食品与饲料基质中真菌毒素检测技术研究进展

真菌毒素是真菌在生长繁殖过程中产生的相对分子质量较小的次生有毒代谢产物,不易被加工或烹调加热所破坏,超过一定摄入量后会引起人的肝肾功能下降、癌变或诱发免疫抑制性疾病。目前尚无绝对有效的措施避免真菌毒素的污染,因此,研究可以准确高效地测定食品和饲料基质中的真菌毒素的检测技术非常重要。本研究通过对真菌毒素提取方法、净化方法及检测技术的研究进展进行综述,发现目前研究方法大多是针对某一种真菌毒素进行检测的常规检测方法或者同时检测多种真菌毒素的检测方法,由于这些方法存在稳定性差、定量不准确或前处理复杂等,因此未来应大力开发检测快速、高灵敏度、高特异性的真菌毒素检测方法。     

Occurrence and significance of mycotoxins in forage crops and silage: a review

Study of mycotoxins in animal feeding stuffs has concentrated on the occurrence of aflatoxins and, to a lesser extent, other mycotoxins in cereals, raw materials and concentrate feeds. However, ruminant diets contain a high proportion of forage crops such as grass or maize silage, hay and straw. Under adverse growing, production or storage conditions, fungal spoilage is likely to occur with some degree of mycotoxin contamination. The mould flora of forage crops is likely to differ significantly from that of cereals and mycotoxin contamination, should it occur, could differ qualitatively and quantitatively. Information relating to forage crops as a potential source of mycotoxins is reviewed. Some field incidents and animal disease which may be mycotoxin related are discussed and analytical methods are reviewed. Information on dose and effect of candidate mycotoxins is given where available. The review suggests areas which the authors consider merit further study. Crown Copyright 1998.     

水果制品中真菌毒素污染及控制研究进展

真菌侵染是造成水果腐烂和水果及其制品中真菌毒素残留的根本原因。全球由于真菌侵染而腐烂的水果损失约占新鲜水果的20%~30%,而真菌毒素也成为危害人体健康的潜在风险。本文首先综述了水果制品中常见的6种真菌毒素的来源、稳定性及毒性,包括展青霉素、赭曲霉毒素A、黄曲霉毒素B1、交链孢酚、交链孢酚单甲醚、细交链孢菌酮酸、赭曲霉毒素A和黄曲霉毒素B1展青霉素,并分析了各种真菌毒素在不同种类水果制品中的污染状况。其次,重点从控制霉菌生长、产毒和毒素消减3个层面,综述了目前在水果采前、采后和加工3个环节真菌毒素的防控措施,并提出真菌毒素防控领域尚需进一步开展的研究,明确安全、无污染和精准的绿色防控手段为防控真菌毒素污染的发展方向,采取传统控制手段结合生物防控等新技术有望实现对真菌毒素的有效控制。     

小麦产毒病原菌及其毒素防控的研究进展

小麦是全球第二大粮食作物,每年因病害造成小麦严重减产,品质下降。一些病原菌还能够产生真菌毒素,进一步危害小麦及其制品的质量安全,对人畜健康造成巨大危害。由镰刀菌引起的小麦赤霉病是我国最主要的小麦病害之一,由交链孢引起的小麦黑胚病也备受关注。这2种真菌既能引起小麦病害,又能产生真菌毒素,故称之为产毒病害。镰刀菌产生的脱氧雪腐镰刀菌烯醇(deoxynivalenol,DON)、玉米赤霉烯酮(zearalenone,ZEN)、伏马菌素等和交链孢产生的交链孢酚(alternariol,AOH)、交链孢酚单甲醚(alternariol monomethyl ether,AME)和细交链格孢酮酸(tenuazonic acid,Te A)等是2类病原菌产生的主要真菌毒素。本文综述了能引起小麦产毒病害的镰刀菌和交链孢的特点、真菌毒素以及病害和毒素的防控技术,尤其是2类病原菌引起的小麦病害和真菌毒素的防控。这将为后期防治小麦产毒病害及控制毒素产生的研究提供有利参考。     

