The AFLATOX® Project: Approaching the Development of New Generation,Natural-Based Compounds for the Containment of the Mycotoxigenic Phytopathogen Aspergillus flavus and Aflatoxin Contamination

The control of the fungal contamination on crops is considered a priority by the sanitary authorities of an increasing number of countries, and this is also due to the fact that the geographic areas interested in mycotoxin outbreaks are widening. Among the different pre- and post-harvest strategies that may be applied to prevent fungal and/or aflatoxin contamination, fungicides still play a prominent role; however, despite of countless efforts, to date the problem of food and feed contamination remains unsolved, since the essential factors that affect aflatoxins production are various and hardly to handle as a whole. In this scenario, the exploitation of bioactive natural sources to obtain new agents presenting novel mechanisms of action may represent a successful strategy to minimize, at the same time, aflatoxin contamination and the use of toxic pesticides. The Aflatox^® Project was aimed at the development of new-generation inhibitors of aflatoxigenic Aspergillus spp. proliferation and toxin production, through the modification of naturally occurring molecules: a panel of 177 compounds, belonging to the thiosemicarbazones class, have been synthesized and screened for their antifungal and anti-aflatoxigenic potential. The most effective compounds, selected as the best candidates as aflatoxin containment agents, were also evaluated in terms of cytotoxicity, genotoxicity and epi-genotoxicity to exclude potential harmful effect on the human health, the plants on which fungi grow and the whole ecosystem.     

我国国家标准和行业标准中黄曲霉毒素测定方法综述

黄曲霉毒素广泛存在于自然界中,能够污染多种农作物和食品,对人类的健康造成极其严重的危害。目前,尚无绝对有效的措施避免黄曲霉毒素的污染,因此,制定相应的标准并进一步规范和加强对黄曲霉毒素的检测和监控显得尤为重要。截至到2015年1月,我国现行的针对黄曲霉毒素的国家标准和行业标准共有21个,其中有11个标准在2010~2015年发布实施。本文全面综述了21个标准中采纳的7种测定方法及其特点,其中高效液相色谱法是目前广泛使用的最权威的确证定量测定方法,而免疫层析法则是最简便的快速筛查定性检测方法。在此基础上,本文进一步讨论了标准的制修订现状,并展望了黄曲霉毒素检测技术的发展趋势。      

稻米中黄曲霉毒素和赭曲霉毒素A的研究进展

真菌毒素是由某些丝状真菌产生的有毒代谢产物,在自然界中普遍存在,严重威胁人畜健康。综述了黄曲霉毒素和赭曲霉毒素A的理化性质及危害,在稻米中的污染现状、限量标准及分析方法等方面进行阐述,希望能给真菌毒素的研究提供一些参考。      

发酵豆制品潜在风险因子分析

采集了59份重庆生产酱油、豆豉、腐乳和豆瓣酱4大类发酵豆制品,检测其理化指标和可能存在的6种风险因子,包括蛋白质降解物挥发性盐基氮、硫化氢和生物胺、防腐剂苯甲酸和丙酸及微生物代谢毒素黄曲霉毒素。结果表明,各类样品的pH值及总酸含量均在正常范围;腐乳及包装酱油样品所含氨基酸态氮含量均满足国家标准;各类生物胺普遍存在于发酵豆制品中,不同类型的样品中所含生物胺的种类和含量不同,主要含有酪胺和腐胺。腐乳中生物胺总量最高;挥发性盐基氮的含量较高,可能对样品安全性有影响;硫化氢含量较少,暂不影响食品的可食性;总黄曲霉毒素的含量均未超过国家限量标准,安全性良好。苯甲酸和丙酸普遍存在于各类发酵豆制品中,个别样品中苯甲酸和丙酸含量远高于国家的限量标准,问题较为突出。有必要追踪发酵豆制品加工及贮藏过程中风险因子的变化及产生规律,提出有效的质量安全控制措施。     

酶联免疫吸附法快速检测黄曲霉毒素

结合间接竞争反应机制,运用酶联免疫吸附技术(enzyme-linked immunosorbent assay,ELISA),从抗原(antigen,Ag)包被时间、体系反应温度、酶标二抗作用时间、显色时间等主要因素开展快速检测黄曲霉毒素改进方法的研究。研究结果表明适宜的优化条件为:采用Ag包被20 h、反应温度24℃左右、酶标二抗作用时间30 min、底物显色时间15min。通过对饲料等11种样本的检测得出:半抑制浓度IC50<0.1μg/L,添加回收率在67%~116%,灵敏度达到0.03μg/L,线性系数>0.99,板内变异小于5%。检测试验结果良好,表明该改进方法具有可行性。     

