H . WILSON...D. K. SOLOMON, AND H . L. JENNINGS MARTIN-MARIETTA ENERGY SYSTEMS OAK RIDGE NATIONAL LABORATORY P. O. BOX 2008 OAK RIDGE TN 37831 OTI AUGUST 1991 S...sificat,cn) (U) In Situ Soil Ver.ting - Full Scale Test Hill AFB, Guidance Document, Literature Review 2 PERSO’.AL AUTH-O’.S, a W ApP li- S_ T’.- erber:. H

  6. Evaluation of a method to determine the natural occurrence of aflatoxins in commercial traditional herbal medicines from Malaysia and Indonesia.

    PubMed

    Ali, N; Hashim, N H; Saad, B; Safan, K; Nakajima, M; Yoshizawa, T

    2005-12-01

    Traditional herbal medicines, popularly known as 'jamu' and 'makjun' in Malaysia and Indonesia, are consumed regularly to promote health. In consideration of their frequent and prolonged consumption, the natural occurrence of aflatoxins (AF) in these products was determined using immunoaffinity column clean-up and high-performance liquid chromatography with pre-column derivatization. The evaluated method, which entails dilution of sample extracts with Tween 20-phosphate buffered saline (1:9, v/v) and a chromatographic system using isocratic mobile phase composed of water-methanol-acetonitrile (70:20:10, v/v/v), was effective in separating AFB1, AFG1 and AFG2 from interference at their retention times. Results were confirmed using post-column derivatization with photochemical reactor. For 23 commercial samples analyzed, mean levels (incidence) of AFB(1), AFB(2) and AFG1 in positive samples were 0.26 (70%), 0.07 (61%) and 0.10 (30%) microg/kg, respectively; one sample was positive for AFG2 at a level of 0.03 (4%) microg/kg. In contrast to the high levels of AF in crude herbal drugs and medicinal plants reported previously by other researchers, the low contamination levels reported in this study may be attributed to the higher selectivity to AF of the method applied. Based on the AFB1 levels and the daily consumption of positive samples, a mean probable daily intake of 0.022 ng/kg body weight was calculated.

  7. Mould and mycotoxin exposure assessment of melon and bush mango seeds, two common soup thickeners consumed in Nigeria.

    PubMed

    Ezekiel, Chibundu N; Sulyok, Michael; Somorin, Yinka; Odutayo, Foluke I; Nwabekee, Stella U; Balogun, Afeez T; Krska, Rudolf

    2016-11-21

    An examination of the mould and fungal metabolite pattern in melon and bush mango seeds locally produced in Nigeria was undertaken in order to understand the mycotoxicological risk posed to consumers of both of these important and commonly consumed soup thickeners. The variation in mycotoxin levels in graded categories of both foodstuffs were also determined. Aspergillus, Fusarium, Penicillium, Mucorales and Trichoderma were the recovered fungi from the foodstuffs with Aspergillus species dominating (melon=97.8%; bush mango=89.9%). Among the Aspergillus species identified Aspergillus section Flavi dominated (melon: 72%; bush mango: 57%) and A. flavus, A. parasiticus, A. parvisclerotigenus and A. tamarii were the recovered species. About 56% and 73% of the A. flavus isolates from melon and bush mango seed samples, respectively were aflatoxigenic. Thirty-four and 59 metabolites including notable mycotoxins were found in the melon and bush mango seeds respectively. Mean aflatoxin levels (μg/kg) in melon (aflatoxin B 1 (AFB 1 )=37.5 and total aflatoxins=142) and bush mango seeds (AFB 1 =68.1 and total aflatoxins=61.7) were higher than other mycotoxins, suggesting potential higher exposure for consumer populations. Significantly (p<0.05) higher levels of mycotoxins were found in hand-peeled melon and discoloured bush mango seeds than in machine-peeled melon and non-discoloured seeds except for HT-2 and T-2 toxins which occurred conversely. All melon and bush mango seeds exceeded the 2μg/kg AFB 1 limit whereas all melon and 55% of bush mango seeds exceeded the 4μg/kg total aflatoxin EU limit adopted in Nigeria. This is the first report of (1) mycotoxin co-occurrence in bush mango seeds, (2) cyclopiazonic acid, HT-2 toxin, moniliformin, mycophenolic acid, T-2 toxin and tenuazonic acid occurrence, and (3) mycotoxin exposure assessment of both foodstuffs. Copyright © 2016 Elsevier B.V. All rights reserved.