梨果实及其制品中真菌毒素的污染、检测及控制方法研究进展

梨果实营养丰富, 水分含量较高, 在生产、采收和贮运过程中易受病原菌侵染, 特别是在贮藏期间发生真菌性病害后腐烂霉变, 产生并积累各种真菌毒素。本文首先介绍了链格孢毒素、展青霉素、橘霉素和黄曲霉毒素的毒性和在梨果实及其制品中的污染状况, 其次, 对在梨和其制品中应用的薄层色谱法和液相色谱-质谱联用法的特点和应用实例进行了综述, 最后, 总结了果品中真菌毒素的降解方法, 并对有效防控真菌毒素的重点研究方向进行了展望。当前国内对于梨果实及其制品中真菌毒素的研究报道很少, 今后应加强这方面研究, 明确当前真菌毒素的种类以及污染水平, 并重点开展有效防控真菌毒素的研究, 提高我国梨果实及其制品的质量安全水平。     

Sensing of Mycotoxin Producing Fungi in the Processing of Grains

Due to the growing demand for rapid, sensitive and inexpensive detection of contaminations in food and feed, both in the field and after harvest, non-invasive and non-destructive spectroscopic methods are reviewed for the direct identification of fungi and mycotoxins on grain and in grain products. Different model samples containing defined contaminations of fungi and/or mycotoxins as well as real-world samples were investigated using spectroscopic methods (steady-state and time-resolved fluorescence spectroscopy, ultraviolet/visible/near infrared (NIR) spectroscopy, two-photon-induced fluorescence (TPIF)). To evaluate the capability of TPIF for on-line and in-line mycotoxin analysis in real-world samples, fundamental parameters of different mycotoxins, such as two-photon cross-sections, were determined in beer and wine. A comparison showed that the disturbance due to background emission originating from matrix constituents is significantly reduced under two-photon excitation conditions, and TPIF can be applied for a qualitative and quantitative analysis of mycotoxins in wine and beer. The combination of data obtained from different spectroscopic methods (such as optimised excitation and emission wavelengths, fluorescence decay times and fluorescence quantum efficiency on the one hand side and NIR spectroscopy on the other side) is promising for the qualitative as well as quantitative identification of fungi and mycotoxins. Moreover, NIR reflection spectra yield additional information on ingredients, moisture content and the presence (or absence) of fungi in the sample. The spectroscopic techniques are complemented by chemometric tools to extract the desired chemical information, e.g. with respect to the presence of contaminations.     

液相色谱-串联质谱法检测茶叶中真菌毒素的研究进展

茶叶由于富含多酚类、儿茶素和多种矿物质等，具有降血脂、抗氧化、减肥、抗菌等多重保健功效而备受欢迎，其质量安全一直广受关注。茶叶在其生产加工和保存过程中有可能被真菌毒素污染，真菌毒素由于对人和动物具有严重的危害（致癌性）更是备受关注。茶叶中毒素残留的高效、高通量检测方法，成为了研究的重点。本文参考国内外大量文献，综述了茶叶中真菌毒素的检测方法，比较了不同检测方法的局限性和优越性，重点综述了检出限低、灵敏度高的液相色谱-串联质谱法(liquid chromatography-tandem mass spectrometry, LC-MS/MS)用于茶叶真菌毒素的研究。对不同前处理提取试剂、净化小柱和检测仪器条件的优劣进行了比较；对该方法用于检测不同品种茶叶时，其基质效应对结果的影响进行了分析；总结了茶叶中真菌毒素的检测结果。对该方法用于茶叶真菌毒素检测的发展进行了展望，为检测茶叶中真菌毒素检测标准和限量标准的建立提供理论依据     

Natural Occurrence,Analysis, and Prevention of Mycotoxins in Fruits and their Processed Products

Mycotoxins are small toxic chemical products formed as the secondary metabolites by fungi that readily contaminate foods with toxins in the field or after harvest. The presence of mycotoxins, such as aflatoxins, ochratoxin A, and patulin, in fruits and their processed products is of high concern for human health due to their properties to induce severe acute and chronic toxicity at low-dose levels. Currently, a broad range of detection techniques used for practical analysis and detection of a wide spectrum of mycotoxins are available. Many analytical methods have been developed for the determination of each group of these mycotoxins in different food matrices, but new methods are still required to achieve higher sensitivity and address other challenges that are posed by these mycotoxins. Effective technologies are needed to reduce or even eliminate the presence of the mycotoxins in fruits and their processed products. Preventive measures aimed at the inhibition of mycotoxin formation in fruits and their processed products are the most effective approach. Detoxification of mycotoxins by different physical, chemical, and biological methods are less effective and sometimes restricted because of concerns of safety, possible losses in nutritional quality of the treated commodities and cost implications. This article reviewed the available information on the major mycotoxins found in foods and feeds, with an emphasis of fruits and their processed products, and the analytical methods used for their determination. Based on the current knowledge, the major strategies to prevent or even eliminate the presence of the mycotoxins in fruits and their processed products were proposed.     

