Different Concentrations Of Aflatoxin B1 Research Articles

A multicolor aptasensor based on target-induced anti-etching gold nanorod for visible detection of aflatoxin B1

Foods contaminated with aflatoxin B1 (AFB1) pose a serious threat to humans, so rapid and sensitive detection of AFB1 plays an important role in public health. In this work, we developed a multicolor aptasensor based on target-mediated anti-etching of gold nanorods (AuNRs) for on-site visible detection of AFB1. The target was used as a competitor that competed with a cDNA probe labeled by horse radish peroxidase (cDNA-HRP) to bind with the aptamer coated in a microplate. The sensing mechanism was based on regulating the etching process of AuNRs upon the addition of different concentrations of AFB1. In the absence of AFB1, the AuNRs solution undergoes a distinct change from red to pink due to the etching effect of TMB2+ generated by a specific redox reaction catalyzed by the cDNA-HRP. However, in the presence of AFB1, the specific binding between AFB1 and aptamer made cDNA-HRP free out of the microplate, inhibiting the etching of AuNRs. Consequently, a redshift of longitudinal localized surface plasmon resonance (LSPR) and color alterations can be observed along with increasing the concentration of AFB1. The LSPR for visual estimation limits were 0.46 ng/mL. A good linear relationship between the wavelength shift and the concentration of AFB1 was achieved from violet to light green in the range of 1–1000 ng/mL. The established aptasensor displayed excellent detection performance, which was suitable for on-site analysis of AFB1 in actual food samples, and provided a new idea of immediate visualization detection for food hazardous factors.

Effect of Different Levels of Aflatoxin B1 in Concentrated Livestock Diets on Some Characteristics of Rumen Fluid in Laboratory with Sodium Bentonite

This study was conducted in the nutrition laboratory of the Department of Animal Production in the College of Agricultural Engineering Sciences at the University of Baghdad, Al-Jadriya. The experiment lasted for 90 days to investigate the effects of adding sodium bentonite (SB) to a concentrated diet contaminated with aflatoxin B1 on some characteristics of rumen fluid in a laboratory setting. Four diets were prepared, each contaminated with different concentrations of aflatoxin B1 (0, 20, 40, 60 ppb), and four different percentages of sodium bentonite (0, 3, 5, 7%) were added to each ration. The results showed a significant increase in pH values among the different treatments, with the diet contaminated with 40 ppb aflatoxin B1 (T3) recording the highest acidity compared to the control treatment at 0% concentration. However, there were no significant differences in pH rates between the treatments. The pH rates of 7% sodium bentonite addition did not differ significantly from 5% and 0%, while 3% had the lowest pH rate. Regarding volatile fatty acids (VFA), there was a significant increase in their percentage with the increase in sodium bentonite concentrations. The second treatment contaminated with 20 ppb aflatoxin B1 recorded the highest VFA percentage among the treatments, while the fourth treatment (T4) at 0% concentration recorded the lowest VFA percentage. Significant differences in VFA percentages were observed among all the treatments, with the highest VFA percentage in T2, followed by T3 and T1, while T4 recorded the lowest percentage. The analysis of ammonia nitrogen values showed a significant increase among the different treatments, with T2 recording the highest value among the treatments, and T4 at 0% concentration recording the lowest ammonia nitrogen value. Significant differences in the average percentage of ammonia nitrogen were observed among all the experimental treatments, with the highest value in T2, followed by T1 and T3, while T4 recorded the lowest percentage. The total number of microorganisms showed a significant increase with the increase in sodium bentonite concentrations, with T2 at 7% concentration having the highest number of microorganisms, while T4 at 0% concentration recorded the lowest number. Significant differences in the average total number of microorganisms were observed among all the experimental treatments, with T2 having the highest number, followed by T1 and T3, while T4 had the lowest number.