  8. Leptin induces CYP1B1 expression in MCF-7 cells through ligand-independent activation of the ERα pathway

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Khanal, Tilak; Kim, Hyung Gyun; Do, Minh Truong

    2014-05-15

    Leptin, a hormone with multiple biological actions, is produced predominantly by adipose tissue. Among its functions, leptin can stimulate tumour cell growth. Oestrogen receptor α (ERα), which plays an essential role in breast cancer development, can be transcriptionally activated in a ligand-independent manner. In this study, we investigated the effect of leptin on CYP1B1 expression and its mechanism in breast cancer cells. Leptin induced CYP1B1 protein, messenger RNA expression and promoter activity in ERα-positive MCF-7 cells but not in ERα-negative MDA-MB-231 cells. Additionally, leptin increased 4-hydroxyoestradiol in MCF-7 cells. Also, ERα knockdown by siRNA significantly blocked the induction of CYP1B1more » expression by leptin, indicating that leptin induced CYP1B1 expression via an ERα-dependent mechanism. Transient transfection with CYP1B1 deletion promoter constructs revealed that the oestrogen response element (ERE) plays important role in the up-regulation of CYP1B1 by leptin. Furthermore, leptin stimulated phosphorylation of ERα at serine residues 118 and 167 and increased ERE-luciferase activity, indicating that leptin induced CYP1B1 expression by ERα activation. Finally, we found that leptin activated ERK and Akt signalling pathways, which are upstream kinases related to ERα phosphorylation induced by leptin. Taken together, our results indicate that leptin-induced CYP1B1 expression is mediated by ligand-independent activation of the ERα pathway as a result of the activation of ERK and Akt in MCF-7 cells. - Highlights: • Leptin increased 4-hydroxyoestradiol in MCF-7 breast cancer cells. • Leptin activated ERK and Akt kinases related to ERα phosphorylation. • Leptin induces phosphorylation of ERα at serine residues 118 and 167. • Leptin induces ERE-luciferase activity.« less

  9. Celastrol ameliorates HIV-1 Tat-induced inflammatory responses via NF-kappaB and AP-1 inhibition and heme oxygenase-1 induction in astrocytes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Youn, Gi Soo; Kwon, Dong-Joo; Ju, Sung Mi

    HIV-1 Tat causes extensive neuroinflammation that may progress to AIDS-related encephalitis and dementia. Celastrol possesses various biological activities such as anti-oxidant, anti-tumor, and anti-inflammatory activities. In this study, we investigated the modulatory effects of celastrol on HIV-1 Tat-induced inflammatory responses and the molecular mechanisms underlying its action in astrocytes. Pre-treatment of CRT-MG human astroglioma cells with celastrol significantly inhibited HIV-1 Tat-induced expression of ICAM-1/VCAM-1 and subsequent monocyte adhesiveness in CRT-MG cells. In addition, celastrol suppressed HIV-1 Tat-induced expression of pro-inflammatory chemokines, such as CXCL10, IL-8, and MCP-1. Celastrol decreased HIV-1 Tat-induced activation of JNK MAPK, AP-1, and NF-κB. Furthermore, celastrolmore » induced mRNA and protein expression of HO-1 as well as Nrf2 activation. Blockage of HO-1 expression using siRNA reversed the inhibitory effect of celastrol on HIV-1 Tat-induced inflammatory responses. These results suggest that celastrol has regulatory effects on HIV-1 Tat-induced inflammatory responses by blocking the JNK MAPK-AP-1/NF-κB signaling pathways and inducing HO-1 expression in astrocytes. - Highlights: • Celastrol suppressed HIV-1 Tat-induced expression of pro-inflammatory genes. • Celastrol inhibited HIV-1 Tat -induced activation of JNK MAPK. • Celastrol inhibited HIV-1 Tat-induced activation of both NF-κB and AP-1. • Celastrol inhibited HIV-1 Tat-induced inflammatory responses via HO-1 induction.« less

  10. Photoinduced discharge of electrons stored in a TiO2-MWCNT composite to an analyte: application to the fluorometric determination of hydrogen peroxide, glucose and aflatoxin B1.