Some major mycotoxins and their mycotoxicoses--an overview

Mycotoxins likely have existed for as long as crops have been grown but recognition of the true chemical nature of such entities of fungal metabolism was not known until recent times. Conjecturally, there is historical evidence of their presence back as far as the time reported in the Dead Sea Scrolls. Evidence of their periodic, historical occurrence exists until the recognition of aflatoxins in the early 1960s. At that time mycotoxins were considered as a storage phenomenon whereby grains becoming moldy during storage allowed for the production of these secondary metabolites proven to be toxic when consumed by man and other animals. Subsequently, aflatoxins and mycotoxins of several kinds were found to be formed during development of crop plants in the field. The determination of which of the many known mycotoxins are significant can be based upon their frequency of occurrence and/or the severity of the disease that they produce, especially if they are known to be carcinogenic. Among the mycotoxins fitting into this major group would be the aflatoxins, deoxynivalenol, fumonisins, zearalenone, T-2 toxin, ochratoxin and certain ergot alkaloids. The diseases (mycotoxicoses) caused by these mycotoxins are quite varied and involve a wide range of susceptible animal species including humans. Most of these diseases occur after consumption of mycotoxin contaminated grain or products made from such grains but other routes of exposure exist. The diagnosis of mycotoxicoses may prove to be difficult because of the similarity of signs of disease to those caused by other agents. Therefore, diagnosis of a mycotoxicoses is dependent upon adequate testing for mycotoxins involving sampling, sample preparation and analysis.     

微芯片技术在农产品及食品真菌毒素快速检测中的应用

真菌毒素污染的农产品及食品会对人类及牲畜的健康产生严重威胁。目前,真菌毒素的检测主要采用液相色谱-质谱联用法(liquid chromatography-mass spectrometry,LC-MS)、气相色谱-质谱联用法(gas chromatography-mass spectrometry,GC-MS)等传统检测方法。然而,这些检测方法的样品前处理过程比较繁琐、费时,并且在分析过程中还会消耗大量的有毒试剂。微芯片技术所需样品的消耗量少并且分析时间短,可实现样品的集成化、微型化以及高通量检测。微芯片技术在真菌毒素检测中的应用,弥补了上述传统检测方法的不足。本文主要综述了微芯片技术在农产品及食品真菌毒素快速检测中的应用研究进展。首先对农产品及食品中常见的真菌毒素及其毒性进行了简单介绍;接着重点对微芯片技术在真菌毒素检测中的应用进行了详细论述;最后对微芯片应用于真菌毒素检测的发展前景和挑战进行了展望。     

果蔬常见病原菌及真菌毒素的防控技术研究进展

果蔬在田间的生长过程以及采摘后的贮存、运输和加工过程中, 易发生真菌性病害, 不仅可引起腐烂或腐败, 带来严重的经济损失, 部分病原菌还可能产生真菌毒素对人体造成潜在危害。真菌毒素是一类由丝状真菌在适宜条件下产生的有毒次级代谢产物, 是继农药残留、重金属污染后, 影响果蔬质量安全的又一类关键风险因子, 具有强毒性。链格孢毒素、赭曲霉毒素、展青霉素是存在于果蔬及其制品中主要的真菌毒素种类。本文综述了近年来果蔬中常见病原真菌及其真菌毒素, 并阐述了真菌毒素防控方法, 包括物理、化学和生物方法, 对未来果蔬中真菌毒素的防控研究方向进行了展望, 以期为该领域研究者提供参考。     

The Use and Effects of Lactic Acid Bacteria in Malting and Brewing with Their Relationships to Antifungal Activity,Mycotoxins and Gushing: A Review

Several metabolic properties of lactic acid bacteria (LAB) serve special functions, which directly or indirectly have impact on processes such as improved quality and safety and flavour development in the malting and brewing industry. LAB are widely distributed in nature and in spontaneous fermentations, often they are found to be the dominating microflora resulting in both the inhibition of spoilage bacteria and organisms. This review describes the applications of LAB in malting and brewing. Mycotoxins are naturally occurring toxic secondary metabolites of fungi that may be present in cereals. Several of these mycotoxins have been associated with human and animal diseases and are known to survive the brewing process. LAB have been shown to restrict the growth of the most important toxigenic fungi thereby reducing the formation of these harmful toxins. The occurrence of mycotoxins in cereals is discussed and their effect in beer is reviewed. The main features of this review are: (I) LAB starter cultures in malting and brewing (II) production of acid malt; (III) biological acidification of mash and wort in brewing; (IV) bacteriocins produced by LAB in brewing; (V) LAB and anti‐fungal activity; (VI) mycotoxins in cereals.     