A Review on Mycotoxins in Food and Feed: Malaysia Case Study

Fungi are distributed worldwide and can be found in various foods and feedstuffs from almost every part of the world. Mycotoxins are secondary metabolites produced by some fungal species and may impose food safety risks to human health. Among all mycotoxins, aflatoxins (AFs), ochratoxin A (OTA), trichothecenes, deoxynivalenol (DON and T‐2 toxin), zearalenone (ZEN), and fumonisins (FMN) have received much attention due to high frequency and severe health effects in humans and animals. Malaysia has heavy rainfall throughout the year, high temperatures (28 to 31 °C), and high relative humidity (70% to 80% during wet seasons). Stored crops under such conditions can easily be contaminated by mycotoxin‐producing fungi. The most important mycotoxins in Malaysian foods are AFs, OTA, DON, ZEN, and FMN that can be found in peanuts, cereal grains, cocoa beans, and spices. AFs have been reported to occur in several cereal grains, feeds, nuts, and nut products consumed in Malaysia. Spices, oilseeds, milk, eggs, and herbal medicines have been reported to be contaminated with AFs (lower than the Malaysian acceptable level of 35 ng/g for total AFs). OTA, a possible human carcinogen, was reported in cereal grains, nuts, and spices in Malaysian market. ZEN was detected in Malaysian rice, oat, barley, maize meal, and wheat at different levels. DON contamination, although at low levels, was reported in rice, maize, barley, oat, wheat, and wheat‐based products in Malaysia. FMN was reported in feed and some cereal grains consumed in Malaysia. Since some food commodities are more susceptible than others to fungal growth and mycotoxin contamination, more stringent prevention and control methods are required.     

Aflatoxin contamination and exposure in processed cereal-based complementary foods for infants and young children in greater Accra,Ghana

In the study, aflatoxin levels were assessed in thirty five (35) cereal-based food products intended for infants and young children. Additionally, the results showed that 71% of the processed foods intended for infants contained AFB₁ (0.18 ± 0.01 to 36.10 ± 0.32 μgkg⁻¹) levels higher than the European Union permissible limits of 0.1 μg kg⁻¹. Aflatoxin intake was estimated using aflatoxin levels in the food products and the estimated individual consumption rates. The study also revealed mixed cereals as having the highest intake of aflatoxin B₁ contaminants (0.005–0.852 μgkg⁻¹bw d⁻¹; 0.004–0.657 μgkg⁻¹bwd⁻¹) with mean estimated daily intake (EDI) of 0.23 ± 0.16 μgkg⁻¹bwd⁻¹ and 0.153 ± 0.13 μgkg⁻¹bwd⁻¹ for infants and young children respectively. The estimated AFT intake recorded for infants and young children for all the cereal-based food ranged from 0.005 to 1.054 μgkg⁻¹bwd⁻¹ and 0.004–0.838 μgkg⁻¹bwd⁻¹ respectively.     

Effect of contamination of foods by Aspergillus flavus on the nutritive value of protein

The effect of aflatoxin was measured on the protein quality of peanut meal (PNM) and fish meal (FM) Total protein efficiency, protein efficiency ratio, net protein utilisation, examination of the histopathology of the liver, ileal digesion of amino acids and plasma amino acid concentration were used as bioassays together with chemical score (CS), dye binding capacity (DBC), essential amino acid index (EAAI), and discriminant computered PER (DC-PER) as chemical methods. In trial 1, aflatoxin-free PNM was compared with infected PNM at graded levels of toxin when fed to chickens and ducklings. In trial 2, various mixtures of PNM and FM at a constant aflatoxin level (280 μg kg^?1) were fed to compare the effects of aflatoxin on proteins of differing quality. Ducks were more sensitive to the toxin than chickens, as indicated by deterioration of protein quality, and the effects on growth and the histological appearance of the liver were magnified on diets of low quality (PNM), but not of high quality (FM). Contamination of PNM resulted in progressive increase in DBC and, to a lesser extent, in DC-PER, while EAAI and CS were not affected. The importance of these findings lies in the problems of mould contamination of animal feedstuffs in humid, tropical conditions, which may affect the more sensitive animals, and may not be detected by chemical methods of measuring protein quality, nor by bioassay on chickens, if the levels of contamination are low.     

Determination of aflatoxin M1 in milk by triple quadrupole liquid chromatography-tandem mass spectrometry

A sensitive and reliable method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with an electrospray-positive ionization method was developed for the determination of aflatoxin M₁ in milk. This method includes simple extraction of the sample with acetonitrile by ultrasonic, separation on an MGIII-C₁₈ column using 0.01% formic acid buffer/acetonitrile (60 : 40, v/v) as mobile phase, and MS/MS detection using multiple reaction-monitoring mode. Average recoveries of aflatoxin M₁ from spiked samples at concentrations of 0.02 and 1 ng ml^?1 ranged from 77% to 94%, with a 6% relative standard deviation. The limit of detection and limit of quantification were 0.006 and 0.02 ng ml^?1, respectively. The standard curve was linear between 0.02 and 20.0 ng ml^?1. The recommended method is simple, rapid, specific and reliable for the routine monitoring of aflatoxin M₁ in milk.     

黄曲霉毒素检测方法研究进展

黄曲霉毒素主要是由黄曲霉和寄生曲霉产生的一类次级代谢产物,经常污染粮油原料及其制品,具有强致癌性.介绍了黄曲霉毒素毒性及一些国家和地区的残留限量标准;针对不同基质间组分存在较大差异,概述了液液萃取、超临界流体萃取、固相萃取及免疫亲和柱净化等样品前处理方法;同时对薄层色谱、高效液相色谱、酶联免疫吸附等测定黄曲霉毒素的方法、原理、应用及特点进行了综述;最后分析了黄曲霉毒素检测技术的发展趋势.