    PubMed

    Rhouati, Amina; Nasir, Muhammad; Marty, Jean-Louis; Hayat, Akhtar

    2017-12-06

    The authors describe an analytical detection scheme based on the use of multiwalled carbon nanotubes (MWCNTs) that accept and store electrons upon contact with photo-irradiated TiO 2 nanoparticles (TiO 2 -NPs). The Fermi level equilibration with photo-irradiated TiO 2 -NPs has a storage value of 0.35 mM of electrons per 120 mg·L -1 of MWCNTs. The stored electrons can be discharged on demand upon addition of electron acceptors to the TiO 2 -NP/MWCNT composite. These findings are applied to detect the quencher hydrogen peroxide. H 2 O 2 also is produced on enzymatic action of glucose oxidase on glucose, and this enables glucose also to be quantified by using the TiO 2 -NP/MWCNT fluorescent nanoprobe. The wide scope of the method also is demonstrated by an assay for aflatoxin B1 that is making use of an FAM-labeled aptamer where the FAM fluorophore on the aptamer quenches the emission of the nanoprobe. The following analytical linear ranges and limits of detection are found: H 2 O 2 : 0.1-100 μM and 15 nM; glucose: 5-200 μM and 0.5 μM; aflatoxin: 0.1-40 ng·mL -1 and 0.02 ng·mL -1 . The method was applied to the determination of glucose in human serum. Graphical abstract The assays demonstrated in (b) and (c) are based on the fluorescence quenching ability of MWCNTs-TiO 2 . In the presence of the target (analyte), the fluorescence is restored and the target concentration is determined from the percentage of fluorescence recovery.

  11. Col1A1 Production and Apoptotic Resistance in TGF-β1-Induced Epithelial-to-Mesenchymal Transition-Like Phenotype of 603B Cells

    PubMed Central

    Liu, Jun; Eischeid, Alex N.; Chen, Xian-Ming

    2012-01-01

    Recent studies have suggested that proliferating cholangiocytes have an important role in the induction of fibrosis, either directly via epithelial-to-mesenchymal transition (EMT), or indirectly via activation of other liver cell types. Transforming growth factor beta 1 (TGF-β1), a critical fibrotic cytokine for hepatic fibrosis, is a potent EMT inducer. This study aimed to clarify the potential contributions of TGF-β1-induced EMT-like cholangiocyte phenotype to collagen production and cell survival of cholangiocytes in vitro. Mouse cholangiocytes (603B cells) were treated with TGF-β1 and EMT-like phenotype alterations were monitored by morphological changes and expression of EMT-associated genes. Alterations in Col1A1 gene, Col1A1-associated miR-29s, and pro-apoptotic genes were measured in TGF-β1-treated 603B cells. Snail1 knockdown was achieved using shRNA to evaluate the contribution of EMT-associated changes to Col1A1 production and cell survival. We found TGF-β1 treatment induced partial EMT-like phenotype transition in 603B cells in a Snail1-dependent manner. TGF-β1 also stimulated collagen α1(I) expression in 603B cells. However, this induction was not parallel to the EMT-like alterations and independent of Snail1 or miR-29 expression. Cells undergoing EMT-like changes showed a modest down-regulation of multiple pro-apoptotic genes and displayed resistance to TNF-α-induced apoptosis. TGF-β1-induced apoptosis resistance was attenuated in Snail1 knockdown 603B cells. TGF-β1-induced Col1A1 production seems to be independent of EMT-like transition and miR-29 expression. Nevertheless, TGF-β1-induced EMT may contribute to the increased survival capacity of cholangiocytes via modulating the expression of pro-apoptotic genes. PMID:23236489

  12. Determination of aflatoxins in by-products of industrial processing of cocoa beans.

    PubMed

    Copetti, Marina V; Iamanaka, Beatriz T; Pereira, José Luiz; Lemes, Daniel P; Nakano, Felipe; Taniwaki, Marta H

    2012-01-01

    This study has examined the occurrence of aflatoxins in 168 samples of different fractions obtained during the processing of cocoa in manufacturing plants (shell, nibs, mass, butter, cake and powder) using an optimised methodology for cocoa by-products. The method validation was based on selectivity, linearity, limit of detection and recovery. The method was shown to be adequate for use in quantifying the contamination of cocoa by aflatoxins B(1), B(2), G(1) and G(2). Furthermore, the method was easier to use than other methods available in the literature. For aflatoxin extraction from cocoa samples, a methanol-water solution was used, and then immunoaffinity columns were employed for clean-up before the determination by high-performance liquid chromatography. A survey demonstrated a widespread occurrence of aflatoxins in cocoa by-products, although in general the levels of aflatoxins present in the fractions from industrial processing of cocoa were low. A maximum aflatoxin contamination of 13.3 ng g(-1) was found in a nib sample. The lowest contamination levels were found in cocoa butter. Continued monitoring of aflatoxins in cocoa by-products is nevertheless necessary because these toxins have a high toxicity to humans and cocoa is widely consumed by children through cocoa-containing products, like candies.