Ameliorated effect of Lactobacillus plantarum SCS2 on the oxidative stress in HepG2 cells induced by AFB1

Aflatoxin B1 (AFB1), a widespread mycotoxin found in food, poses a significant threat to the food industry and production. The primary target of AFB1 is the liver. Probiotic-mediated antitoxicity has been proposed to overcome aflatoxin toxicity. In the present study, to investigate the protective effects and molecular mechanisms of Lactobacillus plantarum SCS2 (L. plantarum SCS2) against HepG2 cells oxidative stress induced by AFB1. HepG2 cells were cultured and treated with different concentrations of AFB1 (0-10 μmol/L) to induce oxidative stress and L. plantarum SCS2 (total protein, 0.4 mg/mL) was pretreated HepG2 for 2 h and then supplemented with AFB1 (10 μmol/L) for 22 h to observe cellular oxidative stress. The cytotoxicity, reactive oxygen species, malondialdehyde, activities and mRNA expressions of antioxidant enzymes and activation of the Nrf2 signaling pathway were measured. The results showed that AFB1 exposure caused oxidative stress in HepG2. Supplementation of L. plantarum SCS2 prevented AFB1-induced HepG2 cells activity decreased and activated antioxidant enzymes activity by inhibiting Nrf2 over expression. The results of in vitro experiments revealed that L. plantarum SCS2 evidently protected against AFB1-induced oxidative stress.

Selenium ameliorates aflatoxin B1-induced uterine injury in female mice and necrosis of human endometrial microvascular endothelial cells.

This study aimed to observe the effects of Selenium (Se) on aflatoxin B1 (AFB1)-induced uterine injury in female mice and necrosis of human endometrial microvascular endothelial cells (HEMECs). Fifty female mice were randomly divided into control group; AFB1 group; Se group (0.2 mg/kg each day); AFB1 + Se group; and positive control group. After continuous treatment for 30 days, uterine tissues were harvested, which were used for hematoxylin and eosin (H&E) staining. Necrosis-related proteins including RIPK1, RIPK3, and MLKL were examined in uterine tissues using western blot and immunohistochemistry. HEMECs were treated with different concentrations of AFB1 or Se, which were used for selecting the optimal concentrations. RIPK1, RIPK3 and MLKL expression was detected in HEMECs exposed to AFB1 and/or Se via western blot and immunofluorescence. H&E staining results showed that AFB1 induced uterine injury of female male, which was ameliorated by Se treatment. According to western blot and immunohistochemistry, RIPK1, RIPK3, and MLKL expression was distinctly increased in uterine tissues of AFB1-treated mice, which was decreased by Se treatment. Cell viability of HEMECs was gradually lowered as the concentrations of AFB1and Se increased. A 10-μM AFB1 exposure significantly increased RIPK1, RIPK3, and MLKL expression in HEMECs, which was improved following co-treatment with 5-μM Se. Thus, our findings revealed that Se may ameliorate AFB1-induced uterine injury in female mice and necrosis of HEMECs.

Design and development of an on-line fluorescence spectroscopy system for detection of aflatoxin in pistachio nuts

The on-line detection for aflatoxins contaminated agricultural products using fluorescence spectroscopy method is still a significant challenge due to its weak signal intensity and limitation of detection mode. Thus, the study for real-time aflatoxin B1 (AFB1) contaminated pistachios detection by a self-built laser induced fluorescence spectroscopy (LIFS) system coupled with three collection probes was investigated. A total of 600 kernels consisting of two pistachio varieties including "Yaoshengji" and "Wanlong" were artificially contaminated with five different concentrations of AFB1 (5, 10, 20, 30, and 50 μg kg−1). Good accuracy (> 91.0%) was obtained for classifying single variety of pistachios contaminated with low concentration of AFB1 even for 5 μg kg−1 by linear discriminant analysis (LDA), irrespective of varietal differences. The partial least square regression analysis (PLS) model based on the variation selection method of competitive adaptive reweighted sampling (CARS) presented a considerable predictive precision (root mean square error of prediction (RMSEP) < 6.20 μg kg−1) for pooled lot using 174–1100 nm at a detection speed of 1 kernel s-1. Despite the preliminary feasibility for detection AFB1 in pistachio kernels, the on-line LIFS system should be validated further using naturally contaminated samples.

