14C‐labeled aflatoxin B1 prepared with yeastlike cultures of Aspergillus parasiticus

Abstract

A simple method was developed to produce ¹⁴C‐labeled aflatoxin B₁ by using the yeastlike phase of Aspergillus parasiticus NRRL 2999. Yeastlike cultures resulted from absence of manganese in a synthetic medium. Sodium acetate‐1‐¹⁴C had a 0.22% average incorporation; sodium acetate‐1,2‐¹⁴C, 0.70%. The average yield of labeled B₁ was 10 mg/500 ml medium with an average specific activity of either 63.3 mCi/mole (C‐l label) or 194.3 mCi/mole (C‐1, 2 label).     

Evaluation of Saccharomyces cerevisiae strains as probiotic agent with aflatoxin B1 adsorption ability for use in poultry feedstuffs

In this study the aflatoxin B₁ (AFB₁) removal capacity, the tolerance to salivary and gastrointestinal conditions, autoaggregation and coaggregation with pathogenic bacteria of Saccharomyces cerevisiae strains isolated from broiler feces, were evaluated. Only four of twelve isolated strains were identified as Saccharomyces cerevisiae using molecular techniques. The results obtained in AFB₁ binding studies indicated that the amount of AFB₁ removed was both strain and mycotoxin-concentration dependent. Therefore, a theoretical model was applied in order to select the most efficient strain to remove AFB₁ in a wide range of mycotoxin concentration. The results indicated that S. cerevisiae 08 and S. cerevisiae 01 strains were the most efficient microorganisms in the mycotoxin removal. Viability on simulated salivary and gastrointestinal conditions was investigated and S. cerevisiae 08 strain showed the best results, achieving 98% of total survival whereas S. cerevisiae 01 reached only 75%. Autoaggregation and coaggregation assays showed S. cerevisiae 08 as the most appropriate strain, mainly because it was the unique strain able to coaggregate with the four bacterial pathogens assayed. Consequently, S. cerevisiae 08 is the best candidate for future in vivo studies useful to prevent aflatoxicosis. Further quantitative in vitro and in vivo studies are required to evaluate the real impact of yeast-binding activity on the bioavailability of AFB₁ in poultry. However, this study could be useful in selecting efficient strains in terms of AFB₁ binding and provide an important contribution to research into microorganisms with potential probiotic effects on the host.     

低分子量腐殖酸改性蒙脱土对黄曲霉素的吸附作用

采用振荡平衡法研究了相对分子质量<3 500的低分子量腐殖酸改性蒙脱土对黄曲霉素(AFB1)的吸附作用及机制.吸附动力学实验结果表明,0～24 h内,蒙脱土原土快速吸附AFB1,在2 h处吸附量即至最大值,而后吸附量持续降低,12 h后趋于稳定、吸附量为1.01μg.g-1;0～24 h内,改性蒙脱土对AFB1的吸附量则持续增加,12 h时吸附量达最大值(1.60μg.g-1),其后吸附达平衡.用腐殖酸改性后,蒙脱土对AFB1的等温吸附曲线由Langmuir型转变为Freundlich型,平衡浓度较高条件下,AFB1在改性蒙脱土上的吸附量明显高于蒙脱土原土.蒙脱土改性后晶层坍塌,出现不规则和絮状松散结构,与其表面聚集的腐殖酸形成较为稳定的"矿物-腐殖酸"复合体,增强了其对疏水性AFB1的吸附能力.随溶液pH值的增大,原土和改性蒙脱土对AFB1的吸附量均增大,但AFB1在改性蒙脱土上吸附量的增加幅度较大,这可能主要与溶液碱性的提高促进了AFB1与蒙脱土矿物的氢键及电性结合有关.研究结果可为进一步阐明AFB1在土壤中的界面过程和环境归趋提供依据.     

ELISA determination of Aflatoxin M1 in milk and dairy products in Pakistan

A highly sensitive competitive ELISA was standardized to determine aflatoxin M₁ (AFM₁) in milk and dairy samples randomly collected from Faisalabad city, using Riedel-de-Haen, Aflatoxin M₁ ELISA Systems, (Art–No. 45169). Standards of 5, 10, 25, 50, and 100 ng L^?1 AFM₁ in sample/standard buffer were used in optimizing anti-AFM₁ antibodies and peroxidase–AFM₁ conjugate. Linear standard curve showed increase in concentration (log C) accompanied by decrease in maximal absorbance (%B/B₀) values (%B/B₀ = 124.3 ? 45.0 log C) with highly significant negative correlation (?0.973). The middle of the test (50% B/B₀) was at 30 ng L^?1 with 6.7 ng L^?1 the lowest detection limit (90% B/B₀). Duplicate analysis of 41 samples showed AFM₁ levels below EC permissible limits (50 ng L^?1) with maximums of 40, 13, and 7.4 ng L^?1 in milk, yogurt and butter, respectively. However, possibility of exceeding limits cannot be excluded, and demands regular monitoring.     

Application of a LC–MS/MS method for multi-mycotoxin analysis in infant formula and milk-based products for young children commercialized in Southern Brazil

ABSTRACT

An analytical method based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) was validated and applied for the analysis of aflatoxin M₁ (AFM₁), ochratoxin A (OTA) and deoxynivalenol (DON) in infant formula and milk-based products for young children commercialized in Brazil. A total of 38 samples were evaluated, including 12 infant formula, 14 follow-on formula and 12 samples of milk-based products. AFM₁ was detected in 12 (32%) samples, and seven (18%) samples contained AFM₁ levels above the method limit of quantification in a concentration range between 0.013 and 0.067 ng mL^?1 (0.026 ± 0.019). Two samples of milk-based products exceeded the maximum level (ML) fixed by the European Union for AFM₁ in baby foods, however, all samples were in agreement with the levels established by the Brazilian regulation. OTA and DON were not detected in any of the analyzed samples.     