  13. Salvianolic Acid B (Sal B) Protects Retinal Pigment Epithelial Cells from Oxidative Stress-Induced Cell Death by Activating Glutaredoxin 1 (Grx1)

    PubMed Central

    Liu, Xiaobin; Xavier, Christy; Jann, Jamieson; Wu, Hongli

    2016-01-01

    Protein glutathionylation, defined as the formation of protein mixed disulfides (PSSG) between cysteine residues and glutathione (GSH), can lead to cell death. Glutaredoxin 1 (Grx1) is a thiol repair enzyme which catalyzes the reduction of PSSG. Therefore, Grx1 exerts strong anti-apoptotic effects by improving the redox state, especially in times of oxidative stress. However, there is currently no compound that is identified as a Grx1 activator. In this study, we identified and characterized Salvianolic acid B (Sal B), a natural compound, as a Grx1 inducer, which potently protected retinal pigment epithelial (RPE) cells from oxidative injury. Our results showed that treatment with Sal B protected primary human RPE cells from H2O2-induced cell damage. Interestingly, we found Sal B pretreatment upregulated Grx1 expression in RPE cells in a time- and dose-dependent manner. Furthermore, NF-E2-related factor 2 (Nrf2), the key transcription factor that regulates the expression of Grx1, was activated in Sal B treated RPE cells. Further investigation showed that knockdown of Grx1 by small interfering RNA (siRNA) significantly reduced the protective effects of Sal B. We conclude that Sal B protects RPE cells against H2O2-induced cell injury through Grx1 induction by activating Nrf2 pathway, thus preventing lethal accumulation of PSSG and reversing oxidative damage. PMID:27827892

  14. Immunoaffinity column cleanup with liquid chromatography using post-column bromination for determination of aflatoxins in peanut butter, pistachio paste, fig paste, and paprika powder: collaborative study.

    PubMed

    Stroka, J; Anklam, E; Jörissen, U; Gilbert, J

    2000-01-01

    A collaborative study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for the determination of aflatoxin B1 and total aflatoxins at European regulatory limits. The test portion is extracted with methanol-water (8 + 2) for dried figs and paprika, and with methanol-water (8 + 2) plus hexane (or cyclohexane) for peanut butter and pistachios. The sample extract is filtered, diluted with phosphate buffer saline, and applied to an immunoaffinity column. The column is washed with water and the aflatoxins are eluted with methanol. Aflatoxins are quantitated by reversed-phase LC with post-column derivatization (PCD) involving bromination. PCD is achieved with either an electrochemical cell (Kobra cell) and addition of bromide to the mobile phase or pyridinium hydrobromide perbromide. Determination is by fluorescence. Peanut butter, pistachio paste, dried fig paste, and paprika powder samples, both naturally contaminated with aflatoxins and containing added aflatoxins, were sent to 16 collaborators in 16 European countries. Test portions of samples were spiked at levels of 2.4 and 9.6 ng/g for total aflatoxins which included 1.0 and 4.0 ng/g aflatoxin B1, respectively. Recoveries for total aflatoxins ranged from 71 to 92% with corresponding recoveries for aflatoxin B1 of 82 to 109%. Based on results for spiked samples (blind duplicates at 2 levels) as well as naturally contaminated samples (blind duplicates at 4 levels, including blank), the relative standard deviation for repeatability ranged from 4.6 to 23.3% for total aflatoxins and from 3.1 to 20.0% for aflatoxin B1. The relative standard deviation for reproducibility ranged from 14.1 to 34.2% for total aflatoxins, and from 9.1 to 32.2% for aflatoxin B1. The method showed acceptable within-laboratory and between-laboratory precision for all 4 matrixes, as evidenced by HORRAT values <1, at the low levels of determination for both total aflatoxins and

  15. Chicken Fetal Liver DNA Damage and Adduct Formation by Activation-Dependent DNA-Reactive Carcinogens and Related Compounds of Several Structural Classes

    PubMed Central

    Williams, Gary M.; Duan, Jian-Dong; Brunnemann, Klaus D.; Iatropoulos, Michael J.; Vock, Esther; Deschl, Ulrich