Novel Enzyme-Linked Aptamer Assay for the Determination of Aflatoxin B1 in Peanuts

A novel enzyme-linked aptamer assay is reported for the determination of aflatoxin B1 (AFB1). AFB1 can competitively bind with the immobilized biotin-aptamer and release biotin complementary DNA, leading to the gradual fading of the detection system color with increasing of AFB1 concentration. In the absence of AFB1, the biotinylated complementary DNA is not be released from the fixed aptamer. Therefore, the enzyme reaction occurs in the detection system. Under the optimized experimental conditions, the proposed method possessed a wide linear range for AFB1 from 1 to 80 ng/mL (R2 of 0.990) with a low detection limit of 0.36 ng/mL. The method was then applied to detect uncontaminated peanuts fortified with different concentrations of AFB1. The recovery values were from 82.60% to 94.43%, which indicated the proposed method may be used to detect AFB1 in food and has potential for the development of test kits.

Detection of Aflatoxin B1 in Pistachio Kernels Using Visible/Near-Infrared Hyperspectral Imaging

Abstract. Pistachios are susceptible to aflatoxin contamination because of their rich nutrient content. Hyperspectral imaging (HSI), a new method for collecting spectral and image information, has been successfully employed in contamination research to classify staple agricultural products, such as maize, that are contaminated with aflatoxins. However, only a few studies have been conducted on the nondestructive discrimination among contaminated nuts using HSI for both qualitative and quantitative purposes. Thus, the feasibility of directly detecting aflatoxin B1 (AFB1) in individual pistachio kernels using visible/near-infrared HSI (VNIR HSI) was explored in this study. A total of 300 pistachio kernels were randomly selected to prepare target samples that were artificially contaminated with 5, 10, 20, 30, or 50 ppb (parts per billion) of AFB1. Principal component analysis (PCA) showed an overall separation trend between the control and all contaminated kernels. Accuracies greater than 90.0% were obtained by linear discriminant analysis (LDA) for samples that were artificially contaminated with different concentrations of AFB1 based on spectra at 694 to 988 nm that had been preprocessed with standard normal variate (SNV) and Savitzky-Golay (SG) smoothing. The correlation coefficients of calibration and validation (rc and rv) from stepwise multiple linear regression (SMLR) models were all &amp;gt;0.9100. Moreover, five key wavelengths (708, 771, 892, 915, and 941 nm) closely associated with AFB1 contamination were identified using principal component spectra analysis. Generally, the results indicated that VNIR HSI could be employed for preliminary screening of pistachio kernels that were artificially contaminated with AFB1, even at the 5 ppb level. However, the quantitative prediction of the specific AFB1 concentration needed to be further improved. Keywords: Aflatoxins, Detection analysis, Hyperspectral information, Pistachios, Visible/near-infrared.

Effects of yeast based toxin binder supplementation on growth performance and intestinal microarchitecture in male buffalo calves (Bubalus bubalis) exposed to different concentrations of Aflatoxin B1.

The purpose of study was to analyze the effects of yeast based toxin binder (YTB) on growth performance and intestinal microarchitecture of buffalo calves exposed to different concentrations of aflatoxin B1 (AFB1). Male buffalo calves (n=72) having no disease exposure in the last four months, aged 10 ± 1 months and weighing 150 ± 12.22 kg (Mean ± standard deviation)were purchased from a buffalo calf fattening farm and divided in 8 equal groups (n=9 animals in each group). Animals that were fed common basal diet and basal diet along with yeast cell wall based toxin binder (YTB) and had no exposure to AFB1 were served as controls. Different concentrations of AFB1 (6mg, 8mg and 10mg per kg of feed) were given to 27 animals, similarly 2mg of YTB (per kg of contaminated feed) was added to concentrate with different AFB1 concentrations and fed to another 27 animals. Average daily gain of each animal was recorded. Animals were slaughtered after 27 days and intestinal segments from duodenum, jejunum, ileum and colon were collected which were stained with hematoxylin/eosin or combined alcian blue and periodic acid schiff stain. Average daily gain and intestinal morphometric parameters (villus height, villus width, crypt depth and surface area) in all the selected intestinal segments decreased (P&lt;0.05) whereas, intraepithelial lymphocytes, goblet cells (having acidic and neutral mucins) and proliferative cells increased (P&lt;0.05) with AFB1 exposure. It was also observed that YTB neutralized (P&lt;0.05) the harmful effects of AFB1 on intestinal morphology, boosted activity of goblet cells and decreased (P&lt;0.05) number of proliferative cells. It was concluded that YTB supplementation is advantageous in eliminating the negative effects of AFB1 on gut of buffalo calves.