Strategies to prevent mycotoxin contamination of food and animal feed: a review

Mycotoxins are fungal secondary metabolites that have been associated with severe toxic effects to vertebrates produced by many important phytopathogenic and food spoilage fungi including Aspergillus, Penicillium, Fusarium, and Alternaria species. The contamination of foods and animal feeds with mycotoxins is a worldwide problem. We reviewed various control strategies to prevent the growth of mycotoxigenic fungi as well as to inhibit mycotoxin biosynthesis including pre-harvest (resistance varieties, field management and the use of biological and chemical agents), harvest management, and post-harvest (improving of drying and storage conditions, the use of natural and chemical agents, and irradiation) applications. While much work in this area has been performed on the most economically important mycotoxins, aflatoxin B(1) and ochratoxin A much less information is available on other mycotoxins such as trichothecenes, fumonisin B(1), zearalenone, citrinin, and patulin. In addition, physical, chemical, and biological detoxification methods used to prevent exposure to the toxic and carcinogenic effect of mycotoxins are discussed. Finally, dietary strategies, which are one of the most recent approaches to counteract the mycotoxin problem with special emphasis on in vivo and in vitro efficacy of several of binding agents (activated carbons, hydrated sodium calcium aluminosilicate, bentonite, zeolites, and lactic acid bacteria) have also been reviewed.     

Occurrence,prevention and remediation of toxigenic fungi and mycotoxins in silage: a review

Ruminants are considered to be less sensitive towards mycotoxins than monogastric animals because rumen microbiota have mycotoxin‐detoxifying capacities. Therefore the effect of mycotoxins towards ruminants has been studied to a lesser extent compared with monogastric animals. Worldwide, a high proportion of the ruminant diet consists of silages made of forage crops (i.e. all parts of the crop above the stubble are harvested). In practice, silages are often contaminated with multiple mycotoxins. Exposure to a cocktail of mycotoxins can hamper animal production and have severe health consequences. In this article the different aspects associated with mycotoxin contamination of silage are reviewed ‘from seed to feed’. An overview is given on the occurrence of toxigenic fungal species and their concomitant mycotoxins in forage crops before and after ensiling. The mycotoxin load of visually non‐mouldy samples and mouldy hot spots within the same silo is also compared. Subsequently, this review delves into different problem‐solving strategies. A logical first step is prevention of mould growth and mycotoxin production in the field, during harvest and during ensiling. If prevention should fail, several remediation strategies are available. These are listed, mainly focusing on the possibilities of microbial degradation of mycotoxins in vivo in silage. © 2015 Society of Chemical Industry     

A review of postharvest approaches to reduce fungal and mycotoxin contamination of foods

Contamination of agricultural and food products by some fungi species that produce mycotoxins can result in unsafe food and feed. Mycotoxins have been demonstrated to have disease‐causing activities, including carcinogenicity, immune toxicity, teratogenicity, neurotoxicity, nephrotoxicity, and hepatotoxicity. Most of mycotoxins are heat stable and cannot be easily destroyed by conventional thermal food processing or domestic cooking methods. Postharvest approaches to prevent growth of mycotoxin‐producing fungi and detoxify mycotoxins from contaminated food are important topics in food safety research. Physical, chemical, and biological methods have been applied to prevent fungal growth or mycotoxin production, or to reduce mycotoxin content in the postharvest period and contribute toward mitigating against the effects of mycotoxins on human health. This literature review aims to evaluate postharvest approaches that have been applied to control both fungi growth and mycotoxin content in food and discuss their potential for upscaling to industrial scale.     