    2014-01-01

    The chicken egg genotoxicity assay (CEGA), which utilizes the liver of an intact and aseptic embryo-fetal test organism, was evaluated using four activation-dependent DNA-reactive carcinogens and four structurally related less potent carcinogens or non-carcinogens. In the assay, three daily doses of test substances were administered to eggs containing 9–11-day-old fetuses and the fetal livers were assessed for two endpoints, DNA breaks using the alkaline single cell gel electrophoresis (comet) assay and DNA adducts using the 32P-nucleotide postlabeling (NPL) assay. The effects of four carcinogens of different structures requiring distinct pathways of bioactivation, i.e., 2-acetylaminofluorene (AAF), aflatoxin B1 (AFB1), benzo[a]pyrene (B[a]P), and diethylnitrosamine (DEN), were compared with structurally related non-carcinogens fluorene (FLU) and benzo[e]pyrene (B[e]P) or weak carcinogens, aflatoxin B2 (AFB2) and N-nitrosodiethanolamine (NDELA). The four carcinogens all produced DNA breaks at microgram or low milligram total doses, whereas less potent carcinogens and non-carcinogens yielded borderline or negative results, respectively, at higher doses. AAF and B[a]P produced DNA adducts, whereas none was found with the related comparators FLU or B[e]P, consistent with comet results. DEN and NDELA were also negative for adducts, as expected in the case of DEN for an alkylating agent in the standard NPL assay. Also, AFB1 and AFB2 were negative in NPL, as expected, due to the nature of ring opened aflatoxin adducts, which are resistant to enzymatic digestion. Thus, the CEGA, using comet and NPL, is capable of detection of the genotoxicity of diverse DNA-reactive carcinogens, while not yielding false positives for non-carcinogens. PMID:24973097

  16. Chicken fetal liver DNA damage and adduct formation by activation-dependent DNA-reactive carcinogens and related compounds of several structural classes.

    PubMed

    Williams, Gary M; Duan, Jian-Dong; Brunnemann, Klaus D; Iatropoulos, Michael J; Vock, Esther; Deschl, Ulrich

    2014-09-01

    The chicken egg genotoxicity assay (CEGA), which utilizes the liver of an intact and aseptic embryo-fetal test organism, was evaluated using four activation-dependent DNA-reactive carcinogens and four structurally related less potent carcinogens or non-carcinogens. In the assay, three daily doses of test substances were administered to eggs containing 9-11-day-old fetuses and the fetal livers were assessed for two endpoints, DNA breaks using the alkaline single cell gel electrophoresis (comet) assay and DNA adducts using the (32)P-nucleotide postlabeling (NPL) assay. The effects of four carcinogens of different structures requiring distinct pathways of bioactivation, i.e., 2-acetylaminofluorene (AAF), aflatoxin B1 (AFB1), benzo[a]pyrene (B[a]P), and diethylnitrosamine (DEN), were compared with structurally related non-carcinogens fluorene (FLU) and benzo[e]pyrene (B[e]P) or weak carcinogens, aflatoxin B2 (AFB2) and N-nitrosodiethanolamine (NDELA). The four carcinogens all produced DNA breaks at microgram or low milligram total doses, whereas less potent carcinogens and non-carcinogens yielded borderline or negative results, respectively, at higher doses. AAF and B[a]P produced DNA adducts, whereas none was found with the related comparators FLU or B[e]P, consistent with comet results. DEN and NDELA were also negative for adducts, as expected in the case of DEN for an alkylating agent in the standard NPL assay. Also, AFB1 and AFB2 were negative in NPL, as expected, due to the nature of ring opened aflatoxin adducts, which are resistant to enzymatic digestion. Thus, the CEGA, using comet and NPL, is capable of detection of the genotoxicity of diverse DNA-reactive carcinogens, while not yielding false positives for non-carcinogens. © The Author 2014. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  17. Dip-strip method for monitoring environmental contamination of aflatoxin in food and feed: use of a portable aflatoxin detection kit.