The effects of aflatoxin B1 on growth hormone regulated gene-1 and interaction between DNA and aflatoxin B1 in broiler chickens during hatching

Many types of aflatoxin cause problems for both public and animal health. Aflatoxin B1 (AFB1) is the most toxic and commonly encountered fungal toxin that appears in poultry feed and in feeds stored under unsuitable conditions. AFB1 decreases feed quality, egg production and fertility of hatching eggs. Also, AFB1 alters the development of embryos by infecting eggs. We investigated using sequence analysis the changes caused by different concentrations of AFB1 on the promoter sequences of the growth hormone regulated gene-1 (GHRG-1) in chick embryo at 13, 17, 19 and 21 days incubation. DNA isolated from the liver of chick embryos treated with different concentrations of AFB1 was separated using agarose gel electrophoresis to detect apoptosis, and DNA interaction with AFB1 was investigated using plasmids to detect changes in electrophoretic mobility and their effects on DNA. Base changes of the promoter sequences of GHRG-1 in 5 ng/egg, 15 ng/egg and 40 ng/egg doses of AFB1 were increased on day 19 compared to base changes of the same AFB1 doses on day 13. We also found that AFB at different concentrations changed the mobility of DNA by binding to it, and that high doses of AFB1 destroyed DNA. The DNA interaction study using plasmid demonstrated that AFB1 at high doses was bound to plasmid DNA, slowed its mobility and inhibited restriction cuts.

Plasmonic ELISA based on enzyme-assisted etching of Au nanorods for the highly sensitive detection of aflatoxin B1 in corn samples

In this work, we reported an advanced enzyme-assisted etching method using gold nanorods (AuNRs) as signal output of plasmonic enzyme-linked immunosorbent assay (pELISA) for the quantitative or qualitative detection of aflatoxin B1 (AFB1) in real corn samples. AFB1-labeled glucose oxidase was adopted as the competing antigen to catalyze glucose into H2O2. Meanwhile, horseradish peroxidase was employed to catalyze H2O2 and hence generate .OH. The AuNRs with the aspect ratio of 2:1 were chemically etched by OH to a rod-like morphology. This occurrence reduced the optical density at 650 nm and resulted in a vivid color response from bluish-green to red easily detectable by a microplate reader for quantitative AFB1 analysis or by the naked eye for qualitative AFB1 detection. Various parameters that may influence the detection performance of pELISA were investigated. The proposed method showed an extremely high sensitivity for qualitative AFB1 detection, with a visible cut-off value of 12.5 pg/mL. The technique also achieved a good linear range of 3.1–150 pg/mL for quantitative AFB1 analysis, with a half-maximal inhibitory concentration of 22.3 pg/mL, which was approximately 32 folds lower than those of conventional ELISA (707 pg/mL). The average recoveries for corn samples spiked with different concentrations of AFB1 ranged from 82% to 115%, with a coefficient of variation that ranged from 2% to 13%. These values corresponded to an acceptable accuracy and precision for the proposed method. In addition, the reliability of the proposed method was further confirmed by the liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. In brief, this work offers an improved screening strategy with high sensitivity and robustness for the qualitative or quantitative detection of mycotoxins or other pollutants in food safety monitoring.

Immunotoxicity of ochratoxin A and aflatoxin B1 in combination is associated with the nuclear factor kappa B signaling pathway in 3D4/21 cells

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.