农产品中黄曲霉素B1的酶联免疫吸附分析法

使用牛血清白蛋白(BSA)与黄曲霉素B_1(AFB_1)的结合物免疫兔子获得了高特异性、高效价的抗AFB_1的抗体,用此抗体建立了测定AFB_1的酶联免疫吸附分析法(ELISA),最低检出限为1.0pg/m。应用于花生、花生酱、玉米和大豆等农产品中AFB_1分析,其回收率在81—116％范围,变异系数在1.51—10.65％之间。此法具有简单、快速和适于大批量样品分析等优点。     

Development of enterosorbents that can be added to food and water to reduce toxin exposures during disasters

Humans and animals can be exposed to mixtures of chemicals from food and water, especially during disasters such as extended droughts, hurricanes and floods. Drought stress facilitates the occurrence of mycotoxins such as aflatoxins B₁ (AfB₁) and zearalenone (ZEN), while hurricanes and floods can mobilize toxic soil and sediments containing important pesticides (such as glyphosate). To address this problem in food, feed and water, we developed broad-acting, clay-based enterosorbents that can reduce toxin exposures when included in the diet. In this study, we processed sodium and calcium montmorillonite clays with high concentrations of sulfuric acid to increase surface areas and porosities, and conducted equilibrium isothermal analyses and dosimetry studies to derive binding parameters and gain insight into: (1) surface capacities and affinities, (2) potential mechanisms of sorption, (3) thermodynamics (enthalpy) of toxin/surface interactions and (4) estimated dose of sorbent required to maintain toxin threshold limits. We have also used a toxin-sensitive living organism (Hydra vulgaris) to predict the safety and efficacy of newly developed sorbents. Our results indicated that acid processed montmorillonites were effective sorbents for AfB₁, ZEN and glyphosate, with high capacity and tight binding, and effectively protected hydra against individual toxins, as well as mixtures of mycotoxins.     

真菌毒素引起的氧化应激及其毒理学意义

真菌毒素是一类由真菌产生的次生代谢产物,由于其在自然界污染的普遍性和广泛的毒性作用,被给予了充分关注。从机理上入手来研究真菌毒素毒性作用的起因具有重要意义。真菌毒素根据结构分类可达400种之多,它们的机理也各有不同,但是越来越多的研究表明真菌毒素与氧化应激有重大关联。真菌毒素引起的氧化应激能引起细胞毒性作用,还参与了其他靶点的毒性反应。同时真菌毒素能干扰细胞抗氧化系统的功能,降低细胞对毒素的抵抗能力。本文主要综述了真菌毒素中与氧化应激有密切联系的几种毒素的毒性作用,重点阐述了其与氧化应激之间的关系,以期对真菌毒素的毒性机理有更进一步的认识。     

Influence of herbicide glyphosate on growth and aflatoxin B1 production by Aspergillus section Flavi strains isolated from soil on in vitro assay

The effect of six glyphosate concentrations on growth rate and aflatoxin B₁ (AFB₁) production by Aspergillus section Flavi strains under different water activity (a_W) on maize-based medium was investigated. In general, the lag phase decreased as glyphosate concentration increased and all the strains showed the same behavior at the different conditions tested. The glyphosate increased significantly the growth of all Aspergillus section Flavi strains in different percentages with respect to control depending on pesticide concentration. At 5.0 and 10 mM this fact was more evident; however significant differences between both concentrations were not observed in most strains. Aflatoxin B₁ production did not show noticeable differences among different pesticide concentrations assayed at all a_W in both strains. This study has shown that these Aspergillus flavus and A. parasiticus strains are able to grow effectively and produce aflatoxins in high nutrient status media over a range of glyphosate concentrations under different water activity conditions.     

Study of aflatoxin B1 as a risk factor that impairs the reproductive performance in females – Egypt

Infertility among couples in Egypt is causing increasing concern. Investigations showed a relationship between ovulatory disorders and the incidence of female factor-mediated fertility difficulties in couples. However, the factors responsible for the sudden increase in ovulatory disorders are not fully understood. The aim of this study was to assess the presence of aflatoxin in sera of infertile women and to correlate this with ovarian functions and hormonal parameters. Blood samples were collected from 70 adult Egyptian females comprising 50 infertile and 20 fertile control individuals, and screened for the presence of aflatoxin B₁ (AFB₁). Ovarian function was examined by measuring mean ovarian volume, and the umber and size of ovarian follicles. Blood hormonal levels were determined. All blood samples (patients and controls) showed negative results as regards AFB₁ analysis. However, there was a significant enlargement in the mean ovarian volume but a significant decrease in follicular size. In addition, there were significant higher levels of lutinizing hormone (LH) and significant lower levels of mid-luteal progesterone in infertile females. Although blood samples were negative with respect to AFB₁, the role of aflatoxin cannot be excluded as a contributing factor to female infertility, since the toxin was found to produce deleterious effects on the reproductive system in animals. More studies including aflatoxin analysis in ovarian biopsies are recommended to ascertain involvement of this toxin.