啤酒及其生产原料中的真菌毒素

真菌毒素是由真菌产生的一类次级代谢产物,而啤酒及其生产原料的真菌毒素感染则对人们饮用啤酒的安全性造成了威胁。本文综述了赭曲毒素A(Ochratoxin,OTA)、脱氧雪腐镰刀菌烯醇(Deoxynivalenol,DON)、玉米赤霉烯酮(Zearalenone,ZEN)、伏马毒素(Fumonisin,FB)和黄曲霉毒素(Aflatoxin,AFB)对啤酒及其生产原料的感染情况、分析方法和限量标准,并简要介绍了谷物中真菌毒素感染的预防措施和脱毒技术。     

粮食中隐蔽型真菌毒素污染的研究进展

真菌毒素污染是影响全球食品安全问题的重要因素之一。真菌毒素是真菌产生的次级代谢产物,对人体、动植物皆可产生危害。自然条件下,真菌毒素分子可与葡萄糖、丙二酸或谷胱甘肽等极性较强的基团结合,生成结合型真菌毒素。由于毒素结构发生改变,现有的常规检测手段不能有效地检测其存在,因此称其为隐蔽型真菌毒素(masked mycotoxins)。已有文献报道,隐蔽型真菌毒素往往与其原型同时存在于粮食中,而隐蔽型毒素在进入人和动物体内后,经水解、氧化还原等代谢过程,可释放出有毒的毒素原型分子,给食品安全、人和动物健康造成严重隐患。因此对隐蔽型真菌毒素的形成机制、污染状况和毒理研究应尽快展开。本文综述了国内外近几年以来几种常见隐蔽型真菌毒素的最新研究进展,为隐蔽型真菌毒素的研究提供有价值的参考。     

Molecular strategies for detection and quantification of mycotoxin‐producing Fusarium species: a review

Fusarium contamination is considered a major agricultural problem, which could not only significantly reduce yield and quality of agricultural products, but produce mycotoxins that are virulence factors responsible for many diseases of humans and farm animals. One strategy to identify toxigenic Fusarium species is the use of modern molecular methods, which include the analysis of DNA target regions for differentiation of the Fusarium species, particularly the mycotoxin‐producing Fusarium species such as F. verticillioides and F. graminearum. Additionally, polymerase chain reaction assays are used to determine the genes involved in the biosynthesis of the toxins in order to facilitate a qualitative and quantitative detection of Fusarium‐producing mycotoxins. Also, it is worth mentioning that some factors that modulate the biosynthesis of mycotoxins are not only determined by their biosynthetic gene clusters, but also by environmental conditions. Therefore, all of the aforementioned factors which may affect the molecular diagnosis of mycotoxins will be reviewed and discussed in this paper. © 2014 Society of Chemical Industry     

色谱法检测真菌毒素的研究进展

色谱法在物质分离和检测方面应用广泛。真菌毒素在低浓度下就有明显毒性,采用色谱法可实现对真菌毒素的分离和微量检测。依据真菌毒素的自身特性采用不同的色谱方法进行检测,但不同的色谱法有不同的应用范围。薄层色谱法用于对真菌毒素的快速定性,气相色谱法需要对真菌毒素衍生化汽化后才可检测,液相色谱法对本身具有或衍生化后具有紫外和荧光吸收的真菌毒素才可检测,对无紫外和荧光吸收的则需质谱检测器。简述了色谱法在真菌毒素进行定性定量的研究情况,为利用色谱法对真菌素含量测定的实际运用和方法学开发提供参考。     

Mycotoxin analysis: an update

Mycotoxin contamination of cereals and related products used for feed can cause intoxication, especially in farm animals. Therefore, efficient analytical tools for the qualitative and quantitative analysis of toxic fungal metabolites in feed are required. Current methods usually include an extraction step, a clean-up step to reduce or eliminate unwanted co-extracted matrix components and a separation step with suitably specific detection ability. Quantitative methods of analysis for most mycotoxins use immunoaffinity clean-up with high-performance liquid chromatography (HPLC) separation in combination with UV and/or fluorescence detection. Screening of samples contaminated with mycotoxins is frequently performed by thin layer chromatography (TLC), which yields qualitative or semi-quantitative results. Nowadays, enzyme-linked immunosorbent assays (ELISA) are often used for rapid screening. A number of promising methods, such as fluorescence polarization immunoassays, dipsticks, and even newer methods such as biosensors and non-invasive techniques based on infrared spectroscopy, have shown great potential for mycotoxin analysis. Currently, there is a strong trend towards the use of multi-mycotoxin methods for the simultaneous analysis of several of the important Fusarium mycotoxins, which is best achieved by LC-MS/MS (liquid chromatography with tandem mass spectrometry). This review focuses on recent developments in the determination of mycotoxins with a special emphasis on LC-MS/MS and emerging rapid methods.