Aflatoxin B1 Induced Compositional Changes in Gut Microbial Communities of Male F344 Rats.

Aflatoxins are a group of potent foodborne toxicants naturally occurring in maize and groundnuts. Differential species-specific sensitivity to aflatoxins has been documented but cannot be fully explained by the differences in metabolism of these toxicants among animal species. Commensal microbial communities (microbiota) are critical to human and animal health, but few studies have assessed interactions between xenobiotic toxins and those microbiota, and its potential effects to humans and animals. Here, an exploratory dosing experiment was conducted to explore effects of Aflatoxin B1 (AFB1) on the gut microbiota in a commonly used rat model. Male F344 rats were randomly divided into groups and treated with different concentrations of AFB1. Microbial communities in fecal samples were assessed using 16S rRNA sequence analysis. We found that samples from the control group had a phylogenetically diverse community, and that increasing AFB1 doses decreased this diversity but increased evenness of community composition. In addition, the gut microbiota from different samples was clustered according to their dosing regimens. There is no community shift at the phylum level but some lactic acid bacteria were significantly depleted by AFB1. These findings suggested that AFB1 could modify the gut microbiota in a dose-dependent manner.

Electrochemical piezoelectric reusable immunosensor for aflatoxin B1 detection

A competitive, reusable, reliable and sensitive immunosensor based on electrochemical quartz crystal microbalance (EQCM) was developed for the detection of aflatoxin B1 (AFB1). The sensing platform was developed using a primary monoclonal anti-aflatoxin antibody (anti AFB1) covalently immobilized on a monolayer of 4-amino thiophenol self assembled on a gold coated quartz crystal electrode (anti AFB1/4-ATP/Au). The reusability was achieved by a sandwiched system using secondary rabbit-immunoglobulin antibodies (r-IgGs) tagged with core shell of gold coated iron oxide nanoparticles (r-IgG–Au-Fe3O4), which can be used to regenerate the bioelectrode. A competitive mode was employed, between free and coated AFB1 for fixed concentration of nanoparticle conjugate (r-IgG–Au–Fe3O4). After the competitive interaction, immunoelectrode was washed with phosphate saline buffer (PBS) at pH 7.4 and examined by electrochemical quartz crystal microbalance-cyclic voltammetry (EQCM-CV). Under the optimized conditions, the fabricated immunosensor can be used to detect AFB1 quantitatively with a linear range of 0.05 to 5ngmL−1. For reliability, the fabricated immuno sensor was tested using cereal samples spiked with different concentrations of AFB1. In addition, this immuno electrode displays good reproducibility, and storage stability. The above immunosensor was regenerated with negligible loss in activity through removal of the r-IgG–Au–Fe3O4 conjugate using a strong magnet. It is shown that theAFB1/BSA/aAFB1/4-ATP/Au immuno sensor is a new approach for developing sensitive reliable and reusable electrochemical piezoelectric immunosensors.

Efficacy of beer fermentation residue containing Saccharomyces cerevisiae cells for ameliorating aflatoxicosis in broilers

This study aimed to determine the aflatoxin B1 (AFB1) binding capacity of a beer fermentation residue (BFR) containing Saccharomyces cerevisiae cells, and the efficacy of BFR to ameliorate the toxic effects of AFB1 on performance, serum biochemistry, and histology of broilers. The BFR was collected from a microbrewery, and the yeast cells were counted, dried, and milled before it was used in the study. In vitro evaluation of the BFR was conducted using different concentrations of AFB1 (2.0, 4.0, 8.0, 16.0, and 32.0 μg AFB1/mL) and 100 mg/10 mL of BFR at pH 3.0 or 6.0. Two hundred 1-day-old male broilers (Ross 308) were assigned to chick batteries and allowed ad libitum access to feed and water. A completely randomized design was used with 5 replicate pens of 5 chicks assigned to each of 4 dietary treatments from hatch to 21 d, which included: 1) basal diet (BD), with no BFR or AFB1; 2) BD supplemented with 1% BFR; 3) BD supplemented with 2 mg AFB1/kg of feed; and 4) BD supplemented with 2 mg AFB1/kg feed and 1% BFR. Performance variables were determined weekly, while serum analyses were performed on d 14 and 21. At the end of the study, chicks were anesthetized with carbon dioxide, euthanized by cervical dislocation, and the kidney, liver, and bursa of Fabricius were removed for determination of relative weights, and for histological evaluation. In vitro assays showed that the higher the initial AFB1 concentration in solution, the greater the AFB1 amount adsorbed by BFR at both pHs tested. Feed intake, BW gain, and concentrations of albumin, total protein, and globulin increased (P < 0.05) in broilers fed BFR+AFB1 (Diet 4), when compared to the birds receiving only AFB1 (Diet 2). Although BFR was not able to reduce or prevent the effects of AFB1 on relative weights of kidneys and liver, it reduced the severity of histological changes in the liver and kidney caused by AFB1.