    PubMed Central

    Sashidhar, R B

    1993-01-01

    Aflatoxin contamination of food and feed have gained global significance due to its deleterious effect on human and animal health and its importance in the international trade. The potential of aflatoxin as a carcinogen, mutagen, teratogen, and immunosuppressive agent is well documented. The problem of aflatoxin contamination of food and feed has led to the enactment of various legislation. However, meaningful strategies for implementation of this legislation is limited by nonavailability of simple, cost-effective method for screening and detection of aflatoxin under field conditions. Keeping in mind the analytical constraints in developing countries, a simple-to-operate, rapid, reliable, and cost-effective portable aflatoxin detection kit has been developed. The important components of the kit include a hand-held UV lamp (365 nm, 4 W output), a solvent blender (12,000 rpm) for toxin extraction, and adsorbent-coated dip-strips (polyester film) for detecting and quantifying aflatoxin. Analysis of variance indicates that there were no significant differences between various batches of dip-strips (p > 0.05). The minimum detection limit for aflatoxin B1 was 10 ppb per spot. The kit may find wide application as a research tool in public health laboratories, environmental monitoring agencies, and in the poultry industry. Images FIGURE 1. PMID:8143644

  18. Aflatoxin contamination in foods and foodstuffs in Tokyo: 1986-1990.

    PubMed

    Tabata, S; Kamimura, H; Ibe, A; Hashimoto, H; Iida, M; Tamura, Y; Nishima, T

    1993-01-01

    Aflatoxins were determined in 3054 samples of foods or foodstuffs, including cereals, nuts, beans, spices, dairy products, dry fruits, and edible oil. Samples were collected in Tokyo from 1986 to 1990. Aflatoxins were found in rice products, adlay, corn, crude sugar, peanut products, pistachio nuts, brazil nuts, sesame products, butter beans, white pepper, red pepper, paprika, nutmeg, and mixed spices. The highest incidence of aflatoxin contamination was observed in nutmeg (80%), and the highest level of aflatoxin B1 was observed in pistachio nuts (1382 ppb).

  19. Analysis of cocoa products for ochratoxin A and aflatoxins.

    PubMed

    Turcotte, Anne-Marie; Scott, Peter M; Tague, Brett

    2013-08-01

    Eighty-five samples of cocoa products sampled in Canada were analysed for ochratoxin A (OTA) and aflatoxins in 2011-2012. Inclusion of the aflatoxins in this survey required additional method development. Chocolate was extracted with methanol-water plus NaCl, while for cocoa two successive extractions with methanol and methanol-water were made. Extracts were cleaned on an AflaOchra immunoaffinity column (IAC). Determination was by reversed phase high performance liquid chromatography (HPLC). Detection of the aflatoxins was with a post-column photochemical reactor and of OTA by fluorescence detection. Mean limits of quantification (LOQ) of chocolate and cocoa powders were 0.16 ng/g (OTA) and 0.07 ng/g (aflatoxin B1), respectively. Survey results showed that the incidences of OTA above the LOQ in natural cocoa were 15/15 (mean 1.17 ng/g), 20/21 for alkalized cocoa (mean 1.06 ng/g), 9/9 for baking chocolate (mean 0.49 ng/g), 20/20 for dark chocolate (mean 0.39 ng/g), 7/10 for milk chocolate (mean 0.19 ng/g), 5/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. These results confirm our previous work with OTA. In the same samples, incidences of aflatoxin B1 above the LOQ were 14/15 for natural cocoa (mean 0.86 ng/g), 20/21 for alkalized cocoa (mean 0.37 ng/g), 7/9 for baking chocolate (mean 0.22 ng/g), 16/20 for dark chocolate (mean 0.19 ng/g), 7/10 for milk chocolate (mean 0.09 ng/g), 4/5 for cocoa liquor (mean 0.43 ng/g), and 0/5 for cocoa butter. Both aflatoxins and OTA were confirmed by HPLC-MS/MS when OTA or aflatoxin levels found were above 2 ng/g in cocoa.

  20. Aflatoxin in detannin coffee and tea and its destruction.

    PubMed

    Hasan, H A H

    2002-05-01

    The aflatoxins produced byAspergillus parasiticus var. globosus IMI 12090 in detannin-caffeinated coffee and black tea were five times more concentrated than in regular coffee and tea. The activity of caffeine and tannin on the fungus growth and aflatoxin production in liquid broth was tested at three levels: viz. 0.1, 0.3, and 0.6%. Tannin and caffeine induced 95% inhibition in aflatoxins at 0.3% and 0.6%, respectively. The antiaflatoxigenic properties of regular coffee and tea appear to be due to tannin, followed by caffeine. The roasting of contaminated coffee beans at 200 degrees C for 20 min is effective in the destruction of aflatoxins.