Aqueous extract of Corchorus olitorius decreases cytotoxicity of aflatoxin B1and fumonisin B1in H4IIE-luc cells

75 ABSTRACT Aim: Afl atoxin B1 (AFB1) and fumonisin B1 (FB1) are important food-borne mycotoxins. Co-contamination of foodstuffs with these two mycotoxins is well-known and has been implicated in a possible development of hepatocellular carcinoma in humans living in regions of the world where exposures to these mycotoxins in grain are greatest. The aim of the current study was to evaluate the potential protective effects of an aqueous extract of Cochorus olitorius (C. olitorius, moroheiya) against cytotoxicity of AFB1 and/or FB1 in H4IIE-luc rat hepatoma cells, using assays to measure cell viability and disruption of DNA integrity. Although this transactivation assay was originally developed to specifi cally respond to aryl hydrocarbon agonists, this cell line was used because of its hepatic origin. Methods: H4IIE-luc cells were incubated with different concentrations of AFB1 and/or FB1 for 24 and 48 h with or without aqueous extract of C. olitorius. Results: Both mycotoxins decreased cell viability and increased DNA damage. Cytotoxicity was more pronounced when cells were exposed simultaneously to AFB1 and FB1. Conclusion: Aqueous extract of C. olitorius protected cells against cytotoxicity of mycotoxins. C. olitorius contains a water-soluble, natural chemo-preventative agent for cancer that should be isolated and identifi ed.

Functionalised Au Coated Iron Oxide Nanocomposites Based Reusable Immunosensor for AFB1 Detection

A reusable sandwiched electrochemical piezoelectric immunosensor has been developed for aflatoxin B1 (AFB1) detection using gold coated iron oxide core‐shell (Au‐Fe3O4) nanostructure. The monoclonal anti‐aflatoxin antibody (aAFB1) was immobilized on self‐assembled monolayer of 4‐aminothiophenol on gold coated quartz crystal to fabricate immunoelectrode (BSA/aAFB1/4‐ATP/Au). In addition, secondary rabbit‐immunoglobulin antibodies (r‐IgGs) functionalized with Au‐Fe3O4 NPs via cysteamine (r‐IgG‐Cys‐Au‐Fe3O4) were allowed to interact with AFB1. Both competitive and noncompetitive strategies were employed and a competition between coated AFB1 and free AFB1 was carried out. The competitive mode shows higher linear range (0.05 to 5 ng mL−1) than the noncompetitive one (0.5 to 5 ng mL−1), high sensitivity 335.7 µA ng−1 mL cm−2, and LOD 0.07 ng mL−1. The fabricated immunosensor has been tested using cereal samples spiked with different concentrations of AFB1. The developed competitive immunoelectrode displays good reproducibility, and storage stability and regenerated with negligible loss in activity through removal of the r‐IgG‐Cys‐Au‐Fe3O4 conjugate using a strong external magnet.

In-vitro cytotoxicity of aflatoxin B1 to broiler lymphocytes of broiler chickens

The aim of the present work was to study the in-vitro cytotoxic effects of different concentrations of aflatoxin B1 (AFB1) on broiler lymphocytes. Lymphocyte-rich mononuclear cells were separated by Ficoll-Histopaque density and cultured in 96-wellplates containing the evaluated AFB1 concentrations in 5% CO2 atmosphere at 39°C. Thereafter, MTT, PicoGreen, and reactive oxygen species assays were performed. Cell viability decreased in the presence of 10 µg/mL AFB1 at 48 h (p < 0.05) and of 10 and 20 µg/mL AFB1 at 72 h (p < 0.01 and p < 0.001, respectively) when compared to the control (0 µg/mL). However, a dose-dependent increase in the cell-free DNA at 24 h was observed at 1, 10 and 20 µg/mL (p < 0.001). ROS formation significantly increased at 24 h at all concentrations (p < 0.001). The in-vitro results demonstrate that AFB1 is cytotoxic and causes biomolecular oxidative damage in broiler lymphocytes.

Efficiency and safety evaluation of photodegradation of Aflatoxin B1 on peanut surface

SummaryA photodegradation study of Aflatoxin B1 (AFB1) on peanut surface was performed under UV irradiation at different UV irradiation intensities. With an initial quantity of 10 ng of AFB1, it can be degraded thoroughly within 80 min under the intensity of 800 μw cm−2. The photodegradation pathway of AFB1 on the peanut surface was proposed. Mutagenesis and cytotoxicity were assessed after exposure to different concentrations of AFB1 and the mixtures of photodegradation products on the peanut surface. The results of the Ames and in vitro cytotoxicity assay, providing clues to the assessment of safety issues of UV method applied in AFB1 decontamination, indicate that photodegradation products on the peanut surface has no toxicity, which can be explained by the differences in the chemical nature of the test compounds before and after UV irradiation.

In vitro toxicity of aflatoxin B1 and its photodegradation products in HepG2 cells

The effects of aflatoxin B1 (AFB1) were studied using the HepG2 cell line. Cytotoxicity, apoptosis and p53 expression were assessed after exposure to different concentrations of AFB1 (0-100 μM) and its two types of degradation products, namely the mixtures of photodegradation products in water (Pw) and the mixtures of photodegradation products in peanut oil (Po) for different time periods (0, 24, and 48 h). After exposure of the HepG2 cells to these compounds for different times and concentrations, the cytotoxicity of Pw and Po decreased approximately 40 and 100% compared with AFB1, respectively. The expression of p53 protein decreased significantly in AFB1-exposed cells, decreased slightly in Pw-treated cells and did not decrease compared to the untreated cells. The results of the in vitro cytotoxicity assay indicate that Pw is less toxic than AFB1, and Po has almost no toxicity, which can be explained by the differences in the chemical nature of the various kinds of the test compounds.

Effects of Different Concentrations of Aflatoxin B on Ram Epididymal and Ejaculatory Sperm Viability and Motility in vitro

The present study was carried to observe the effect of aflatoxin on ovine sperm in vitro. Ram epididymal and ejaculatory sperm cells were added into media containing different concentrations of aflatoxin B1. When ram epididymal sperm were put in different concentrations of aflatoxin, one-hour post incubation in control group, 81.25% of sperm cells were alive of which, 82.88% were motile. Significantly (p < 0.05) higher than medium with 31.25 ppb and higher concentration. The lowest motility (15.93%) was observed in 62.5 ppb aflatoxin. Sperm viability did not significantly changed after 2nd and 3rd h incubation but significantly decreased in 4th and 5th h post incubation. However, motility was not altered in the 2nd, 3rd, 4th and 5th h post incubation. In ejaculatory sperm, one hour post-incubation, in control group, 83.24% of sperm were alive of which 93.98% were motile, significantly (p < 0.05) higher than motility in medium with 7.81 ppb and higher concentrations of aflatoxin but not different with 1.96 ppb aflatoxin. In 62.5 ppb aflatoxin, the lowest motility was observed (18.9%). Sperm motility patterns for both epididymal and ejaculatory sperm cells were different after incubation in different concentrations of aflatoxin. The results of the present experiment showed that aflatoxin could decrease sperm motility obtained from ejaculation or epididymis.